Sample records for fresh leaf tissue

  1. Storage effects on genomic DNA in rolled and mature coca leaves.

    PubMed

    Johnson, Emanuel L; Kim, Soo-Hyung; Emche, Stephen D

    2003-08-01

    Rolled and mature leaf tissue was harvested from Erythroxylum coca var. coca Lam. (coca) to determine a method for storage that would maintain DNA with high quality and content up to 50 days. Harvesting coca leaf tissue under Andean field conditions often requires storage from 3 to 10 days before extraction where tissue integrity is lost. All samples of rolled and mature coca leaf tissue were harvested and separately stored fresh in RNAlater for 50 days at 4 degrees, -20 degrees, and 23 degrees C, while similar samples were air-dried for 72 h at 23 degrees C or oven-dried for 72 h at 40 degrees C after storage, before extraction. Triplicate samples of each tissue type were extracted for DNA at 10-day intervals and showed that DNA integrity and content were preserved in leaf tissue stored at 4 degrees and -20 degrees C for 50 days. Rolled and mature leaf tissue stored at 4 degrees, -20 degrees, and 23 degrees C showed insignificant degradation of DNA after 10 days, and by day 50, only leaf tissue stored at 4 degrees and -20 degrees C had not significantly degraded. All air- and oven-dried leaf tissue extracts showed degradation upon drying (day 0) and continuous degradation up to day 50, despite storage conditions. Amplified fragment length polymorphism analysis of DNA from rolled and mature leaf tissue of coca stored at 4 degrees and -20 degrees C for 0, 10, and 50 days showed that DNA integrity and content were preserved. We recommend that freshly harvested rolled or mature coca leaf tissue be stored at 4 degrees, -20 degrees, and 23 degrees C for 10 days after harvest, and if a longer storage is required, then store at 4 degrees or -20 degrees C.

  2. 9 CFR 319.702 - Lard, leaf lard.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... lard. (a) Lard is the fat rendered from clean and sound edible tissues from swine. The tissues may be... misbranding of the lard. The tissues shall be reasonably free from blood, and shall not include stomachs, livers, spleens, kidneys, and brains, or settlings and skimmings. “Leaf Lard” is lard prepared from fresh...

  3. 9 CFR 319.702 - Lard, leaf lard.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... lard. (a) Lard is the fat rendered from clean and sound edible tissues from swine. The tissues may be... misbranding of the lard. The tissues shall be reasonably free from blood, and shall not include stomachs, livers, spleens, kidneys, and brains, or settlings and skimmings. “Leaf Lard” is lard prepared from fresh...

  4. 9 CFR 319.702 - Lard, leaf lard.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... lard. (a) Lard is the fat rendered from clean and sound edible tissues from swine. The tissues may be... misbranding of the lard. The tissues shall be reasonably free from blood, and shall not include stomachs, livers, spleens, kidneys, and brains, or settlings and skimmings. “Leaf Lard” is lard prepared from fresh...

  5. 9 CFR 319.702 - Lard, leaf lard.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... lard. (a) Lard is the fat rendered from clean and sound edible tissues from swine. The tissues may be... misbranding of the lard. The tissues shall be reasonably free from blood, and shall not include stomachs, livers, spleens, kidneys, and brains, or settlings and skimmings. “Leaf Lard” is lard prepared from fresh...

  6. 9 CFR 319.702 - Lard, leaf lard.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... lard. (a) Lard is the fat rendered from clean and sound edible tissues from swine. The tissues may be... misbranding of the lard. The tissues shall be reasonably free from blood, and shall not include stomachs, livers, spleens, kidneys, and brains, or settlings and skimmings. “Leaf Lard” is lard prepared from fresh...

  7. Protoplast isolation and genetically true-to-type plant regeneration from leaf- and callus-derived protoplasts of Albizia julibrissin

    Treesearch

    Mohammad-Shafie Rahmani; Paula M. Pijut; Naghi Shabanian

    2016-01-01

    Protoplast isolation and subsequent plant regeneration of Albizia julibrissin was achieved from leaf and callus explants. Leaf tissue from 4 to 5-week-old in vitro seedlings was the best source for high-yield protoplast isolation. This approach produced 7.77 × 105 protoplasts (Pp) per gram fresh weight with 94 % viability;...

  8. Rapid and simple procedure for homogenizing leaf tissues suitable for mini-midi-scale DNA extraction in rice.

    PubMed

    Yi, Gihwan; Choi, Jun-Ho; Lee, Jong-Hee; Jeong, Unggi; Nam, Min-Hee; Yun, Doh-Won; Eun, Moo-Young

    2005-01-01

    We describe a rapid and simple procedure for homogenizing leaf samples suitable for mini/midi-scale DNA preparation in rice. The methods used tungsten carbide beads and general vortexer for homogenizing leaf samples. In general, two samples can be ground completely within 11.3+/-1.5 sec at one time. Up to 20 samples can be ground at a time using a vortexer attachment. The yields of the DNA ranged from 2.2 to 7.6 microg from 25-150 mg of young fresh leaf tissue. The quality and quantity of DNA was compatible for most of PCR work and RFLP analysis.

  9. Tissue-specific changes of glutamine synthetase activity in oats after rhizosphere infestation by Pseudomonas syringae pv. tabaci. Final report

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Knight, T.J.; Temple, S.; Sengupta-Gopalan, C.

    1996-05-15

    Oats (Avena sativa L. lodi) tolerant of rhizosphere infestation by Pseudomonas syringae pv. tabaci when challenged by the pathogen experience tissue-specific alterations of ammonia assimilatory capabilities. Altered ammonia assimilatory potentials between root and leaf tissue result from selective inactivation of glutamine synthetase (GS) by the toxin Tabtoxinine-B-lactam (TBL). Root GS is sensitive and leaf GSs are resistant to TBL inactivation. With prolonged challenge by the pathogen root GS activity decreases but leaf GS specific activity increase. Higher leaf GS activity is due to decreased rates of degradation rather than increased GS synthesis. Higher leaf GS activity and elevated levels ofmore » GS polypeptide appear to result from a limited interaction between GS and TBL leading to the accumulation of a less active but more stable GS holoenzyme. Tolerant challenged oats besides surviving rhizosphere infestation, experience enhanced growth. A strong correlation exists between leaf GS activity and whole plant fresh weight, suggesting that tissue-specific changes in ammonia assimilatory capability provides the plant a more efficient mechanism for uptake and utilization of nitrogen.« less

  10. Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf

    NASA Astrophysics Data System (ADS)

    Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; An Nguyen, Thien; Alfano, Robert R.

    2014-06-01

    Two-photon (2P) excitation of the second singlet (S) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S2 state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

  11. Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf.

    PubMed

    Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; Nguyen, Thien An; Alfano, Robert R

    2014-06-01

    Two-photon (2P) excitation of the second singlet (S₂) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S₂ state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S₂ state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

  12. Isolation and characterization of allelopathic volatiles from mugwort (Artemisia vulgaris).

    PubMed

    Barney, Jacob N; Hay, Anthony G; Weston, Leslie A

    2005-02-01

    Several volatile allelochemicals were identified and characterized from fresh leaf tissue of three distinct populations of the invasive perennial weed, mugwort (Artemisia vulgaris). A unique bioassay was used to demonstrate the release of volatile allelochemicals from leaf tissues. Leaf volatiles were trapped and analyzed via gas chromatography coupled with mass spectrometry. Some of the components identified were terpenes, including camphor, eucalyptol, alpha-pinene, and beta-pinene. Those commercially available were tested individually to determine their phytotoxicity. Concentrations of detectable volatiles differed in both absolute and relative proportions among the mugwort populations. The three mugwort populations consisted of a taller, highly branched population (ITH-1); a shorter, lesser-branched population (ITH-2) (both grown from rhizome fragments from managed landscapes); and a population grown from seed with lobed leaves (VT). Considerable interspecific variation existed in leaf morphology and leaf surface chemistry. Bioassays revealed that none of the individual monoterpenes could account for the observed phytotoxicity imparted by total leaf volatiles, suggesting a synergistic effect or activity of a component not tested. Despite inability to detect a single dominant phytotoxic compound, decreases in total terpene concentration with increase in leaf age correlated with decreases in phytotoxicity. The presence of bioactive terpenoids in leaf surface chemistry of younger mugwort tissue suggests a potential role for terpenoids in mugwort establishment and proliferation in introduced habitats.

  13. Ambient Molecular Analysis of Biological Tissue Using Low-Energy, Femtosecond Laser Vaporization and Nanospray Postionization Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Shi, Fengjian; Flanigan, Paul M.; Archer, Jieutonne J.; Levis, Robert J.

    2016-03-01

    Direct analysis of plant and animal tissue samples by laser electrospray mass spectrometry (LEMS) was investigated using low-energy, femtosecond duration laser vaporization at wavelengths of 800 and 1042 nm followed by nanospray postionization. Low-energy (<50 μJ), fiber-based 1042 nm LEMS (F-LEMS) allowed interrogation of the molecular species in fresh flower petal and leaf samples using 435 fs, 10 Hz bursts of 20 pulses from a Ytterbium-doped fiber laser and revealed comparable results to high energy (75-1120 μJ), 45 fs, 800 nm Ti:Sapphire-based LEMS (Ti:Sapphire-LEMS) measurements. Anthocyanins, sugars, and other metabolites were successfully detected and revealed the anticipated metabolite profile for the petal and leaf samples. Phospholipids, especially phosphatidylcholine, were identified from a fresh mouse brain section sample using Ti:Sapphire-LEMS without the application of matrix. These lipid features were suppressed in both the fiber-based and Ti:Sapphire-based LEMS measurements when the brain sample was prepared using the optimal cutting temperature compounds that are commonly used in animal tissue cryosections.

  14. Internalization of fresh produce by foodborne pathogens.

    PubMed

    Erickson, Marilyn C

    2012-01-01

    Recent studies addressing the internalization of fresh produce by foodborne pathogens arose in response to the growing number of recent and high profile outbreaks involving fresh produce. Because chemical sanitizing agents used during harvest and minimal processing are unlikely to reach enteric pathogens residing within plant tissue, it is imperative that paths for pathogen entry be recognized and minimized. Using both microscopy and microbial enumeration tools, enteric pathogens have been shown to enter plant tissues through both natural apertures (stomata, lateral junctions of roots, flowers) and damaged (wounds, cut surfaces) tissue. In studies revealing preharvest internalization via plant roots or leaf stomata, experimental conditions have primarily involved exposure of plants to high pathogen concentrations (≥ 6 log g⁻¹ soil or 6 log ml⁻¹ water), but those pathogens internalized appear to have short-term persistence. Postharvest internalization of pathogens via cut surfaces may be minimized by maintaining effective levels of sanitizing agents in waters during harvesting and minimal processing.

  15. Studies on the nitrate reductase activities of the fruit and the source leaf in pepper

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Achhireddy, N.R.; Beevers, L.; Fletcher, J.S.

    Nitrate reductase (NR) activity (NO/sub 2//sup -/ produced in the dark and under anaerobic conditions) of 30-day-old fruit of Capsicum annuum L. was 2.2% that in tissues of a single leaf adjacent to each fruit (33 vs. 1500 nmoles/hr-g fresh weight). The optimal NR activity in one source leaf could only account for about 17% of the fruit's total nitrogen accumulation, while the fruit's own NR activity was almost negligible. Covered and uncovered fruits did not differ significantly in NR activities. 19 references, 1 figure, 1 table.

  16. Epidemiology and control of spinach downy mildew in coastal California

    USDA-ARS?s Scientific Manuscript database

    The most serious threat to global fresh market spinach production is spinach downy mildew, caused by the obligate biotrophic pathogen, Peronospora effusa. Downy mildew causes yellow chlorotic lesions on spinach leaf tissue, often accompanied by abundant sporulation on the undersides of leaves. Very ...

  17. High biological variability of plastids, photosynthetic pigments and pigment forms of leaf primordia in buds.

    PubMed

    Solymosi, Katalin; Morandi, Dominique; Bóka, Károly; Böddi, Béla; Schoefs, Benoît

    2012-05-01

    To study the formation of the photosynthetic apparatus in nature, the carotenoid and chlorophyllous pigment compositions of differently developed leaf primordia in closed and opening buds of common ash (Fraxinus excelsior L.) and horse chestnut (Aesculus hippocastanum L.) as well as in closed buds of tree of heaven (Ailanthus altissima P. Mill.) were analyzed with HPLC. The native organization of the chlorophyllous pigments was studied using 77 K fluorescence spectroscopy, and plastid ultrastructure was investigated with electron microscopy. Complete etiolation, i.e., accumulation of protochlorophyllide, and absence of chlorophylls occurred in the innermost leaf primordia of common ash buds. The other leaf primordia were partially etiolated in the buds and contained protochlorophyllide (0.5-1 μg g(-1) fresh mass), chlorophyllides (0.2-27 μg g(-1) fresh mass) and chlorophylls (0.9-643 μg g(-1) fresh mass). Etio-chloroplasts with prolamellar bodies and either regular or only low grana were found in leaves having high or low amounts of chlorophyll a and b, respectively. After bud break, etioplast-chloroplast conversion proceeded and the pigment contents increased in the leaves, similarly to the greening processes observed in illuminated etiolated seedlings under laboratory conditions. The pigment contents and the ratio of the different spectral forms had a high biological variability that could be attributed to (i) various light conditions due to light filtering in the buds resulting in differently etiolated leaf primordia, (ii) to differences in the light-exposed and inner regions of the same primordia in opening buds due to various leaf folding, and (iii) to tissue-specific slight variations of plastid ultrastructure.

  18. Differences in Leaf Flammability, Leaf Traits and Flammability-Trait Relationships between Native and Exotic Plant Species of Dry Sclerophyll Forest

    PubMed Central

    Murray, Brad R.; Hardstaff, Lyndle K.; Phillips, Megan L.

    2013-01-01

    The flammability of plant leaves influences the spread of fire through vegetation. Exotic plants invading native vegetation may increase the spread of bushfires if their leaves are more flammable than native leaves. We compared fresh-leaf and dry-leaf flammability (time to ignition) between 52 native and 27 exotic plant species inhabiting dry sclerophyll forest. We found that mean time to ignition was significantly faster in dry exotic leaves than in dry native leaves. There was no significant native-exotic difference in mean time to ignition for fresh leaves. The significantly higher fresh-leaf water content that was found in exotics, lost in the conversion from a fresh to dry state, suggests that leaf water provides an important buffering effect that leads to equivalent mean time to ignition in fresh exotic and native leaves. Exotic leaves were also significantly wider, longer and broader in area with significantly higher specific leaf area–but not thicker–than native leaves. We examined scaling relationships between leaf flammability and leaf size (leaf width, length, area, specific leaf area and thickness). While exotics occupied the comparatively larger and more flammable end of the leaf size-flammability spectrum in general, leaf flammability was significantly correlated with all measures of leaf size except leaf thickness in both native and exotic species such that larger leaves were faster to ignite. Our findings for increased flammability linked with larger leaf size in exotics demonstrate that exotic plant species have the potential to increase the spread of bushfires in dry sclerophyll forest. PMID:24260169

  19. Oxygen radical absorbance capacity (ORAC) and phenolic and anthocyanin concentrations in fruit and leaf tissues of highbush blueberry.

    PubMed

    Ehlenfeldt, M K; Prior, R L

    2001-05-01

    Antioxidant capacity, as measured by oxygen radical absorbance capacity (ORAC), and total phenolic and total anthocyanin contents were evaluated in fruit tissues of 87 highbush blueberry (Vacciniumcorymbosum L.) and species-introgressed highbush blueberry cultivars. ORAC and phenolic levels were evaluated in leaf tissues of the same materials. Average values for ORAC, phenolics, and anthocyanins in fruit were 15.9 ORAC units, 1.79 mg/g (gallic acid equivalents), and 0.95 mg/g (cyanidin-3-glucoside equivalents), respectively. Cv. Rubel had the highest ORAC per gram of fresh weight values, at 31.1 units, and cv. Elliott had the highest values on the basis of ORAC per square centimeter of surface area. In leaf tissue, values for both ORAC and phenolics were significantly higher than in fruit tissue, with mean values of 490 ORAC units and 44.80 mg/g (gallic acid equivalents), respectively. Leaf ORAC had a low, but significant, correlation with fruit phenolics and anthocyanins, but not with fruit ORAC. An analysis of ORAC values versus calculated midparent values in 11 plants from the 87-cultivar group in which all parents were tested suggested that, across cultivars, ORAC inheritance is additive. An investigation of ORAC values in a family of 44 cv. Rubel x Duke seedlings showed negative epistasis for ORAC values, suggesting Rubel may have gene combinations contributing to ORAC that are broken up during hybridization.

  20. Effect of nitrogen sources on some morphological characteristics of in vitro stevia rebaudiana Bertoni.

    PubMed

    Akbari, F; Arminian, A; Kahrizi, D; Fazeli, A

    2017-02-28

    Stevia rebaudiana Bertoni belongs to Asteraceae family that leaves 200-300 times sweeter than sugar. Low seed fertility is one of the most important problems in Stevia production. So, Plant tissue culture is an efficient method for mass propagation of Stevia. In this research, we studied the effect of various concentrations of nitrogen on some morphological traits of stevia under in vitro conditions. We used axillary nodes as explants and they were cultured on Murashige and Skoog (MS) medium containing inorganic nitrogen sources i.e. NH4NO3(0, 825 and 1650 mg/l), KNO3(0, 950 and 1900 mg/l) were observed. The cultures were kept for 4 weeks at a temperature of 25±2°C with a photoperiod of 16/8 hour low light/dark each day. Maximum shoot length (89.33 mm), dry weight of plants (0.10 mg) and leaf fresh weight (0.42 mg) was observed on MS medium with 1650 mg/l NH4NO3 and 950 mg/l KNO3. Minimum shoot length (6.13 mm), root length (6.60 mm), leaf number (4.26), leaf dry weight (0.01 mg), leaf fresh weight (0.05 mg), total dry and fresh weight (0.02 and 0.15 mg) and growth rate was observed on a MS medium without nitrogen sources. Moreover, presence of nitrogen sources increases both shooting and rooting in Stevia rebaudiana Bertoni.

  1. Determination of total phenolic content and antioxidant activitity of methanol extract of Maranta arundinacea L fresh leaf and tuber

    NASA Astrophysics Data System (ADS)

    Kusbandari, A.; Susanti, H.

    2017-11-01

    Maranta arundinacea L is one of herbaceous plants in Indonesia which have flavonoid content. Flavonoids has antioxidants activity by inhibition of free radical oxidation reactions. The study aims were to determination total phenolic content and antioxidant activity of methanol extract of fresh leaf and tuber of M. arundinacea L by UV-Vis spectrophotometer. The methanol extracts were obtained with maceration and remaseration method of fresh leaves and tubers. The total phenolic content was assayed with visible spectrophotometric using Folin Ciocalteau reagent. The antioxidant activity was assayed with 1,1-diphenyl-2-picrilhidrazil (DPPH) compared to gallic acid. The results showed that methanol extract of tuber and fresh leaf of M. arundinacea L contained phenolic compound with total phenolic content (TPC) in fresh tuber of 3.881±0.064 (% GAE) and fresh leaf is 6.518±0.163 (% b/b GAE). IC50 value from fresh tuber is 1.780±0.0005 μg/mL and IC50 fresh leaf values of 0.274±0.0004 μg/mL while the standard gallic acid is IC50 of 0.640±0.0002 μg/mL.

  2. Recycling of urea associated with the host plant urease in the silkworm larvae, Bombyx mori.

    PubMed

    Hirayama, C; Sugimura, M; Shinbo, H

    1999-01-01

    Urea concentration and urease activity in the midgut content were compared between larvae of the silkworm, Bombyx mori fed an artificial diet and those fed fresh mulberry leaves. A considerable amount of urea was found in the midgut content of the both larvae, however it was significantly lower in the larvae fed fresh mulberry leaves than in the larvae fed the artificial diet; average urea concentrations in the midgut content of the larvae fed fresh mulberry leaves and the artificial diet were 2.9 and 4.6 &mgr;mol/g, respectively. Urea in the midgut content seems to be secreted from the insect itself since the amount of urea in both diets were negligibly small. Urease activity was detected only in the midgut content of the larvae fed fresh mulberry leaves but not in other tissues of the larvae. On the other hand, no urease activity was detected in the midgut content of the larvae fed the artificial diet. Subsequently, to elucidate the role of mulberry leaf urease in the midgut lumen, larvae that had been reared on the artificial diet were switched to fresh mulberry leaves. The diet switch caused a rapid decrease in urea concentration in the midgut content and an increase in ammonia concentration in the midgut content, suggesting that secreted urea could be hydrolyzed to ammonia by mulberry leaf urease in the midgut lumen. Furthermore, to investigate the physiological significance of mulberry leaf urease on urea metabolism of the silkworm, (15)N-urea was injected into the hemocoel, and after 12 h the larvae were dissected for (15)N analysis. A considerable amount of (15)N was found to be incorporated into the silk-protein of the larvae fed fresh mulberry leaves, but there was little incorporation of (15)N into the silk-protein of the larvae fed the artificial diet. These data indicate that urea is converted into ammonia by the action of mulberry leaf urease in the midgut lumen and used as a nitrogen source in larvae fed mulberry leaves.

  3. Developmental validation of a Cannabis sativa STR multiplex system for forensic analysis.

    PubMed

    Howard, Christopher; Gilmore, Simon; Robertson, James; Peakall, Rod

    2008-09-01

    A developmental validation study based on recommendations of the Scientific Working Group on DNA Analysis Methods (SWGDAM) was conducted on a multiplex system of 10 Cannabis sativa short tandem repeat loci. Amplification of the loci in four multiplex reactions was tested across DNA from dried root, stem, and leaf sources, and DNA from fresh, frozen, and dried leaf tissue with a template DNA range of 10.0-0.01 ng. The loci were amplified and scored consistently for all DNA sources when DNA template was in the range of 10.0-1.0 ng. Some allelic dropout and PCR failure occurred in reactions with lower template DNA amounts. Overall, amplification was best using 10.0 ng of template DNA from dried leaf tissue indicating that this is the optimal source material. Cross species amplification was observed in Humulus lupulus for three loci but there was no allelic overlap. This is the first study following SWGDAM validation guidelines to validate short tandem repeat markers for forensic use in plants.

  4. Balancing photosynthetic light-harvesting and light-utilization capacities in potato leaf tissue during acclimation to different growth temperatures

    NASA Technical Reports Server (NTRS)

    Steffen, K. L.; Wheeler, R. M.; Arora, R.; Palta, J. P.; Tibbitts, T. W.

    1995-01-01

    We investigated the effect of temperature during growth and development on the relationship between light-harvesting capacity, indicated by chlorophyll concentration, and light-utilization potential, indicated by light- and bicarbonate-saturated photosynthetic oxygen evolution, in Solanum tuberosum L. cv. Norland. Clonal plantlets were transplanted and grown at 20 degrees C for 2 weeks before transfer to 12, 16, 20, 24 and 28 degrees C for 6 weeks. After 4 weeks of the temperature treatments, leaf tissue fresh weights per area were one-third higher in plants grown at 12 degrees C vs those grown at 28 degrees C. Conversely, chlorophyll content per area in tissue grown at 12 degrees C was less than one-half of that of tissue grown at 28 degrees C at 4 weeks. Photosynthetic capacity measured at a common temperature of 20 degrees C and expressed on a chlorophyll basis was inversely proportional to growth temperature. Leaf tissue from plants grown at 12 degrees C for 4 weeks had photosynthetic rates that were 3-fold higher on a chlorophyll basis than comparable tissue from plants grown at 28 degrees C. These results suggest that the relationship between light-harvesting capacity and light-utilization potential varies 3-fold in response to the growth temperatures examined. The role of this response in avoidance of photoinhibition is discussed.

  5. Effects of transgenic Bt rice on growth, reproduction, and superoxide dismutase activity of Folsomia candida (Collembola: Isotomidae) in laboratory studies.

    PubMed

    Bai, Yaoyu; Yan, Ruihong; Ke, Xin; Ye, Gongyin; Huang, Fangneng; Luo, Yongming; Cheng, Jiaan

    2011-12-01

    Transgenic rice expressing Bacillus thuringiensis (Bt) CrylAb protein is expected to be commercialized in China in the near future. The use of Bt rice for controlling insect pests sparks intensive debates regarding its biosafety. Folsomia candida is an euedaphic species and is often used as a "standard" test organism in assessing effects of environmental pollutants on soil organisms. In this study, growth, development, reproduction, and superoxide dismutase activity (SOD) of F. candida were investigated in the laboratory for populations reared on leaf tissue or leaf-soil mixtures of two CrylAb rice lines and a non-Bt rice isoline. Two independent tests were performed: 1) a 35-d test using petri dishes containing yeast diet (positive control) or fresh rice leaf tissue, and 2) a 28-d test in soil-litter microcosms containing yeast or a mixture of soil and rice leaf tissue. Biological parameters measured in both tests were number of progeny production, population growth rate, and SOD activity. For the petri dish test, data measured also included insect body length and number of exuviation. There were no significant differences between the populations reared on Bt and non-Bt rice leaf tissue in all measured parameters in both tests and for both Bt rice lines, suggesting no significant effects of the CrylAb protein in Bt rice on F. candida in the laboratory studies. Results of this study should add additional biosafety proofs for use of Bt rice to manage rice pests in China.

  6. Stimulating productivity of hydroponic lettuce in controlled environments with triacontanol

    NASA Technical Reports Server (NTRS)

    Knight, S. L.; Mitchell, C. A.

    1987-01-01

    Triacontanol (1-triacontanol) applied as a foliar spray at 10(-7) M to 4-day-old, hydroponically grown leaf lettuce (Lactuca sativa L.) seedlings in a controlled environment increased leaf fresh and dry weight 13% to 20% and root fresh and dry weight 13% to 24% 6 days after application, relative to plants sprayed with water. When applied at 8 as well as 4 days after seeding, triacontanol increased plant fresh and dry weight, leaf area, and mean relative growth rate 12% to 37%. There was no benefit of repeating application of triacontanol in terms of leaf dry weight gain.

  7. Metabolic engineering of biomass for high energy density: oilseed-like triacylglycerol yields from plant leaves

    PubMed Central

    Vanhercke, Thomas; El Tahchy, Anna; Liu, Qing; Zhou, Xue-Rong; Shrestha, Pushkar; Divi, Uday K; Ral, Jean-Philippe; Mansour, Maged P; Nichols, Peter D; James, Christopher N; Horn, Patrick J; Chapman, Kent D; Beaudoin, Frederic; Ruiz-López, Noemi; Larkin, Philip J; de Feyter, Robert C; Singh, Surinder P; Petrie, James R

    2014-01-01

    High biomass crops have recently attracted significant attention as an alternative platform for the renewable production of high energy storage lipids such as triacylglycerol (TAG). While TAG typically accumulates in seeds as storage compounds fuelling subsequent germination, levels in vegetative tissues are generally low. Here, we report the accumulation of more than 15% TAG (17.7% total lipids) by dry weight in Nicotiana tabacum (tobacco) leaves by the co-expression of three genes involved in different aspects of TAG production without severely impacting plant development. These yields far exceed the levels found in wild-type leaf tissue as well as previously reported engineered TAG yields in vegetative tissues of Arabidopsis thaliana and N. tabacum. When translated to a high biomass crop, the current levels would translate to an oil yield per hectare that exceeds those of most cultivated oilseed crops. Confocal fluorescence microscopy and mass spectrometry imaging confirmed the accumulation of TAG within leaf mesophyll cells. In addition, we explored the applicability of several existing oil-processing methods using fresh leaf tissue. Our results demonstrate the technical feasibility of a vegetative plant oil production platform and provide for a step change in the bioenergy landscape, opening new prospects for sustainable food, high energy forage, biofuel and biomaterial applications. PMID:24151938

  8. A comparison of leaf crystal macropatterns in the two sister genera Piper and Peperomia (Piperaceae).

    PubMed

    Horner, Harry T; Wanke, Stefan; Samain, Marie-Stéphanie

    2012-06-01

    This is the first large-scale study comparing leaf crystal macropatterns of the species-rich sister genera Piper and Peperomia. It focuses on identifying types of calcium oxalate crystals and their macropatterns in leaves of both genera. The Piper results are placed in a phylogenetic context to show evolutionary patterns. This information will expand knowledge about crystals and provide specific examples to help study their form and function. One example is the first-time observation of Piper crystal sand tumbling in chlorenchyma vacuoles. Herbarium and fresh leaves were cleared of cytoplasmic content and examined with polarizing microscopy to identify types of crystals and their macropatterns. Selected hydrated herbarium and fresh leaf punches were processed for scanning electron microscopy and x-ray elemental analysis. Vibratome sections of living Piper and Peperomia leaves were observed for anatomical features and crystal movement. Both genera have different leaf anatomies. Piper displays four crystal types in chlorenchyma-crystal sand, raphides, styloids, and druses, whereas Peperomia displays three types-druses, raphides, and prisms. Because of different leaf anatomies and crystal types between the genera, macropatterns are completely different. Crystal macropattern evolution in both is characterized by increasing complexity, and both may use their crystals for light gathering and reflection for efficient photosynthesis under low-intensity light environments. Both genera have different leaf anatomies, types of crystals and crystal macropatterns. Based on Piper crystals associated with photosynthetic tissues and low-intensity light, further study of their function and association with surrounding chloroplasts is warranted, especially active crystal movement.

  9. Using the conservative nature of fresh leaf surface density to measure foliar area

    NASA Astrophysics Data System (ADS)

    Castillo, Omar S.; Zaragoza, Esther M.; Alvarado, Carlos J.; Barrera, Maria G.; Dasgupta-Schubert, Nabanita

    2014-10-01

    For a herbaceous species, the inverse of the fresh leaf surface density, the Hughes constant, is nearly conserved. We apply the Hughes constant to develop an absolute method of leafarea measurement that requires no regression fits, prior calibrations or oven-drying. The Hughes constant was determined in situ using a known geometry and weights of a sub-set obtained from the fresh leaves whose areas are desired. Subsequently, the leaf-areas (at any desired stratification level), were derived by utilizing the Hughes constant and the masses of the fresh leaves. The proof of concept was established for leaf-discs of the plants Mandevilla splendens and Spathiphyllum wallisii. The conservativeness of the Hughes constant over individual leaf-zones and different leaftypes from the leaves of each species was quantitatively validated. Using the globally averaged Hughes constant for each species, the leaf-area of these and additional co-species plants, were obtained. The leaf-area-measurement-by-mass was cross-checked with standard digital image analysis. There were no statistically significant differences between the leaf-area-measurement-by-mass and the digital image analysis measured leaf-areas and the linear correlation between the two methods was very good. Leaf-areameasurement- by-mass was found to be rapid and simple with accuracies comparable to the digital image analysis method. The greatly reduced cost of leaf-area-measurement-by-mass could be beneficial for small agri-businesses in developing countries.

  10. Simultaneous qualitative assessment and quantitative analysis of flavonoids in various tissues of lotus (Nelumbo nucifera) using high performance liquid chromatography coupled with triple quad mass spectrometry.

    PubMed

    Chen, Sha; Fang, Linchuan; Xi, Huifen; Guan, Le; Fang, Jinbao; Liu, Yanling; Wu, Benhong; Li, Shaohua

    2012-04-29

    Flavonoid composition and concentration were investigated in 12 different tissues of 'Ti-1' lotus (Nelumbo nucifera) by high performance liquid chromatography equipped with photodiode array detection tandem electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS(n)). A total of 20 flavonoids belonging to six groups (myricetin, quercetin, kaempferol, isohamnetin, diosmetin derivatives) were separated and identified. Myricetin 3-O-galactoside, myricetin 3-O-glucuronide, isorhamnetin 3-O-glucuronide and free aglycone diometin (3',5,7-trihydroxy-4'-methoxyflavone) were first reported in lotus. Flavonoid composition varied largely with tissue type, and diverse compounds (5-15) were found in leaf and flower stalks, flower pistils, seed coats and embryos. Flower tissues including flower petals, stamens, pistils, and, especially, reproductive tissue fruit coats had more flavonoid compounds (15-17) than leaves (12), while no flavonoids were detectable in seed kernels. The flavonoid content of seed embryos was high, 730.95 mg 100g(-1) DW (dry weight). As regards the other tissues, mature leaf pulp (771.79 mg 100 g(-1) FW (fresh weight)) and young leaves (650.67 mg 100 g(-1) FW) had higher total flavonoid amount than flower stamens (359.45 mg 100 g(-1) FW) and flower petals (342.97 mg 100g(-1) FW), while leaf stalks, flower stalks and seed coats had much less total flavonoid (less than 40 mg 100 g(-1) FW). Copyright © 2012 Elsevier B.V. All rights reserved.

  11. Variation of hyperforin in Hypericum montbretii during its phenological cycle.

    PubMed

    Cirak, C; Radusiene, J

    2007-11-01

    Hypericum montbretii, a perennial herbaceous plant from Turkish flora has a great pharmaceutical potential with its well-documented chemical content. In the present study, morphogenetic and phenological variations of hyperforin were investigated in this species for the first time. Wild growing plants were harvested at vegetative, floral budding, full flowering, fresh fruiting, and mature fruiting stages and dissected into stem, leaf and reproductive tissues and assayed for hyperforin by HPLC method. Phenological changes in hyperforin content were found to be significant. After decreasing at floral budding slightly, hyperforin concentration in whole shoots increased with advancing of plant development and the highest level was reached at fresh fruiting. Among different parts of the plant, reproductive tissues namely green capsules and full opened flowers accumulated significantly higher amount of hyperforin when compared to stems and leaves. Such kind of data could be useful for elucidation of the chemotaxonomical significance of hyperforin and phytochemical evaluation of H. montbretii.

  12. Metabolic engineering of biomass for high energy density: oilseed-like triacylglycerol yields from plant leaves.

    PubMed

    Vanhercke, Thomas; El Tahchy, Anna; Liu, Qing; Zhou, Xue-Rong; Shrestha, Pushkar; Divi, Uday K; Ral, Jean-Philippe; Mansour, Maged P; Nichols, Peter D; James, Christopher N; Horn, Patrick J; Chapman, Kent D; Beaudoin, Frederic; Ruiz-López, Noemi; Larkin, Philip J; de Feyter, Robert C; Singh, Surinder P; Petrie, James R

    2014-02-01

    High biomass crops have recently attracted significant attention as an alternative platform for the renewable production of high energy storage lipids such as triacylglycerol (TAG). While TAG typically accumulates in seeds as storage compounds fuelling subsequent germination, levels in vegetative tissues are generally low. Here, we report the accumulation of more than 15% TAG (17.7% total lipids) by dry weight in Nicotiana tabacum (tobacco) leaves by the co-expression of three genes involved in different aspects of TAG production without severely impacting plant development. These yields far exceed the levels found in wild-type leaf tissue as well as previously reported engineered TAG yields in vegetative tissues of Arabidopsis thaliana and N. tabacum. When translated to a high biomass crop, the current levels would translate to an oil yield per hectare that exceeds those of most cultivated oilseed crops. Confocal fluorescence microscopy and mass spectrometry imaging confirmed the accumulation of TAG within leaf mesophyll cells. In addition, we explored the applicability of several existing oil-processing methods using fresh leaf tissue. Our results demonstrate the technical feasibility of a vegetative plant oil production platform and provide for a step change in the bioenergy landscape, opening new prospects for sustainable food, high energy forage, biofuel and biomaterial applications. © 2013 CSIRO. Plant Biotechnology Journal published by Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  13. Spectrometric Estimation of Total Nitrogen Concentration in Douglas-Fir Foliage

    NASA Technical Reports Server (NTRS)

    Johnson, Lee F.; Billow, Christine R.; Peterson, David L. (Technical Monitor)

    1995-01-01

    Spectral measurements of fresh and dehydrated Douglas-fir foliage, from trees cultivated under three fertilization treatments, were acquired with a laboratory spectrophotometer. The slope (first-derivative) of the fresh- and dry-leaf absorbance spectra at locations near known protein absorption features was strongly correlated with total nitrogen (TN) concentration of the foliage samples. Particularly strong correlation was observed between the first-derivative spectra in the 2150-2170 nm region and TN, reaching a local maximum in the fresh-leaf spectra of -0.84 at 2 160 nm. Stepwise regression was used to generate calibration equations relating first derivative spectra from fresh, dry/intact, and dry/ground samples to TN concentration. Standard errors of calibration were 1.52 mg g-1 (fresh), 1.33 (dry/intact), and 1.20 (dry/ground), with goodness-of-fit 0.94 and greater. Cross-validation was performed with the fresh-leaf dataset to examine the predictive capability of the regression method; standard errors of prediction ranged from 1.47 - 2.37 mg g(exp -1) across seven different validation sets, prediction goodness of fit ranged from .85-.94, and wavelength selection was fairly insensitive to the membership of the calibration set. All regressions in this study tended to select wavelengths in the 2100-2350 nm region, with the primary selection in the 2142-2172 nm region. The study provides positive evidence concerning the feasibility of assessing TN status of fresh-leaf samples by spectrometric means. We assert that the ability to extract biochemical information from fresh-leaf spectra is a necessary but insufficient condition regarding the use of remote sensing for canopy-level biochemical estimation.

  14. ANTI-ULCER ACTIVITY OF THE ALKALI PREPARATION OF THE ROOT AND FRESH LEAF JUICE OF MORINGA OLEIFERA LAM

    PubMed Central

    Ruckmani, K.; Kavimani, S.; Jayakar, B.; Anandan, R.

    1998-01-01

    The alkali preparation of the root and fresh leaf juice of Moringa oleifera possessed significant dose –depen-dent anti-ulcer activity in experimentally induced acute gastric ulcers with aspirin, the anti-ulcer effect of the alkali preparation of the root seems to be more pronounced than that of the fresh leaf juice. Te anti-ulcer activity of the alkali preparation of the root could be due to its content of alkaloids or its anticholinergic and antihistaminic activities, or a combination of these factors. PMID:22556845

  15. Quality of fresh organic matter affects priming of soil organic matter and substrate utilization patterns of microbes

    PubMed Central

    Wang, Hui; Boutton, Thomas W.; Xu, Wenhua; Hu, Guoqing; Jiang, Ping; Bai, Edith

    2015-01-01

    Changes in biogeochemical cycles and the climate system due to human activities are expected to change the quantity and quality of plant litter inputs to soils. How changing quality of fresh organic matter (FOM) might influence the priming effect (PE) on soil organic matter (SOM) mineralization is still under debate. Here we determined the PE induced by two 13C-labeled FOMs with contrasting nutritional quality (leaf vs. stalk of Zea mays L.). Soils from two different forest types yielded consistent results: soils amended with leaf tissue switched faster from negative PE to positive PE due to greater microbial growth compared to soils amended with stalks. However, after 16 d of incubation, soils amended with stalks had a higher PE than those amended with leaf. Phospholipid fatty acid (PLFA) results suggested that microbial demand for carbon and other nutrients was one of the major determinants of the PE observed. Therefore, consideration of both microbial demands for nutrients and FOM supply simultaneously is essential to understand the underlying mechanisms of PE. Our study provided evidence that changes in FOM quality could affect microbial utilization of substrate and PE on SOM mineralization, which may exacerbate global warming problems under future climate change. PMID:25960162

  16. Quality of fresh organic matter affects priming of soil organic matter and substrate utilization patterns of microbes.

    PubMed

    Wang, Hui; Boutton, Thomas W; Xu, Wenhua; Hu, Guoqing; Jiang, Ping; Bai, Edith

    2015-05-11

    Changes in biogeochemical cycles and the climate system due to human activities are expected to change the quantity and quality of plant litter inputs to soils. How changing quality of fresh organic matter (FOM) might influence the priming effect (PE) on soil organic matter (SOM) mineralization is still under debate. Here we determined the PE induced by two (13)C-labeled FOMs with contrasting nutritional quality (leaf vs. stalk of Zea mays L.). Soils from two different forest types yielded consistent results: soils amended with leaf tissue switched faster from negative PE to positive PE due to greater microbial growth compared to soils amended with stalks. However, after 16 d of incubation, soils amended with stalks had a higher PE than those amended with leaf. Phospholipid fatty acid (PLFA) results suggested that microbial demand for carbon and other nutrients was one of the major determinants of the PE observed. Therefore, consideration of both microbial demands for nutrients and FOM supply simultaneously is essential to understand the underlying mechanisms of PE. Our study provided evidence that changes in FOM quality could affect microbial utilization of substrate and PE on SOM mineralization, which may exacerbate global warming problems under future climate change.

  17. Quality of fresh organic matter affects priming of soil organic matter and substrate utilization patterns of microbes

    NASA Astrophysics Data System (ADS)

    Wang, Hui; Boutton, Thomas W.; Xu, Wenhua; Hu, Guoqing; Jiang, Ping; Bai, Edith

    2015-05-01

    Changes in biogeochemical cycles and the climate system due to human activities are expected to change the quantity and quality of plant litter inputs to soils. How changing quality of fresh organic matter (FOM) might influence the priming effect (PE) on soil organic matter (SOM) mineralization is still under debate. Here we determined the PE induced by two 13C-labeled FOMs with contrasting nutritional quality (leaf vs. stalk of Zea mays L.). Soils from two different forest types yielded consistent results: soils amended with leaf tissue switched faster from negative PE to positive PE due to greater microbial growth compared to soils amended with stalks. However, after 16 d of incubation, soils amended with stalks had a higher PE than those amended with leaf. Phospholipid fatty acid (PLFA) results suggested that microbial demand for carbon and other nutrients was one of the major determinants of the PE observed. Therefore, consideration of both microbial demands for nutrients and FOM supply simultaneously is essential to understand the underlying mechanisms of PE. Our study provided evidence that changes in FOM quality could affect microbial utilization of substrate and PE on SOM mineralization, which may exacerbate global warming problems under future climate change.

  18. USE OF NATIVE PLANTS FOR REMEDIATION OF TRICHLOROETHYLENE: I. DECIDUOUS TREES.

    PubMed

    Strycharz, S; Newman, L

    2009-02-01

    Phytoremediation of trichloroethylene (TCE) can be accomplished using fast-growing, deep-rooting trees. The most commonly used tree for phytoremediation of TCE has been the hybrid poplar. This study looks at native southeastern trees of the United States as alternatives to the use of hybrid poplar. The use of native trees for phytoremediation allows for simultaneous restoration of contaminated sites. A 2-mo, greenhouse-based study was conducted to determine if sycamore (Plantanus L.), eastern cottonwood (Populus deltoides), sweetgum (Liquidambar styraciflua L.), and willow (Salix sachalinensis) trees possess the ability to degrade TCE by assessing TCE metabolite formation in the plant tissue. In addition to the metabolic capabilities of each tree species, growth parameters were measured including change in height, water usage, total fresh weight of each tissue type, and calculated total leaf surface area. Willow trees had the greatest increase in height among all trees tested; however, at higher concentrations TCE inhibits growth. Sycamore trees had the highest overall leaf surface area and total biomass, which correlated with sycamore trees also having the highest average water usage over the course of the experiment. Carbon tubes used to sample transpiration gases from sycamore, sweetgum, and cottonwood trees did not contain detectable levels of TCE. Tenex sample collection tubes used to sample willow trees during TCE exposure showed average TCE concentrations of up to 0.354 ng TCE cm -2 leaf tissue. All exposed trees contained TCE in the root, stem, and leaf tissues. The concentration of TCE remaining in tissues at the conclusion of the experiment varied, with the highest levels found in the roots and the lowest levels found in the leaves. Metabolites were also observed in different tissue types of all trees tested. The highest concentrations of trichloroacetic acid were observed in the leaves of the sycamore trees and cottonwood trees. Based on the growth parameters tested and the ability to metabolize TCE, sycamore and native cottonwood species are the best candidates for phytoremediation from this study.

  19. Induction of wound-periderm-like tissue in Kalanchoe pinnata (Lam.) Pers. (Crassulaceae) leaves as a defence response to high UV-B radiation levels

    PubMed Central

    Nascimento, Luana Beatriz dos Santos; Moreira, Nattacha dos Santos; Leal-Costa, Marcos Vinícius; Costa, Sônia Soares; Tavares, Eliana Schwartz

    2015-01-01

    Background and Aims UV-B radiation can be stressful for plants and cause morphological and biochemical changes. Kalanchoe pinnata is a CAM leaf-succulent species distributed in hot and dry regions, and is rich in flavonoids, which are considered to be protective against UV-B radiation. This study aims to verify if K. pinnata has morphological or anatomical responses as a strategy in response to high UV-B levels. Methods Kalanchoe pinnata plants of the same age were grown under white light (control) or white light plus supplemental UV-B radiation (5 h d–1). The plants were treated with the same photoperiod, photosynthetically active radiation, temperature and daily watering system. Fragments of the middle third of the leaf blade and petiole were dehydrated and then embedded in historesin and sectioned in a rotary microtome. Sections were stained with toluidine blue O and mounted in Entellan®. Microchemical analyses by optical microscopy were performed on fresh material with Sudan III, Sudan IV and phloroglucinol, and analysed using fluorescence microscopy. Key Results Supplemental UV-B radiation caused leaf curling and the formation of brown areas on the leaves. These brown areas developed into a protective tissue on the adaxial side of the leaf, but only in directly exposed regions. Anatomically, this protective tissue was similar to a wound-periderm, with outer layer cell walls impregnated with suberin and lignin. Conclusions This is the first report of wound-periderm formation in leaves in response to UV-B radiation. This protective tissue could be important for the survival of the species in desert regions under high UV-B stress conditions. PMID:26346722

  20. Potato leaf explants as a spaceflight plant test system

    NASA Technical Reports Server (NTRS)

    Wheeler, R. M.

    1986-01-01

    The use of explant tissues or organs may circumvent limitations facing whole-plant experimentation during spaceflight. In the case of potato, a crop currently being studied for application to bioregenerative life support systems, excised leaves and their subtended axillary buds can be used to test a variety of stem growth and development phases ranging from tubers through stolons (horizontal stems) to upright leafy shoots. The leaves can be fit well into small-volume test packages and sustained under relatively low irradiance levels using light-weight growing media. Tubers formed on potato leaf cuttings can yield up from 0.5 to 1.0 g fresh mass 10 days after excision and up to 2.0 g or more, 14 days from excision.

  1. Extraction of high-quality DNA from ethanol-preserved tropical plant tissues.

    PubMed

    Bressan, Eduardo A; Rossi, Mônica L; Gerald, Lee T S; Figueira, Antonio

    2014-04-24

    Proper conservation of plant samples, especially during remote field collection, is essential to assure quality of extracted DNA. Tropical plant species contain considerable amounts of secondary compounds, such as polysaccharides, phenols, and latex, which affect DNA quality during extraction. The suitability of ethanol (96% v/v) as a preservative solution prior to DNA extraction was evaluated using leaves of Jatropha curcas and other tropical species. Total DNA extracted from leaf samples stored in liquid nitrogen or ethanol from J. curcas and other tropical species (Theobroma cacao, Coffea arabica, Ricinus communis, Saccharum spp., and Solanum lycopersicon) was similar in quality, with high-molecular-weight DNA visualized by gel electrophoresis. DNA quality was confirmed by digestion with EcoRI or HindIII and by amplification of the ribosomal gene internal transcribed spacer region. Leaf tissue of J. curcas was analyzed by light and transmission electron microscopy before and after exposure to ethanol. Our results indicate that leaf samples can be successfully preserved in ethanol for long periods (30 days) as a viable method for fixation and conservation of DNA from leaves. The success of this technique is likely due to reduction or inactivation of secondary metabolites that could contaminate or degrade genomic DNA. Tissue conservation in 96% ethanol represents an attractive low-cost alternative to commonly used methods for preservation of samples for DNA extraction. This technique yields DNA of equivalent quality to that obtained from fresh or frozen tissue.

  2. Extraction of high-quality DNA from ethanol-preserved tropical plant tissues

    PubMed Central

    2014-01-01

    Background Proper conservation of plant samples, especially during remote field collection, is essential to assure quality of extracted DNA. Tropical plant species contain considerable amounts of secondary compounds, such as polysaccharides, phenols, and latex, which affect DNA quality during extraction. The suitability of ethanol (96% v/v) as a preservative solution prior to DNA extraction was evaluated using leaves of Jatropha curcas and other tropical species. Results Total DNA extracted from leaf samples stored in liquid nitrogen or ethanol from J. curcas and other tropical species (Theobroma cacao, Coffea arabica, Ricinus communis, Saccharum spp., and Solanum lycopersicon) was similar in quality, with high-molecular-weight DNA visualized by gel electrophoresis. DNA quality was confirmed by digestion with EcoRI or HindIII and by amplification of the ribosomal gene internal transcribed spacer region. Leaf tissue of J. curcas was analyzed by light and transmission electron microscopy before and after exposure to ethanol. Our results indicate that leaf samples can be successfully preserved in ethanol for long periods (30 days) as a viable method for fixation and conservation of DNA from leaves. The success of this technique is likely due to reduction or inactivation of secondary metabolites that could contaminate or degrade genomic DNA. Conclusions Tissue conservation in 96% ethanol represents an attractive low-cost alternative to commonly used methods for preservation of samples for DNA extraction. This technique yields DNA of equivalent quality to that obtained from fresh or frozen tissue. PMID:24761774

  3. Evaluation of cycloate followed by two-leaf stage phenmedipham application in fresh market spinach

    USDA-ARS?s Scientific Manuscript database

    Fresh market spinach has one primary herbicide, cycloate, which does not control all weeds. Previous studies demonstrated that cycloate PRE followed by (fb) phenmedipham at the four-leaf spinach stage is a safe and effective treatment. However, this treatment is not useful for the main growing seaso...

  4. Variations in the chemical composition of cassava ( Manihot esculenta Crantz) leaves and roots as affected by genotypic and environmental variation.

    PubMed

    Burns, Anna Elizabeth; Gleadow, Roslyn Margaret; Zacarias, Anabela M; Cuambe, Constantino Estevão; Miller, Rebecca Elizabeth; Cavagnaro, Timothy Richard

    2012-05-16

    The purpose of this study was to assess the quality of cassava cultivars, in terms of cyanogenic potential and composition of macro- and micronutrients, sampled from different locations in rural Mozambique. Total cyanide concentrations in fresh cassava tissues were measured using portable cyanide testing kits, and elemental nutrients were later analyzed from dried plant tissue. Variation in cyanogenic potential and nutrient composition occurred both among cultivars and across locations. The majority of cultivars contained >100 ppm total cyanide, fresh weight, and are therefore considered to be dangerously poisonous unless adequately processed before consumption. Leaf cyanogenic and nutrient content varied with plant water status, estimated using carbon isotope discrimination (δ(13)C). The colonization of roots of all cultivars by arbuscular mycorrhizal fungi was also quantified and found to be high, indicating that mycorrhizas could play a key role in plant nutrient acquisition in these low-input farming systems.

  5. New uses for calcium chloride solution as a mounting medium.

    PubMed

    Herr, J M

    1992-01-01

    Fresh cross sections of stems (Psilotum nudum, Coleus blumei, and Pelargonium peltatum) and roots (Setcreasea purpurea) 120 microns thick were fixed in FPA50 (formalin: propionic acid: 50% ethanol, 5:5:90, v/v) for 24 hr and stored in 70% ethanol. The sections were transferred to water and then to 1% phloroglucin in 20% calcium chloride solution plus either hydrochloric, nitric, or lactic acid in the following ratios of phloroglucin-CaCl2 solution:acid: 25:4, 20:2, or 15:5. The sections were mounted on slides either in one of the three mixtures or in fresh 20% calcium chloride solution. A rapid reaction of the acid-phloroglucin with lignin produced a deep red color in tracheary elements and an orange-red color in sclerenchyma. Fixed and stored leaf pieces from Nymphaea odorata were autoclaved in lactic acid, washed in two changes of 95% ethanol, transferred to water, and treated with the three acid-phloroglucin-calcium chloride mixtures. The abundant astrosclereids stained an orange-red color similar to that of sclerenchyma in the sections. In addition, a new method is reported for specifically staining lignified tissues. When sections or leaf pieces are stained in aqueous 0.05% toluidine blue O, then placed in 20% calcium chloride solution, all tissues destain except those with lignified or partially lignified cell walls. Thus, toluidine blue O applied as described becomes a reliable specific test for lignin comparable to the acid-phloroglucin test.

  6. Evaluation of in vitro enzymatic and non-enzymatic antioxidant properites of leaf extract from Alpinia Purpurata (Vieill.) K. Schum.

    PubMed

    Raj, Chinthamony Arul; Ragavendran, Paramasivam; Sophia, Dominic; Starlin, Thangarajan; Rathi, Muthian Ahalliya; Gopalakrishnan, Velliyur Kanniappan

    2016-09-01

    To evaluate the enzymatic and non-enzymatic antioxidants of leaf extract from Alpinia purpurata. One gram of fresh leaf of Alpinia purpurata was grinded in 2 mL of 50% ethanol and centrifuged at 10,000×g at 4°C for 10 min. The supernatant obtained was used within 4 h for various enzymatic antioxidants assays like superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), ascorbate oxidase, peroxidase, polyphenol oxidase (PPO) and non-enzymatic antioxidants such as vitamin C, total reduced glutathione (TRG) and lipid peroxidation (LPO). The leaf extract of Alpinia purpurata possess antioxidants like vitamin C 472.92±6.80 μg/mg protein, GST 372.11±5.70 μmol of 1-chloro 2,4 dinitrobenzene (CDNB)-reduced glutathione (GSH) conjugate formed/min/mg protein, GPx 281.69±6.43 μg of glutathione oxidized/min/mg protein, peroxidases 173.12±9.40 μmol/g tissue, TRG 75.27±3.55 μg/mg protein, SOD 58.03±2.11 U/mg protein, CAT 46.70±2.35 μmol of H2O2 consumed/min/mg protein in high amount whereas ascorbate oxidase 17.41±2.46 U/g tissue, LPO 2.71±0.14 nmol/L of malondialdehyde formed/min/mg protein and PPO 1.14±0.11 μmol/g tissue in moderate amount. Alpinia purpurata has the potential to scavenge the free radicals and protect against oxidative stress causing diseases. In future, Alpinia purpurata may serve as a good pharmacotherapeutic agent.

  7. 18O Spatial Patterns of Vein Xylem Water, Leaf Water, and Dry Matter in Cotton Leaves

    PubMed Central

    Gan, Kim Suan; Wong, Suan Chin; Yong, Jean Wan Hong; Farquhar, Graham Douglas

    2002-01-01

    Three leaf water models (two-pool model, Péclet effect, and string-of-lakes) were assessed for their robustness in predicting leaf water enrichment and its spatial heterogeneity. This was achieved by studying the 18O spatial patterns of vein xylem water, leaf water, and dry matter in cotton (Gossypium hirsutum) leaves grown at different humidities using new experimental approaches. Vein xylem water was collected from intact transpiring cotton leaves by pressurizing the roots in a pressure chamber, whereas the isotopic content of leaf water was determined without extracting it from fresh leaves with the aid of a purpose-designed leaf punch. Our results indicate that veins have a significant degree of lateral exchange with highly enriched leaf water. Vein xylem water is thus slightly, but progressively enriched in the direction of water flow. Leaf water enrichment is dependent on the relative distances from major veins, with water from the marginal and intercostal regions more enriched and that next to veins and near the leaf base more depleted than the Craig-Gordon modeled enrichment of water at the sites of evaporation. The spatial pattern of leaf water enrichment varies with humidity, as expected from the string-of-lakes model. This pattern is also reflected in leaf dry matter. All three models are realistic, but none could fully account for all of the facets of leaf water enrichment. Our findings acknowledge the presence of capacitance in the ground tissues of vein ribs and highlight the essential need to incorporate Péclet effects into the string-of-lakes model when applying it to leaves. PMID:12376664

  8. Canopy and seasonal profiles of nitrate reductase in soybeans

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Harper, J.E.; Hageman, R.H.

    1972-01-01

    Nitrate reductase activity of soybeans (Glycine max L. Merr.) was evaluated in soil plots and outdoor hydroponic gravel culture systems throughout the growing season. Nitrate reductase profiles within the plant canopy were also established. Mean activity per gram fresh weight per hour of the entire plant canopy was highest in the seedling stage while total activity (activity per gram fresh weight per hour times the total leaf weight) reached a maximum when plants were in the full bloom to midpod fill stage. Nitrate reductase activity per gram fresh weight per hour was highest in the uppermost leaf just prior tomore » full expansion and declined with leaf positions lower in the canopy. Total nitrate reductase activity per leaf was also highest in the uppermost fully expanded leaf during early growth stages. Maximum total activity shifted to leaf positions lower in the plant canopy with later growth stages. Nitrate reductase activity of soybeans grown in hydroponic systems was significantly higher than activity of adjacent soil grown plants at later growth stages, which suggested that under normal field conditions the potential for nitrate utilization may not be realized. Nitrate reductase activity per gram fresh weight per hour and nitrate content were positively correlated over the growing season with plants grown in either soil or solution culture. Computations based upon the nitrate reductase assay of plants grown in hydroponics indicated that from 1.7 to 1.8 grams N could have been supplied to the plant via the nitrate reductase process. 11 references, 9 figures, 3 tables.« less

  9. Induction of wound-periderm-like tissue in Kalanchoe pinnata (Lam.) Pers. (Crassulaceae) leaves as a defence response to high UV-B radiation levels.

    PubMed

    Nascimento, Luana Beatriz dos Santos; Moreira, Nattacha dos Santos; Leal-Costa, Marcos Vinícius; Costa, Sônia Soares; Tavares, Eliana Schwartz

    2015-10-01

    UV-B radiation can be stressful for plants and cause morphological and biochemical changes. Kalanchoe pinnata is a CAM leaf-succulent species distributed in hot and dry regions, and is rich in flavonoids, which are considered to be protective against UV-B radiation. This study aims to verify if K. pinnata has morphological or anatomical responses as a strategy in response to high UV-B levels. Kalanchoe pinnata plants of the same age were grown under white light (control) or white light plus supplemental UV-B radiation (5 h d(-1)). The plants were treated with the same photoperiod, photosynthetically active radiation, temperature and daily watering system. Fragments of the middle third of the leaf blade and petiole were dehydrated and then embedded in historesin and sectioned in a rotary microtome. Sections were stained with toluidine blue O and mounted in Entellan®. Microchemical analyses by optical microscopy were performed on fresh material with Sudan III, Sudan IV and phloroglucinol, and analysed using fluorescence microscopy. Supplemental UV-B radiation caused leaf curling and the formation of brown areas on the leaves. These brown areas developed into a protective tissue on the adaxial side of the leaf, but only in directly exposed regions. Anatomically, this protective tissue was similar to a wound-periderm, with outer layer cell walls impregnated with suberin and lignin. This is the first report of wound-periderm formation in leaves in response to UV-B radiation. This protective tissue could be important for the survival of the species in desert regions under high UV-B stress conditions. © The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  10. Use of a SPAD-502 meter to measure leaf chlorophyll concentration in Arabidopsis thaliana.

    PubMed

    Ling, Qihua; Huang, Weihua; Jarvis, Paul

    2011-02-01

    The SPAD-502 meter is a hand-held device that is widely used for the rapid, accurate and non-destructive measurement of leaf chlorophyll concentrations. It has been employed extensively in both research and agricultural applications, with a range of different plant species. However, its utility has not been fully exploited in relation to the most intensively studied model organism for plant science research, Arabidopsis thaliana. Measurements with the SPAD-502 meter produce relative SPAD meter values that are proportional to the amount of chlorophyll present in the leaf. In order to convert these values into absolute units of chlorophyll concentration, calibration curves must be derived and utilized. Here, we present calibration equations for Arabidopsis that can be used to convert SPAD values into total chlorophyll per unit leaf area (nmol/cm(2); R(2) = 0.9960) or per unit fresh weight of leaf tissue (nmol/mg; R(2) = 0.9809). These relationships were derived using a series of Arabidopsis chloroplast biogenesis mutants that exhibit chlorophyll deficiencies of varying severity, and were verified by the subsequent analysis of senescent or light-stressed leaves. Our results revealed that the converted SPAD values differ from photometric measurements of solvent-extracted chlorophyll by just ~6% on average.

  11. Do we Underestimate the Importance of Leaf Size in Plant Economics? Disproportional Scaling of Support Costs Within the Spectrum of Leaf Physiognomy

    PubMed Central

    Niinemets, Ülo; Portsmuth, Angelika; Tena, David; Tobias, Mari; Matesanz, Silvia; Valladares, Fernando

    2007-01-01

    Background Broad scaling relationships between leaf size and function do not take into account that leaves of different size may contain different fractions of support in petiole and mid-rib. Methods The fractions of leaf biomass in petiole, mid-rib and lamina, and the differences in chemistry and structure among mid-ribs, petioles and laminas were investigated in 122 species of contrasting leaf size, life form and climatic distribution to determine the extent to which differences in support modify whole-lamina and whole-leaf structural and chemical characteristics, and the extent to which size-dependent support investments are affected by plant life form and site climate. Key Results For the entire data set, leaf fresh mass varied over five orders of magnitude. The percentage of dry mass in mid-rib increased strongly with lamina size, reaching more than 40 % in the largest laminas. The whole-leaf percentage of mid-rib and petiole increased with leaf size, and the overall support investment was more than 60 % in the largest leaves. Fractional support investments were generally larger in herbaceous than in woody species and tended to be lower in Mediterranean than in cool temperate and tropical plants. Mid-ribs and petioles had lower N and C percentages, and lower dry to fresh mass ratio, but greater density (mass per unit volume) than laminas. N percentage of lamina without mid-rib was up to 40 % higher in the largest leaves than the total-lamina (lamina and mid-rib) N percentage, and up to 60 % higher than whole-leaf N percentage, while lamina density calculated without mid-rib was up to 80 % less than that with the mid-rib. For all leaf compartments, N percentage was negatively associated with density and dry to fresh mass ratio, while C percentage was positively linked to these characteristics, reflecting the overall inverse scaling between structural and physiological characteristics. However, the correlations between N and C percentages and structural characteristics differed among mid-ribs, petioles and laminas, implying that the mass-weighted average leaf N and C percentage, density, and dry to fresh mass ratio can have different functional values depending on the importance of within-leaf support investments. Conclusions These data demonstrate that variation in leaf size is associated with major changes in within-leaf support investments and in large modifications in integrated leaf chemical and structural characteristics. These size-dependent alterations can importantly affect general leaf structure vs. function scaling relationships. These data further demonstrate important life-form effects on and climatic differentiation in foliage support costs. PMID:17586597

  12. Prediction of leaf area in individual leaves of cherrybark oak seedlings (Quercus pagoda Raf.)

    Treesearch

    Yanfei Guo; Brian Lockhart; John Hodges

    1995-01-01

    The prediction of leaf area for cherrybark oak (Quercus pagoda Raf.) seedlings is important for studying the physiology of the species. Linear and polynomial models involving leaf length, width, fresh weight, dry weight, and internodal length were tested independently and collectively to predict leaf area. Twenty-nine cherrybark oak seedlings were...

  13. Tibiofemoral Contact Mechanics with Horizontal Cleavage Tear and Resection of the Medial Meniscus in the Human Knee.

    PubMed

    Koh, Jason L; Yi, Seung Jin; Ren, Yupeng; Zimmerman, Todd A; Zhang, Li-Qun

    2016-11-02

    The meniscus is known to increase the contact area and decrease contact pressure in the tibiofemoral compartments of the knee. Radial tears of the meniscal root attachment along with partial resections of the torn meniscal tissue decrease the contact area and increase pressure; however, there is a lack of information on the effects of a horizontal cleavage tear (HCT) and partial leaf meniscectomy of such tears on tibiofemoral contact pressure and contact area. Twelve fresh-frozen human cadaveric knees were tested under 10 conditions: 5 serial conditions of posterior medial meniscectomy (intact meniscus, HCT, repaired HCT, inferior leaf resection, and resection of both inferior and superior leaves), each at 2 knee flexion angles (0° and 60°) under an 800-N axial load. Tekscan sensors (model 4000) were used to measure the contact pressure and contact area. HCT and HCT repair resulted in small changes in the contact area and an increase in contact pressure compared with the intact condition. Resection of the inferior leaf resulted in significantly decreased contact area (to a mean 82.3% of the intact condition at 0° of flexion and 81.8% at 60° of flexion; p < 0.05) and increased peak contact pressure (a mean 36.3% increase at 0° flexion and 43.2% increase at 60° flexion; p < 0.05) in the medial compartment. Further resection of the remaining superior leaf resulted in additional significant decreases in contact area (to a mean 60.1% of the intact condition at 0° of flexion and 49.7% at 60° of flexion; p < 0.05) and increases in peak contact pressure (a mean 79.2% increase at 0° of flexion and 74.9% increase at 60° of flexion; p < 0.05). Resection of meniscal tissue forming the inferior leaf of an HCT resulted in substantially decreased contact area and increased contact pressure. Additional resection of the superior leaf resulted in a further significant decrease in contact area and increase in contact pressure in the medial compartment. Repair or minimal resection of meniscal tissue of an HCT may be preferred to complete leaf resection to maintain knee tibiofemoral contact mechanics. Copyright © 2016 by The Journal of Bone and Joint Surgery, Incorporated.

  14. [Allelopathy of grape root aqueous extracts].

    PubMed

    Li, Kun; Guo, Xiu-wu; Guo, Yin-shan; Li, Cheng-xiang; Xie, Hong-gang; Hu, Xi-xi; Zhang, Li-heng; Sun, Ying-ni

    2010-07-01

    Taking the tissue-cultured seedlings of grape cultivar Red Globe as test objects, this paper examined the effects of their root aqueous extracts on seedling's growth, with the allelochemicals identified by LC-MS. The results showed that 0.02 g x ml(-1) (air-dried root mass in aqueous extracts volume; the same below), 0.1 g x ml(-1), and 0.2 g x ml(-1) of the aqueous extracts inhibited the growth of the seedlings significantly, and the inhibition effect increased with increasing concentration of the extracts. The identified allelochemicals of the extracts included p-hydroxybenzoic acid, salicylic acid, phenylpropionic acid, and coumaric acid. Pot experiment showed that different concentration (0.1, 1, and 10 mmol x L(-1)) salicylic acid and phenylpropionic acid inhibited the seedling' s growth remarkably. With the increasing concentration of the two acids, the plant height, stem diameter, shoot- and root fresh mass, leaf net photosynthetic rate and starch content, and root activity of the seedlings decreased, while the leaf soluble sugar and MDA contents increased. No obvious change pattern was observed in leaf protein content.

  15. Ozone Sensitivity in Sweet Corn (Zea mays L.) Plants: A Possible Relationship to Water Balance

    PubMed Central

    Harris, Michael J.; Heath, Robert L.

    1981-01-01

    Stomatal characteristics affecting gas exchange were compared in two sweet corn hybrids (var. Bonanza and Monarch Advance) which differ in foliar ozone sensitivity. No significant differences were observed in stomatal frequencies, guard cell lengths, or conductances to water vapor diffusion. When plant water status was compared, leaf-water potentials for the ozone-resistant cultivar, Bonanza, were lower. A comparison of the relationship between soil- and leaf-water potential indicated that the leaf-water potential of Bonanza was more sensitive to declines in soil-water potential. Additionally, a comparison of stomatal conductance to water vapor diffusion as soil moisture declined and following root detachment indicated that stomata of Bonanza were, likewise, more sensitive to increasing water stress. Data suggest that these differences are attributable to a greater shoot-to-root fresh weight ratio and higher resistance to water movement in the water-conducting tissues for Bonanza. Our observations suggest that root and water delivery system characteristics play a major contributory role in the determination of foliar ozone sensitivity in this species. PMID:16662019

  16. The impact of hydration changes in fresh bio-tissue on THz spectroscopic measurements.

    PubMed

    Png, G M; Choi, J W; Ng, B W-H; Mickan, S P; Abbott, D; Zhang, X-C

    2008-07-07

    We present a study of how residual hydration in fresh rat tissue samples can vastly alter their extracted terahertz (THz) optical properties and influence their health assessment. Fresh (as opposed to preserved) tissue most closely mimics in vivo conditions, but high water content creates many challenges for tissue handling and THz measurement. Our THz measurements of fresh tissue over time highlight the effect of tissue hydration on tissue texture and dimension, the latter directly influencing the accuracy of calculated optical properties. We then introduce lyophilization (freeze drying) as a viable solution for overcoming hydration and freshness problems. Lyophilization removes large amounts of water while retaining sample freshness. In addition, lyophilized tissue samples are easy to handle and their textures and dimensions do not vary over time, allowing for consistent and stable THz measurements. A comparison of lyophilized and fresh tissue shows for the first time that freeze drying may be one way of overcoming tissue hydration issues while preserving tissue cellular structure. Finally, we compare THz measurements from fresh tissue against necrotic tissue to verify freshness over time. Indeed, THz measurements from fresh and necrotic tissues show marked differences.

  17. Cloning and characterization of a serotonin N-acetyltransferase from a gymnosperm, loblolly pine (Pinus taeda).

    PubMed

    Park, Sangkyu; Byeon, Yeong; Lee, Hyoung Yool; Kim, Young-Soon; Ahn, Taeho; Back, Kyoungwhan

    2014-10-01

    Serotonin N-acetyltransferase (SNAT) is the penultimate enzyme in melatonin biosynthesis in both animals and plants. SNAT catalyzes serotonin into N-acetylserotonin, an immediate precursor for melatonin biosynthesis by N-acetylserotonin methyltransferase (ASMT). We cloned the SNAT gene from a gymnosperm loblolly pine (Pinus teada). The loblolly pine SNAT (PtSNAT) gene encodes 255 amino acids harboring a transit sequence with 67 amino acids and shows 67% amino acid identity with rice SNAT when comparing the mature polypeptide regions. Purified recombinant PtSNAT showed peak activity at 55°C with the K(m) (428 μM) and Vmax (3.9 nmol/min/mg protein) values. As predicted, PtSNAT localized to chloroplasts. The SNAT mRNA was constitutively expressed in all tissues, including leaf, bud, flower, and pinecone, whereas the corresponding protein was detected only in leaf. In accordance with the exclusive SNAT protein expression in leaf, melatonin was detected only in leaf at 0.45 ng per gram fresh weight. Sequence and phylogenetic analysis indicated that the gymnosperm PtSNAT had high homology with SNATs from all plant phyla (even with cyanobacteria), and formed a clade separated from the angiosperm SNATs, suggestive of direct gene transfer from cyanobacteria via endosymbiosis. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Effect of fertigation through drip and micro sprinkler on plant biometric characters in cocoa (Theobroma cacao L.).

    PubMed

    Krishnamoorthy, C; Rajamani, K

    2013-12-15

    A field experiment to study the influence of fertigation of N, P and K fertilizers on biometric characters of cocoa (Theobroma cacao L.) was conducted at the Department of Spices and Plantation Crops, Horticultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore during January 2010 to December 2011. The experiment was laid out with thirteen treatments replicated three times in a randomized block design. A phenomenal increase in growth parameters such as trunk girth, canopy spread and weight of the pruned branches removed, leaf fresh weight and leaf dry weight was observed with increasing levels of NPK as well as methods of fertilizer application in this study. Among the various treatments, fertigation with 125% 'Recommended Dose of Fertilizers' (125:50:175 g NPK plant year(-1)) as Water Soluble Fertilizers (WSF) through drip irrigation increased all vegetative growth parameters like trunk girth increment (1.62 cm), canopy spread increment (66.79 cm), leaf fresh weight (3.949 g), leaf dry weight (2.039 g), weight of the pruned branches removed (fresh weight 7.628 kg plant(-1)) and dry weight (4.650 kg plant(-1)).

  19. Is There a Relation between the Microscopic Leaf Morphology and the Association of Salmonella and Escherichia coli O157:H7 with Iceberg Lettuce Leaves?

    PubMed

    VAN der Linden, Inge; Eriksson, Markus; Uyttendaele, Mieke; Devlieghere, Frank

    2016-10-01

    To prevent contamination of fresh produce with enteric pathogens, more insight into mechanisms that may influence the association of these pathogens with fresh produce is needed. In this study, Escherichia coli O157:H7 and Salmonella were chosen as model pathogens, and fresh cut iceberg lettuce was chosen as a model fresh produce type. The morphological structure of iceberg lettuce leaves (stomatal density and length of cell margins per leaf area) was quantified by means of leaf peels and light microscopy of leaves at different stages of development (outer, middle, and inner leaves of the crop) on both leaf sides (abaxial and adxial) and in three leaf regions (top, center, and bottom). The morphology of the top region of the leaves was distinctly different from that of the center and base, with a significantly higher stomatal density (up to five times more stomata), different cell shape, and longer cell margins (two to three times longer). Morphological differences between the same regions of the leaves at different stages of development were smaller or nonsignificant. An attachment assay with two attenuated E. coli O157:H7 strains (84-24h11-GFP and BRMSID 188 GFP) and two Salmonella strains (serovars Thompson and Typhimurium) was performed on different regions of the middle leaves. Our results confirmed earlier reports that these pathogens have a higher affinity for the base of the lettuce leaf than the top. Differences of up to 2.12 log CFU/g were seen ( E. coli O157:H7 86-24h11-GFP). Intermediate attachment occurred in the central region. The higher incidence of preferential bacterial attachment sites such as stomata and cell margins or grooves could not explain the differences observed in the association of the tested pathogens with different regions of iceberg lettuce leaves.

  20. Confirmation of 1-Phenylethane-1-thiol as the Character Impact Aroma Compound in Curry Leaves and Its Behavior during Tissue Disruption, Drying, and Frying.

    PubMed

    Steinhaus, Martin

    2017-03-15

    The most odor-active compounds previously identified by application of an aroma extract dilution analysis were quantitated in freshly picked curry leaves, either by stable isotope dilution assays in combination with GC-GC-MS or by GC-FID after simultaneous extraction/fractionation. Odor activity values (OAVs) were calculated as ratios of concentrations to odor threshold values. The topmost OAVs were obtained for (3Z)-hex-3-enal (grassy; OAV 180 000), (1S)-1-phenylethane-1-thiol (sulfury, burnt; OAV 150 000), (1R)-1-phenylethane-1-thiol (sulfury, burnt; OAV 120 000), (3R)-linalool (citrusy; OAV 58 000), and myrcene (geranium leaf-like; OAV 23 000). The high OAVs calculated for its enantiomers confirmed 1-phenylethane-1-thiol as character impact compound of the typical sulfury and burnt aroma of curry leaves. The 1-phenylethane-1-thiol concentration in curry leaves decreased upon tissue disruption and drying, as well as upon frying of fresh leaves. By contrast, frying of dried leaves led to an increase of 1-phenylethane-1-thiol, indicating a yet unknown thermolabile precursor.

  1. Quantitative study of Xanthosoma violaceum leaf surfaces using RIMAPS and variogram techniques.

    PubMed

    Favret, Eduardo A; Fuentes, Néstor O; Molina, Ana M

    2006-08-01

    Two new imaging techniques (rotated image with maximum averaged power spectrum (RIMAPS) and variogram) are presented for the study and description of leaf surfaces. Xanthosoma violaceum was analyzed to illustrate the characteristics of both techniques. Both techniques produce a quantitative description of leaf surface topography. RIMAPS combines digitized images rotation with Fourier transform, and it is used to detect patterns orientation and characteristics of surface topography. Variogram relates the mathematical variance of a surface with the area of the sample window observed. It gives the typical scale lengths of the surface patterns. RIMAPS detects the morphological variations of the surface topography pattern between fresh and dried (herbarium) samples of the leaf. The variogram method finds the characteristic dimensions of the leaf microstructure, i.e., cell length, papillae diameter, etc., showing that there are not significant differences between dry and fresh samples. The results obtained show the robustness of RIMAPS and variogram analyses to detect, distinguish, and characterize leaf surfaces, as well as give scale lengths. Both techniques are tools for the biologist to study variations of the leaf surface when different patterns are present. The use of RIMAPS and variogram opens a wide spectrum of possibilities by providing a systematic, quantitative description of the leaf surface topography.

  2. Eucalypt smoke and wildfires

    NASA Astrophysics Data System (ADS)

    Maleknia, Simin D.; Bell, Tina L.; Adams, Mark A.

    2009-01-01

    Eucalypt contributions to biogenic sources of volatile organic compounds (VOCs) in Australia are estimated at teragram (Tg = 1012 g) amounts each year. Biogenic VOCs include plant-specific isoprenoids (isoprene and a range of terpenes) and other reactive organic compounds (i.e., acids, aldehydes and ketones). Atmospheric reactions of VOCs are numerous and many have significant environmental impact. Wildfires increase both the amounts of VOCs released and the complexity of their reactions. Proton-transfer reaction mass spectrometry (PTR-MS), gas chromatography mass spectrometry (GCMS) and direct analysis in real time (DART) mass spectrometry were applied to analyze release of VOCs as a function of temperatures ranging from ambient to combustion. PTR-MS enabled trace level analysis of VOCs from a complex forest atmosphere and revealed the release of terpenes associated with leaf damage during a storm. Temperature profile studies revealed ion abundances (i.e., emissions of VOCs) could be correlated with boiling points and vapor pressures of specific compounds. PTR-MS analysis of VOCs resulting from heating fresh leaf (E. grandis) material suggested that emissions of protonated methanol (m/z 33) and protonated acetaldehyde (m/z 45) were greatest at ~60 °C while m/z 137 and 153 (associated with a series of terpenes) showed monotonic increases in ion abundance over a wide temperature range from ambient to 200 °C. GCMS analysis of fresh and senescent leaves of E. grandis showed that a series of VOCs (ethylvinylketone, diethylketone, 2-ethylfuran, hexanal and hexenals) are present only in fresh leaves while several terpenes ([alpha] and [beta] pinenes, [alpha]-phellandrene, eucalyptol, [gamma]-terpinene) were common in both. DART analysis of fresh leaf and stem of E. sideroxylon identified tissue-specific VOCs (e.g., methanol and ethanol were more abundant in stems). PTR-MS combustion studies of senescent leaves (E. grandis) identified two distinct, temperature-dependent VOC compositions. Before the appearance of smoke, the composition of VOCs remained consistent and correlated well with various naturally occurring isoprenoids, as observed in temperature profile studies. Sampling of eucalypt smoke suggested ions (m/z 75, 85, 87, 99, 111 and 125) correlated with protonated mass of oxygenated aldehydes, ketones, furans and substituted benzenes, and were due to pyrolysis of polycarbohydrates (cellulose and lignin) that are common in many types of wood.

  3. Antioxidant effect of olive leaf powder on fresh Atlantic horse mackerel (Trachurus trachurus) minced muscle

    USDA-ARS?s Scientific Manuscript database

    There is a growing interest in the replacement of synthetic food antioxidants by natural extracts. Olive leaf is an abundant by-product of the olive oil industry. The aim of this study was to investigate the antioxidant capacity of olive leaf powder (OLP) in chilled minced muscle. Concentrations of ...

  4. Iodine uptake by spinach (Spinacia oleracea L.) plants grown in solution culture: effects of iodine species and solution concentrations.

    PubMed

    Zhu, Y-G; Huang, Y-Z; Hu, Y; Liu, Y-X

    2003-04-01

    A hydroponic experiment was carried out to investigate the effects of iodine species and solution concentrations on iodine uptake by spinach (Spinacia oleracea L.). Five iodine concentrations (0, 1, 10, 50 and 100 microM) for iodate (IO(3)(-)) and iodide (I(-)) were used. Results show that higher concentrations of I(-) (> or =10 microM) had some detrimental effect on plant growth, while IO(3)(-) had little effect on the biomass production of spinach plants. Increases in iodine concentration in the growth solution significantly enhanced I concentrations in plant tissues. The detrimental effect of I(-) on plant growth was probably due to the excessively high accumulation of I in plant tissues. The solution-to-spinach leaf transfer factors (TF(leaf), fresh weight basis) for plants treated with iodide were between 14.2 and 20.7 at different solution concentrations of iodide; TF(leaf) for plants treated with iodate decreased gradually from 23.7 to 2.2 with increasing solution concentrations of iodate. The distribution coefficients (DCs) of I between leaves and roots were constantly higher for plants treated with iodate than those treated with iodide. DCs for plants treated with iodide increased with increasing solution concentrations of iodide, while DCs for plants treated with iodate (around 5.5) were similar across the range of solution concentrations of iodate used in this experiment. The implications of iodine accumulation in leafy vegetables in human iodine nutrition are also discussed. Copyright 2002 Elsevier Science Ltd.

  5. Whole-leaf sanitizing wash improves chlorine efficacy for microbial reduction and prevents pathogen cross contamination during fresh-cut lettuce processing

    USDA-ARS?s Scientific Manuscript database

    Currently, nearly all fresh-cut lettuce processing facilities in the United States use chlorinated water or other sanitizer solutions for microbial reduction after lettuce is cut. It is believed that freshly cut lettuce releases significant amounts of organic matters that negatively impact the effec...

  6. Regulation of Purine Metabolism in Intact Leaves of Coffea arabica.

    PubMed

    Nazario, G. M.; Lovatt, C. J.

    1993-12-01

    The capacity of Coffea arabica leaves (5- x 5-mm pieces) to synthesize de novo and catabolize purine nucleotides to provide precursors for caffeine (1,3,7-trimethylxanthine) was investigated. Consistent with de novo synthesis, glycine, bicarbonate, and formate were incorporated into the purine ring of inosine 5[prime]-monophosphate (IMP) and adenine nucleotides ([sigma]Ade); azaserine, a known inhibitor of purine de novo synthesis, inhibited incorporation. Activity of the de novo pathway in C. arabica per g fresh weight of leaf tissue during a 3-h incubation period was 8 [plus or minus] 4 nmol of formate incorporated into IMP, 61 [plus or minus] 7 nmol into [sigma]Ade, and 150 nmol into caffeine (the latter during a 7-h incubation). Coffee leaves exhibited classical purine catabolism. Radiolabeled formate, inosine, adenosine, and adenine were incorporated into hypoxanthine and xanthine, which were catabolized to allantoin and urea. Urease activity was demonstrated. Per g fresh weight, coffee leaf squares incorporated 90 [plus or minus] 22 nmol of xanthine into caffeine in 7 h but degraded 102 [plus or minus] 1 nmol of xanthine to allantoin in 3 h. Feedback control of de novo purine biosynthesis was contrasted in C. arabica and Cucurbita pepo, a species that does not synthesize purine alkaloids. End-product inhibition was demonstrated to occur in both species but at different enzyme reactions.

  7. Carbon and nitrogen balance of leaf-eating sesarmid crabs ( Neoepisesarma versicolor) offered different food sources

    NASA Astrophysics Data System (ADS)

    Thongtham, Nalinee; Kristensen, Erik

    2005-10-01

    Carbon and nitrogen budgets for the leaf-eating crab, Neoepisesarma versicolor, were established for individuals living on pure leaf diets. Crabs were fed fresh (green), senescent (yellow) and partly degraded (brown) leaves of the mangrove tree Rhizophora apiculata. Ingestion, egestion and metabolic loss of carbon and nitrogen were determined from laboratory experiments. In addition, bacterial abundance in various compartments of the crabs' digestive tract was enumerated after dissection of live individuals. Ingestion and egestion rates (in terms of dry weight) were highest, while the assimilation efficiency was poorest for crabs fed on brown leaves. The low assimilation efficiency was more than counteracted by the high ingestion rate providing more carbon for growth than for crabs fed green and yellow leaves. In any case, the results show that all types of leaves can provide adequate carbon while nitrogen was insufficient to support both maintenance (yellow leaves) and growth (green, yellow and brown leaves). Leaf-eating crabs must therefore obtain supplementary nitrogen by other means in order to meet their nitrogen requirement. Three hypotheses were evaluated: (1) crabs supplement their diet with bacteria and benthic microalgae by ingesting own faeces and/or selective grazing at the sediment surface; (2) assimilation of symbiotic nitrogen-fixing bacteria in the crabs' own intestinal system; and (3) nitrogen storage following occasional feeding on animal tissues (e.g. meiofauna and carcasses). It appears that hypothesis 1 is of limited importance for N. versicolor since faeces and sediment can only supply a minor fraction of the missing nitrogen due to physical constraints on the amount of material the crabs can consume. Hypothesis 2 can be ruled out because tests showed no nitrogen fixation activity in the intestinal system of N. versicolor. It is therefore likely that leaf-eating crabs provide most of their nitrogen requirement from intracellular deposits following occasionally ingestion of animal tissue (hypothesis 3).

  8. Phenotypic plasticity of Neonotonia wightii and Pueraria phaseoloides grown under different light intensities.

    PubMed

    Santos, Leonardo D T; Da Cruz, Leandro R; Dos Santos, Samuel A; Sant'anna-Santos, Bruno F; Dos Santos, Izabela T; De Oliveira, Ariane M; Barros, Rodrigo E; Santos, Márcia V; Faria, Rodrigo M

    2015-03-01

    Plants have the ability to undergo morphophysiological changes based on availability of light. The present study evaluated biomass accumulation, leaf morphoanatomy and physiology of Neonotonia wightii and Pueraria phaseoloides grown in full sunlight, as well as in 30% and 50% shade. Two assays were performed, one for each species, using a randomized block design with 10 replicates. A higher accumulation of fresh mass in the shoot of the plants was observed for both species under cultivation in 50% shade, while no differences were detected between the full sunlight and 30% shade. N. wightii and P. phaseoloides showed increase in area and reduction in thickness leaf when cultivated in 50% shade. There were no changes in photosynthetic rate, stomatal conductance, water use efficiency and evapotranspiration of P. phaseoloides plants because growth environment. However, the shade treatments caused alterations in physiological parameters of N. wightii. In both species, structural changes in the mesophyll occurred depending on the availability of light; however, the amount of leaf blade tissue remained unaltered. Despite the influence of light intensity variation on the morphophysiological plasticity of N. wightii and P. phaseoloides, no effects on biomass accumulation were observed in response to light.

  9. Growth promoting potential of fresh and stored Moringa oleifera leaf extracts in improving seedling vigor, growth and productivity of wheat crop.

    PubMed

    Khan, Shahbaz; Basra, Shahzad Maqsood Ahmed; Afzal, Irfan; Nawaz, Muhammad; Rehman, Hafeez Ur

    2017-12-01

    Wheat is staple food of region, as it contributes 60% of daily caloric intake, but its delayed sowing reduces yield due to short life span. Moringa leaf extract (MLE) is considered to improve growth and development of field crops. Study comprised of two experiments. First experiment, freshly extracted MLE and in combination with growth-promoting substances were stored at two temperature regimes. Chemical analysis, after 1, 2, and 3 months' storage period, showed that phenolics and ascorbic acid concentrations decreased with increasing storage period. Fresh extracts improved speed and spread of emergence and seedling vigor. Effectiveness of MLE in terms of phenolics and ascorbate concentrations was highest up to 1 month which decreased with prolonged storage. Growth enhancing potential of MLE also reduced with increasing storage duration. Under field conditions, the bio-efficacy of these fresh and stored MLE was compared when applied as foliar spray at tillering and booting stages of wheat. Foliar applied fresh MLE was the most effective in improving growth parameters. Fresh MLE enhanced biochemical and yield attributes in late sown wheat. This growth-promoting potential of MLE decreased with storage time. Application of fresh MLE helped to achieve higher economic yield.

  10. Comparison study on biosynthesis of silver nanoparticles using fresh and hot air oven dried IMPERATA CYLINDRICA leaf

    NASA Astrophysics Data System (ADS)

    Najmi Bonnia, Noor; Fairuzi, Afiza Ahmad; Akhir, Rabiatuladawiyah Md.; Yahya, Sabrina M.; Rani, Mohd Azri Ab; Ratim, Suzana; Rahman, Norafifah A.; Akil, Hazizan Md

    2018-01-01

    The perennial rhizomatous grass; Imperata cylindrica (I. cylindrica) has been reported rich in various phytochemicals. In present study, silver nanoparticles were synthesized from aqueous leaf extract of I. cylindrica at two different leaf conditions; fresh leaves and hot-air oven dried leaves. Biosynthesized silver nanoparticles were characterized by UV-visible spectroscopy, field emission scanning electron microscopy (FESEM) and Fourier transform infrared spectroscopy (FTIR). Maximum absorption was recorded between 400 nm to 500 nm. FESEM analysis revealed that the silver nanoparticles predominantly form spherical shapes. The particles sizes were ranging from 22-37 nm. The elemental composition of the synthesized silver nanoparticles was confirmed by using energy dispersive X-ray spectroscopy (EDX) analysis. Fourier transform infrared spectroscopy (FTIR) confirmed the reducing and stabilizing actions came from biomolecules associated with I. cylindrica leaf extract. Thus in this investigation, an environmentally safe method to synthesized silver nanoparticles using local plant extract was successfully established.

  11. Effects of nitrogen fertilization on growth and reflectance characteristics of winter wheat

    NASA Technical Reports Server (NTRS)

    Hinzman, L. D.; Bauer, M. E.; Daughtry, C. S. T.

    1986-01-01

    The use of remote sensing to determine seasonal changes in agronomic and spectral properties of winter wheat canopies with different levels of N fertilization is investigated. Field experiments were conducted at Purdue Agronomy Farm, West Lafayette, IN during the 1978-1979 and 1979-1980 growing season. Spectral reflectance, total leaf N concentration, leaf chlorophyll concentration, leaf are index (LAI), and fresh and dry phytomass are measured and analyzed. Three distinct wheat canopies are detected for the O, 60, and 120 kg N/ha levels of fertilization; it is observed that with an increase in N the reflectance in the visible, and middle IR wavelengths decrease, and the IR reflectance is increased. The canopies with 120 kg N/ha display the highest LAI, maintain green leaf area the longest, and increase in fresh and dry phytomass. The relationship between spectral and agronomic variables is examined; the effect of changing chlorophyll concentration and LAI on the reflectance is studied.

  12. Simplified methods for screening cowpea cultivars for manganese leaf-tissue tolerance. [Vigna unguiculata

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wissemeier, A.H.; Horst, W.J.

    In cowpea (Vigna unguiculata (L.) Walp.) dark brown speckles on old leaves are typical symptoms of Mn toxicity and indicate Mn sensitivity of leaf tissue. Induction and subsequent quantification of brown Mn speckles in leaf tissues were used to screen cowpea cultivars for Mn leaf-tissue tolerance using three different techniques: (i) leaf cuttings cultured for 22 days in solution culture with 20 {mu}M MnSO{sub 4}, (ii) leaf rings mounted on leaves of intact plants and filled with 500 {mu}M MnSO{sub 4} for 5 days, and (iii) leaf disks floated for 3 days on 500 {mu}M MnSO{sub 4}. Density of brownmore » speckles differed considerably among the six cultivars tested, and was not related to the Mn concentrations of the leaf tissues. There were close relationships between genotypic Mn-toxicity symptom expression and depression of dry matter production of the cultivars at high Mn supply in a long-term sand culture experiment. The floating leaf-disk method is particularly suited for screening large numbers of cowpea cultivars for Mn leaf-tissue tolerance because it requires only 3 days. The ranking of the cultivars for Mn tolerance was highly correlated to Mn tolerance of intact plants.« less

  13. Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virus-derived transgenes from plant tissues.

    PubMed

    Ndunguru, Joseph; Taylor, Nigel J; Yadav, Jitender; Aly, Haytham; Legg, James P; Aveling, Terry; Thompson, Graham; Fauquet, Claude M

    2005-05-18

    Plant viral diseases present major constraints to crop production. Effective sampling of the viruses infecting plants is required to facilitate their molecular study and is essential for the development of crop protection and improvement programs. Retaining integrity of viral pathogens within sampled plant tissues is often a limiting factor in this process, most especially when sample sizes are large and when operating in developing counties and regions remote from laboratory facilities. FTA is a paper-based system designed to fix and store nucleic acids directly from fresh tissues pressed into the treated paper. We report here the use of FTA as an effective technology for sampling and retrieval of DNA and RNA viruses from plant tissues and their subsequent molecular analysis. DNA and RNA viruses were successfully recovered from leaf tissues of maize, cassava, tomato and tobacco pressed into FTA Classic Cards. Viral nucleic acids eluted from FTA cards were found to be suitable for diagnostic molecular analysis by PCR-based techniques and restriction analysis, and for cloning and nucleotide sequencing in a manner equivalent to that offered by tradition isolation methods. Efficacy of the technology was demonstrated both from sampled greenhouse-grown plants and from leaf presses taken from crop plants growing in farmer's fields in East Africa. In addition, FTA technology was shown to be suitable for recovery of viral-derived transgene sequences integrated into the plant genome. Results demonstrate that FTA is a practical, economical and sensitive method for sampling, storage and retrieval of viral pathogens and plant genomic sequences, when working under controlled conditions and in the field. Application of this technology has the potential to significantly increase ability to bring modern analytical techniques to bear on the viral pathogens infecting crop plants.

  14. Impairment of leaf photosynthesis after insect herbivory or mechanical injury on common milkweed, Asclepias syriaca.

    PubMed

    Delaney, K J; Haile, F J; Peterson, R K D; Higley, L G

    2008-10-01

    Insect herbivory has variable consequences on plant physiology, growth, and reproduction. In some plants, herbivory reduces photosynthetic rate (Pn) activity on remaining tissue of injured leaves. We sought to better understand the influence of leaf injury on Pn of common milkweed, Asclepias syriaca (Asclepiadaceae), leaves. Initially, we tested whether Pn reductions occurred after insect herbivory or mechanical injury. We also (1) examined the duration of photosynthetic recovery, (2) compared mechanical injury with insect herbivory, (3) studied the relationship between leaf Pn with leaf injury intensity, and (4) considered uninjured leaf compensatory Pn responses neighboring an injured leaf. Leaf Pn was significantly reduced on mechanically injured or insect-fed leaves in all reported experiments except one, so some factor(s) (cardiac glycoside induction, reproductive investment, and water stress) likely interacts with leaf injury to influence whether Pn impairment occurs. Milkweed tussock moth larval herbivory, Euchaetes egle L. (Arctiidae), impaired leaf Pn more severely than mechanical injury in one experiment. Duration of Pn impairment lasted > 5 d to indicate high leaf Pn sensitivity to injury, but Pn recovery occurred within 13 d in one experiment. The degree of Pn reduction was more severe from E. egle herbivory than similar levels of mechanical tissue removal. Negative linear relationships characterized leaf Pn with percentage tissue loss from single E. egle-fed leaves and mechanically injured leaves and suggested that the signal to trigger leaf Pn impairment on remaining tissue of an injured leaf was amplified by additional tissue loss. Finally, neighboring uninjured leaves to an E. egle-fed leaf had a small (approximately 10%) degree of compensatory Pn to partly offset tissue loss and injured leaf Pn impairment.

  15. Prospecting for Energy-Rich Renewable Raw Materials: Agave Leaf Case Study.

    PubMed

    Corbin, Kendall R; Byrt, Caitlin S; Bauer, Stefan; DeBolt, Seth; Chambers, Don; Holtum, Joseph A M; Karem, Ghazwan; Henderson, Marilyn; Lahnstein, Jelle; Beahan, Cherie T; Bacic, Antony; Fincher, Geoffrey B; Betts, Natalie S; Burton, Rachel A

    2015-01-01

    Plant biomass from different species is heterogeneous, and this diversity in composition can be mined to identify materials of value to fuel and chemical industries. Agave produces high yields of energy-rich biomass, and the sugar-rich stem tissue has traditionally been used to make alcoholic beverages. Here, the compositions of Agave americana and Agave tequilana leaves are determined, particularly in the context of bioethanol production. Agave leaf cell wall polysaccharide content was characterized by linkage analysis, non-cellulosic polysaccharides such as pectins were observed by immuno-microscopy, and leaf juice composition was determined by liquid chromatography. Agave leaves are fruit-like--rich in moisture, soluble sugars and pectin. The dry leaf fiber was composed of crystalline cellulose (47-50% w/w) and non-cellulosic polysaccharides (16-22% w/w), and whole leaves were low in lignin (9-13% w/w). Of the dry mass of whole Agave leaves, 85-95% consisted of soluble sugars, cellulose, non-cellulosic polysaccharides, lignin, acetate, protein and minerals. Juice pressed from the Agave leaves accounted for 69% of the fresh weight and was rich in glucose and fructose. Hydrolysis of the fructan oligosaccharides doubled the amount of fermentable fructose in A. tequilana leaf juice samples and the concentration of fermentable hexose sugars was 41-48 g/L. In agricultural production systems such as the tequila making, Agave leaves are discarded as waste. Theoretically, up to 4000 L/ha/yr of bioethanol could be produced from juice extracted from waste Agave leaves. Using standard Saccharomyces cerevisiae strains to ferment Agave juice, we observed ethanol yields that were 66% of the theoretical yields. These data indicate that Agave could rival currently used bioethanol feedstocks, particularly if the fermentation organisms and conditions were adapted to suit Agave leaf composition.

  16. Prospecting for Energy-Rich Renewable Raw Materials: Agave Leaf Case Study

    PubMed Central

    Corbin, Kendall R.; Byrt, Caitlin S.; Bauer, Stefan; DeBolt, Seth; Chambers, Don; Holtum, Joseph A. M.; Karem, Ghazwan; Henderson, Marilyn; Lahnstein, Jelle; Beahan, Cherie T.; Bacic, Antony; Fincher, Geoffrey B.; Betts, Natalie S.; Burton, Rachel A.

    2015-01-01

    Plant biomass from different species is heterogeneous, and this diversity in composition can be mined to identify materials of value to fuel and chemical industries. Agave produces high yields of energy-rich biomass, and the sugar-rich stem tissue has traditionally been used to make alcoholic beverages. Here, the compositions of Agave americana and Agave tequilana leaves are determined, particularly in the context of bioethanol production. Agave leaf cell wall polysaccharide content was characterized by linkage analysis, non-cellulosic polysaccharides such as pectins were observed by immuno-microscopy, and leaf juice composition was determined by liquid chromatography. Agave leaves are fruit-like—rich in moisture, soluble sugars and pectin. The dry leaf fiber was composed of crystalline cellulose (47–50% w/w) and non-cellulosic polysaccharides (16–22% w/w), and whole leaves were low in lignin (9–13% w/w). Of the dry mass of whole Agave leaves, 85–95% consisted of soluble sugars, cellulose, non-cellulosic polysaccharides, lignin, acetate, protein and minerals. Juice pressed from the Agave leaves accounted for 69% of the fresh weight and was rich in glucose and fructose. Hydrolysis of the fructan oligosaccharides doubled the amount of fermentable fructose in A. tequilana leaf juice samples and the concentration of fermentable hexose sugars was 41–48 g/L. In agricultural production systems such as the tequila making, Agave leaves are discarded as waste. Theoretically, up to 4000 L/ha/yr of bioethanol could be produced from juice extracted from waste Agave leaves. Using standard Saccharomyces cerevisiae strains to ferment Agave juice, we observed ethanol yields that were 66% of the theoretical yields. These data indicate that Agave could rival currently used bioethanol feedstocks, particularly if the fermentation organisms and conditions were adapted to suit Agave leaf composition. PMID:26305101

  17. Determination of total flavonoids content in fresh Ginkgo biloba leaf with different colors using near infrared spectroscopy

    NASA Astrophysics Data System (ADS)

    Shi, Ji-yong; Zou, Xiao-bo; Zhao, Jie-wen; Mel, Holmes; Wang, Kai-liang; Wang, Xue; Chen, Hong

    Total flavonoids content is often considered an important quality index of Ginkgo biloba leaf. The feasibility of using near infrared (NIR) spectra at the wavelength range of 10,000-4000 cm-1 for rapid and nondestructive determination of total flavonoids content in G. biloba leaf was investigated. 120 fresh G. biloba leaves in different colors (green, green-yellowish and yellow) were used to spectra acquisition and total flavonoids determination. Partial least squares (PLS), interval partial least squares (iPLS) and synergy interval partial least squares (SiPLS) were used to develop calibration models for total flavonoids content in two colors leaves (green-yellowish and yellow) and three colors leaves (green, green-yellowish and yellow), respectively. The level of total flavonoids content for green, green-yellowish and yellow leaves was in an increasing order. Two characteristic wavelength regions (5840-6090 cm-1 and 6620-6880 cm-1), which corresponded to the absorptions of two aromatic rings in basic flavonoid structure, were selected by SiPLS. The optimal SiPLS model for total flavonoids content in the two colors leaves (r2 = 0.82, RMSEP = 2.62 mg g-1) had better performance than PLS and iPLS models. It could be concluded that NIR spectroscopy has significant potential in the nondestructive determination of total flavonoids content in fresh G. biloba leaf.

  18. Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters

    PubMed Central

    Martínez-Rodríguez, Julia del C.; la Mora-Amutio, Marcela De; Plascencia-Correa, Luis A.; Audelo-Regalado, Esmeralda; Guardado, Francisco R.; Hernández-Sánchez, Elías; Peña-Ramírez, Yuri J.; Escalante, Adelfo; Beltrán-García, Miguel J.; Ogura, Tetsuya

    2014-01-01

    Agave tequilana Weber var. ‘Azul’ is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB) by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI). Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost. PMID:25763038

  19. Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters.

    PubMed

    Martínez-Rodríguez, Julia del C; De la Mora-Amutio, Marcela; Plascencia-Correa, Luis A; Audelo-Regalado, Esmeralda; Guardado, Francisco R; Hernández-Sánchez, Elías; Peña-Ramírez, Yuri J; Escalante, Adelfo; Beltrán-García, Miguel J; Ogura, Tetsuya

    2014-01-01

    Agave tequilana Weber var. 'Azul' is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB) by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI). Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost.

  20. Does Carica papaya leaf-extract increase the platelet count? An experimental study in a murine model.

    PubMed

    Dharmarathna, Sinhalagoda Lekamlage Chandi Asoka; Wickramasinghe, Susiji; Waduge, Roshitha Nilmini; Rajapakse, Rajapakse Peramune Veddikkarage Jayanthe; Kularatne, Senanayake Abeysinghe Mudiyanselage

    2013-09-01

    To investigate the potential role of fresh Carica papaya (C. papaya) leaf extract on haematological and biochemical parameters and toxicological changes in a murine model. In total 36 mice were used for the trial. Fresh C. papaya leaf extract [0.2 mL (2 g)/mouse] was given only to the test group (18 mice). General behavior, clinical signs and feeding patterns were recorded. Blood and tissue samples were collected at intervals. Haematological parameters including platelet, red blood cell (RBC), white blood cell (WBC), packed cell volume (PCV), serum biochemistry including serum creatinine, serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic-pyruvic transaminase (SGPT) were determined. Organs for possible histopathological changes were examined. Neither group exhibited alteration of behavior or reduction in food and water intake. Similarly, no significant changes in SGOT, SGPT and serum creatinine levels were detected in the test group. Histopathological organ changes were not observed in either group of mice except in three liver samples of the test group which had a mild focal necrosis. The platelet count (11.33±0.35)×10⁵/µL (P=0.00004) and the RBC count (7.97±0.61)×10⁶/µL (P=0.00003) were significantly increased in the test group compared to that of the controls. However, WBC count and PCV (%) values were not changed significantly in the test group. The platelet count in the test group started to increase significantly from Day 3 (3.4±0.18×10⁵/µL), reaching almost a fourfold higher at Day 21 (11.3×10⁵/µL), while it was 3.8×10⁵/µL and 5.5×10⁵/µL at Day 3 and Day 21 respectively in the control. Likewise, the RBC count in the test group increased from 6×10⁶/µL to 9×10⁶/ µL at Day 21 while it remained near constant in the control group (6×10⁶/µL). Fresh C. papaya leaf extract significantly increased the platelet and RBC counts in the test group as compared to controls. Therefore, it is very important to identify those chemicals of C. papaya leaves as it can be recommended to be used as a medication to boost thrombopoiesis and erythropoiesis in humans and in animals in which these cell lineages have been compromised.

  1. Does Carica papaya leaf-extract increase the platelet count? An experimental study in a murine model

    PubMed Central

    Dharmarathna, Sinhalagoda Lekamlage Chandi Asoka; Wickramasinghe, Susiji; Waduge, Roshitha Nilmini; Rajapakse, Rajapakse Peramune Veddikkarage Jayanthe; Kularatne, Senanayake Abeysinghe Mudiyanselage

    2013-01-01

    Objective To investigate the potential role of fresh Carica papaya (C. papaya) leaf extract on haematological and biochemical parameters and toxicological changes in a murine model. Methods In total 36 mice were used for the trial. Fresh C. papaya leaf extract [0.2 mL (2 g)/mouse] was given only to the test group (18 mice). General behavior, clinical signs and feeding patterns were recorded. Blood and tissue samples were collected at intervals. Haematological parameters including platelet, red blood cell (RBC), white blood cell (WBC), packed cell volume (PCV), serum biochemistry including serum creatinine, serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic-pyruvic transaminase (SGPT) were determined. Organs for possible histopathological changes were examined. Results Neither group exhibited alteration of behavior or reduction in food and water intake. Similarly, no significant changes in SGOT, SGPT and serum creatinine levels were detected in the test group. Histopathological organ changes were not observed in either group of mice except in three liver samples of the test group which had a mild focal necrosis. The platelet count (11.33±0.35)×105/µL (P=0.000 04) and the RBC count (7.97±0.61)×106/µL (P=0.000 03) were significantly increased in the test group compared to that of the controls. However, WBC count and PCV (%) values were not changed significantly in the test group. The platelet count in the test group started to increase significantly from Day 3 (3.4±0.18×105/µL), reaching almost a fourfold higher at Day 21 (11.3×105/µL), while it was 3.8×105/µL and 5.5×105/µL at Day 3 and Day 21 respectively in the control. Likewise, the RBC count in the test group increased from 6×106/µL to 9×106/ µL at Day 21 while it remained near constant in the control group (6×106/µL). Conclusions Fresh C. papaya leaf extract significantly increased the platelet and RBC counts in the test group as compared to controls. Therefore, it is very important to identify those chemicals of C. papaya leaves as it can be recommended to be used as a medication to boost thrombopoiesis and erythropoiesis in humans and in animals in which these cell lineages have been compromised. PMID:23998013

  2. Acid-soluble nucleotides of pinto bean leaves at different stages of development.

    PubMed

    Weinstein, L H; McCune, D C; Mancini, J F; van Leuken, P

    1969-11-01

    Acid-soluble nucleotides of unifoliate leaves of Pinto bean plants (Phaseolus vulgaris L.) were determined at young, mature, and senescent stages of development. At least 25 components could be distinguished on the basis of inorganic phosphorus determinations and 37 or more fractions on the basis of (32)P labeling, with adenosine di- and triphosphates accounting for 60% of the total moles of nucleotide. The total nucleotide P and inorganic P, on a fresh weight basis, decreased about 44% between each stage of leaf development, but decrements in the levels of individual nucleotides varied from this over-all pattern.Minor changes in the relative abundance of the individual nucleotides accompanied aging although the percentage of purine-containing nucleotides decreased with age. Total (32)P activity per leaf in the nucleotide pool increased about 3-fold between the young and mature leaves and decreased slightly as leaves became senescent. In general, the specific activities of the nucleotides increased with increased age and adenosine-, guanosine-, uridine-, and cytidine triphosphates and adenosine diphosphate accounted for approximately 90% of the total activity. The changes in the relative sizes and energy status of the nucleotide pools were not so obvious as the changes in other metabolites that have been reported to accompany aging in leaf tissue.

  3. Role of aquaporins in determining transpiration and photosynthesis in water-stressed plants: crop water-use efficiency, growth and yield.

    PubMed

    Moshelion, Menachem; Halperin, Ofer; Wallach, Rony; Oren, Ram; Way, Danielle A

    2015-09-01

    The global shortage of fresh water is one of our most severe agricultural problems, leading to dry and saline lands that reduce plant growth and crop yield. Here we review recent work highlighting the molecular mechanisms allowing some plant species and genotypes to maintain productivity under water stress conditions, and suggest molecular modifications to equip plants for greater production in water-limited environments. Aquaporins (AQPs) are thought to be the main transporters of water, small and uncharged solutes, and CO2 through plant cell membranes, thus linking leaf CO2 uptake from the intercellular airspaces to the chloroplast with water loss pathways. AQPs appear to play a role in regulating dynamic changes of root, stem and leaf hydraulic conductivity, especially in response to environmental changes, opening the door to using AQP expression to regulate plant water-use efficiency. We highlight the role of vascular AQPs in regulating leaf hydraulic conductivity and raise questions regarding their role (as well as tonoplast AQPs) in determining the plant isohydric threshold, growth rate, fruit yield production and harvest index. The tissue- or cell-specific expression of AQPs is discussed as a tool to increase yield relative to control plants under both normal and water-stressed conditions. © 2014 John Wiley & Sons Ltd.

  4. Fluorescent Staining of Tea Pathogenic Fungi in Tea Leaves Using Fluorescein-labeled Lectin

    NASA Astrophysics Data System (ADS)

    Yamada, Kengo; Yoshida, Katsuyuki; Sonoda, Ryoichi

    Fluorochrome-labeled lectin, fluorescein conjugated wheat germ agglutinin (F-WGA) was applied to stain tea pathogenic fungi in tea leaf tissue. Infected leaves were fixed and decolorized with a mixture of ethanol and acetic acid, and cleared with 10% KOH for whole mount before staining with F-WGA. Hyphae of Pestalotiopsis longiseta, Pseudocercospora ocellata, Botrytis cinerea and Colletotrichum theae-sinensis fluoresced brightly in whole mount and sectioned samples of infected leaf tissue. In browned tissue, hyphae did not fluoresce frequently in whole mount sample. Autofluorescence of leaf tissue was strong in browned tissue of sections, it was removed by 10% KOH treatment before staining. Penetration hyphae of C. theae-sinensis in cell wall of trichome and hyphae in basal part of trichome did not fluoresced frequently. In whole mount samples of tea leaf infected with Exobasidium vexans and E. reticulatum, hymenia appeared on leaf surface fluoresced, but hyphae in leaf tissue did not fluoresce. In sectioned samples, hyphae fluoresced brightly when sections were treated with 10% KOH before staining.

  5. Predicting the presence of whiteflies and tomato yellow leaf curl virus in Florida tomato fields

    USDA-ARS?s Scientific Manuscript database

    Florida is one of the leading states for production of fresh market tomatoes. Production is severely affected by Tomato yellow leaf curl virus (TYLCV). The objective of this study was to identify landscape and climatic factors that drive whitefly populations and TYLCV incidence in commercial tomato ...

  6. Expression of red-shifted green fluorescent protein by Escherichia coli O157:H7 as a marker for the detection of cells on fresh produce.

    PubMed

    Takeuchi, K; Frank, J F

    2001-03-01

    Escherichia coli O157:H7 was transformed with a plasmid vector red-shifted green fluorescence protein (pEGFP) to express red-shifted green fluorescence protein (EGFP) from Aequorea victoria. The EGFP expression among total cells and nonviable cells was determined at the cellular level by microscopic observation of immunostained and membrane-impermeable, dye-stained cultures, respectively. E. coli O157:H7 retained pEGFP during frozen storage at -80 degrees C. The percentage of EGFP expression was improved by repeated subculturing, reaching 83.4 +/- 0.1%, although the fluorescence intensity varied among cells. A low percentage of EGFP-expressing cells was nonviable. The percentage of EGFP decreased when the culture plate was kept at 4 degrees C, suggesting that some cells lost pEGFP during refrigeration. The storage of the culture suspension in sterile deionized water at 4 degrees C for 24 h reduced the percentage of EGFP expression, indicating that some EGFP was denatured. The application of EGFP as a marker for E. coli O157:H7 on green leaf lettuce, cauliflower, and tomato was evaluated using confocal scanning laser microscopy. EGFP-transformed cells were readily visible under confocal scanning laser microscopy on all produce types. The numbers of E. coli O157:H7 cells detected with EGFP were equivalent to those detected with immunostaining for green leaf lettuce and cauliflower but less for tomato. E. coli O157:H7 attached preferentially to damaged tissues of green leaf lettuce and tomato over intact tissue surfaces and to flowerets of cauliflower than to stem surfaces. EGFP can serve as a marker to characterize E. coli O157:H7 attachment on green leaf lettuce and cauliflower but may not be suitable on tomato.

  7. Whole-leaf wash improves chlorine efficacy for microbial reduction and prevents pathogen cross-contamination during fresh-cut lettuce processing.

    PubMed

    Nou, Xiangwu; Luo, Yaguang

    2010-06-01

    Currently, most fresh-cut processing facilities in the United States use chlorinated water or other sanitizer solutions for microbial reduction after lettuce is cut. Freshly cut lettuce releases significant amounts of organic matter that negatively impacts the effectiveness of chlorine or other sanitizers for microbial reduction. The objective of this study is to evaluate whether a sanitizer wash before cutting improves microbial reduction efficacy compared to a traditional postcutting sanitizer wash. Romaine lettuce leaves were quantitatively inoculated with E. coli O157:H7 strains and washed in chlorinated water before or after cutting, and E. coli O157:H7 cells that survived the washing process were enumerated to determine the effectiveness of microbial reduction for the 2 cutting and washing sequences. Whole-leaf washing in chlorinated water improved pathogen reduction by approximately 1 log unit over traditional cut-leaf sanitization. Similar improvement in the reduction of background microflora was also observed. Inoculated "Lollo Rossa" red lettuce leaves were mixed with noninoculated Green-Leaf lettuce leaves to evaluate pathogen cross-contamination during processing. High level (96.7% subsamples, average MPN 0.6 log CFU/g) of cross-contamination of noninoculated green leaves by inoculated red leaves was observed when mixed lettuce leaves were cut prior to washing in chlorinated water. In contrast, cross-contamination of noninoculated green leaves was significantly reduced (3.3% of subsamples, average MPN

  8. Carbon Dioxide Metabolism in Leaf Epidermal Tissue 1

    PubMed Central

    Willmer, C. M.; Pallas, J. E.; Black, C. C.

    1973-01-01

    A number of plant species were surveyed to obtain pure leaf epidermal tissue in quantity. Commelina communis L. and Tulipa gesnariana L. (tulip) were chosen for further work. Chlorophyll a/b ratios of epidermal tissues were 2.41 and 2.45 for C. communis and tulip, respectively. Phosphoenolpyruvate carboxylase, ribulose-1,5-diphosphate carboxylase, malic enzyme, and NAD+ and NADP+ malate dehydrogenases were assayed with epidermal tissue and leaf tissue minus epidermal tissue. In both species, there was less ribulose 1,5-diphosphate than phosphoenolpyruvate carboxylase activity in epidermal tissue whether expressed on a protein or chlorophyll basis whereas the reverse was true for leaf tissue minus epidermal tissue. In both species, malic enzyme activities were higher in epidermal tissue than in the remaining leaf tissue when expressed on a protein or chlorophyll basis. In both species, NAD+ and NADP+ malate dehydrogenase activities were higher in the epidermal tissue when expressed on a chlorophyll basis; however, on a protein basis, the converse was true. Microautoradiography of C. communis epidermis and histochemical tests for keto acids suggested that CO2 fixation occurred predominantly in the guard cells. The significance and possible location of the enzymes are discussed in relation to guard cell metabolism. Images PMID:16658581

  9. Endophytic Ability of Different Isolates of Entomopathogenic Fungi Beauveria bassiana (Balsamo) Vuillemin in Stem and Leaf Tissues of Maize (Zea mays L.).

    PubMed

    Renuka, S; Ramanujam, B; Poornesha, B

    2016-06-01

    The present study was conducted to examine the ability of six promising indigenous isolates of Beauveria bassiana (NBAII-Bb-5a, 7, 14, 19, 23 and 45) as an endophyte in maize stem and leaf tissues. Maize seedlings (var. Nithyashree) were inoculated with conidial suspensions and were examined for endophytic establishment in leaf and stems at different intervals during 15-90 days after treatment. All six isolates showed colonization in stem and leaf tissues with varying abilities of colonization and persistence. The mean percent colonization ranged from 7.41 to 20.37 % in older stem tissues and 3.70 to 21.29 % in young stem tissues and in leaf, it ranged from 6.46 to 27.78 % in older leaf tissues and 11.11 to 26.85 % in young leaf tissues. Among six isolates tested, Bb-23 isolate recorded the maximum mean colonization in older stem (20.37 %), older leaf (27.78 %) and in young stem (21.29 %). Bb-5a isolate showed maximum mean colonization in young leaf tissues (26.85 %). Persistence of inoculated fungal isolates decreased with increase in age of the plant. No physical symptoms of damage were observed in any of the B. bassiana treated plants. No colonization of B. bassiana was observed in the untreated control maize plants. The results obtained in plating and PCR techniques were similar with regard to the confirmation of endophytic establishment of B. bassiana. This study indicated the possibility of using B. bassiana as an endophyte in maize for management of maize stem borer, Chilo partellus.

  10. [Effects of stereoscopic cultivation on photosynthetic characteristics and growth of Tulipa edulis].

    PubMed

    Sun, Yuan; Guo, Qiao-Sheng; Zhu, Zai-Biao; Lin, Jian-Luo; Zhou, Bo-Ya; Zhao, Min-Jie

    2016-06-01

    The effect of stereoscopic cultivation on the growth, photosynthetic characteristics and yield of Tulipa edulis was studied to explore the feasibility of stereoscopic cultivation on efficient cultivation of T.edulis. Total leaf area and photosynthetic parameters of T.edulis under stereoscopic cultivation (the upper, middle and the lower layers ) and the control were measured using LI-3100 leaf area meter and LI-6400XT photosynthesis system in the growing peak period of T.edulis.Plant biomass and biomass allocation were also determined.In addition, the bulb regeneration and yield of T.edulis were measured in the harvesting time.The results indicated that in the middle layer of stereoscopic cultivation, leaf biomass proportion was the highest, but total bulb fresh and dry weight and output growth (fresh weight) were the lowest among the treatments.And total bulb fresh weight in the middle of stereoscopic cultivation reduced significantly, by 22.84%, compared with the control.Light intensity in the lower layer of stereoscopic cultivation was moderate, in which T.edulis net photosynthetic rate and water use efficiency were higher than those of the other layers of stereoscopic cultivation, and bulb biomass proportion was the highest in all the treatments.No significant difference was detected in the total bulb fresh weight, dry weight and output growth (fresh weight) between the middle layer of stereoscopic cultivation and the control.In general, there was no significant difference in the growth status of T.edulis between stereoscopic cultivation and the control.Stereoscopic cultivation increased the yield of T.edulis by 161.66% in fresh weight and 141.35% in dry weight compared with the control in the condition of the same land area, respectively.In conclusion, stereoscopic cultivation can improve space utilization, increase the production, and achieve the high density cultivation of T.edulis. Copyright© by the Chinese Pharmaceutical Association.

  11. Contrasting cDNA-AFLP profiles between crown and leaf tissues of cold-acclimated wheat plants indicate differing regulatory circuitries for low temperature tolerance.

    PubMed

    Ganeshan, Seedhabadee; Sharma, Pallavi; Young, Lester; Kumar, Ashwani; Fowler, D Brian; Chibbar, Ravindra N

    2011-03-01

    Low-temperature (LT) tolerance in winter wheat (Triticum aestivum L.) is an economically important but complex trait. Four selected wheat genotypes, a winter hardy cultivar, Norstar, a tender spring cultivar, Manitou and two near-isogenic lines with Vrn-A1 (spring Norstar) and vrn-A1 (winter Manitou) alleles of Manitou and Norstar were cold-acclimated at 6°C and crown and leaf tissues were collected at 0, 2, 14, 21, 35, 42, 56 and 70 days of cold acclimation. cDNA-AFLP profiling was used to determine temporal expression profiles of transcripts during cold-acclimation in crown and leaf tissues, separately to determine if LT regulatory circuitries in crown and leaf tissues could be delineated using this approach. Screening 64 primer combinations identified 4,074 and 2,757 differentially expressed transcript-derived fragments (TDFs) out of which 38 and 16% were up-regulated as compared to 3 and 6% that were down-regulated in crown and leaf tissues, respectively. DNA sequencing of TDFs revealed sequences common to both tissues including genes coding for DEAD-box RNA helicase, choline-phosphate cytidylyltransferase and delta-1-pyrroline carboxylate synthetase. TDF specific to crown tissues included genes coding for phospahtidylinositol kinase, auxin response factor protein and brassinosteroid insensitive 1-associated receptor kinase. In leaf, genes such as methylene tetrahydrofolate reductase, NADH-cytochrome b5 reductase and malate dehydrogenase were identified. However, 30 and 14% of the DNA sequences from the crown and leaf tissues, respectively, were hypothetical or unknown proteins. Cluster analysis of up-, down-regulated and unique TDFs, DNA sequence and real-time PCR validation, infer that mechanisms operating in crown and leaf tissue in response to LT are differently regulated and warrant further studies.

  12. Growth responses of the mangrove Avicennia marina to salinity: development and function of shoot hydraulic systems require saline conditions

    PubMed Central

    Nguyen, Hoa T.; Stanton, Daniel E.; Schmitz, Nele; Farquhar, Graham D.; Ball, Marilyn C.

    2015-01-01

    Background and Aims Halophytic eudicots are characterized by enhanced growth under saline conditions. This study combines physiological and anatomical analyses to identify processes underlying growth responses of the mangrove Avicennia marina to salinities ranging from fresh- to seawater conditions. Methods Following pre-exhaustion of cotyledonary reserves under optimal conditions (i.e. 50 % seawater), seedlings of A. marina were grown hydroponically in dilutions of seawater amended with nutrients. Whole-plant growth characteristics were analysed in relation to dry mass accumulation and its allocation to different plant parts. Gas exchange characteristics and stable carbon isotopic composition of leaves were measured to evaluate water use in relation to carbon gain. Stem and leaf hydraulic anatomy were measured in relation to plant water use and growth. Key Results Avicennia marina seedlings failed to grow in 0–5 % seawater, whereas maximal growth occurred in 50–75 % seawater. Relative growth rates were affected by changes in leaf area ratio (LAR) and net assimilation rate (NAR) along the salinity gradient, with NAR generally being more important. Gas exchange characteristics followed the same trends as plant growth, with assimilation rates and stomatal conductance being greatest in leaves grown in 50–75 % seawater. However, water use efficiency was maintained nearly constant across all salinities, consistent with carbon isotopic signatures. Anatomical studies revealed variation in rates of development and composition of hydraulic tissues that were consistent with salinity-dependent patterns in water use and growth, including a structural explanation for low stomatal conductance and growth under low salinity. Conclusions The results identified stem and leaf transport systems as central to understanding the integrated growth responses to variation in salinity from fresh- to seawater conditions. Avicennia marina was revealed as an obligate halophyte, requiring saline conditions for development of the transport systems needed to sustain water use and carbon gain. PMID:25600273

  13. Ultrasensitive quantification of endogenous brassinosteroids in milligram fresh plant with a quaternary ammonium derivatization reagent by pipette-tip solid-phase extraction coupled with ultra-high-performance liquid chromatography tandem mass spectrometry.

    PubMed

    Deng, Ting; Wu, Dapeng; Duan, Chunfeng; Guan, Yafeng

    2016-07-22

    Determination of endogenous brassinosteroids (BRs) in limited sample amount is vital to elucidating their tissue- and even local tissue-specific signaling pathway and physiological effects on plant growth and development. In this work, an ultra-sensitive quantification method was established for endogenous BRs in milligram fresh plant by using pipette-tip solid-phase extraction coupled with ultra-performance liquid chromatography tandem mass spectrometry (PT-SPE-UPLC-MS/MS), in which a quaternary ammonium phenyl boronic acid, 4-borono-N,N,N-trimethylbenzenaminium iodide (BTBA) was first developed for chemical derivatization of BRs. Due to the cationic quaternary ammonium group of BTBA, the ionization efficiencies of the BRs chelates with BTBA (BTBA-BRs) were enhanced by 1190-448785 times, which is the highest response enhancement factor among all derivatization reagents reported for BRs. In addition, PT-SPE packed with C18 sorbent was first used for purifying BRs from plant extracts, so the required sample amount was minimized, and recoveries higher than 91% were achieved. Under the optimized conditions, the minimal detectable amounts (MDA) of five target BRs were in the range of 27-94 amol, and the correlation coefficients (R(2)) were >0.9985 over four orders of magnitude. The relative recoveries of 75.8-104.9% were obtained with the intra- and inter-day relative standard deviations (RSDs) less than 18.7% and 19.6%, respectively. Finally, three BRs were successfully quantified in only 5mg fresh rice plant samples, and 24-epiBL can even be detected in only 0.5mg FW rice leaf segments. It is the first time that the BRs content in sub-milligram fresh plant sample has been quantified. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Cyanide in the chemical arsenal of garlic mustard, Alliaria petiolata.

    PubMed

    Cipollini, Don; Gruner, Bill

    2007-01-01

    Cyanide production has been reported from over 2500 plant species, including some members of the Brassicaceae. We report that the important invasive plant, Alliaria petiolata, produces levels of cyanide in its tissues that can reach 100 ppm fresh weight (FW), a level considered toxic to many vertebrates. In a comparative study, levels of cyanide in leaves of young first-year plants were 25 times higher than in leaves of young Arabidopsis thaliana plants and over 150 times higher than in leaves of young Brassica kaber, B. rapa, and B. napus. In first-year plants, cyanide levels were highest in young leaves of seedlings and declined with leaf age on individual plants. Leaves of young plants infested with green peach aphids (Myzus persicae) produced just over half as much cyanide as leaves of healthy plants, suggesting that aphid feeding led to loss of cyanide from intact tissues before analysis, or that aphid feeding inhibited cyanide precursor production. In a developmental study, levels of cyanide in the youngest and oldest leaf of young garlic mustard plants were four times lower than in the youngest and oldest leaf of young Sorghum sudanense (cv. Cadan 97) plants, but cyanide levels did not decline in these leaves with plant age as in S. sudanense. Different populations of garlic mustard varied moderately in the constitutive and inducible expression of cyanide in leaves, but no populations studied were acyanogenic. Although cyanide production could result from breakdown products of glucosinolates, no cyanide was detected in vitro from decomposition of sinigrin, the major glucosinolate of garlic mustard. These studies indicate that cyanide produced from an as yet unidentified cyanogenic compound is a part of the battery of chemical defenses expressed by garlic mustard.

  15. Allelopathic effects of the aqueous extract of the leaf and seed of Leucaena leucocephala on three selected weed species

    NASA Astrophysics Data System (ADS)

    Ishak, Muhamad Safwan; Sahid, Ismail

    2014-09-01

    A laboratory experiment was conducted to study the allelopathic effects of the aqueous extract of the leaf and seed of Leucaena leucocephala. The aqueous extracts were individually tested on three selected weed species, namely goatweed (Ageratum conyzoides), coat buttons (Tridax procumbens) and lilac tasselflower (Emilia sonchifolia). The allelopathic effects of the leaf and seed extracts on germination, shoot length, root length and fresh weight of each of the selected weed species were determined. Germination of goatweed, coat buttons and lilac tasselflower were inhibited by the aqueous extracts of both the leaf and seed of L. leucocephala and was concentration dependent. Different concentrations of the aqueous extracts showed various germination patterns on the selected weeds species. Seedling length and fresh weight of goatweed, coat buttons and lilac tasselflower were reduced in response to respective increasing concentrations of the seed extracts. Maximum inhibition by the aqueous seed extract was observed more on the root rather than the shoot growth. The aqueous seed extract at T3 concentration reduced root length of goatweed, coat buttons and lilac tasselflower by 95%, 86% and 91% (of the control) respectively. The aqueous seed extract showed greater inhibitory effects than that of the aqueous leaf extract.

  16. In vitro erythrocyte membrane stabilization properties of Carica papaya L. leaf extracts

    PubMed Central

    Ranasinghe, Priyanga; Ranasinghe, Pathmasiri; Abeysekera, W. P. Kaushalya M.; Premakumara, G. A. Sirimal; Perera, Yashasvi S.; Gurugama, Padmalal; Gunatilake, Saman B.

    2012-01-01

    Background: Carica papaya L. fruit juice and leaf extracts are known to have many beneficial medical properties. Recent reports have claimed possible beneficial effects of C. papaya L. leaf juice in treating patients with dengue viral infections. This study aims to evaluate the membrane stabilization potential of C. papaya L. leaf extracts using an in vitro hemolytic assay. Materials and Methods: The study was conducted in between June and August 2010. Two milliliters of blood from healthy volunteers and patients with serologically confirmed current dengue infection were freshly collected and used in the assays. Fresh papaya leaves at three different maturity stages (immature, partly matured, and matured) were cleaned with distilled water, crushed, and the juice was extracted with 10 ml of cold distilled water. Freshly prepared cold water extracts of papaya leaves (1 ml containing 30 μl of papaya leaf extracts, 20 μl from 40% erythrocytes suspension, and 950 μl of phosphate buffered saline) were used in the heat-induced and hypotonic-induced hemolytic assays. In dose response experiments, six different concentrations (9.375, 18.75, 37.5, 75, 150, and 300 μg/ml) of freeze dried extracts of the partly matured leaves were used. Membrane stabilization properties were investigated with heat-induced and hypotonicity-induced hemolysis assays. Results: Extracts of papaya leaves of all three maturity levels showed a significant reduction in heat-induced hemolysis compared to controls (P < 0.05). Papaya leaf extracts of all three maturity levels showed more than 25% inhibition at a concentration of 37.5 μg/ml. The highest inhibition of heat-induced hemolysis was observed at 37.5 μg/ml. Inhibition activity of different maturity levels was not significantly (P < 0.05) different from one another. Heat-induced hemolysis inhibition activity did not demonstrate a linear dose response relationship. At 37.5 μg/ml concentration of the extract, a marked inhibition of hypotonicity-induced hemolysis was observed. Conclusion: C. papaya L. leaf extracts showed a significant inhibition of hemolysis in vitro and could have a potential therapeutic effect on disease processes causing destabilization of biological membranes. PMID:23225962

  17. In vitro erythrocyte membrane stabilization properties of Carica papaya L. leaf extracts.

    PubMed

    Ranasinghe, Priyanga; Ranasinghe, Pathmasiri; Abeysekera, W P Kaushalya M; Premakumara, G A Sirimal; Perera, Yashasvi S; Gurugama, Padmalal; Gunatilake, Saman B

    2012-10-01

    Carica papaya L. fruit juice and leaf extracts are known to have many beneficial medical properties. Recent reports have claimed possible beneficial effects of C. papaya L. leaf juice in treating patients with dengue viral infections. This study aims to evaluate the membrane stabilization potential of C. papaya L. leaf extracts using an in vitro hemolytic assay. The study was conducted in between June and August 2010. Two milliliters of blood from healthy volunteers and patients with serologically confirmed current dengue infection were freshly collected and used in the assays. Fresh papaya leaves at three different maturity stages (immature, partly matured, and matured) were cleaned with distilled water, crushed, and the juice was extracted with 10 ml of cold distilled water. Freshly prepared cold water extracts of papaya leaves (1 ml containing 30 μl of papaya leaf extracts, 20 μl from 40% erythrocytes suspension, and 950 μl of phosphate buffered saline) were used in the heat-induced and hypotonic-induced hemolytic assays. In dose response experiments, six different concentrations (9.375, 18.75, 37.5, 75, 150, and 300 μg/ml) of freeze dried extracts of the partly matured leaves were used. Membrane stabilization properties were investigated with heat-induced and hypotonicity-induced hemolysis assays. Extracts of papaya leaves of all three maturity levels showed a significant reduction in heat-induced hemolysis compared to controls (P < 0.05). Papaya leaf extracts of all three maturity levels showed more than 25% inhibition at a concentration of 37.5 μg/ml. The highest inhibition of heat-induced hemolysis was observed at 37.5 μg/ml. Inhibition activity of different maturity levels was not significantly (P < 0.05) different from one another. Heat-induced hemolysis inhibition activity did not demonstrate a linear dose response relationship. At 37.5 μg/ml concentration of the extract, a marked inhibition of hypotonicity-induced hemolysis was observed. C. papaya L. leaf extracts showed a significant inhibition of hemolysis in vitro and could have a potential therapeutic effect on disease processes causing destabilization of biological membranes.

  18. Effects of supplementary lighting by natural light for growth of Brassica chinensis

    NASA Astrophysics Data System (ADS)

    Yeh, Shih-Chuan; Lee, Hui-Ping; Kao, Shih-Tse; Lu, Ju-Lin

    2016-04-01

    This paper present a model of cultivated chamber with supplementary natural colour light. We investigate the effects of supplementary natural red light and natural blue light on growth of Brassica chinensis under natural white light illumination. After 4 weeks of supplementary colour light treatment, the experiment results shown that the weight of fresh leaf were not affected by supplementary natural blue light. However, those Brassica chinensis were cultivated in the chambers with supplementary natural red light obtained a significant increasing of fresh weight of leaf under both white light illuminate models. The combination of natural white light with supplementary natural red light illumination will be benefits in growth for cultivation and energy saving.

  19. Leaf biomechanics, morphology, and anatomy of the deciduous mesophyte Prunus serrulata (Rosaceae) and the evergreen sclerophyllous shrub Heteromeles arbutifolia (Rosaceae).

    PubMed

    Balsamo, Ronald A; Bauer, Aaron M; Davis, Stephen D; Rice, Benita M

    2003-01-01

    Leaf tensile properties were compared between the mesic deciduous tree Prunus serrulata (var. "Kwanzan") and the xeric and sclerophyllous chaparral evergreen shrub Heteromeles arbutifolia (M. Roem). All values for biomechanical parameters for H. arbutifolia were significantly greater than those of P. serrulata. The fracture planes also differed between the two species with P. serrulata fracturing along the secondary veins, while H. arbutifolia most often fractured across the leaf irrespective of the vein or mesophyll position, thus yielding qualitative differences in the stress-strain curves of the two species. Anatomically, P. serrulata exhibits features typical for a deciduous mesophytic leaf such as a thin cuticle, a single layer of palisade mesophyll, isodiametric spongy mesophyll, and extensive reticulation of the laminar veins. Heteromeles arbutifolia leaves, however, are typically two- to three-fold thicker with a 35% higher dry mass/fresh mass ratio. The vascular tissue is restricted to the interface of the palisade and spongy mesophyll near the center of the leaf. Both epidermal layers have a thick cuticle. The palisade mesophyll is tightly packed and two to three layers thick. The spongy mesophyll cells are ameboid in shape and tightly interlinked both to other spongy cells as well as to the overlying palisade layer. We conclude that the qualitative and quantitative biomechanical differences between the leaves of these two species are likely due to a complex interaction of internal architectural arrangement and the physical/chemical differences in the properties of their respective cell walls. These studies illustrate the importance that morphological and anatomical correlates play with mechanical behavior in plant material and ultimately reflect adaptations present in the leaves of chaparral shrubs that are conducive to surviving in arid environments.

  20. Effect of route of introduction and host cultivar on the colonization, internalization, and movement of the human pathogen Escherichia coli O157:H7 in spinach.

    PubMed

    Mitra, R; Cuesta-Alonso, E; Wayadande, A; Talley, J; Gilliland, S; Fletcher, J

    2009-07-01

    Human pathogens can contaminate leafy produce in the field by various routes. We hypothesized that interactions between Escherichia coli O157:H7 and spinach are influenced by the route of introduction and the leaf microenvironment. E. coli O157:H7 labeled with green fluorescent protein was dropped onto spinach leaf surfaces, simulating bacteria-laden raindrops or sprinkler irrigation, and survived on the phylloplane for at least 14 days, with increasing titers and areas of colonization over time. The same strains placed into the rhizosphere by soil infiltration remained detectable on very few plants and in low numbers (10(2) to 10(6) CFU/g fresh tissue) that decreased over time. Stem puncture inoculations, simulating natural wounding, rarely resulted in colonization or multiplication. Bacteria forced into the leaf interior survived for at least 14 days in intercellular spaces but did not translocate or multiply. Three spinach cultivars with different leaf surface morphologies were compared for colonization by E. coli O157:H7 introduced by leaf drop or soil drench. After 2 weeks, cv. Bordeaux hosted very few bacteria. More bacteria were seen on cv. Space and were dispersed over an area of up to 0.3 mm2. The highest bacterial numbers were observed on cv. Tyee but were dispersed only up to 0.15 mm2, suggesting that cv. Tyee may provide protected niches or more nutrients or may promote stronger bacterial adherence. These findings suggest that the spinach phylloplane is a supportive niche for E. coli O157:H7, but no conclusive evidence was found for natural entry into the plant interior. The results are relevant for interventions aimed at minimizing produce contamination by human pathogens.

  1. Quantitative Assessment of Contamination of Fresh Food Produce of Various Retail Types by Human-Virulent Microsporidian Spores▿

    PubMed Central

    Jedrzejewski, Szymon; Graczyk, Thaddeus K.; Slodkowicz-Kowalska, Anna; Tamang, Leena; Majewska, Anna C.

    2007-01-01

    This study demonstrated that fresh food produce, such as berries, sprouts, and green-leafed vegetables, sold at the retail level can contain potentially viable microsporidian spores of human-virulent species, such as Enterocytozoon bieneusi, Encephalitozoon intestinalis, and Encephalitozoon cuniculi, at quantities representing a threat of food-borne infection. PMID:17449682

  2. Distinct palisade tissue development processes promoted by leaf autonomous signalling and long-distance signalling in Arabidopsis thaliana.

    PubMed

    Munekage, Yuri Nakajima; Inoue, Shio; Yoneda, Yuki; Yokota, Akiho

    2015-06-01

    Plants develop palisade tissue consisting of cylindrical mesophyll cells located at the adaxial side of leaves in response to high light. To understand high light signalling in palisade tissue development, we investigated leaf autonomous and long-distance signal responses of palisade tissue development using Arabidopsis thaliana. Illumination of a developing leaf with high light induced cell height elongation, whereas illumination of mature leaves with high light increased cell density and suppressed cell width expansion in palisade tissue of new leaves. Examination using phototropin1 phototropin2 showed that blue light signalling mediated by phototropins was involved in cell height elongation of the leaf autonomous response rather than the cell density increase induced by long-distance signalling. Hydrogen peroxide treatment induced cylindrical palisade tissue cell formation in both a leaf autonomous and long-distance manner, suggesting involvement of oxidative signals. Although constitutive expression of transcription factors involved in systemic-acquired acclimation to excess light, ZAT10 and ZAT12, induced cylindrical palisade tissue cell formation, knockout of these genes did not affect cylindrical palisade tissue cell formation. We conclude that two distinct signalling pathways - leaf autonomous signalling mostly dependent on blue light signalling and long-distance signalling from mature leaves that sense high light and oxidative stress - control palisade tissue development in A. thaliana. © 2014 John Wiley & Sons Ltd.

  3. Regulation of leaf hydraulics: from molecular to whole plant levels.

    PubMed

    Prado, Karine; Maurel, Christophe

    2013-01-01

    The water status of plant leaves is dependent on both stomatal regulation and water supply from the vasculature to inner tissues. The present review addresses the multiple physiological and mechanistic facets of the latter process. Inner leaf tissues contribute to at least a third of the whole resistance to water flow within the plant. Physiological studies indicated that leaf hydraulic conductance (K leaf) is highly dependent on the anatomy, development and age of the leaf and can vary rapidly in response to physiological or environmental factors such as leaf hydration, light, temperature, or nutrient supply. Differences in venation pattern provide a basis for variations in K leaf during development and between species. On a short time (hour) scale, the hydraulic resistance of the vessels can be influenced by transpiration-induced cavitations, wall collapses, and changes in xylem sap composition. The extravascular compartment includes all living tissues (xylem parenchyma, bundle sheath, and mesophyll) that transport water from xylem vessels to substomatal chambers. Pharmacological inhibition and reverse genetics studies have shown that this compartment involves water channel proteins called aquaporins (AQPs) that facilitate water transport across cell membranes. In many plant species, AQPs are present in all leaf tissues with a preferential expression in the vascular bundles. The various mechanisms that allow adjustment of K leaf to specific environmental conditions include transcriptional regulation of AQPs and changes in their abundance, trafficking, and intrinsic activity. Finally, the hydraulics of inner leaf tissues can have a strong impact on the dynamic responses of leaf water potential and stomata, and as a consequence on plant carbon economy and leaf expansion growth. The manipulation of these functions could help optimize the entire plant performance and its adaptation to extreme conditions over short and long time scales.

  4. Laboratory and Airborne BRDF Analysis of Vegetation Leaves and Soil Samples

    NASA Technical Reports Server (NTRS)

    Georgiev, Georgi T.; Gatebe, Charles K.; Butler, James J.; King, Michael D.

    2008-01-01

    Laboratory-based Bidirectional Reflectance Distribution Function (BRDF) analysis of vegetation leaves, soil, and leaf litter samples is presented. The leaf litter and soil samples, numbered 1 and 2, were obtained from a site located in the savanna biome of South Africa (Skukuza: 25.0degS, 31.5degE). A third soil sample, number 3, was obtained from Etosha Pan, Namibia (19.20degS, 15.93degE, alt. 1100 m). In addition, BRDF of local fresh and dry leaves from tulip tree (Liriodendron tulipifera) and acacia tree (Acacia greggii) were studied. It is shown how the BRDF depends on the incident and scatter angles, sample size (i.e. crushed versus whole leaf,) soil samples fraction size, sample status (i.e. fresh versus dry leaves), vegetation species (poplar versus acacia), and vegetation s biochemical composition. As a demonstration of the application of the results of this study, airborne BRDF measurements acquired with NASA's Cloud Absorption Radiometer (CAR) over the same general site where the soil and leaf litter samples were obtained are compared to the laboratory results. Good agreement between laboratory and airborne measured BRDF is reported.

  5. NMR-based metabolomics study of the biochemical relationship between sugarcane callus tissues and their respective nutrient culture media

    PubMed Central

    Mahmud, Iqbal; Thapaliya, Monica; Boroujerdi, Arezue; Chowdhury, Kamal

    2014-01-01

    The culture of sugarcane leaf explant onto culture induction medium triggers the stimulation of cell metabolism into both embryogenic and non-embryogenic callus tissues. Previous analyses demonstrated that embryogenic and nonembryogenic callus tissues have distinct metabolic profiles. This study is the follow-up to understand the biochemical relationship between the nutrient media and callus tissues using one-dimensional (1D 1H) and two-dimensional (2D 1H–13C) NMR spectroscopy followed by principal component analysis (PCA). 1D 1H spectral comparisons of fresh unspent media (FM), embryogenic callus media (ECM), non-embryogenic callus media (NECM), embryogenic callus (EC), and non-embryogenic callus (NEC), showed different metabolic relationships between callus tissues and media. Based on metabolite fold change analysis, significantly changing sugar compounds such as glucose, fructose, sucrose, and maltose were maintained in large quantities by EC only. Significantly different amino acid compounds such as valine, leucine, alanine, threonine, asparagine, and glutamine and different organic acid derivatives such as lactate, 2-hydroxyisobutyrate, 4-aminobutyrate, malonate, and choline were present in EC, NEC, and NECM, which indicates that EC maintained these nutrients, while NEC either maintained or secreted the metabolites. These media and callus-specific results suggest that EC and NEC utilize and/or secrete media nutrients differently. PMID:25012359

  6. A comparison of embalming fluids for use in surgical workshops.

    PubMed

    Jaung, Rebekah; Cook, Peter; Blyth, Phil

    2011-03-01

    There is a growing need to learn surgical skills without risk to patients. One of the major determining factors on the suitability of specimens for surgical workshops is the fluid used for embalming. This study sought to compare three different arterial embalming preparations to a single fresh cadaver. Eleven cadavers embalmed using Graz (single cadaver), Dodge (four cadavers) and Genelyn (five cadavers) preparations were compared using four criteria; joint flexibility measured with a goniometer, tissue pliability rated on standardized videos of instrument handling, tissue color analyzed on standardized photographs and resistance to fungal growth identified by inoculation and observation of tissue blocks. The cadaver embalmed according to the Graz method had joint flexibility comparable to fresh tissue while the Dodge and Genelyn cadavers were less flexible. Tissue pliability was significantly affected by the Dodge and Genelyn methods while the Graz method tissue remained most like fresh tissue. The Graz method cadaver had color that was most akin to fresh tissue and the Dodge method cadavers were relatively more like fresh than the Genelyn. The Dodge and Genelyn method had quite similar fungicidal properties (3/11 Dodge and 2/9 Genelyn embalmed cadavers susceptible) while the Graz method cadaver did not grow mould. Variation exists between cadavers; however, the Graz method produced a cadaver with more flexible joints, better tissue quality and muscle color closest to the fresh specimen. The Dodge and Genelyn methods are similar with the exception of tissue color where the Dodge method was more similar to fresh tissue. Copyright © 2011 Wiley-Liss, Inc.

  7. Leaf water dynamics of Arabidopsis thaliana monitored in-vivo using terahertz time-domain spectroscopy

    NASA Astrophysics Data System (ADS)

    Castro-Camus, E.; Palomar, M.; Covarrubias, A. A.

    2013-10-01

    The declining water availability for agriculture is becoming problematic for many countries. Therefore the study of plants under water restriction is acquiring extraordinary importance. Botanists currently follow the dehydration of plants comparing the fresh and dry weight of excised organs, or measuring their osmotic or water potentials; these are destructive methods inappropriate for in-vivo determination of plants' hydration dynamics. Water is opaque in the terahertz band, while dehydrated biological tissues are partially transparent. We used terahertz spectroscopy to study the water dynamics of Arabidopsis thaliana by comparing the dehydration kinetics of leaves from plants under well-irrigated and water deficit conditions. We also present measurements of the effect of dark-light cycles and abscisic acid on its water dynamics. The measurements we present provide a new perspective on the water dynamics of plants under different external stimuli and confirm that terahertz can be an excellent non-contact probe of in-vivo tissue hydration.

  8. Effect of girdling at various positions of 'Huang Zhi Xiang' tea tree (Camellia sinensis) on the contents of catechins and starch in fresh leaf.

    PubMed

    Chen, Yulong; Jiang, Yueming; Shi, John; Xu, Xinlan; Yang, Shaoyu; Duan, Jun

    2011-06-01

    The contents of starch and catechins in the fresh leaf of 'Huang Zhi Xiang' Oolong tea trees girdled at the bottom, middle (on the big branches) and top (on the small branches) were determined. The study demonstrated that the starch contents from girdled trees were significantly higher (p<0.05) than that from non-girdled ones. Furthermore, the contents of (-)-epicatechin (EC), (-)-epigallocatechin (EGC), total catechins (TC) and simple catechins (SC) from girdled trees were significantly higher (p<0.05) than those from non-girdled ones. Especially, the contents of (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG) and catechin gallate (CG) from girdled at the middle were also significantly higher (p<0.05) than those from the non-girdled. The starch contents were negatively correlated with the contents of (-)-gallocatechin (GC), EC, SC, TC and EGC, while positively correlated with the contents of EGCG and CG in fresh shoots.

  9. Comparative Study of Lettuce and Radish Grown Under Red and Blue LEDs and White Fluorescent Lamps

    NASA Technical Reports Server (NTRS)

    Mickens, Matthew A.; Massa, Gioia; Newsham, Gerard; Wheeler, Raymond; Birmele, Michele

    2016-01-01

    Growing vegetable crops in space will be an essential part of sustaining astronauts during long-range missions. To drive photosynthesis, red and blue light-emitting diodes (LEDs) have attracted attention because of their efficiency, longevity, small size, and safety. In efforts to optimize crop yield, there is also recent interest in analyzing the subtle effects of additional wavelengths on plant growth. For instance, since plants often look purplish gray under red and blue LEDs, the addition of green light allows easy recognition of disease and the assessment of plant health status. However, it is important to know if wavelengths outside the traditional red and blue wavebands have a direct effect on enhancing or hindering the mechanisms involved in plant growth. In this experiment, a comparative study was performed on two short cycle crops of red romaine lettuce (Lactuca sativa cv. "Outredgeous") and radish (Raphanus sativa cv. 'Cherry Bomb'), which were grown under two light treatments. The first treatment being red (630 nm) and blue (450 nm) LEDs alone, while the second treatment consisted of daylight tri-phosphor fluorescent lamps (CCT approximately 5000 K) at equal photosynthetic photon flux (PPF). The treatment effects were evaluated by measuring the fresh biomass produced, plant morphology and leaf dimensions, leaf chlorophyll content, and adenosine triphosphate (ATP) within plant leaf/storage root tissues.

  10. Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virus-derived transgenes from plant tissues

    PubMed Central

    Ndunguru, Joseph; Taylor, Nigel J; Yadav, Jitender; Aly, Haytham; Legg, James P; Aveling, Terry; Thompson, Graham; Fauquet, Claude M

    2005-01-01

    Background Plant viral diseases present major constraints to crop production. Effective sampling of the viruses infecting plants is required to facilitate their molecular study and is essential for the development of crop protection and improvement programs. Retaining integrity of viral pathogens within sampled plant tissues is often a limiting factor in this process, most especially when sample sizes are large and when operating in developing counties and regions remote from laboratory facilities. FTA is a paper-based system designed to fix and store nucleic acids directly from fresh tissues pressed into the treated paper. We report here the use of FTA as an effective technology for sampling and retrieval of DNA and RNA viruses from plant tissues and their subsequent molecular analysis. Results DNA and RNA viruses were successfully recovered from leaf tissues of maize, cassava, tomato and tobacco pressed into FTA® Classic Cards. Viral nucleic acids eluted from FTA cards were found to be suitable for diagnostic molecular analysis by PCR-based techniques and restriction analysis, and for cloning and nucleotide sequencing in a manner equivalent to that offered by tradition isolation methods. Efficacy of the technology was demonstrated both from sampled greenhouse-grown plants and from leaf presses taken from crop plants growing in farmer's fields in East Africa. In addition, FTA technology was shown to be suitable for recovery of viral-derived transgene sequences integrated into the plant genome. Conclusion Results demonstrate that FTA is a practical, economical and sensitive method for sampling, storage and retrieval of viral pathogens and plant genomic sequences, when working under controlled conditions and in the field. Application of this technology has the potential to significantly increase ability to bring modern analytical techniques to bear on the viral pathogens infecting crop plants. PMID:15904535

  11. Exogenous Methyl Jasmonate Treatment Increases Glucosinolate Biosynthesis and Quinone Reductase Activity in Kale Leaf Tissue

    PubMed Central

    Ku, Kang-Mo; Jeffery, Elizabeth H.; Juvik, John A.

    2014-01-01

    Methyl jasmonate (MeJA) spray treatments were applied to the kale varieties ‘Dwarf Blue Curled Vates’ and ‘Red Winter’ in replicated field plantings in 2010 and 2011 to investigate alteration of glucosinolate (GS) composition in harvested leaf tissue. Aqueous solutions of 250 µM MeJA were sprayed to saturation on aerial plant tissues four days prior to harvest at commercial maturity. The MeJA treatment significantly increased gluconasturtiin (56%), glucobrassicin (98%), and neoglucobrassicin (150%) concentrations in the apical leaf tissue of these genotypes over two seasons. Induction of quinone reductase (QR) activity, a biomarker for anti-carcinogenesis, was significantly increased by the extracts from the leaf tissue of these two cultivars. Extracts of apical leaf tissues had greater MeJA mediated increases in phenolics, glucosinolate concentrations, GS hydrolysis products, and QR activity than extracts from basal leaf tissue samples. The concentration of the hydrolysis product of glucoraphanin, sulforphane was significantly increased in apical leaf tissue of the cultivar ‘Red Winter’ in both 2010 and 2011. There was interaction between exogenous MeJA treatment and environmental conditions to induce endogenous JA. Correlation analysis revealed that indole-3-carbanol (I3C) generated from the hydrolysis of glucobrassicin significantly correlated with QR activity (r = 0.800, P<0.001). Concentrations required to double the specific QR activity (CD values) of I3C was calculated at 230 µM, which is considerably weaker at induction than other isothiocyanates like sulforphane. To confirm relationships between GS hydrolysis products and QR activity, a range of concentrations of MeJA sprays were applied to kale leaf tissues of both cultivars in 2011. Correlation analysis of these results indicated that sulforaphane, NI3C, neoascorbigen, I3C, and diindolylmethane were all significantly correlated with QR activity. Thus, increased QR activity may be due to combined increases in phenolics (quercetin and kaempferol) and GS hydrolysis product concentrations rather than by individual products alone. PMID:25084454

  12. Antinociceptive, anti-inflammatory and antidiabetic effects of Bryophyllum pinnatum (Crassulaceae) leaf aqueous extract.

    PubMed

    Ojewole, John A O

    2005-05-13

    In order to scientifically appraise some of the ethnomedical uses of Bryophyllum pinnatum leaves, the present study was undertaken to investigate the antinociceptive, anti-inflammatory and antidiabetic properties of the plant's leaf aqueous extract in experimental animal models. The antinociceptive effect of the herb's leaf extract was evaluated by the 'hot-plate' and 'acetic acid' test models of pain in mice. The anti-inflammatory and antidiabetic effects of the plant's extract were investigated in rats, using fresh egg albumin-induced pedal (paw) oedema, and streptozotocin (STZ)-induced diabetes mellitus. Diclofenac (DIC, 100 mg/kg) and chlorpropamide (250 mg/kg) were used respectively as reference drugs for comparison. Bryophyllum pinnatum leaf aqueous extract (BPE, 25-800 mg/kg i.p.) produced significant (P<0.05-0.001) antinociceptive effects against thermally- and chemically-induced nociceptive pain stimuli in mice. The plant extract (BPE, 25-800 mg/kg p.o. or i.p.) also significantly (P<0.05-0.001) inhibited fresh egg albumin-induced acute inflammation and caused significant (P<0.05-0.001) hypoglycaemia in rats. The results of this experimental animal study suggest that Bryophyllum pinnatum leaf aqueous extract possesses antinociceptive, anti-inflammatory and hypoglycaemic properties. The different flavonoids, polyphenols, triterpenoids and other chemical constituents of the herb are speculated to account for the observed antinociceptive, anti-inflammatory and antidiabetic properties of the plant.

  13. Transgenic cotton expressing Cry10Aa toxin confers high resistance to the cotton boll weevil.

    PubMed

    Ribeiro, Thuanne Pires; Arraes, Fabricio Barbosa Monteiro; Lourenço-Tessutti, Isabela Tristan; Silva, Marilia Santos; Lisei-de-Sá, Maria Eugênia; Lucena, Wagner Alexandre; Macedo, Leonardo Lima Pepino; Lima, Janaina Nascimento; Santos Amorim, Regina Maria; Artico, Sinara; Alves-Ferreira, Márcio; Mattar Silva, Maria Cristina; Grossi-de-Sa, Maria Fatima

    2017-08-01

    Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation. Quantitative PCR (qPCR) assays revealed high transcription levels of cry10Aa in both T 0 GM cotton leaf and flower bud tissues. Southern blot and qPCR-based 2 -ΔΔCt analyses revealed that T 0 GM plants had either one or two transgene copies. Quantitative and qualitative analyses of Cry10Aa protein expression showed variable protein expression levels in both flower buds and leaves tissues of T 0 GM cotton plants, ranging from approximately 3.0 to 14.0 μg g -1 fresh tissue. CBW susceptibility bioassays, performed by feeding adults and larvae with T 0 GM cotton leaves and flower buds, respectively, demonstrated a significant entomotoxic effect and a high level of CBW mortality (up to 100%). Molecular analysis revealed that transgene stability and entomotoxic effect to CBW were maintained in T 1 generation as the Cry10Aa toxin expression levels remained high in both tissues, ranging from 4.05 to 19.57 μg g -1 fresh tissue, and the CBW mortality rate remained around 100%. In conclusion, these Cry10Aa GM cotton plants represent a great advance in the control of the devastating CBW insect pest and can substantially impact cotton agribusiness. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  14. Remote sensing of forest canopy and leaf biochemical contents

    NASA Technical Reports Server (NTRS)

    Peterson, David L.; Matson, Pamela A.; Card, Don H.; Aber, John D.; Wessman, Carol; Swanberg, Nancy; Spanner, Michael

    1988-01-01

    Recent research on the remote sensing of forest leaf and canopy biochemical contents suggests that the shortwave IR region contains this information; laboratory analyses of dry ground leaves have yielded reliable predictive relationships between both leaf nitrogen and lignin with near-IR spectra. Attention is given to the application of these laboratory techniques to a limited set of spectra from fresh, whole leaves of conifer species. The analysis of Airborne Imaging Spectrometer data reveals that total water content variations in deciduous forest canopies appear as overall shifts in the brightness of raw spectra.

  15. Cryopreservation and hypothermic storage of lacrimal gland: towards enabling delivery of regenerative medicine therapies for treatment of dry eye syndrome.

    PubMed

    Massie, I; Spaniol, K; Geerling, G; Schrader, S

    2017-12-01

    Severe dry eye syndrome (DES) can cause painful loss of vision and may result from lacrimal gland dysfunction. Current treatments are palliative, so a causative therapy is desirable. The ability to (cryo)preserve lacrimal gland tissue or epithelial cells would simplify this. Here, lacrimal gland tissue was cryopreserved in 10% dimethylsulphoxide in liquid nitrogen, or stored at 4 °C in culture medium for up to 7 days, and compared with fresh tissue using immunohistochemistry. Cultures were initiated from fresh and stored tissue, and cells characterised in P1 for proliferation (WST-1), colony-forming efficiency (CFE) and secretory capacity (immunocytochemistry and β-hexosaminidase activity assay). Tissue stored for > 3 days at 4 °C displayed grossly altered tissue architecture when compared with fresh tissue, decreased acinus density and increased caspase-3 activity. Cryopreserved tissue showed less obvious signs of damage without caspase-3 activation. Storage at 4 °C and cryopreservation delayed epithelial outgrowth compared with that from fresh tissue initially (p  <  0.05) but, by day 9, all explants showed comparable outgrowth (~90%), except tissue stored at 4 °C for 3 or 7 days (p  <  0.05 compared with fresh tissue). Epithelial cell yields per explant were similar from fresh and stored tissue, apart from tissue stored at 4 °C for 7 days (p  <  0.01). In P1, epithelial cells from fresh and stored tissue were largely equivalent in terms of: proliferation; CFE (~21%); Rab3D, HexA and lysozyme expression; mucin production; and β-hexosaminidase activity. These data demonstrate that cryo(preservation) of lacrimal gland tissue and cells is possible, which may enable use of autologous cells in regenerative medicine approaches to treating DES. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  16. Protein profiling in potato (Solanum tuberosum L.) leaf tissues by differential centrifugation.

    PubMed

    Lim, Sanghyun; Chisholm, Kenneth; Coffin, Robert H; Peters, Rick D; Al-Mughrabi, Khalil I; Wang-Pruski, Gefu; Pinto, Devanand M

    2012-04-06

    Foliar diseases, such as late blight, result in serious threats to potato production. As such, potato leaf tissue becomes an important substrate to study biological processes, such as plant defense responses to infection. Nonetheless, the potato leaf proteome remains poorly characterized. Here, we report protein profiling of potato leaf tissues using a modified differential centrifugation approach to separate the leaf tissues into cell wall and cytoplasmic fractions. This method helps to increase the number of identified proteins, including targeted putative cell wall proteins. The method allowed for the identification of 1484 nonredundant potato leaf proteins, of which 364 and 447 were reproducibly identified proteins in the cell wall and cytoplasmic fractions, respectively. Reproducibly identified proteins corresponded to over 70% of proteins identified in each replicate. A diverse range of proteins was identified based on their theoretical pI values, molecular masses, functional classification, and biological processes. Such a protein extraction method is effective for the establishment of a highly qualified proteome profile.

  17. Callus induction of leaf explant Piper betle L. Var Nigra with combination of plant growth regulators indole-3-acetic acid (IAA), benzyl amino purin (BAP) and kinetin

    NASA Astrophysics Data System (ADS)

    Junairiah, Zuraidassanaaz, Nabilah Istighfari; Izdihar, Fairuz Nabil; Manuhara, Yosephine Sri Wulan

    2017-09-01

    The purpose of this research was to determine the combination of plant growth regulators IAA, BAP and kinetin towards callus induction and growth of leaf explants Piper betle L. VarNigra. Explants from leaf of Piper betle L. VarNigra was cultured on MS medium with 24 treatment combinations of plant growth regulators IAA and BAP and 24 treatment combinations of plant growth regulators IAA and kinetin with 0.0;0.5;1.0;1.5;2.0 mg/L concentration respectively, the observed variable were the length of time the formation of callus, callus morphology, fresh and dry weight of callus. The results of this research showed that the combination of growth regulators IAA with BAP and kinetin had effects on leaf growth of Piper betle L. VarNigra. During 8 weeks observation, it indicated that the combination of concentration IAA 0.5 mg/L and BAP 2.0 mg/L showed fastest callus formation at 8.5 days. Combination of concentration IAA 1.0 mg/L and BAP 1.5 mg/L showed the highest of fresh weight at 0.6596 grams, and the highest dry weight was obtained from the combination of concentration IAA 0.5 mg/L and BAP 0.5 mg/L at 0.0727 grams. Combination of concentration IAA 1.0 mg/L and kinetin 1.5 mg/L had the highest of fresh weight at 0.2972 grams and the highest dry weight at 0.1660 grams. Callus of Piper betle L. VarNigra had two textures, that were compact and friable, and also showed various kind of colors, like white, greenish white, yellowish white, tanned white, brown and black. Based on this research, that concentration IAA 1.0 mg/L and 1.5 mg/L kinetin was the best combination for induction of callus from leaf of Piper betle L. Var Nigra.

  18. Plant phenolics and absorption features in vegetation reflectance spectra near 1.66 μm

    NASA Astrophysics Data System (ADS)

    Kokaly, Raymond F.; Skidmore, Andrew K.

    2015-12-01

    Past laboratory and field studies have quantified phenolic substances in vegetative matter from reflectance measurements for understanding plant response to herbivores and insect predation. Past remote sensing studies on phenolics have evaluated crop quality and vegetation patterns caused by bedrock geology and associated variations in soil geochemistry. We examined spectra of pure phenolic compounds, common plant biochemical constituents, dry leaves, fresh leaves, and plant canopies for direct evidence of absorption features attributable to plant phenolics. Using spectral feature analysis with continuum removal, we observed that a narrow feature at 1.66 μm is persistent in spectra of manzanita, sumac, red maple, sugar maple, tea, and other species. This feature was consistent with absorption caused by aromatic Csbnd H bonds in the chemical structure of phenolic compounds and non-hydroxylated aromatics. Because of overlapping absorption by water, the feature was weaker in fresh leaf and canopy spectra compared to dry leaf measurements. Simple linear regressions of feature depth and feature area with polyphenol concentration in tea resulted in high correlations and low errors (% phenol by dry weight) at the dry leaf (r2 = 0.95, RMSE = 1.0%, n = 56), fresh leaf (r2 = 0.79, RMSE = 2.1%, n = 56), and canopy (r2 = 0.78, RMSE = 1.0%, n = 13) levels of measurement. Spectra of leaves, needles, and canopies of big sagebrush and evergreens exhibited a weak absorption feature centered near 1.63 μm, short ward of the phenolic compounds, possibly consistent with terpenes. This study demonstrates that subtle variation in vegetation spectra in the shortwave infrared can directly indicate biochemical constituents and be used to quantify them. Phenolics are of lesser abundance compared to the major plant constituents but, nonetheless, have important plant functions and ecological significance. Additional research is needed to advance our understanding of the spectral influences of plant phenolics and terpenes relative to dominant leaf biochemistry (water, chlorophyll, protein/nitrogen, cellulose, and lignin).

  19. Anti Microbial activity of the various leaf extracts of Vitex negundo Linn.

    PubMed Central

    Devi, P. Renuka; KokilaVani, R.; Poogotha, S. Gnana

    2008-01-01

    The antibacterial activity of the leaves of Vitex negundo was tested against three bacteria Viz., Staphylococcus aureus Escherichia coli and Klebsiella Peumoniae, The fresh aqueous, heated aqueous extract, chloroform and methanolic extract of leaves were used for screening their antibacterial potential. The fresh and aqueous extracts of leaves in various dilutions were found to have antibacterial activity against the three bacteria. PMID:22557306

  20. Comparison of human papillomavirus detection between freshly frozen tissue and paraffin embedded tissue of invasive cervical cancer

    PubMed Central

    2010-01-01

    Background Human Papillomavirus (HPV) detection results comparing paraffin embedded cervical tissue and other cervical specimens have been done with varying degrees of agreement. However, studies comparing freshly frozen specimens and paraffin embedded specimens of invasive cervical carcinomas are lacking. The aim of the study was to compare HPV detection using SPF10 broad-spectrum primers PCR followed by DEIA and genotyping by LiPA25 (version 1) between freshly frozen cervical tissue samples and paraffin embedded blocks of cervical tissue from the same patient. There were 171 pairs of paraffin embedded and freshly frozen samples analyzed from cervical carcinoma cases from Kampala, Uganda. Results 88.9% (95% CI: 83.2%-93.2%) of paraffin embedded samples were HPV positive compared with 90.1% (95% CI: 84.6%-94.1%) of freshly frozen samples, giving an overall agreement in HPV detection between fresh tissue and paraffin embedded tissue at 86.0% (95% CI: 79.8%-90.8%). Although the proportion of HPV positive cases in freshly frozen tissue was higher than those in paraffin blocks, the difference was not statistically significant (p > 0.05). In both types of tissues, single HPV infections were predominant, with HPV16 accounting for 47% of positive cases. Comparison in the overall agreement, taking into accounts not only positivity in general, but also HPV types, showed a 65% agreement (complete agreement of 59.7%, partial agreement of 5.3%) and complete disagreement of 35.0%. HPV detection in squamous cell carcinomas (SCC) and adenocarcinomas (ADC) was similar in fresh tissue or paraffin blocks (p ≥ 0.05). p16 immunostaining in samples that had at least one HPV negative results showed that 24 out of 25 cases had an over-expressed pattern. Conclusions HPV DNA detection was lower among ADC as compared to SCC. However, such differences were minimized when additional p16 testing was added, suggesting that the technical issues may largely explain the HPV negative cases. PMID:20846370

  1. Enhancement of germination, growth, and photosynthesis in soybean by pre-treatment of seeds with magnetic field.

    PubMed

    Shine, M B; Guruprasad, K N; Anand, Anjali

    2011-09-01

    Experiments were conducted to study the effect of static magnetic fields on the seeds of soybean (Glycine max (L.) Merr. var: JS-335) by exposing the seeds to different magnetic field strengths from 0 to 300 mT in steps of 50 mT for 30, 60, and 90 min. Treatment with magnetic fields improved germination-related parameters like water uptake, speed of germination, seedling length, fresh weight, dry weight and vigor indices of soybean seeds under laboratory conditions. Improvement over untreated control was 5-42% for speed of germination, 4-73% for seedling length, 9-53% for fresh weight, 5-16% for dry weight, and 3-88% and 4-27% for vigor indices I and II, respectively. Treatment of 200 mT (60 min) and 150 mT (60 min), which were more effective than others in increasing most of the seedling parameters, were further explored for their effect on plant growth, leaf photosynthetic efficiency, and leaf protein content under field conditions. Among different growth parameters, leaf area, and leaf fresh weight showed maximum enhancement (more than twofold) in 1-month-old plants. Polyphasic chlorophyll a fluorescence (OJIP) transients from magnetically treated plants gave a higher fluorescence yield at the J-I-P phase. The total soluble protein map (SDS-polyacrylamide gel) of leaves showed increased intensities of the bands corresponding to a larger subunit (53 KDa) and smaller subunit (14 KDa) of Rubisco in the treated plants. We report here the beneficial effect of pre-sowing magnetic treatment for improving germination parameters and biomass accumulation in soybean. Copyright © 2011 Wiley-Liss, Inc.

  2. In situ analysis of plant tissue underivatized carbohydrates and on-probe enzymatic degraded starch by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry by using carbon nanotubes as matrix.

    PubMed

    Gholipour, Yousef; Nonami, Hiroshi; Erra-Balsells, Rosa

    2008-12-15

    Underivatized carbohydrates of tulip bulb and leaf tissues were characterized in situ by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) by using carbon nanotubes (CNTs) as matrix. Two sample preparation methods--(i) depositing CNTs on the fresh tissue slices placed on the probe and (ii) locating semitransparent tissues on a dried layer of CNTs on the probe--were examined. Furthermore, practicability of in situ starch analysis by MALDI-TOF MS was examined by detection of glucose originated from on-probe amyloglucosidase-catalyzed degradation of starch on the tissue surface. Besides, CNTs could efficiently desorb/ionize natural mono-, di-, and oligosaccharides extracted from tulip bulb tissues as well as glucose resulting from starch enzymatic degradation in vitro. These results were compared with those obtained by in situ MALDI-TOF MS analysis of similar tissues. Positive ion mode showed superior signal reproducibility. CNTs deposited under semitransparent tissue could also desorb/ionize neutral carbohydrates, leading to nearly complete elimination of matrix cluster signals but with an increase in tissue-originated signals. Furthermore, several experiments were carried out to compare the efficiency of 2,5-dihydroxybenzoic acid, nor-harmane, alpha-cyano-4-hydroxycinnamic acid, and CNTs as matrices for MALDI of neutral carbohydrates from the intact plant tissue surface and for enzymatic tissue starch degradation; these results are discussed in brief. Among matrices studied, the lowest laser power was needed to acquire carbohydrate signals with high signal-to-noise ratio and resolution when CNTs were used.

  3. Leaves from banana (Musa nana) and maize (Zea mays) have no phyto-prophylactic effects on the susceptibility of grass carp (Ctenopharyngodon idella) to Aeromonas hydrophila infection.

    PubMed

    Mayrhofer, Richard; Menanteau-Ledouble, Simon; Pucher, Johannes; Focken, Ulfert; El-Matbouli, Mansour

    2017-11-10

    The ubiquitous and opportunistic bacterial pathogen Aeromonas hydrophila has been associated with ulcerative dermatitis in fish, especially under stressful conditions. It can cause severe losses in fresh water aquaculture and is particularly prevalent in tropical and subtropical regions. Fresh leaves from maize and bananas have been used as feed supplement by fish farmers in Vietnam and it has been reported that they may have phyto-prophylactic benefits. In the present study, a feeding trial was conducted to investigate the benefits of providing maize and banana leaves as feed supplement: to determine if they were taken up and digested by grass carp (Ctenopharyngodon idella), if this uptake resulted in improved growth performance, and if leaf supplementation protected fish when challenged with A. hydrophila by intramuscular injection. All fish were fed an identical ratio of commercial pelleted feed relative to biomass. However, in 12/18 tanks, this diet was supplemented with either fresh banana leaves or fresh maize leaves; offered ad libitum. Addition of leaves increased the overall feed conversion ratio (FCR) significantly. However, if only the pellet were taken into account, then no difference was found between treatments. Changes to the isotopic composition of the fish showed leaf nutrient uptake occurred. No prophylactic effects of feeding banana or maize leaves were detected against infection with A. hydrophila, and the diet did not induce changes in the fish haematocrit. However, addition of the maize leaves was associated with significantly reduced severity of the skin lesions, which could improve the market value of the fish. Addition of the leaf supplement did not result in significantly improved growth performance. Similarly, the effect of the supplement on the fish survival to infection was not significant.

  4. Water Potential in Excised Leaf Tissue

    PubMed Central

    Nelsen, Charles E.; Safir, Gene R.; Hanson, Andrew D.

    1978-01-01

    Leaf water potential (Ψleaf) determinations were made on excised leaf samples using a commercial dew point hygrometer (Wescor Inc., Logan, Utah) and a thermocouple psychrometer operated in the isopiestic mode. With soybean leaves (Glycine max L.), there was good agreement between instruments; equilibration times were 2 to 3 hours. With cereals (Triticum aestivum L. and Hordeum vulgare L.), agreement between instruments was poor for moderately wilted leaves when 7-mm-diameter punches were used in the hygrometer and 20-mm slices were used in the psychrometer, because the Ψleaf values from the dew point hygrometer were too high. Agreement was improved by replacing the 7-mm punch samples in the hygrometer by 13-mm slices, which had a lower cut edge to volume ratio. Equilibration times for cereals were normally 6 to 8 hours. Spuriously high Ψleaf values obtained with 7-mm leaf punches may be associated with the ion release and reabsorption that occur upon tissue excision; such errors evidently depend both on the species and on tissue water status. PMID:16660227

  5. Microscopic evaluation and physiochemical analysis of Dillenia indica leaf

    PubMed Central

    Kumar, S; Kumar, V; Prakash, Om

    2011-01-01

    Objective To study detail microscopic evaluation and physiochemical analysis of Dillenia indica (D. indica) leaf. Methods Fresh leaf sample and dried power of the leaf were studied macroscopically and microscopically. Preliminary phytochemical investigation of plant material was done. Other WHO recommended parameters for standardizations were also performed. Results The detail microscopy revealed the presence of anomocytic stomata, unicellular trichome, xylem fibres, calcium oxalate crystals, vascular bundles, etc. Leaf constants such as stomatal number, stomatal index, vein-islet number and veinlet termination numbers were also measured. Physiochemical parameters such as ash values, loss on drying, extractive values, percentage of foreign matters, swelling index, etc. were also determined. Preliminary phytochemical screening showed the presence of steroids, terpenoids, glycosides, fatty acids, flavonoids, phenolic compounds and carbohydrates. Conclusions The microscopic and physiochemical analysis of the D. indica leaf is useful in standardization for quality, purity and sample identification. PMID:23569789

  6. Advantages and pitfalls of using free-hand sections of frozen needles for three-dimensional analysis of mesophyll by stereology and confocal microscopy

    PubMed Central

    LHOTÁKOVÁ, Z; ALBRECHTOVÁ, J; JANÁČEK, J; KUBÍNOVÁ, L

    2008-01-01

    The anatomical structure of mesophyll tissue in the leaf is tightly connected with many physiological processes in plants. One of the most important mesophyll parameters related to photosynthesis is the internal leaf surface area, i.e. the surface area of mesophyll cell walls exposed to intercellular spaces. An efficient design-based stereological method can be applied for estimation of this parameter, using software-randomized virtual fakir test probes in stacks of optical sections acquired by a confocal microscope within thick physical free-hand sections (i.e. acquired using a hand microtome), as we have shown in the case of fresh Norway spruce needles recently. However, for wider practical use in plant ecophysiology, a suitable form of sample storage and other possible technical constraints of this methodology need to be checked. We tested the effect of freezing conifer needles on their anatomical structure as well as the effect of possible deformations due to the cutting of unembedded material by a hand microtome, which can result in distortions of cutting surfaces. In the present study we found a higher proportion of intercellular spaces in mesophyll in regions near to the surface of a physical section, which means that the measurements should be restricted only to the middle region of the optical section series. On the other hand, the proportion of intercellular spaces in mesophyll as well as the internal needle surface density in mesophyll did not show significant difference between fresh and frozen needles; therefore, we conclude that freezing represents a suitable form of storage of sampled material for proposed stereological evaluation. PMID:19017201

  7. Advantages and pitfalls of using free-hand sections of frozen needles for three-dimensional analysis of mesophyll by stereology and confocal microscopy.

    PubMed

    Lhotáková, Z; Albrechtová, J; Janácek, J; Kubínová, L

    2008-10-01

    The anatomical structure of mesophyll tissue in the leaf is tightly connected with many physiological processes in plants. One of the most important mesophyll parameters related to photosynthesis is the internal leaf surface area, i.e. the surface area of mesophyll cell walls exposed to intercellular spaces. An efficient design-based stereological method can be applied for estimation of this parameter, using software-randomized virtual fakir test probes in stacks of optical sections acquired by a confocal microscope within thick physical free-hand sections (i.e. acquired using a hand microtome), as we have shown in the case of fresh Norway spruce needles recently. However, for wider practical use in plant ecophysiology, a suitable form of sample storage and other possible technical constraints of this methodology need to be checked. We tested the effect of freezing conifer needles on their anatomical structure as well as the effect of possible deformations due to the cutting of unembedded material by a hand microtome, which can result in distortions of cutting surfaces. In the present study we found a higher proportion of intercellular spaces in mesophyll in regions near to the surface of a physical section, which means that the measurements should be restricted only to the middle region of the optical section series. On the other hand, the proportion of intercellular spaces in mesophyll as well as the internal needle surface density in mesophyll did not show significant difference between fresh and frozen needles; therefore, we conclude that freezing represents a suitable form of storage of sampled material for proposed stereological evaluation.

  8. Physics of Fresh Produce Safety: Role of Diffusion and Tissue Reaction in Sanitization of Leafy Green Vegetables with Liquid and Gaseous Ozone-Based Sanitizers.

    PubMed

    Shynkaryk, Mykola V; Pyatkovskyy, Taras; Mohamed, Hussein M; Yousef, Ahmed E; Sastry, Sudhir K

    2015-12-01

    Produce safety has received much recent attention, with the emphasis being largely on discovery of how microbes invade produce. However, the sanitization operation deserves more attention than it has received. The ability of a sanitizer to reach the site of pathogens is a fundamental prerequisite for efficacy. This work addresses the transport processes of ozone (gaseous and liquid) sanitizer for decontamination of leafy greens. The liquid sanitizer was ineffective against Escherichia coli K-12 in situations where air bubbles may be trapped within cavities. A model was developed for diffusion of sanitizer into the interior of produce. The reaction rate of ozone with the surface of a lettuce leaf was determined experimentally and was used in a numerical simulation to evaluate ozone concentrations within the produce and to determine the time required to reach different locations. For aqueous ozone, the penetration depth was limited to several millimeters by ozone self-decomposition due to the significant time required for diffusion. In contrast, gaseous sanitizer was able to reach a depth of 100 mm in several minutes without depletion in the absence of reaction with surfaces. However, when the ozone gas reacted with the produce surface, gas concentration was significantly affected. Simulation data were validated experimentally by measuring ozone concentrations at the bottom of a cylinder made of lettuce leaf. The microbiological test confirmed the relationship between ozone transport, its self-decomposition, reaction with surrounding materials, and the degree of inactivation of E. coli K-12. Our study shows that decontamination of fresh produce, through direct contact with the sanitizer, is more feasible with gaseous than with aqueous sanitizers. Therefore, sanitization during a high-speed washing process is effective only for decontaminating the wash water.

  9. Biotechnology and genetic optimization of fast-growing hardwoods

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Garton, S.; Syrkin-Wurtele, E.; Griffiths, H.

    1991-02-01

    A biotechnology research program was initiated to develop new clones of fast-growing Populus clones resistant to the herbicide glyphosate and resistant to the leaf-spot and canker disease caused by the fungus Septoria musiva. Glyphosate-resistant callus was selected from stem segments cultured in vitro on media supplemented with the herbicide. Plants were regenerated from the glyphosate-resistant callus tissue. A portion of plants reverted to a glyphosate susceptible phenotype during organogenesis. A biologically active filtrate was prepared from S. musiva and influenced fresh weight of Populus callus tissue. Disease-resistant plants were produced through somaclonal variation when shoots developed on stem internodes culturedmore » in vitro. Plantlets were screened for disease symptoms after spraying with a suspension of fungal spores. A frequency of 0.83 percent variant production was observed. Genetically engineered plants were produced after treatment of plant tissue with Agrobacterium tumefasciens strains carrying plasmid genes for antibiotic resistance. Transformers were selected on media enriched with the antibiotic, kanamycin. Presence of foreign DNA was confirmed by Southern blot analysis. Protoplasts of popular were produced but did not regenerate into plant organs. 145 refs., 12 figs., 36 tabs.« less

  10. Pharmacodynamic correlations using fresh and cryopreserved tissue following use of vaginal rings containing dapivirine and/or maraviroc in a randomized, placebo controlled trial

    PubMed Central

    Dezzutti, Charlene S.; Richardson-Harman, Nicola; Rohan, Lisa C.; Marzinke, Mark A.; Hoesley, Craig J.; Panther, Lori; Johnson, Sherri; Nuttall, Jeremy P.; Nel, Annalene; Chen, Beatrice A.

    2016-01-01

    Abstract Background: The ex vivo challenge assay is a bio-indicator of drug efficacy and was utilized in this randomized, placebo controlled trial as one of the exploratory endpoints. Fresh and cryopreserved tissues were evaluated for human immunodeficiency virus (HIV) infection and pharmacokinetic (PK)/pharmacodynamic (PD) relationships. Methods: HIV-negative women used vaginal rings containing 25 mg dapivirine (DPV)/100 mg maraviroc (MVC) (n = 12), DPV only (n = 12), MVC only (n = 12), or placebo (n = 12) for 28 days. Blood plasma, cervicovaginal fluid (CVF), and cervical biopsies were collected for drug quantification and the ex vivo challenge assay; half (fresh) were exposed immediately to HIV while the other half were cryopreserved, thawed, then exposed to HIV. HIV replication was monitored by p24 enzyme-linked immunosorbent assay from culture supernatant. Data were log-transformed and analyzed by linear least squared regression, nonlinear Emax dose–response model and Satterthwaite t test. Results: HIV replication was greater in fresh compared to cryopreserved tissue (P = 0.04). DPV was detected in all compartments, while MVC was consistently detected only in CVF. Significant negative correlations between p24 and DPV levels were observed in fresh cervical tissue (P = 0.01) and CVF (P = 0.03), but not plasma. CVF MVC levels showed a significant negative correlation with p24 levels (P = 0.03); drug levels in plasma and tissue were not correlated with HIV suppression. p24 levels from cryopreserved tissue did not correlate to either drug from any compartment. Conclusion: Fresh tissue replicated HIV to greater levels and defined PK/PD relationships while cryopreserved tissue did not. The ex vivo challenge assay using fresh tissue could prioritize drugs being considered for HIV prevention. PMID:27428211

  11. Pharmacodynamic correlations using fresh and cryopreserved tissue following use of vaginal rings containing dapivirine and/or maraviroc in a randomized, placebo controlled trial.

    PubMed

    Dezzutti, Charlene S; Richardson-Harman, Nicola; Rohan, Lisa C; Marzinke, Mark A; Hoesley, Craig J; Panther, Lori; Johnson, Sherri; Nuttall, Jeremy P; Nel, Annalene; Chen, Beatrice A

    2016-07-01

    The ex vivo challenge assay is a bio-indicator of drug efficacy and was utilized in this randomized, placebo controlled trial as one of the exploratory endpoints. Fresh and cryopreserved tissues were evaluated for human immunodeficiency virus (HIV) infection and pharmacokinetic (PK)/pharmacodynamic (PD) relationships. HIV-negative women used vaginal rings containing 25 mg dapivirine (DPV)/100 mg maraviroc (MVC) (n = 12), DPV only (n = 12), MVC only (n = 12), or placebo (n = 12) for 28 days. Blood plasma, cervicovaginal fluid (CVF), and cervical biopsies were collected for drug quantification and the ex vivo challenge assay; half (fresh) were exposed immediately to HIV while the other half were cryopreserved, thawed, then exposed to HIV. HIV replication was monitored by p24 enzyme-linked immunosorbent assay from culture supernatant. Data were log-transformed and analyzed by linear least squared regression, nonlinear Emax dose-response model and Satterthwaite t test. HIV replication was greater in fresh compared to cryopreserved tissue (P = 0.04). DPV was detected in all compartments, while MVC was consistently detected only in CVF. Significant negative correlations between p24 and DPV levels were observed in fresh cervical tissue (P = 0.01) and CVF (P = 0.03), but not plasma. CVF MVC levels showed a significant negative correlation with p24 levels (P = 0.03); drug levels in plasma and tissue were not correlated with HIV suppression. p24 levels from cryopreserved tissue did not correlate to either drug from any compartment. Fresh tissue replicated HIV to greater levels and defined PK/PD relationships while cryopreserved tissue did not. The ex vivo challenge assay using fresh tissue could prioritize drugs being considered for HIV prevention.

  12. Fresh, dried or smoked? repellent properties of volatiles emitted from ethnomedicinal plant leaves against malaria and yellow fever vectors in Ethiopia

    PubMed Central

    2011-01-01

    Background In the search for plant-based mosquito repellents, volatile emanations were investigated from five plant species, Corymbia citriodora, Ocimum suave, Ocimum lamiifolium, Olea europaea and Ostostegia integrifolia, traditionally used in Ethiopia as protection against mosquitoes. Methods The behaviour of two mosquitoes, the malaria vector Anopheles arabiensis and the arbovirus vector Aedes aegypti, was assessed towards volatiles collected from the headspace of fresh and dried leaves, and the smoke from burning the dried leaves in a two-choice landing bioassay and in the background of human odour. Results Volatile extracts from the smoke of burning dried leaves were found to be more repellent than those from fresh leaves, which in turn were more repellent to mosquitoes than volatiles from dried leaves. Of all smoke and fresh volatile extracts, those from Co. citriodora (52-76%) and Oc. suave (58-68%) were found to be the most repellent, Os. integrifolia (29-56%) to be intermediate while Ol. europaea (23-40%) and Os. integrifolia (19-37%) were the least repellent. One volatile present in each of the fresh leaf extracts of Co. citriodora, Oc. suave and Os. integrifolia was ß-ocimene. The levels of ß-ocimene reflected the mosquito repellent activity of these three fresh leaf extracts. Female host-seeking mosquitoes responded dose-dependently to ß-ocimene, both physiologically and behaviourally, with a maximal behavioural repulsion at 14% ß-ocimene. ß-ocimene (14%) repels mosquitoes in our 6-minute landing assays comparable to the synthetic insect repellent N,N-diethyl-m-toluamide (10% DEET). Conclusions Volatiles in the smoke of burning as well as fresh leaves of Co. citriodora and Oc. suave have significant repellent properties against host seeking An. arabiensis and Ae. aegypti mosquitoes. ß-ocimene, present in the fresh leaf headspace of Co. citriodora, Oc. suave and Os. integrifolia, is a significantly effective volatile mosquito repellent in the laboratory. In addition to its repellent properties, ß-ocimene has long approved safe for use in food and cosmetics, making this volatile an intriguing compound to pursue in further tests in the laboratory and field to validate its mosquito repellent activity and potential for use in a commercial product. Also, the landing bioassay with humanised membranes is a potentially useful repellent screening technique that does not require the exposure of humans to the vectors, however further tests in parallel with conventional techniques are advised. PMID:22182798

  13. Fresh, dried or smoked? Repellent properties of volatiles emitted from ethnomedicinal plant leaves against malaria and yellow fever vectors in Ethiopia.

    PubMed

    Dube, Fitsum Fikru; Tadesse, Kassahun; Birgersson, Göran; Seyoum, Emiru; Tekie, Habte; Ignell, Rickard; Hill, Sharon R

    2011-12-19

    In the search for plant-based mosquito repellents, volatile emanations were investigated from five plant species, Corymbia citriodora, Ocimum suave, Ocimum lamiifolium, Olea europaea and Ostostegia integrifolia, traditionally used in Ethiopia as protection against mosquitoes. The behaviour of two mosquitoes, the malaria vector Anopheles arabiensis and the arbovirus vector Aedes aegypti, was assessed towards volatiles collected from the headspace of fresh and dried leaves, and the smoke from burning the dried leaves in a two-choice landing bioassay and in the background of human odour. Volatile extracts from the smoke of burning dried leaves were found to be more repellent than those from fresh leaves, which in turn were more repellent to mosquitoes than volatiles from dried leaves. Of all smoke and fresh volatile extracts, those from Co. citriodora (52-76%) and Oc. suave (58-68%) were found to be the most repellent, Os. integrifolia (29-56%) to be intermediate while Ol. europaea (23-40%) and Os. integrifolia (19-37%) were the least repellent. One volatile present in each of the fresh leaf extracts of Co. citriodora, Oc. suave and Os. integrifolia was ß-ocimene. The levels of ß-ocimene reflected the mosquito repellent activity of these three fresh leaf extracts. Female host-seeking mosquitoes responded dose-dependently to ß-ocimene, both physiologically and behaviourally, with a maximal behavioural repulsion at 14% ß-ocimene. ß-ocimene (14%) repels mosquitoes in our 6-minute landing assays comparable to the synthetic insect repellent N,N-diethyl-m-toluamide (10% DEET). Volatiles in the smoke of burning as well as fresh leaves of Co. citriodora and Oc. suave have significant repellent properties against host seeking An. arabiensis and Ae. aegypti mosquitoes. ß-ocimene, present in the fresh leaf headspace of Co. citriodora, Oc. suave and Os. integrifolia, is a significantly effective volatile mosquito repellent in the laboratory. In addition to its repellent properties, ß-ocimene has long approved safe for use in food and cosmetics, making this volatile an intriguing compound to pursue in further tests in the laboratory and field to validate its mosquito repellent activity and potential for use in a commercial product. Also, the landing bioassay with humanised membranes is a potentially useful repellent screening technique that does not require the exposure of humans to the vectors, however further tests in parallel with conventional techniques are advised.

  14. Regulation of leaf hydraulics: from molecular to whole plant levels

    PubMed Central

    Prado, Karine; Maurel, Christophe

    2013-01-01

    The water status of plant leaves is dependent on both stomatal regulation and water supply from the vasculature to inner tissues. The present review addresses the multiple physiological and mechanistic facets of the latter process. Inner leaf tissues contribute to at least a third of the whole resistance to water flow within the plant. Physiological studies indicated that leaf hydraulic conductance (Kleaf) is highly dependent on the anatomy, development and age of the leaf and can vary rapidly in response to physiological or environmental factors such as leaf hydration, light, temperature, or nutrient supply. Differences in venation pattern provide a basis for variations in Kleaf during development and between species. On a short time (hour) scale, the hydraulic resistance of the vessels can be influenced by transpiration-induced cavitations, wall collapses, and changes in xylem sap composition. The extravascular compartment includes all living tissues (xylem parenchyma, bundle sheath, and mesophyll) that transport water from xylem vessels to substomatal chambers. Pharmacological inhibition and reverse genetics studies have shown that this compartment involves water channel proteins called aquaporins (AQPs) that facilitate water transport across cell membranes. In many plant species, AQPs are present in all leaf tissues with a preferential expression in the vascular bundles. The various mechanisms that allow adjustment of Kleaf to specific environmental conditions include transcriptional regulation of AQPs and changes in their abundance, trafficking, and intrinsic activity. Finally, the hydraulics of inner leaf tissues can have a strong impact on the dynamic responses of leaf water potential and stomata, and as a consequence on plant carbon economy and leaf expansion growth. The manipulation of these functions could help optimize the entire plant performance and its adaptation to extreme conditions over short and long time scales. PMID:23874349

  15. Antiinflammatory and analgesic effects of Psidium guajava Linn. (Myrtaceae) leaf aqueous extract in rats and mice.

    PubMed

    Ojewole, J A O

    2006-09-01

    In many parts of Africa, the leaf, stem-bark, and roots of Psidium guajava Linn. (Family: Myrtaceae) are used traditionally for the management, control, and/or treatment of an array of human disorders. In an effort to scientifically appraise some of the ethnomedical properties of P. guajava leaf, and probe its efficacy and safety, the present study was undertaken to examine the antiinflammatory and analgesic properties of the plant's leaf aqueous extract in some experimental animal paradigms. The antiinflammatory property of the aqueous leaf extract was investigated in rats, using fresh egg albumin-induced pedal (paw) edema, while the analgesic effect of the plant extract was evaluated by the "hot-plate" and "acetic acid" test models of pain in mice. Diclofenac (100 mg/kg, i.p.) and morphine (10 mg/kg, i.p.) were used respectively as standard, reference antiinflammatory and analgesic agents for comparison. P. guajava leaf aqueous extract (PGE, 50-800 mg/kg, i.p.) produced dose-dependent and significant (p < 0.05-0.001) inhibition of fresh egg albumin-induced acute inflammation (edema) in rats. The plant extract (PGE, 50-800 mg/kg, i.p.) also produced dose-dependent and significant (p < 0.05-0.001) analgesic effects against thermally and chemically induced nociceptive pain in mice. The numerous tannins, polyphenolic compounds, flavonoids, ellagic acid, triterpenoids, guiajaverin, quercetin, and other chemical compounds present in the plant are speculated to account for the observed antiinflammatory and analgesic effects of the plant's leaf extract. In summary, the findings of this experimental animal study indicate that the leaf aqueous extract of P. guajava possesses analgesic and antiinflammatory properties, and thus lend pharmacological credence to the suggested ethnomedical, folkloric uses of the plant in the management and/or control of painful, arthritic and other inflammatory conditions in some rural communities of Africa. (c) 2006 Prous Science. All rights reserved.

  16. Response of Grape Leaf Spectra to Phylloxera Infestation

    NASA Technical Reports Server (NTRS)

    Johnson, Lee F.

    1999-01-01

    During the 1993 growing season, leaf reflectance and chlorophyll concentrations were monitored with respect to phylloxera (root-louse) infestation in a Napa Valley (California) vineyard. Study plots were established in areas of severely infested, mildly infested, and uninfested sections of the vineyard. A handheld chlorophyll meter, measuring leaf transmittance of near-infrared and red light, confirmed that reduced foliar chlorophyll concentrations were symptomatic of phylloxera stress in the sample vines. Bidirectional reflectance measurements of green and near-infrared light, taken on fresh leaves with a laboratory spectrophotometer, were related to chlorophyll concentration but did not allow discrimination of mildly infested from uninfested vines.

  17. Anti microbial activity of the various leaf extracts of Vitex negundo Linn.

    PubMed Central

    Devi, P. Renuka; Kokilavani, R.; Poongothai, S. Gnana

    2008-01-01

    The antibacterial activity of the leaves of Vitex negundo was tested against three types of bacteria Viz., Staphylococcus aureus, Escherichia coli and Klebsiella Pneumoniae. The fresh, aqueous, heated aqueous extract, chloroform and methanolic extract of leaves were used for screening their antibacterial potential. The fresh and aqueous extracts of leaves in various dilutions were found to have antibacterial activity against the three bacteria. PMID:22557282

  18. Response of hydroponically grown head lettuce on residual monomer from polyacrylamide.

    PubMed

    Mroczek, E; Konieczny, P; Kleiber, T; Waśkiewicz, A

    2014-01-01

    The aim was to assess acrylamide monomer (AMD) uptake by hydroponically grown lettuce. Lettuce was cultivated by applying plant tissue testing in a recycled system by the use of nutrient solutions prepared with two water-soluble flocculants F3 and F4 containing 176 and 763 mg kg(-1) of AMD, respectively. The effects on growth, fresh weight and plant leaf quality were evaluated by comparing these treatments and one control standard nutrient solution typically recommended for lettuce hydroponic cultivation. To assess the nutritional status of lettuce, samples were collected and lyophilised before determination of the selected micro- and macro-element contents. An HPLC with photodiode array detector method was applied to determine AMD in both selected flocculants and dried plant samples. Results show that lettuces cultivated under the conditions described above absorb AMD from nutrient solutions into their leaves. The AMD presence in recycled nutrient solutions has a negative influence on the growth of lettuce, reducing their average fresh weight and average number of leaves. The study confirmed that the problem of AMD mobility and its accumulation risk in plants should to be an important topic with respect to safe polyacrylamide (PAM) handling in the agro food area.

  19. The influence of some environmental factors on cytological and biometric parameters and chlorophyll content of Deschampsia antarctica Desv. in the maritime Antarctic.

    PubMed

    Parnikoza, I Yu; Loro, P; Miryuta, N Yu; Kunakh, V A; Kozeretska, I A

    2011-01-01

    Under the environmental conditions of the Point Thomas Oasis (King George Island, the South Shetland Islands), we studied the influence of month-long artificial treatment with fresh water, salt water, and guano solution on the biometric characteristics, chlorophyll content, as well as the nuclear area of leaf parenchymal cells and nuclear DNA content, in a maritime Antarctic aboriginal plant Deschampsia antarctica. The modeled factors induced an increase in the generative shoot height and the length of the largest leaf, but did not influence the number of flowers. Treatment with guano caused an increase in the chlorophyll a and b contents, while fresh water treatment only led to some increase in chlorophyll a. Fluctuations of physiologically significant traits, such as the nuclear area and DNA content in the leaf parenchyma cells of D. antarctica, have been traced under the influence of the studied factors. Understanding of the hierarchy of influence of these factors as well as and sensitivity of plants of this species to external agents require further investigation.

  20. Wound healing effects of noni (Morinda citrifolia L.) leaves: a mechanism involving its PDGF/A2A receptor ligand binding and promotion of wound closure.

    PubMed

    Palu, Afa; Su, Chen; Zhou, Bing-Nan; West, Brett; Jensen, Jarakae

    2010-10-01

    Morinda citrifolia L. (Rubiaceae) commonly known as noni, has been used in Polynesia by traditional healers for the treatment of cuts, bruises and wounds. Our objective was to investigate the wound-healing mechanisms of the noni leaf. The investigations of its wound-healing mechanisms were carried out using fresh noni leaf juice (NLJ), noni leaf ethanol extract (NLEE) and its methanol (MFEE) and hexane (HFEE) fractions on the PDGF and A(2A) receptors in vitro and topically in mice. Fresh noni leaf juice showed significant affinity to PDGF receptors, and displayed 166% binding inhibition of the ligand binding to its receptors, while at the same concentration, it only had 7% inhibition of the ligand binding to the A(2A) receptors. NLEE, HFEE and MFEE showed significant affinity to A(2A) receptors, concentration dependently, with IC(50) values of 34.1, 42.9 and 86.7 μg/mL, respectively. However, MFEE significantly increased wound closure and reduced the half closure time in mice with a CT(50) of 5.4 ± 0.2 days compared with control (p < 0.05). These results suggest that noni leaf significantly accelerated wound healing in mice via its ligand binding to the PDGF and A(2A) receptors as its probable mechanisms of wound-healing and also support its traditional usage for wound-healing in Polynesia. Copyright © 2010 John Wiley & Sons, Ltd.

  1. Comparative Transcriptomics Unravel Biochemical Specialization of Leaf Tissues of Stevia for Diterpenoid Production1

    PubMed Central

    Kim, Mi Jung; Jin, Jingjing; Zheng, Junshi

    2015-01-01

    Stevia (Stevia rebaudiana) produces not only a group of diterpenoid glycosides known as steviol glycosides (SGs), but also other labdane-type diterpenoids that may be spatially separated from SGs. However, their biosynthetic routes and spatial distribution in leaf tissues have not yet been elucidated. Here, we integrate metabolome and transcriptome analyses of Stevia to explore the biosynthetic capacity of leaf tissues for diterpenoid metabolism. Tissue-specific chemical analyses confirmed that SGs were accumulated in leaf cells but not in trichomes. On the other hand, Stevia leaf trichomes stored other labdane-type diterpenoids such as oxomanoyl oxide and agatholic acid. RNA sequencing analyses from two different tissues of Stevia provided a comprehensive overview of dynamic metabolic activities in trichomes and leaf without trichomes. These metabolite-guided transcriptomics and phylogenetic and gene expression analyses clearly identified specific gene members encoding enzymes involved in the 2-C-methyl-d-erythritol 4-phosphate pathway and the biosynthesis of steviol or other labdane-type diterpenoids. Additionally, our RNA sequencing analysis uncovered copalyl diphosphate synthase (SrCPS) and kaurene synthase1 (SrKS1) homologs, SrCPS2 and KS-like (SrKSL), which were specifically expressed in trichomes. In vitro and in planta assays showed that unlike SrCPS and SrKS1, SrCPS2 synthesized labda-13-en-8-ol diphosphate and successively catalyzed the formation of manoyl oxide and epi-manoyl oxide in combination with SrKSL. Our findings suggest that Stevia may have evolved to use distinct metabolic pathways to avoid metabolic interferences in leaf tissues for efficient production of diverse secondary metabolites. PMID:26438788

  2. The relationships between leaf economics and hydraulic traits of woody plants depend on water availability.

    PubMed

    Yin, Qiulong; Wang, Lei; Lei, Maolin; Dang, Han; Quan, Jiaxin; Tian, Tingting; Chai, Yongfu; Yue, Ming

    2018-04-15

    Leaf economics and hydraulic traits are simultaneously involved in the process of trading water for CO 2 , but the relationships between these two suites of traits remain ambiguous. Recently, Li et al. (2015) reported that leaf economics and hydraulic traits were decoupled in five tropical-subtropical forests in China. We tested the hypothesis that the relationships between economics and hydraulic traits may depend on water availability. We analysed five leaf economics traits, four hydraulic traits and anatomical structures of 47 woody species on the Loess Plateau with poor water availability and compared those data with Li et al. (2015) obtained in tropical-subtropical regions with adequate water. The results showed that plants on the Loess Plateau tend to have higher leaf tissue density (TD), leaf nitrogen concentrations and venation density (VD) and lower stomatal guard cell length (SL) and maximum stomatal conductance to water vapour (g wmax ). VD showed positive correlations with leaf nitrogen concentrations, palisade tissue thickness (PT) and ratio of palisade tissue thickness to spongy tissue thickness (PT/ST). Principal component analysis (PCA) showed a result opposite from those of tropical-subtropical regions: leaf economics and hydraulic traits were coupled on the Loess Plateau. A stable correlation between these two suites of traits may be more cost-effective on the Loess Plateau, where water availability is poor. The correlation of leaf economics and hydraulic traits may be a type of adaptation mechanism in arid conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Errors in Measuring Water Potentials of Small Samples Resulting from Water Adsorption by Thermocouple Psychrometer Chambers 1

    PubMed Central

    Bennett, Jerry M.; Cortes, Peter M.

    1985-01-01

    The adsorption of water by thermocouple psychrometer assemblies is known to cause errors in the determination of water potential. Experiments were conducted to evaluate the effect of sample size and psychrometer chamber volume on measured water potentials of leaf discs, leaf segments, and sodium chloride solutions. Reasonable agreement was found between soybean (Glycine max L. Merr.) leaf water potentials measured on 5-millimeter radius leaf discs and large leaf segments. Results indicated that while errors due to adsorption may be significant when using small volumes of tissue, if sufficient tissue is used the errors are negligible. Because of the relationship between water potential and volume in plant tissue, the errors due to adsorption were larger with turgid tissue. Large psychrometers which were sealed into the sample chamber with latex tubing appeared to adsorb more water than those sealed with flexible plastic tubing. Estimates are provided of the amounts of water adsorbed by two different psychrometer assemblies and the amount of tissue sufficient for accurate measurements of leaf water potential with these assemblies. It is also demonstrated that water adsorption problems may have generated low water potential values which in prior studies have been attributed to large cut surface area to volume ratios. PMID:16664367

  4. Errors in measuring water potentials of small samples resulting from water adsorption by thermocouple psychrometer chambers.

    PubMed

    Bennett, J M; Cortes, P M

    1985-09-01

    The adsorption of water by thermocouple psychrometer assemblies is known to cause errors in the determination of water potential. Experiments were conducted to evaluate the effect of sample size and psychrometer chamber volume on measured water potentials of leaf discs, leaf segments, and sodium chloride solutions. Reasonable agreement was found between soybean (Glycine max L. Merr.) leaf water potentials measured on 5-millimeter radius leaf discs and large leaf segments. Results indicated that while errors due to adsorption may be significant when using small volumes of tissue, if sufficient tissue is used the errors are negligible. Because of the relationship between water potential and volume in plant tissue, the errors due to adsorption were larger with turgid tissue. Large psychrometers which were sealed into the sample chamber with latex tubing appeared to adsorb more water than those sealed with flexible plastic tubing. Estimates are provided of the amounts of water adsorbed by two different psychrometer assemblies and the amount of tissue sufficient for accurate measurements of leaf water potential with these assemblies. It is also demonstrated that water adsorption problems may have generated low water potential values which in prior studies have been attributed to large cut surface area to volume ratios.

  5. 7 CFR 51.1221 - Damage.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... FRESH FRUITS, VEGETABLES AND OTHER PRODUCTS 1,2 (INSPECTION, CERTIFICATION, AND STANDARDS) United States...) Hail injury which is unhealed, or deep, or when aggregating more than 1/4 inch in diameter; (e) Leaf or...

  6. Fresh broad (Vicia faba) tissue homogenate-based biosensor for determination of phenolic compounds.

    PubMed

    Ozcan, Hakki Mevlut; Sagiroglu, Ayten

    2014-08-01

    In this study, a novel fresh broad (Vicia faba) tissue homogenate-based biosensor for determination of phenolic compounds was developed. The biosensor was constructed by immobilizing tissue homogenate of fresh broad (Vicia faba) on to glassy carbon electrode. For the stability of the biosensor, general immobilization techniques were used to secure the fresh broad tissue homogenate in gelatin-glutaraldehyde cross-linking matrix. In the optimization and characterization studies, the amount of fresh broad tissue homogenate and gelatin, glutaraldehyde percentage, optimum pH, optimum temperature and optimum buffer concentration, thermal stability, interference effects, linear range, storage stability, repeatability and sample applications (Wine, beer, fruit juices) were also investigated. Besides, the detection ranges of thirteen phenolic compounds were obtained with the help of the calibration graphs. A typical calibration curve for the sensor revealed a linear range of 5-60 μM catechol. In reproducibility studies, variation coefficient (CV) and standard deviation (SD) were calculated as 1.59%, 0.64×10(-3) μM, respectively.

  7. Factors contributing to deep supercooling capability and cold survival in dwarf bamboo (Sasa senanensis) leaf blades.

    PubMed

    Ishikawa, Masaya; Oda, Asuka; Fukami, Reiko; Kuriyama, Akira

    2014-01-01

    Wintering Sasa senanensis, dwarf bamboo, is known to employ deep supercooling as the mechanism of cold hardiness in most of its tissues from leaves to rhizomes. The breakdown of supercooling in leaf blades has been shown to proceed in a random and scattered manner with a small piece of tissue surrounded by longitudinal and transverse veins serving as the unit of freezing. The unique cold hardiness mechanism of this plant was further characterized using current year leaf blades. Cold hardiness levels (LT20: the lethal temperature at which 20% of the leaf blades are injured) seasonally increased from August (-11°C) to December (-20°C). This coincided with the increases in supercooling capability of the leaf blades as expressed by the initiation temperature of low temperature exotherms (LTE) detected in differential thermal analyses (DTA). When leaf blades were stored at -5°C for 1-14 days, there was no nucleation of the supercooled tissue units either in summer or winter. However, only summer leaf blades suffered significant injury after prolonged supercooling of the tissue units. This may be a novel type of low temperature-induced injury in supercooled state at subfreezing temperatures. When winter leaf blades were maintained at the threshold temperature (-20°C), a longer storage period (1-7 days) increased lethal freezing of the supercooled tissue units. Within a wintering shoot, the second or third leaf blade from the top was most cold hardy and leaf blades at lower positions tended to suffer more injury due to lethal freezing of the supercooled units. LTE were shifted to higher temperatures (2-5°C) after a lethal freeze-thaw cycle. The results demonstrate that the tissue unit compartmentalized with longitudinal and transverse veins serves as the unit of supercooling and temperature- and time-dependent freezing of the units is lethal both in laboratory freeze tests and in the field. To establish such supercooling in the unit, structural ice barriers such as development of sclerenchyma and biochemical mechanisms to increase the stability of supercooling are considered important. These mechanisms are discussed in regard to ecological and physiological significance in winter survival.

  8. Leaf Pressure Volume Data in Caxiuana and Tapajos National Forest, Para, Brazil (2011)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Powell, Thomas; Moorcroft, Paul

    Pressure volume curve measurements on leaves of canopy trees from the from the Caxiuana and Tapajos National Forests, Para, Brazil. Tapajos samples were harvested from the km 67 forested area, which is adjacent to the decommissioned throughfall exclusion drought experimental plot. Caxiuana samples were harvested from trees growing in the throughfall exclusion plots. Data were collected in 2011. Dataset includes: date of measurement, site ID, plot ID, tree ID (species, tree tag #), leaf area, fresh weight, relative weight, leaf water potential, and leaf water loss. P-V curve parameters (turgor loss point, osmotic potential, and bulk modulus of elasticity) canmore » be found in Powell et al. (2017) Differences in xylem cavitation resistance and leaf hydraulic traits explain differences in drought tolerance among mature Amazon rainforest trees. Global Change Biology.« less

  9. Cultivation and phenotypic characterization of rabbit epithelial cells expanded ex vivo from fresh and cryopreserved limbal and oral mucosal explants.

    PubMed

    Promprasit, Daranee; Bumroongkit, Kanokkan; Tocharus, Chainarong; Mevatee, Umnat; Tananuvat, Napaporn

    2015-03-01

    To compare the morphology of cultured rabbit epithelial sheets and the expression of stem cells with differentiated cell markers of cultivated epithelial cells from fresh and cryopreserved limbal and oral mucosal biopsies. Six New Zealand white rabbits were divided into two groups of three, from which limbal and oral mucosal biopsies were taken. Harvested tissues from each rabbit were brought to immediate cultivation, while another set of tissues was cryopreserved. Cultivation was performed by the explant culture technique using human amniotic membrane as a culture substrate, co-culturing with 3T3 fibroblasts and using the air-lifting method. Cells were cultured for three weeks; then cultured epithelial sheets were stained with hematoxylin-eosin and examined for expression patterns of p63, keratin 3 (K3) and connexin 43 (Cx43). Cryopreservation was carried out using the vitrification method. Tissues were preserved in liquid nitrogen using 25% dimethyl sulfoxide combined with 25% propylene glycol in Dulbecco's Modified Eagle's Medium containing 20% fetal bovine serum. After two months, the tissues were warmed, cultured and stained using the same processes as for fresh tissue cultures. Cultivation of fresh limbal and fresh oral mucosal tissues showed epithelial stratification, with two to five cell layers. Immunohistochemical staining showed p63-positive cells in basal and intermediate cell layers. K3 staining was observed in cells in the suprabasal layer, while expression of Cx43 was scattered throughout all layers of the epithelia. All culture sheets expressed p63, K3 and Cx43 with the exception of one sheet from the oral mucosal culture that was p63-negative. Cultured epithelial sheets from cryopreserved tissues showed results similar to those from fresh tissue culture. This study found that cells in cultivated fresh limbal and oral mucosal tissues had similar morphology to cells in cultivated cryopreserved limbal and oral mucosal tissues, both containing a heterogeneous population of cells including stem cells and differentiated cells.

  10. Influence of Vacuum Cooling on Escherichia coli O157:H7 Infiltration in Fresh Leafy Greens via a Multiphoton-Imaging Approach

    PubMed Central

    Vonasek, Erica

    2015-01-01

    Microbial pathogen infiltration in fresh leafy greens is a significant food safety risk factor. In various postharvest operations, vacuum cooling is a critical process for maintaining the quality of fresh produce. The overall goal of this study was to evaluate the risk of vacuum cooling-induced infiltration of Escherichia coli O157:H7 into lettuce using multiphoton microscopy. Multiphoton imaging was chosen as the method to locate E. coli O157:H7 within an intact lettuce leaf due to its high spatial resolution, low background fluorescence, and near-infrared (NIR) excitation source compared to those of conventional confocal microscopy. The variables vacuum cooling, surface moisture, and leaf side were evaluated in a three-way factorial study with E. coli O157:H7 on lettuce. A total of 188 image stacks were collected. The images were analyzed for E. coli O157:H7 association with stomata and E. coli O157:H7 infiltration. The quantitative imaging data were statistically analyzed using analysis of variance (ANOVA). The results indicate that the low-moisture condition led to an increased risk of microbial association with stomata (P < 0.05). Additionally, the interaction between vacuum cooling levels and moisture levels led to an increased risk of infiltration (P < 0.05). This study also demonstrates the potential of multiphoton imaging for improving sensitivity and resolution of imaging-based measurements of microbial interactions with intact leaf structures, including infiltration. PMID:26475109

  11. Genome-Wide Identification of miRNAs Responsive to Drought in Peach (Prunus persica) by High-Throughput Deep Sequencing

    PubMed Central

    Eldem, Vahap; Çelikkol Akçay, Ufuk; Ozhuner, Esma; Bakır, Yakup; Uranbey, Serkan; Unver, Turgay

    2012-01-01

    Peach (Prunus persica L.) is one of the most important worldwide fresh fruits. Since fruit growth largely depends on adequate water supply, drought stress is considered as the most important abiotic stress limiting fleshy fruit production and quality in peach. Plant responses to drought stress are regulated both at transcriptional and post-transcriptional level. As post-transcriptional gene regulators, miRNAs (miRNAs) are small (19–25 nucleotides in length), endogenous, non-coding RNAs. Recent studies indicate that miRNAs are involved in plant responses to drought. Therefore, Illumina deep sequencing technology was used for genome-wide identification of miRNAs and their expression profile in response to drought in peach. In this study, four sRNA libraries were constructed from leaf control (LC), leaf stress (LS), root control (RC) and root stress (RS) samples. We identified a total of 531, 471, 535 and 487 known mature miRNAs in LC, LS, RC and RS libraries, respectively. The expression level of 262 (104 up-regulated, 158 down-regulated) of the 453 miRNAs changed significantly in leaf tissue, whereas 368 (221 up-regulated, 147 down-regulated) of the 465 miRNAs had expression levels that changed significantly in root tissue upon drought stress. Additionally, a total of 197, 221, 238 and 265 novel miRNA precursor candidates were identified from LC, LS, RC and RS libraries, respectively. Target transcripts (137 for LC, 133 for LS, 148 for RC and 153 for RS) generated significant Gene Ontology (GO) terms related to DNA binding and catalytic activites. Genome-wide miRNA expression analysis of peach by deep sequencing approach helped to expand our understanding of miRNA function in response to drought stress in peach and Rosaceae. A set of differentially expressed miRNAs could pave the way for developing new strategies to alleviate the adverse effects of drought stress on plant growth and development. PMID:23227166

  12. Effect of pest management system on 'Empire' apple leaf phyllosphere populations

    USDA-ARS?s Scientific Manuscript database

    The phyllosphere of plant tissues is varied and dynamic. Pest management, time of sampling, proximity to immigration sources, tissue and tissue status such as leaf/fruit age and location within the canopy, and other environmental and biological factors interact to influence the composition and abun...

  13. Water potential in excised leaf tissue: comparison of a commercial dew point hygrometer and a thermocouple psychrometer on soybean, wheat, and barley.

    PubMed

    Nelsen, C E; Safir, G R; Hanson, A D

    1978-01-01

    Leaf water potential (Psi(leaf)) determinations were made on excised leaf samples using a commercial dew point hygrometer (Wescor Inc., Logan, Utah) and a thermocouple psychrometer operated in the isopiestic mode. With soybean leaves (Glycine max L.), there was good agreement between instruments; equilibration times were 2 to 3 hours. With cereals (Triticum aestivum L. and Hordeum vulgare L.), agreement between instruments was poor for moderately wilted leaves when 7-mm-diameter punches were used in the hygrometer and 20-mm slices were used in the psychrometer, because the Psi(leaf) values from the dew point hygrometer were too high. Agreement was improved by replacing the 7-mm punch samples in the hygrometer by 13-mm slices, which had a lower cut edge to volume ratio. Equilibration times for cereals were normally 6 to 8 hours. Spuriously high Psi(leaf) values obtained with 7-mm leaf punches may be associated with the ion release and reabsorption that occur upon tissue excision; such errors evidently depend both on the species and on tissue water status.

  14. Leaf blade versus petiole nutrient tests as predictors of nitrogen, phosphorus, and potassium status of ‘Pinot noir’ grapevines

    USDA-ARS?s Scientific Manuscript database

    Grape growers rely on tissues tests of leaf blades or petioles for routine monitoring of vine nutritional health and for diagnosing potential nutrient deficiency or toxicity. There has been a long standing debate as to which tissue better reflects the nutrient status of vines. A comparison of leaf b...

  15. Comparative Transcriptomics Unravel Biochemical Specialization of Leaf Tissues of Stevia for Diterpenoid Production.

    PubMed

    Kim, Mi Jung; Jin, Jingjing; Zheng, Junshi; Wong, Limsoon; Chua, Nam-Hai; Jang, In-Cheol

    2015-12-01

    Stevia (Stevia rebaudiana) produces not only a group of diterpenoid glycosides known as steviol glycosides (SGs), but also other labdane-type diterpenoids that may be spatially separated from SGs. However, their biosynthetic routes and spatial distribution in leaf tissues have not yet been elucidated. Here, we integrate metabolome and transcriptome analyses of Stevia to explore the biosynthetic capacity of leaf tissues for diterpenoid metabolism. Tissue-specific chemical analyses confirmed that SGs were accumulated in leaf cells but not in trichomes. On the other hand, Stevia leaf trichomes stored other labdane-type diterpenoids such as oxomanoyl oxide and agatholic acid. RNA sequencing analyses from two different tissues of Stevia provided a comprehensive overview of dynamic metabolic activities in trichomes and leaf without trichomes. These metabolite-guided transcriptomics and phylogenetic and gene expression analyses clearly identified specific gene members encoding enzymes involved in the 2-C-methyl-d-erythritol 4-phosphate pathway and the biosynthesis of steviol or other labdane-type diterpenoids. Additionally, our RNA sequencing analysis uncovered copalyl diphosphate synthase (SrCPS) and kaurene synthase1 (SrKS1) homologs, SrCPS2 and KS-like (SrKSL), which were specifically expressed in trichomes. In vitro and in planta assays showed that unlike SrCPS and SrKS1, SrCPS2 synthesized labda-13-en-8-ol diphosphate and successively catalyzed the formation of manoyl oxide and epi-manoyl oxide in combination with SrKSL. Our findings suggest that Stevia may have evolved to use distinct metabolic pathways to avoid metabolic interferences in leaf tissues for efficient production of diverse secondary metabolites. © 2015 American Society of Plant Biologists. All Rights Reserved.

  16. Colonization and Movement of Xanthomonas fragariae in Strawberry Tissues.

    PubMed

    Wang, Hehe; McTavish, Christine; Turechek, William W

    2018-06-01

    Xanthomonas fragariae causes angular leaf spot of strawberry, an important disease in strawberry growing regions worldwide. To better understand how X. fragariae multiplies and moves in strawberry plants, a green fluorescent protein (GFP)-labeled strain was constructed and used to monitor the pathogen's presence in leaf, petiole, and crown tissue with fluorescence microscopy following natural and wound inoculation in three strawberry cultivars. Taqman PCR was used to quantify bacterial densities in these same tissues regardless of the presence of GFP signal. Results showed X. fragariae colonized leaf mesophyll, the top 1 cm portion of the petiole adjacent to the leaf blade, and was occasionally found colonizing xylem vessels down to the middle of the petioles. The colonization of vascular bundles and the limited systemic movement that was observed appeared to be a passive process, of which the frequency increased with wounding and direct infiltration of bacteria into leaf veins. X. fragariae was able to directly enter petioles and colonize the space under the epidermis. Systemic movement of the bacteria into crown and other uninoculated tissues was not detected visually by GFP. However, X. fragariae was occasionally detected in these tissues by qPCR, but at quantities very near the qPCR detection limit. Petiole tissue harboring bacteria introduced either by direct entry through natural openings or wounds, or by systemic movement from infected foliar tissue, likely serves as a main source of initial inoculum in field plantings.

  17. Comparative efficacy of Zataria multiflora Boiss., Origanum compactum and Eugenia caryophyllus essential oils against E. coli O157:H7, feline calicivirus and endogenous microbiota in commercial baby-leaf salads.

    PubMed

    Azizkhani, Maryam; Elizaquível, Patricia; Sánchez, Gloria; Selma, María Victoria; Aznar, Rosa

    2013-09-02

    Ready-to-eat salads using baby-leaf and multi-leaf mixes are one of the most promising developments in the fresh-cut food industry. There is great interest in developing novel decontamination treatments, which are both safe for consumers and more efficient against foodborne pathogens. In this study, emulsions of essential oils (EOs) from Origanum compactum (oregano), Eugenia caryophyllus (clove), and Zataria multiflora Boiss (zataria) were applied by spray (0.8 ml) after the sanitizing washing step. The aim was to investigate their ability to control the growth of potentially cross-contaminating pathogens and endogenous microbiota in commercial baby leaves, processed in a fresh-cut produce company. Zataria EO emulsions of 3%, 5% and 10% reduced Escherichia coli O157:H7 by 1.7, 2.2 and 3.5 log cfu/g in baby-leaf salads after 5 days of storage at 7°C. By contrast, reductions in E. coli O157:H7 counts remained the same when clove was applied at concentrations of 5% and 10% (2.5 log cfu/g reduction). Oregano (10%) reduced inoculated E. coli O157:H7 counts in baby-leaf salads by a maximum of 0.5 log cfu/g after 5 days of storage. Zataria showed strong antimicrobial efficacy against E. coli O157:H7 and also against the endogenous microbiota of baby-leaf salads stored for 9 days. Feline calicivirus (FCV), a norovirus surrogate, survived on inoculated baby-leaf salads during refrigerated storage (9 days at 7°C) regardless of treatment. Refrigeration temperatures completely annulled the effectiveness of the EOs against FCV inoculated in baby-leaf salads as occurred in FCV cultures. This study shows that EOs, and zataria in particular, have great potential use as an additional barrier to reduce contamination-related risks in baby-leaf salads. However, further research should be done into foodborne viruses in order to improve food safety. © 2013.

  18. Responses of Atriplex spongiosa and Suaeda monoica to Salinity

    PubMed Central

    Storey, Richard; Jones, R. Gareth Wyn

    1979-01-01

    The growth and tissue water, K+, Na+, Cl−, proline and glycinebetaine contents of the shoots and roots of two Chenopodiaceae, Atriplex spongiosa and Suaeda monoica have been measured over a range of external NaCl salinities. Both species showed some fresh weight response to low salinity mainly due to increased succulence. S. monoica showed both a greater increase in succulence (at low salinities) and tolerance of high salinities than A. spongiosa. Both species had high affinities for Na+ and maintained constant but low shoot K+ contents with increasing salinity. These trends were more marked with S. monoica in which Na+ stimulated the accumulation of K+ in roots. An association between high leaf Na+ accumulation, high osmotic pressure, succulence, and a positive growth response at low salinities was noted. Proline accumulation was observed in shoot tissues with suboptimal water contents. High glycinebetaine contents were found in the shoots of both species. These correlated closely with the sap osmotic pressure and it is suggested that glycinebetaine is the major cytoplasmic osmoticum (with K+ salts) in these species at high salinities. Na+ salts may be preferentially utilized as vacuolar osmotica. PMID:16660671

  19. [Concentration and risk assessment of DEHP in vegetables around plastic industrial area].

    PubMed

    Wang, Jia-Wen; Du, Qi-Zhen; Song, Ying-Qi

    2010-10-01

    Concentration of di-(2-ethylhexyl) phthalate (DEHP)in the inner tissue of various vegetable species and their growing environment (soil and atmosphere) around plastic industrial area were investigated and determined by gas chromatography-mass spectrum (GC/MS). The results showed that concentrations of DEHP in 5 kinds of vegetable were 0.23-9.11 mg/kg, 3.82 mg/kg in average (fresh weight). Of the various vegetable species determined, the highest burden was observed in the leafy vegetables, followed by melon and root vegetables. Statistical analysis of variance showed that environment and species are the factors that significantly affect DEHP concentrations in inner vegetable tissue and soil, respectively. Atmosphere deposition is the principal pathway for the accumulation of DEHP. The ability of the plant accumulating DEHP was mainly influenced by the lipid content of the plant. Leaf with pubescence or rough surface was found to have higher DEHP than the other, when the lipid contents were similar. Evaluation of the vegetable around plastic industrial area with the acceptable daily intake (ADI) by OEHHA, concentrations of DEHP has exceeded the safety standard.

  20. Vertical leaf mass per area gradient of mature sugar maple reflects both height-driven increases in vascular tissue and light-driven increases in palisade layer thickness.

    PubMed

    Coble, Adam P; Cavaleri, Molly A

    2017-10-01

    A key trait used in canopy and ecosystem function modeling, leaf mass per area (LMA), is influenced by changes in both leaf thickness and leaf density (LMA = Thickness × Density). In tall trees, LMA is understood to increase with height through two primary mechanisms: (i) increasing palisade layer thickness (and thus leaf thickness) in response to light and/or (ii) reduced cell expansion and intercellular air space in response to hydrostatic constraints, leading to increased leaf density. Our objective was to investigate within-canopy gradients in leaf anatomical traits in order to understand environmental factors that influence leaf morphology in a sugar maple (Acer saccharum Marshall) forest canopy. We teased apart the effects of light and height on anatomical traits by sampling at exposed and closed canopies that had different light conditions at similar heights. As expected, palisade layer thickness responded strongly to cumulative light exposure. Mesophyll porosity, however, was weakly and negatively correlated with light and height (i.e., hydrostatic gradients). Reduced mesophyll porosity was not likely caused by limitations on cell expansion; in fact, epidermal cell width increased with height. Palisade layer thickness was better related to LMA, leaf density and leaf thickness than was mesophyll porosity. Vein diameter and fraction of vascular tissue also increased with height and LMA, density and thickness, revealing that greater investment in vascular and support tissue may be a third mechanism for increased LMA with height. Overall, decreasing mesophyll porosity with height was likely due to palisade cells expanding into the available air space and also greater investments in vascular and support tissue, rather than a reduction of cell expansion due to hydrostatic constraints. Our results provide evidence that light influences both palisade layer thickness and mesophyll porosity and indicate that hydrostatic gradients influence leaf vascular and support tissues in mature Acer saccharum trees. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  1. Leaf-morphology-assisted selection for resistance to two-spotted spider mite Tetranychus urticae Koch (Acari: Tetranychidae) in carnations (Dianthus caryophyllus L).

    PubMed

    Seki, Kousuke

    2016-10-01

    The development of a cultivar resistant to the two-spotted spider mite has provided both ecological and economic benefits to the production of cut flowers. This study aimed to clarify the mechanism of resistance to mites using an inbred population of carnations. In the resistant and susceptible plants selected from an inbred population, a difference was recognised in the thickness of the abaxial palisade tissue by microscopic examination of the damaged leaf. Therefore, it was assumed that mites displayed feeding preferences within the internal leaf structure of the carnation leaf. The suitability of the host plant for mites was investigated using several cultivars selected using an index of the thickness from the abaxial leaf surface to the spongy tissue. The results suggested that the cultivar associated with a thicker abaxial tissue lowered the intrinsic rate of natural increase of the mites. The cultivars with a thicker abaxial tissue of over 120 µm showed slight damage in the field test. The ability of mites to feed on the spongy tissue during an early life stage from hatching to adult emergence was critical. It was possible to select a cultivar that is resistant to mites under a real cultivation environment by observing the internal structure of the leaf. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  2. Plant phenolics and absorption features in vegetation reflectance spectra near 1.66 μm

    USGS Publications Warehouse

    Kokaly, Raymond F.; Skidmore, Andrew K

    2015-01-01

    Past laboratory and field studies have quantified phenolic substances in vegetative matter from reflectance measurements for understanding plant response to herbivores and insect predation. Past remote sensing studies on phenolics have evaluated crop quality and vegetation patterns caused by bedrock geology and associated variations in soil geochemistry. We examined spectra of pure phenolic compounds, common plant biochemical constituents, dry leaves, fresh leaves, and plant canopies for direct evidence of absorption features attributable to plant phenolics. Using spectral feature analysis with continuum removal, we observed that a narrow feature at 1.66 μm is persistent in spectra of manzanita, sumac, red maple, sugar maple, tea, and other species. This feature was consistent with absorption caused by aromatic C-H bonds in the chemical structure of phenolic compounds and non-hydroxylated aromatics. Because of overlapping absorption by water, the feature was weaker in fresh leaf and canopy spectra compared to dry leaf measurements. Simple linear regressions of feature depth and feature area with polyphenol concentration in tea resulted in high correlations and low errors (% phenol by dry weight) at the dry leaf (r2 = 0.95, RMSE = 1.0%, n = 56), fresh leaf (r2 = 0.79, RMSE = 2.1%, n = 56), and canopy (r2 = 0.78, RMSE = 1.0%, n = 13) levels of measurement. Spectra of leaves, needles, and canopies of big sagebrush and evergreens exhibited a weak absorption feature centered near 1.63 μm, short ward of the phenolic compounds, possibly consistent with terpenes. This study demonstrates that subtle variation in vegetation spectra in the shortwave infrared can directly indicate biochemical constituents and be used to quantify them. Phenolics are of lesser abundance compared to the major plant constituents but, nonetheless, have important plant functions and ecological significance. Additional research is needed to advance our understanding of the spectral influences of plant phenolics and terpenes relative to dominant leaf biochemistry (water, chlorophyll, protein/nitrogen, cellulose, and lignin).

  3. Characterization of E coli biofim formations on baby spinach leaf surfaces using hyperspectral fluorescence imaging

    NASA Astrophysics Data System (ADS)

    Cho, Hyunjeong; Baek, Insuck; Oh, Mirae; Kim, Sungyoun; Lee, Hoonsoo; Kim, Moon S.

    2017-05-01

    Bacterial biofilm formed by pathogens on fresh produce surfaces is a food safety concern because the complex extracellular matrix in the biofilm structure reduces the reduction and removal efficacies of washing and sanitizing processes such as chemical or irradiation treatments. Therefore, a rapid and nondestructive method to identify pathogenic biofilm on produce surfaces is needed to ensure safe consumption of fresh, raw produce. This research aimed to evaluate the feasibility of hyperspectral fluorescence imaging for detecting Escherichia.coli (ATCC 25922) biofilms on baby spinach leaf surfaces. Samples of baby spinach leaves were immersed and inoculated with five different levels (from 2.6x104 to 2.6x108 CFU/mL) of E.coli and stored at 4°C for 24 h and 48 h to induce biofilm formation. Following the two treatment days, individual leaves were gently washed to remove excess liquid inoculums from the leaf surfaces and imaged with a hyperspectral fluorescence imaging system equipped with UV-A (365 nm) and violet (405 nm) excitation sources to evaluate a spectral-image-based method for biofilm detection. The imaging results with the UV-A excitation showed that leaves even at early stages of biofilm formations could be differentiated from the control leaf surfaces. This preliminary investigation demonstrated the potential of fluorescence imaging techniques for detection of biofilms on leafy green surfaces.

  4. Correlation between plant physiology and CO2 removable

    NASA Astrophysics Data System (ADS)

    Leman, A. M.; Shamsuri, Mohd Mahathir Suhaimi; Hariri, Azian; Kadir, Aeslina Abdul; Idris, Ahmad Fu'ad; Afandi, Azizi

    2017-09-01

    Certain plants that are able to live in the building are known as indoor plants. Plants have tolerance with indoor environment in order to survive. Usually these plants are able to improve indoor air quality (IAQ). Absorption of carbon dioxide (CO2) by plants is one of the indicators that plants are still alive during photosynthesis process. The possibility of plants structure (plant physiology) to affect CO2 absorption had been the concerns of former researchers. This research intends to study the significant of plant structure (leaf area, fresh weight, and dry weight) that leads to reducing the concentration of CO2 by seven plant species (Anthurium, Dumb Cane, Golden Pothos, Kadaka Fern, Prayer Plants, Spider Plants, and Syngonium). The data of CO2 reduction by plants has been obtained from previous studies. Based on results show that, the leaf area is the most contributing the significant effect to the plant absorb CO2 compare to fresh weight and dry weight. It can be prove by Pearson Correlation, where only the value of leaf area is more than 0.5 for every four conditions. This study can be conclude that the leaf area is quite plays an important role to the plant treat air from CO2, while concentration of light and CO2 will become catalytic factor for the plants improve their photosynthesis process.

  5. Frozen and fresh ovarian tissue require different culture media to promote in vitro development of bovine preantral follicles.

    PubMed

    Castro, Simone Vieira; Carvalho, Adeline Andrade; Silva, Cleidson Manoel Gomes; Santos, Francielli Weber; Campello, Cláudio Cabral; de Figueiredo, José Ricardo; Rodrigues, Ana Paula Ribeiro

    2014-10-01

    The aim of this study was to evaluate the efficiency of different media in the in vitro culture of bovine preantral follicles that were used either fresh or following slow freezing treatment. Frozen and fresh noncultured or cultured ovarian fragments were processed for histological, viability, and cell proliferation analyses. For cryopreservation, a solution containing 1.5 M ethylene glycol was frozen in a programmable biological freezer. After thawing, a portion of the samples was destined for frozen controls. The remainder were cultured in vitro for 5 days in three media: α-MEM, McCoy, or M199. Samples from these culture media were collected on days 1 and 5 for quantification of reactive oxygen species (ROS) and for hormonal assays. In fresh-cultured tissues, the percentage of morphologically normal follicles was significantly higher when cultured in M199 compared to that in the other media. In frozen-cultured tissues, McCoy medium was significantly superior to the other media, and was the only treatment that helped in maintaining the viability similar to fresh and frozen controls. Upon quantification of the nucleolus organizer region, we observed greater proliferation of granulosa cells in the frozen-cultured tissues with McCoy medium, and lesser proliferation in fresh-cultured tissues only with α-MEM. In frozen-cultured tissues, ROS levels were highest at day 1 and progressively reduced during culture, independent of the media used. In conclusion, under the conditions used in this study, the M199 and McCoy media are recommended for the culture of follicles derived from fresh and frozen ovarian tissues, respectively.

  6. Specimen Sample Preservation for Cell and Tissue Cultures

    NASA Technical Reports Server (NTRS)

    Meeker, Gabrielle; Ronzana, Karolyn; Schibner, Karen; Evans, Robert

    1996-01-01

    The era of the International Space Station with its longer duration missions will pose unique challenges to microgravity life sciences research. The Space Station Biological Research Project (SSBRP) is responsible for addressing these challenges and defining the science requirements necessary to conduct life science research on-board the International Space Station. Space Station will support a wide range of cell and tissue culture experiments for durations of 1 to 30 days. Space Shuttle flights to bring experimental samples back to Earth for analyses will only occur every 90 days. Therefore, samples may have to be retained for periods up to 60 days. This presents a new challenge in fresh specimen sample storage for cell biology. Fresh specimen samples are defined as samples that are preserved by means other than fixation and cryopreservation. The challenge of long-term storage of fresh specimen samples includes the need to suspend or inhibit proliferation and metabolism pending return to Earth-based laboratories. With this challenge being unique to space research, there have not been any ground based studies performed to address this issue. It was decided hy SSBRP that experiment support studies to address the following issues were needed: Fixative Solution Management; Media Storage Conditions; Fresh Specimen Sample Storage of Mammalian Cell/Tissue Cultures; Fresh Specimen Sample Storage of Plant Cell/Tissue Cultures; Fresh Specimen Sample Storage of Aquatic Cell/Tissue Cultures; and Fresh Specimen Sample Storage of Microbial Cell/Tissue Cultures. The objective of these studies was to derive a set of conditions and recommendations that can be used in a long duration microgravity environment such as Space Station that will permit extended storage of cell and tissue culture specimens in a state consistent with zero or minimal growth, while at the same time maintaining their stability and viability.

  7. A Novel Approach for Ovine Primary Alveolar Epithelial Type II Cell Isolation and Culture from Fresh and Cryopreserved Tissue Obtained from Premature and Juvenile Animals.

    PubMed

    Marcinkiewicz, Mariola M; Baker, Sandy T; Wu, Jichuan; Hubert, Terrence L; Wolfson, Marla R

    2016-01-01

    The in vivo ovine model provides a clinically relevant platform to study cardiopulmonary mechanisms and treatments of disease; however, a robust ovine primary alveolar epithelial type II (ATII) cell culture model is lacking. The objective of this study was to develop and optimize ovine lung tissue cryopreservation and primary ATII cell culture methodologies for the purposes of dissecting mechanisms at the cellular level to elucidate responses observed in vivo. To address this, we established in vitro submerged and air-liquid interface cultures of primary ovine ATII cells isolated from fresh or cryopreserved lung tissues obtained from mechanically ventilated sheep (128 days gestation-6 months of age). Presence, abundance, and mRNA expression of surfactant proteins was assessed by immunocytochemistry, Western Blot, and quantitative PCR respectively on the day of isolation, and throughout the 7 day cell culture study period. All biomarkers were significantly greater from cells isolated from fresh than cryopreserved tissue, and those cultured in air-liquid interface as compared to submerged culture conditions at all time points. Surfactant protein expression remained in the air-liquid interface culture system while that of cells cultured in the submerged system dissipated over time. Despite differences in biomarker magnitude between cells isolated from fresh and cryopreserved tissue, cells isolated from cryopreserved tissue remained metabolically active and demonstrated a similar response as cells from fresh tissue through 72 hr period of hyperoxia. These data demonstrate a cell culture methodology using fresh or cryopreserved tissue to support study of ovine primary ATII cell function and responses, to support expanded use of biobanked tissues, and to further understanding of mechanisms that contribute to in vivo function of the lung.

  8. Abscisic Acid Content, Transpiration, and Stomatal Conductance As Related to Leaf Age in Plants of Xanthium strumarium L.

    PubMed

    Raschke, K; Zeevaart, J A

    1976-08-01

    Among the four uppermost leaves of greenhouse-grown plants of Xanthium strumarium L. the content of abscisic acid per unit fresh or dry weight was highest in the youngest leaf and decreased gradually with increasing age of the leaves. Expressed per leaf, the second youngest leaf was richest in ABA; the amount of ABA per leaf declined only slightly as the leaves expanded. Transpiration and stomatal conductance were negatively correlated with the ABA concentration in the leaves; the youngest leaf lost the least amount of water. This correlation was always very good if the youngest leaf was compared with the older leaves but not always good among the older leaves. Since stomatal sensitivity to exogenous (+/-)-ABA was the same in leaves of all four age groups ABA may be in at least two compartments in the leaf, one of which is isolated from the guard cells.The ability to synthesize ABA in response to wilting or chilling was strongly expressed in young leaves and declined with leaf age. There was no difference between leaves in their content of the metabolites of ABA, phaseic, and dihydrophaseic acid, expressed per unit weight.

  9. Abscisic Acid Content, Transpiration, and Stomatal Conductance As Related to Leaf Age in Plants of Xanthium strumarium L. 1

    PubMed Central

    Raschke, Klaus; Zeevaart, Jan A. D.

    1976-01-01

    Among the four uppermost leaves of greenhouse-grown plants of Xanthium strumarium L. the content of abscisic acid per unit fresh or dry weight was highest in the youngest leaf and decreased gradually with increasing age of the leaves. Expressed per leaf, the second youngest leaf was richest in ABA; the amount of ABA per leaf declined only slightly as the leaves expanded. Transpiration and stomatal conductance were negatively correlated with the ABA concentration in the leaves; the youngest leaf lost the least amount of water. This correlation was always very good if the youngest leaf was compared with the older leaves but not always good among the older leaves. Since stomatal sensitivity to exogenous (±)-ABA was the same in leaves of all four age groups ABA may be in at least two compartments in the leaf, one of which is isolated from the guard cells. The ability to synthesize ABA in response to wilting or chilling was strongly expressed in young leaves and declined with leaf age. There was no difference between leaves in their content of the metabolites of ABA, phaseic, and dihydrophaseic acid, expressed per unit weight. PMID:16659640

  10. Nitrate Reductase Activity and Polyribosomal Content of Corn (Zea mays L.) Having Low Leaf Water Potentials 1

    PubMed Central

    Morilla, Camila A.; Boyer, J. S.; Hageman, R. H.

    1973-01-01

    Desiccation of 8- to 13-day-old seedlings, achieved by withholding nutrient solution from the vermiculite root medium, caused a reduction in nitrate reductase activity of the leaf tissue. Activity declined when leaf water potentials decreased below −2 bars and was 25% of the control at a leaf water potential of −13 bars. Experiments were conducted to determine whether the decrease in nitrate reductase activity was due to reduced levels of nitrate in the tissue, direct inactivation of the enzyme by low leaf water potentials, or to changes in rates of synthesis or decay of the enzyme. Although tissue nitrate content decreased with the onset of desiccation, it did not continue to decline with tissue desiccation and loss of enzyme activity. Nitrate reductase activity recovered when the plants were rewatered with nitrate-free medium, suggesting that the nitrate in the plant was adequate for high nitrate reductase activity. The rate of decay of nitrate reductase activity from desiccated tissue was essentially identical to that of the control, in vivo or in vitro, regardless of the rapidity of desiccation of the tissue. Direct inactivation of the enzyme by the low water potentials was not detected. Polyribosomal content of the tissue declined with the decrease in water potential, prior to the decline in nitrate reductase activity. Changes in ribosomal profiles occurred during desiccation, regardless of whether the tissue had been excised or not and whether desiccation was rapid or slow. Reduction in polyribosomal content did not appear to be associated with changes in ribonuclease activity. Nitrate reductase activity and the polyribosomal content of the tissue recovered upon rewatering, following the recovery in water potential. The increase in polyribosomal content preceded the increase in nitrate reductase activity. Recovery of enzyme activity was prevented by cycloheximide. Based on these results, it appears that nitrate reductase activity was affected primarily by a decrease in the rate of enzyme synthesis at low leaf water potentials. PMID:16658419

  11. Nondestructive and Real-time Measurement of Moisture in Plant

    NASA Astrophysics Data System (ADS)

    Ogawa, Yuichi; Kawase, Kodo; Mizuno, Maya; Yamashita, Masatsugu; Otani, Chiko

    We constructed a THz transillumination system for water content monitoring, and we succeeded in measuring the moisture level in plants. Our measurement system uses a widely tunable coherent THz parametric oscillator source. As target we chose for this experiment a leaf of Japanese basil. The time variation of the water content in the leaf was monitored in two situations: a leaf freshly cut which is left to dry out, and the leaf of a water stressed plant. We found by real-time measurements that the water content of a cut leaf does not decrease uniformly in time. Also, the response to water stress is delayed by about 5-10 minutes. Furthermore, we demonstrated a moisture measurement using a transillumination THz imaging system. As target we chose for this experiment a leaf of Hedera helix held between two thin plastic sheets. The change of the moisture distribution is clearly visible. These results show that the method described here can be applied to nondestructive and real-time monitoring of water content in plants.

  12. Supplemental Upward Lighting from Underneath to Obtain Higher Marketable Lettuce (Lactuca sativa) Leaf Fresh Weight by Retarding Senescence of Outer Leaves

    PubMed Central

    Zhang, Geng; Shen, Shanqi; Takagaki, Michiko; Kozai, Toyoki; Yamori, Wataru

    2015-01-01

    Recently, the so-called “plant factory with artificial lighting” (PFAL) approach has been developed to provide safe and steady food production. Although PFALs can produce high-yielding and high-quality plants, the high plant density in these systems accelerates leaf senescence in the bottom (or outer) leaves owing to shading by the upper (or inner) leaves and by neighboring plants. This decreases yield and increases labor costs for trimming. Thus, the establishment of cultivation methods to retard senescence of outer leaves is an important research goal to improve PFAL yield and profitability. In the present study, we developed an LED lighting apparatus that would optimize light conditions for PFAL cultivation of a leafy vegetable. Lettuce (Lactuca sativa L.) was hydroponically grown under white, red, or blue LEDs, with light provided from above (downward), with or without supplemental upward lighting from underneath the plant. White LEDs proved more appropriate for lettuce growth than red or blue LEDs, and the supplemental lighting retarded the senescence of outer leaves and decreased waste (i.e., dead or low-quality senescent leaves), leading to an improvement of the marketable leaf fresh weight. PMID:26697055

  13. [Comparative analysis of agronomic and qualitative characters in different lines of Dendrobium denneanum].

    PubMed

    He, Tao; Deng, Li; Lin, Yuan; Li, Bo; Yang, Xiaofan; Wang, Fang; Chun, Ze

    2010-08-01

    To provide theoretical basis for breeding good variety of Dendrobium denneanum, agronomic and qualitative characters of 4 different lines and relationships among them were studied. The stem length, stem diameter, leaf length, leaf width, length/ width ratio and leaf area were measured. The single fresh and dry stem was weighed and drying rate was calculated. The contents of polysaccharides and total alkaloids were determined by sulfuric acid-phenol colorimetry and acid-dye colorimetry, respectively. The correlations between characters were analyzed. The results showed that differences in major agronomic characters between four lines were significant. The plant types of dq-1 and dq-2 were higher, dq-3 was medium and dq-4 was lower. The fresh weigh of stem and content of polysaccharides were the highest in dq-2, 7.81 g and 14.33%. While the highest content of total alkaloids and was 0. 486% in dq-3. There were significant correlations between agronomic characters, but these characters had low or non correlations with qualitative characters such as polysaccharides and total alkaloids. It was shown that the content of polysaccharides and total alkaloids were significantly different among 4 lines of D. denneanum, which could be selected for different uses.

  14. Supplemental Upward Lighting from Underneath to Obtain Higher Marketable Lettuce (Lactuca sativa) Leaf Fresh Weight by Retarding Senescence of Outer Leaves.

    PubMed

    Zhang, Geng; Shen, Shanqi; Takagaki, Michiko; Kozai, Toyoki; Yamori, Wataru

    2015-01-01

    Recently, the so-called "plant factory with artificial lighting" (PFAL) approach has been developed to provide safe and steady food production. Although PFALs can produce high-yielding and high-quality plants, the high plant density in these systems accelerates leaf senescence in the bottom (or outer) leaves owing to shading by the upper (or inner) leaves and by neighboring plants. This decreases yield and increases labor costs for trimming. Thus, the establishment of cultivation methods to retard senescence of outer leaves is an important research goal to improve PFAL yield and profitability. In the present study, we developed an LED lighting apparatus that would optimize light conditions for PFAL cultivation of a leafy vegetable. Lettuce (Lactuca sativa L.) was hydroponically grown under white, red, or blue LEDs, with light provided from above (downward), with or without supplemental upward lighting from underneath the plant. White LEDs proved more appropriate for lettuce growth than red or blue LEDs, and the supplemental lighting retarded the senescence of outer leaves and decreased waste (i.e., dead or low-quality senescent leaves), leading to an improvement of the marketable leaf fresh weight.

  15. Permeability of tritiated water through human cervical and vaginal tissue.

    PubMed

    Sassi, Alexandra B; McCullough, Kristy D; Cost, Marilyn R; Hillier, Sharon L; Rohan, Lisa Cencia

    2004-08-01

    The increased incidence of human immunodeficiency virus infection in women has identified an urgent need to develop a female-controlled method to prevent acquisition of human immunodeficiency virus and other sexually transmitted diseases. Women would apply the product intravaginally before intercourse. Development of such a product requires a better understanding of the permeability characteristics of the tissues with which such products would come into contact. However, limited studies have been performed in this area. In the present study, water permeability of fresh human cervical and vaginal tissue was evaluated. The average apparent permeability coefficient was found to be 8 x 10(-5) cm/s for fresh human cervical tissue and 7 x 10(-5) cm/s for fresh human vaginal tissue. Considering the lack of regularity in obtaining cervical and vaginal tissue from surgical specimens, additional tests were performed to evaluate the effect of freezing on tritiated water permeability. No statistically significant differences were observed in the permeability values obtained when comparing fresh versus frozen tissues. Copyright 2004 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 93:2009-2016, 2004

  16. Water potential in excised leaf tissue. Comparison of a commercial dew point hygrometer and a thermocouple psychrometer on soybean, wheat, and barley

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nelsen, C.E.; Safir, G.R.; Hanson, A.D.

    1978-01-01

    Leaf water potential (Psi/sub leaf/) determinations were made on excised leaf samples using a commercial dew point hygrometer (Wescor Inc., Logan, Utah) and a thermocouple psychrometer operated in the isopiestic mode. With soybean leaves (Glycine max L.), there was good agreement between instruments; equilibration times were 2 to 3 hours. With cereals (Triticum aestivum L. and Hordeum vulgare L.), agreement between instruments was poor for moderately wilted leaves when 7-mm-diameter punches were used in the hygrometer and 20-mm slices were used in the psychrometer, because the Psi/sub leaf/ values from the dew point hygrometer were too high. Agreement was improvedmore » by replacing the 7-mm punch samples in the hygrometer by 13-mm slices, which had a lower cut edge to volume ratio. Equilibration times for cereals were normally 6 to 8 hours. Spuriously high Psi/sub leaf/ values obtained with 7-mm leaf punches may be associated with the ion release and reabsorption that occur upon tissue excision; such errors evidently depend both on the species and on tissue water status.« less

  17. RNA isolation from loquat and other recalcitrant woody plants with high quality and yield.

    PubMed

    Morante-Carriel, Jaime; Sellés-Marchart, Susana; Martínez-Márquez, Ascensión; Martínez-Esteso, María José; Luque, Ignacio; Bru-Martínez, Roque

    2014-05-01

    RNA isolation is difficult in plants that contain large amounts of polysaccharides and polyphenol compounds. To date, no commercial kit has been developed for the isolation of high-quality RNA from tissues with these characteristics, especially for fruit. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to recalcitrant plant tissues. Here an efficient RNA isolation protocol based on cetyltrimethylammonium bromide (CTAB) and two successive precipitations with 10 M lithium chloride (LiCl) was developed specifically for loquat fruits, but it was proved to work efficiently in other tissues of loquat and woody plants. The RNA isolated by this improved protocol was not only of high purity and integrity (A260/A280 ratios ranged from 1.90 to 2.04 and A260/A230 ratios were>2.0) but also of high yield (up to 720 μg on average [coefficient of variation=21%] total RNA per gram fresh tissue). The protocol was tested on loquat fruit (different stages of development, postharvest, ripening, and bruising), leaf, root, flower, stem, and bud; quince fruit and root; grapevine cells in liquid culture; and rose petals. The RNA obtained with this method is amenable to enzymatic treatments and can be efficiently applied for research on gene characterization, expression, and function. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Effects of deoxynivalenol on content of chloroplast pigments in barley leaf tissues.

    PubMed

    Bushnell, W R; Perkins-Veazie, P; Russo, V M; Collins, J; Seeland, T M

    2010-01-01

    To understand further the role of deoxynivalenol (DON) in development of Fusarium head blight (FHB), we investigated effects of the toxin on uninfected barley tissues. Leaf segments, 1 to 1.2 cm long, partially stripped of epidermis were floated with exposed mesophyll in contact with DON solutions. In initial experiments with the leaf segments incubated in light, DON at 30 to 90 ppm turned portions of stripped tissues white after 48 to 96 h. The bleaching effect was greatly enhanced by addition of 1 to 10 mM Ca(2+), so that DON at 10 to 30 ppm turned virtually all stripped tissues white within 48 h. Content of chlorophylls a and b and of total carotenoid pigment was reduced. Loss of electrolytes and uptake of Evans blue indicated that DON had a toxic effect, damaging plasmalemmas in treated tissues before chloroplasts began to lose pigment. When incubated in the dark, leaf segments also lost electrolytes, indicating DON was toxic although the tissues remained green. Thus, loss of chlorophyll in light was due to photobleaching and was a secondary effect of DON, not required for toxicity. In contrast to bleaching effects, some DON treatments that were not toxic kept tissues green without bleaching or other signs of injury, indicating senescence was delayed compared with slow yellowing of untreated leaf segments. Cycloheximide, which like DON, inhibits protein synthesis, also bleached some tissues and delayed senescence of others. Thus, the effects of DON probably relate to its ability to inhibit protein synthesis. With respect to FHB, the results suggest DON may have multiple roles in host cells of infected head tissues, including delayed senescence in early stages of infection and contributing to bleaching and death of cells in later stages.

  19. Phenotypic and biochemical profile changes in calendula (Calendula officinalis L.) plants treated with two chemical mutagenesis.

    PubMed

    El-Nashar, Y I; Asrar, A A

    2016-05-06

    Chemical mutagenesis is an efficient tool used in mutation-breeding programs to improve the vital characters of the floricultural crops. This study aimed to estimate the effects of different concentrations of two chemical mutagens; sodium azide (SA) and diethyl sulfate (DES). The vegetative growth and flowering characteristics in two generations (M1 and M2) of calendula plants were investigated. Seeds were treated with five different concentrations of SA and DES (at the same rates) of 1000, 2000, 3000, 4000, and 5000 ppm, in addition to a control treatment of 0 ppm. Results showed that lower concentrations of SA mutagen had significant effects on seed germination percentage, plant height, leaf area, plant fresh weight, flowering date, inflorescence diameter, and gas-exchange measurements in plants of both generations. Calendula plants tended to flower earlier under low mutagen concentrations (1000 ppm), whereas higher concentrations delayed flowering significantly. Positive results on seed germination, plant height, number of branches, plant fresh weight, and leaf area were observed in the M2-generation at lower concentrations of SA (1000 ppm), as well as at 4000 ppm DES on number of leaves and inflorescences. The highest total soluble protein was detected at the concentrations of 1000 ppm SA and 2000 ppm DES. DES showed higher average of acid phosphatase activity than SA. Results indicated that lower concentrations of SA and DES mutagens had positive effects on seed germination percentage, plant height, leaf area, plant fresh weight, flowering date, inflorescence diameter, and gas-exchange measurements. Thus, lower mutagen concentrations could be recommended for better floral and physio-chemical performance.

  20. Boron accumulation and toxicity in hybrid poplar (Populus nigra × euramericana).

    PubMed

    Rees, Rainer; Robinson, Brett H; Menon, Manoj; Lehmann, Eberhard; Günthardt-Goerg, Madeleine S; Schulin, Rainer

    2011-12-15

    Poplars accumulate high B concentrations and are thus used for the phytomanagement of B contaminated soils. Here, we performed pot experiments in which Populus nigra × euramericana were grown on a substrate with B concentrations ranging from 13 to 280 mg kg(-1) as H(3)BO(3). Salix viminalis, Brassica juncea, and Lupinus albus were grown under some growing conditions for comparison. Poplar growth was unaffected at soil B treatment levels up to 93 mg kg(-1). Growth was progressively reduced at levels of 168 and 280 mg kg(-1). None of the other species survived at these substrate B levels. At leaf B concentrations <900 mg kg(-1) only <10% of the poplar leaf area showed signs of toxicity. Neutron radiography revealed that chlorotic leaf tissues had B concentrations of 1000-2000 mg kg(-1), while necrotic tissues had >2000 mg kg(-1). Average B concentrations of up to 3500 mg kg(-1) were found in leaves, while spots within leaves had concentrations >7000 mg kg(-1), showing that B accumulation in leaf tissue continued even after the onset of necrosis. The B accumulation ability of P. nigra × euramericana is associated with B hypertolerance in the living tissue and storage of B in dead leaf tissue.

  1. Direct observation of the photodegradation of anthracene and pyrene adsorbed onto mangrove leaves.

    PubMed

    Wang, Ping; Wu, Tun-Hua; Zhang, Yong

    2014-01-01

    An established synchronous fluorimetry method was used for in situ investigation of the photodegradation of pyrene (PYR) and anthracene (ANT) adsorbed onto fresh leaves of the seedlings of two mangrove species, Aegiceras corniculatum (L.) Blanco (Ac) and Kandelia obovata (Ko) in multicomponent mixtures (mixture of the ANT and PYR). Experimental results indicated that photodegradation was the main transformation pathway for both ANT and PYR in multicomponent mixtures. The amount of the PAHs volatilizing from the leaf surfaces and entering the inner leaf tissues was negligible. Over a certain period of irradiation time, the photodegradation of both PYR and ANT adsorbed onto the leaves of Ac and Ko followed first-order kinetics, with faster rates being observed on Ac leaves. In addition, the photodegradation rate of PYR on the leaves of the mangrove species in multicomponent mixtures was much slower than that of adsorbed ANT. Compared with the PAHs adsorbed as single component, the photodegradation rate of ANT adsorbed in multicomponent mixtures was slower, while that of PYR was faster. Moreover, the photodegradation of PYR and ANT dissolved in water in multicomponent mixtures was investigated for comparison. The photodegradation rate on leaves was much slower than in water. Therefore, the physical-chemical properties of the substrate may strongly influence the photodegradation rate of adsorbed PAHs.

  2. Direct Observation of the Photodegradation of Anthracene and Pyrene Adsorbed onto Mangrove Leaves

    PubMed Central

    Wang, Ping; Wu, Tun-Hua; Zhang, Yong

    2014-01-01

    An established synchronous fluorimetry method was used for in situ investigation of the photodegradation of pyrene (PYR) and anthracene (ANT) adsorbed onto fresh leaves of the seedlings of two mangrove species, Aegiceras corniculatum (L.) Blanco (Ac) and Kandelia obovata (Ko) in multicomponent mixtures (mixture of the ANT and PYR). Experimental results indicated that photodegradation was the main transformation pathway for both ANT and PYR in multicomponent mixtures. The amount of the PAHs volatilizing from the leaf surfaces and entering the inner leaf tissues was negligible. Over a certain period of irradiation time, the photodegradation of both PYR and ANT adsorbed onto the leaves of Ac and Ko followed first-order kinetics, with faster rates being observed on Ac leaves. In addition, the photodegradation rate of PYR on the leaves of the mangrove species in multicomponent mixtures was much slower than that of adsorbed ANT. Compared with the PAHs adsorbed as single component, the photodegradation rate of ANT adsorbed in multicomponent mixtures was slower, while that of PYR was faster. Moreover, the photodegradation of PYR and ANT dissolved in water in multicomponent mixtures was investigated for comparison. The photodegradation rate on leaves was much slower than in water. Therefore, the physical-chemical properties of the substrate may strongly influence the photodegradation rate of adsorbed PAHs. PMID:25144741

  3. A hyper-thermostable α-amylase from Pyrococcus furiosus accumulates in Nicotiana tabacum as functional aggregates.

    PubMed

    Zhu, Hong; Reynolds, L Bruce; Menassa, Rima

    2017-06-19

    Alpha amylase hydrolyzes α-bonds of polysaccharides such as starch and produces malto-oligosaccharides. Its starch saccharification applications make it an essential enzyme in the textile, food and brewing industries. Commercially available α-amylase is mostly produced from Bacillus or Aspergillus. A hyper-thermostable and Ca 2++ independent α-amylase from Pyrococcus furiosus (PFA) expressed in E.coli forms insoluble inclusion bodies and thus is not feasible for industrial applications. We expressed PFA in Nicotiana tabacum and found that plant-produced PFA forms functional aggregates with an accumulation level up to 3.4 g/kg FW (fresh weight) in field conditions. The aggregates are functional without requiring refolding and therefore have potential to be applied as homogenized plant tissue without extraction or purification. PFA can also be extracted from plant tissue upon dissolution in a mild reducing buffer containing SDS. Like the enzyme produced in P. furiosus and in E. coli, plant produced PFA preserves hyper-thermophilicity and hyper-thermostability and has a long shelf life when stored in lyophilized leaf tissue. With tobacco's large biomass and high yield, hyper-thermostable α-amylase was produced at a scale of 42 kg per hectare. Tobacco may be a suitable bioreactor for industrial production of active hyperthermostable alpha amylase.

  4. Outside-Xylem Vulnerability, Not Xylem Embolism, Controls Leaf Hydraulic Decline during Dehydration.

    PubMed

    Scoffoni, Christine; Albuquerque, Caetano; Brodersen, Craig R; Townes, Shatara V; John, Grace P; Bartlett, Megan K; Buckley, Thomas N; McElrone, Andrew J; Sack, Lawren

    2017-02-01

    Leaf hydraulic supply is crucial to maintaining open stomata for CO 2 capture and plant growth. During drought-induced dehydration, the leaf hydraulic conductance (K leaf ) declines, which contributes to stomatal closure and, eventually, to leaf death. Previous studies have tended to attribute the decline of K leaf to embolism in the leaf vein xylem. We visualized at high resolution and quantified experimentally the hydraulic vulnerability of xylem and outside-xylem pathways and modeled their respective influences on plant water transport. Evidence from all approaches indicated that the decline of K leaf during dehydration arose first and foremost due to the vulnerability of outside-xylem tissues. In vivo x-ray microcomputed tomography of dehydrating leaves of four diverse angiosperm species showed that, at the turgor loss point, only small fractions of leaf vein xylem conduits were embolized, and substantial xylem embolism arose only under severe dehydration. Experiments on an expanded set of eight angiosperm species showed that outside-xylem hydraulic vulnerability explained 75% to 100% of K leaf decline across the range of dehydration from mild water stress to beyond turgor loss point. Spatially explicit modeling of leaf water transport pointed to a role for reduced membrane conductivity consistent with published data for cells and tissues. Plant-scale modeling suggested that outside-xylem hydraulic vulnerability can protect the xylem from tensions that would induce embolism and disruption of water transport under mild to moderate soil and atmospheric droughts. These findings pinpoint outside-xylem tissues as a central locus for the control of leaf and plant water transport during progressive drought. © 2017 The author(s). All Rights Reserved.

  5. Tissue culture of surgically prepared temporalis fascia.

    PubMed

    Walby, A P; Kerr, A G; Nevin, N C; Woods, G

    1982-10-01

    Temporalis fascia which is used to graft the tympanic membrane has been shown to be viable in tissue culture by a previous pilot study. This present study reports the effect on the viability of the fascia by scraping loose connective tissue from it and allowing it to dry. Pieces of fascia from 30 patients were each divided in 4 and prepared to give explants, fresh, fresh and scraped, dried, and dried and scraped. The fascia grew from 17 patients when cultured fresh, 5 when fresh and scraped, 1 when dried, and none when dried and scraped. These results are significantly different and show that the fascia is devitilized when prepared by the normal method for use in tympanoplasty.

  6. Apricot (Prunus armeniaca L.).

    PubMed

    Petri, César; Alburquerque, Nuria; Burgos, Lorenzo

    2015-01-01

    A protocol for Agrobacterium-mediated stable transformation of whole leaf explants of the apricot (Prunus armeniaca) cultivars 'Helena' and 'Canino' is described. Regenerated buds were selected using a two-step selection strategy with paromomycin sulfate and transferred to bud multiplication medium 1 week after they were detected for optimal survival. After buds were transferred to bud multiplication medium, antibiotic was changed to kanamycin and concentration increased gradually at each transfer to fresh medium in order to eliminate possible escapes and chimeras. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines, was 5.6%. Green and healthy buds, surviving high kanamycin concentration, were transferred to shoot multiplication medium where they elongated in shoots and proliferated. Elongated transgenic shoots were rooted in a medium containing 70 μM kanamycin. Rooted plants were acclimatized following standard procedures. This constitutes the only transformation protocol described for apricot clonal tissues and one of the few of Prunus.

  7. Systematic evaluation of RNA quality, microarray data reliability and pathway analysis in fresh, fresh frozen and formalin-fixed paraffin-embedded tissue samples.

    PubMed

    Wimmer, Isabella; Tröscher, Anna R; Brunner, Florian; Rubino, Stephen J; Bien, Christian G; Weiner, Howard L; Lassmann, Hans; Bauer, Jan

    2018-04-20

    Formalin-fixed paraffin-embedded (FFPE) tissues are valuable resources commonly used in pathology. However, formalin fixation modifies nucleic acids challenging the isolation of high-quality RNA for genetic profiling. Here, we assessed feasibility and reliability of microarray studies analysing transcriptome data from fresh, fresh-frozen (FF) and FFPE tissues. We show that reproducible microarray data can be generated from only 2 ng FFPE-derived RNA. For RNA quality assessment, fragment size distribution (DV200) and qPCR proved most suitable. During RNA isolation, extending tissue lysis time to 10 hours reduced high-molecular-weight species, while additional incubation at 70 °C markedly increased RNA yields. Since FF- and FFPE-derived microarrays constitute different data entities, we used indirect measures to investigate gene signal variation and relative gene expression. Whole-genome analyses revealed high concordance rates, while reviewing on single-genes basis showed higher data variation in FFPE than FF arrays. Using an experimental model, gene set enrichment analysis (GSEA) of FFPE-derived microarrays and fresh tissue-derived RNA-Seq datasets yielded similarly affected pathways confirming the applicability of FFPE tissue in global gene expression analysis. Our study provides a workflow comprising RNA isolation, quality assessment and microarray profiling using minimal RNA input, thus enabling hypothesis-generating pathway analyses from limited amounts of precious, pathologically significant FFPE tissues.

  8. Delayed Senescence

    NASA Technical Reports Server (NTRS)

    2004-01-01

    Researcher Dr. Yi Li developed a technique to manipulate certain characteristics of plant growth such as anit-senescence. For example, the tobacco leaf was clipped from a transgenic plant (right), and a wildtype plant (left). During ground-based laboratory studies, both leaves were left in a darkened area for 4 months. When retrieved, the wildtype plant leaf was dried-out and the transgenic leaf remained fresh and green. A variation of this technology that involves manipulating plant hormones has been conducted in space-based studies on tomato plants through BioServe Space Technologies. The transport and distribution of auxin, an important plant hormone has shown to be influenced by microgravity, which could lead to improving the quality of fruits and vegetables grown on Earth.

  9. Microgravity

    NASA Image and Video Library

    2004-04-15

    Researcher Dr. Yi Li developed a technique to manipulate certain characteristics of plant growth such as anit-senescence. For example, the tobacco leaf was clipped from a transgenic plant (right), and a wildtype plant (left). During ground-based laboratory studies, both leaves were left in a darkened area for 4 months. When retrieved, the wildtype plant leaf was dried-out and the transgenic leaf remained fresh and green. A variation of this technology that involves manipulating plant hormones has been conducted in space-based studies on tomato plants through BioServe Space Technologies. The transport and distribution of auxin, an important plant hormone has shown to be influenced by microgravity, which could lead to improving the quality of fruits and vegetables grown on Earth.

  10. Effects on growth and oxidative stress status of rice plants (Oryza sativa) exposed to two extracts of toxin-producing cyanobacteria (Aphanizomenon ovalisporum and Microcystis aeruginosa).

    PubMed

    Prieto, Ana; Campos, Alexandre; Cameán, Ana; Vasconcelos, Vitor

    2011-10-01

    Toxic cyanobacteria are considered emerging world threats, being responsible for the degradation of the aquatic ecosystems. Aphanizomenon ovalisporum produces the toxin Cylindrospermopsin (CYN) being a concern in fresh water habitats. This work aims to increase our knowledge on the effects of this toxic cyanobacterium in plants by studying the alterations in growth parameters and oxidative stress status of rice (Oriza sativa) exposed to the cyanobacteria cell extracts containing CYN. Significant increases in glutathione S-transferase (GST) and glutathione peroxidase (GPx) activities were detected in the different experiments performed. The roots showed to be more sensitive than leaves regarding the enzyme activities. A reduction in the leaf tissue fresh weight was observed after 9 days of plant treatment suggesting a major physiological stress. The exposure of rice plants to a mixture of A. ovalisporum and Microcystis aeruginosa cell extracts containing CYN and microcystins including microcystin-LR, resulted in a significant increase in the GST and GPx activities, suggesting a synergistic effect of both extracts. Together these results point out the negative effects of cyanotoxins on plant growth and oxidative status, induced by A. ovalisporum cell extracts, raising also concerns in the accumulation of CYN. Copyright © 2011 Elsevier Inc. All rights reserved.

  11. Comparison of Photoacoustic Signals in Photosynthetic and Nonphotosynthetic Leaf Tissues of Variegated Pelargonium zonale

    NASA Astrophysics Data System (ADS)

    Veljović-Jovanović, S.; Vidović, M.; Morina, F.; Prokić, Lj.; Todorović, D. M.

    2016-09-01

    Green-white variegated leaves of Pelargonium zonale were studied using the photoacoustic method. Our aim was to characterize photosynthetically active green tissue and nonphotosynthetically active white tissue by the photoacoustic amplitude signals. We observed lower stomatal conductance and higher leaf temperature in white tissue than in green tissue. Besides these thermal differences, significantly higher absorbance in green tissue was based on chlorophyll and carotenoids which were absent in white tissue. However, optical properties of epidermal layers of both tissues were equal. The photoacoustic amplitude of white tissue was over four times higher compared to green tissue, which was correlated with lower stomatal conductance. In addition, at frequencies >700 Hz, the significant differences between the photoacoustic signals of green and white tissue were obtained. We identified the photoacoustic signal deriving from photosynthetic oxygen evolution in green tissue, using high intensity of red light modulated at 10 Hz. Moreover, the photoacoustic amplitude of green tissue increased progressively with time which corresponded to the period of induction of photosynthetic oxygen evolution. For the first time, very high frequencies (1 kHz to 5 kHz) were applied on leaf material.

  12. Histologic and ultrastructural evaluation of fresh and frozen-thawed human ovarian xenografts in nude mice.

    PubMed

    Nisolle, M; Casanas-Roux, F; Qu, J; Motta, P; Donnez, J

    2000-07-01

    To compare histologic and ultrastructural characteristics of fresh and frozen-thawed human ovarian cortical tissue grafted into nude mice. Experimental prospective study. An academic research environment. Ovarian biopsy specimens were obtained from 13 women undergoing laparoscopy for tubal ligation or infertility. Forty nude mice. A minilaparotomy was performed to place fresh and frozen-thawed ovarian grafts subcutaneously (sc) or intraperitoneally (ip). Removal of the ovarian grafts was performed at 24 days. [1] the follicular population, [2] fibrosis, [3] vascularization of the grafted tissue, and [4] ultrastructural evaluation. A greater fibrosis relative surface area was noted in frozen-thawed transplanted tissue than in fresh transplants. Regardless of this fibrosis, a similar follicular density was observed in fresh and frozen-thawed ovarian tissue 24 days after transplantation. Active angiogenesis was proved by both immunohistochemical study of the vascular endothelial growth factor and morphometric study of the vascular network. Normal ultrastructural characteristics were noted in frozen-thawed ovarian biopsies. Angiogenesis allows implantation of the graft even if it has been cryopreserved and thawed similarly to implantation of fresh tissue. The greater fibrosis observed in grafts after cryopreservation and implantation does not seem to affect the primordial and primary ovocyte population and their ultrastructural characteristics, but further studies must be conducted to prove that after cryopreservation and transplantation, ovocytes may achieve full maturation and fertilization.

  13. Variation in levels of some leaf enzymes.

    PubMed

    Downton, J; Slatyer, R O

    1971-03-01

    Several procedures were compared for efficiency in the extraction of certain leaf enzymes (phosphoenolpyruvate carboxylase, ribulose 1,5-diphosphate carboxylase and malate dehydrogenase) in Atriplex hastata (a "C3" species exhibiting conventional photosynthetic metabolism), and in A. spongiosa (a "C4" species in which the initial photosynthetic products are C4 dicarboxylic acids). Glycolate oxidase was also assayed in some cases, and Atriplex nummularia and Sorghum bicolor were also used as test material. A simple procedure, involving a mortar and pestle grind with carborundum added to the grinding mixture, was found to be as effective as glass bead grind procedures. In addition, it was more rapid and showed less variability with different operations.Using the carborundum grind procedure, sources of variability in enzyme activity in apparently uniform leaves were compared, as were effects of time of day, leaf age and storage procedure. In general, if apparently uniform leaves could be selected, variability in levels of enzyme activity appeared to be relatively small, not exceeding about 12%. Time of day also appeared to be relatively unimportant for the enzymes examined. However, the ontogentic status of the plant was found to be an important source of variability. Leaf age was also a major source of variability where the activity was expressed on a fresh weight basis, but specific activity (i.e. activity expressed on a protein basis) was relatively constant, at least with the range of species and leaf ages examined here.Storage of fresh samples in liquid nitrogen for 24 h, prior to extraction and assay, led to only a small reduction in activity, but substantial changes occurred if storage was in dry ice or in ice and also where extracts were stored in a deep freeze.

  14. Effects of irrigation moisture regimes on yield and quality of paprika ( Capsicum annuum L)

    NASA Astrophysics Data System (ADS)

    Shongwe, Victor D.; Magongo, Bekani N.; Masarirambi, Michael T.; Manyatsi, Absalom M.

    Although paprika ( Capsicum annuum L) is not widely grown in Swaziland it is becoming increasingly popular as a spice and food colourant. It is a crop that requires irrigation at specific stages of growth as this affects not only the yield but most importantly the quality of the crop. Yield of paprika has been found to increase with relative increase in moisture whereas the quality of fruits has not followed the same trend. The objective of this study was to find the effect of varying irrigation water regimes on the yield and quality of paprika at uniform fertiliser levels. The study was carried out in the 2006/2007 cropping season at the Luyengo campus of the University of Swaziland in a greenhouse. A randomised complete block design was used with four water treatments (0.40, 0.60, 0.80, and 1.00 × Field Capacity). Parameters measured included leaf number per plant, plant height, chlorophyll content, canopy size, leaf width, leaf length, stem girth, dry mass, fresh mass, fruit length, and brix content. There were significant ( P < 0.05) increases in leaf number, plant height, chlorophyll content, canopy size, fresh and dry mass tops and fruit length at the highest moisture level (1.00 × FC) followed by the second highest regime (0.80 × FC) whilst the lower water regimes resulted in lower increases in each of the parameters. Leaf area index did not differ significantly across all treatments. In increasing order the treatments 0.80 × FC and 1.00 × FC gave higher yields but in decreasing order lower brix and thus subsequent lower paprika quality. It is recommended that growers who are aiming for optimum yield and high quality of paprika may use the 0.8 × FC treatment when irrigating.

  15. A nature-based approach for managing the invasive weed species Gutenbergia cordifolia for sustainable rangeland management.

    PubMed

    Ngondya, Issakwisa B; Munishi, Linus K; Treydte, Anna C; Ndakidemi, Patrick A

    2016-01-01

    The invasive weed species Gutenbergia cordifolia has been observed to suppress native plants and to dominate more than half of the entire crater floor (250 km 2 ) in the Ngorongoro Conservation Area (NCA). As this species has been found to be toxic to ruminants it might strongly impact animal populations in this ecologically diverse ecosystem. Hence, a nature-based approach is urgently needed to manage its spread. We tested two Desmodium spp extracts applied to G. cordifolia and assessed the latter's germination rate, height, fresh weight and leaf total chlorophyll content after 30 days in both laboratory and screen house experiments. Seedling germination rate was halved by Desmodium uncinatum leaf extract (DuL), particularly under higher concentrations (≥75 %) rather than lower concentrations (≤62.5 %). Likewise, in both laboratory and screen house experiments, germination rate under DuL treatments declined with increasing concentrations. Seedling height, fresh weight and leaf total chlorophyll content (Chl) were also most strongly affected by DuL treatments rather than D. uncinatum root extract, Desmodium intortum leaf extract or D. intortum root extract treatments. Generally, seedlings treated with higher DuL concentrations were half as tall, had one-third the weight and half the leaf Chl content compared to those treated with lower concentrations. Our study shows a novel technique that can be applied where G. cordifolia may be driving native flora and fauna to local extinction. Our data further suggest that this innovative approach is both ecologically safe and effective and that D. uncinatum can be sustainably used to manage invasive plants, and thus, to improve rangeland productivity.

  16. Leaf hydraulics II: vascularized tissues.

    PubMed

    Rockwell, Fulton E; Holbrook, N Michele; Stroock, Abraham D

    2014-01-07

    Current models of leaf hydration employ an Ohm's law analogy of the leaf as an ideal capacitor, neglecting the resistance to flow between cells, or treat the leaf as a plane sheet with a source of water at fixed potential filling the mid-plane, neglecting the discrete placement of veins as well as their resistance. We develop a model of leaf hydration that considers the average conductance of the vascular network to a representative areole (region bounded by the vascular network), and represent the volume of tissue within the areole as a poroelastic composite of cells and air spaces. Solutions to the 3D flow problem are found by numerical simulation, and these results are then compared to 1D models with exact solutions for a range of leaf geometries, based on a survey of temperate woody plants. We then show that the hydration times given by these solutions are well approximated by a sum of the ideal capacitor and plane sheet times, representing the time for transport through the vasculature and tissue respectively. We then develop scaling factors relating this approximate solution to the 3D model, and examine the dependence of these scaling factors on leaf geometry. Finally, we apply a similar strategy to reduce the dimensions of the steady state problem, in the context of peristomatal transpiration, and consider the relation of transpirational gradients to equilibrium leaf water potential measurements. © 2013 Published by Elsevier Ltd. All rights reserved.

  17. Silica distribution in various bamboos species and its effects on plant growth

    NASA Astrophysics Data System (ADS)

    Collin, B.; Meunier, J.; Keller, C.; Doelsch, E.; Panfili, F.

    2010-12-01

    Bamboos are distributed throughout the world’s temperate, tropical and subtropical regions. They are widely used in industry, as fresh edible shoots, paper maker, building and even in medicine. Bamboos also play multiple ecologic functions such as soil and water conservation and erosion control. Bamboos have generally high silicon (Si) content. Silicon is known to have beneficial effects on plants and alleviate various stresses. The aim of this study is to quantify the Si uptake and distribution in various bamboos species and to investigate the effects of Si on the plant growth. Two complementary studies were carried out, one under natural conditions and one under controlled conditions. First of all, we performed an inventory of Si tissue content in 16 bamboos species growing in a non-polluted tropical soil at the Reunion Island (France, Indian ocean). We determined Si content in leaf and in stem tissues sampled at several heights for each plant. One of these species Gigantocloa sp « Malay Dwarf » was grown for 3 months in nutrient solution at five Si concentrations (0, 0.25, 0.75, 1.15, 1.5 mM Si). Silica deposition was examined in leaves using a cryo-SEM equipped with EDS. The Si concentration varies significantly between species, depending on rhizome morphology. Bamboos having leptomorph rhizomes show significantly higher leaf and stem Si content than that of species having pachymorph rhizomes. The distribution of Si in the plant has the same trends for all species. Leaves are the most concentrated organs (10.9 %), and within the stem Si concentration significantly increases from the bottom (0.32%) to the top of the plant (2.1%). Plant Si content increases with the Si supply. Leaves of Gigantocloa sp « Malay Dwarf » accumulate 15.2 % of Si under natural conditions and up to 24 % when exposed to the highest Si treatment. Unlike previous studies, our experiment shows that the concentration of Si had no significant effect on nutrient uptake and biomass production. Leaf silica deposition occurs mainly in epidermal cells, micro-hairs, prickles hairs and bulliform cells. A better knowledge of Si distribution in various species and in various tissues into the plant is promising to understand the mechanisms of absorption, transportation and precipitation of Si, and thus its role on the stress tolerance.

  18. Leaf density explains variation in leaf mass per area in rice between cultivars and nitrogen treatments.

    PubMed

    Xiong, Dongliang; Wang, Dan; Liu, Xi; Peng, Shaobing; Huang, Jianliang; Li, Yong

    2016-05-01

    Leaf mass per area (LMA) is an important leaf trait; however, correlations between LMA and leaf anatomical features and photosynthesis have not been fully investigated, especially in cereal crops. The objectives of this study were (a) to investigate the correlations between LMA and leaf anatomical traits; and (b) to clarify the response of LMA to nitrogen supply and its effect on photosynthetic nitrogen use efficiency (PNUE). In the present study, 11 rice varieties were pot grown under sufficient nitrogen (SN) conditions, and four selected rice cultivars were grown under low nitrogen (LN) conditions. Leaf anatomical traits, gas exchange and leaf N content were measured. There was large variation in LMA across selected rice varieties. Regression analysis showed that the variation in LMA was more closely related to leaf density (LD) than to leaf thickness (LT). LMA was positively related to the percentage of mesophyll tissue area (%mesophyll), negatively related to the percentage of epidermis tissue area (%epidermis) and unrelated to the percentage of vascular tissue area (%vascular). The response of LMA to N supplementation was dependent on the variety and was also mainly determined by the response of LD to N. Compared with SN, photosynthesis was significantly decreased under LN, while PNUE was increased. The increase in PNUE was more critical in rice cultivars with a higher LMA under SN supply. Leaf density is the major cause of the variation in LMA across rice varieties and N treatments, and an increase in LMA under high N conditions would aggravate the decrease in PNUE. © The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  19. Caffeine biosynthesis and degradation in tea [Camellia sinensis (L.) O. Kuntze] is under developmental and seasonal regulation.

    PubMed

    Mohanpuria, Prashant; Kumar, Vinay; Joshi, Robin; Gulati, Ashu; Ahuja, Paramvir Singh; Yadav, Sudesh Kumar

    2009-10-01

    To study caffeine biosynthesis and degradation, here we monitored caffeine synthase gene expression and caffeine and allantoin content in various tissues of four Camellia sinensis (L.) O. Kuntze cultivars during non-dormant (ND) and dormant (D) growth phases. Caffeine synthase expression as well as caffeine content was found to be higher in commercially utilized tissues like apical bud, 1st leaf, 2nd leaf, young stem, and was lower in old leaf during ND compared to D growth phase. Among fruit parts, fruit coats have higher caffeine synthase expression, caffeine content, and allantoin content. On contrary, allantoin content was found lower in the commercially utilized tissues and higher in old leaf. Results suggested that caffeine synthesis and degradation in tea appears to be under developmental and seasonal regulation.

  20. Laser-enhanced thermal effect of moderate intensity focused ultrasound on bio-tissues

    NASA Astrophysics Data System (ADS)

    Zhao, JinYu; Zhang, ShuYi; Shui, XiuJi; Fan, Li

    2017-09-01

    For avoiding extra-damage to healthy tissues surrounding the focal point during high intensity focused ultrasound (HIFU) treatment in medical therapy, to reduce the ultrasonic intensity outside the focal point is expected. Thus, the heating processes induced by moderate intensity focused ultrasound (MIFU) and enhanced by combined irradiation of laser pulses for bio-tissues are studied in details. For fresh bio-tissues, the enhanced thermal effects by pulsed laser combined with MIFU irradiation are observed experimentally. To explore the mechanisms of these effects, several tissue-mimicking materials composed of agar mixed with graphite powders are prepared and studied for comparison, but the laser-enhanced thermal effects in these mimicking materials are much less than that in the fresh bio-tissues. Therefore, it is suggested that the laser-enhanced thermal effects may be mainly attributed to bio-activities and related photo-bio-chemical effects of fresh tissues.

  1. Glycine betaine accumulation, ionic and water relations of red-beet at contrasting levels of sodium supply

    NASA Technical Reports Server (NTRS)

    Subbarao, G. V.; Wheeler, R. M.; Levine, L. H.; Stutte, G. W.; Sager, J. C. (Principal Investigator)

    2001-01-01

    Exposure of plants to sodium (Na) and salinity may increase glycine betaine accumulation in tissues. To study this, red-beet cvs. Scarlet Supreme and Ruby Queen, were grown for 42 days in a growth chamber using a re-circulating nutrient film technique with 0.25 mmol/L K and either 4.75 mmol/L (control) or 54.75 mmol/L (saline) Na (as NaCl). Plants were harvested at weekly intervals and measurements were taken on leaf water relations, leaf photosynthetic rates, chlorophyll fluorescence, chlorophyll levels, glycine betaine levels, and tissue elemental composition. Glycine betaine accumulation increased under salinity and this accumulation correlated with higher tissue levels of Na in both cultivars. Na accounted for 80 to 90% of the total cation uptake under the saline treatment. At final harvest (42 days), K concentrations in laminae ranged from approximately 65-95 micromoles g-1 dry matter (DM), whereas Na in shoot tissue ranged from approximately 3000-4000 micromoles g-1. Leaf sap osmotic potential at full turgor [psi(s100)] increased as lamina Na content increased. Glycine betaine levels of leaf laminae showed a linear relationship with leaf sap [psi(s100)]. Chlorophyll levels, leaf photosynthetic rates, and chlorophyll fluorescence were not affected by Na levels. These results suggest that the metabolic tolerance to high levels of tissue Na in red-beet could be due to its ability to synthesize and regulate glycine betaine production, and to control partitioning of Na and glycine betaine between the vacuole and the cytoplasm.

  2. Connecting infrared spectra with plant traits to identify species

    NASA Astrophysics Data System (ADS)

    Buitrago, Maria F.; Skidmore, Andrew K.; Groen, Thomas A.; Hecker, Christoph A.

    2018-05-01

    Plant traits are used to define species, but also to evaluate the health status of forests, plantations and crops. Conventional methods of measuring plant traits (e.g. wet chemistry), although accurate, are inefficient and costly when applied over large areas or with intensive sampling. Spectroscopic methods, as used in the food industry and mineralogy, are nowadays applied to identify plant traits, however, most studies analysed visible to near infrared, while infrared spectra of longer wavelengths have been little used for identifying the spectral differences between plant species. This study measured the infrared spectra (1.4-16.0 μm) on individual, fresh leaves of 19 species (from herbaceous to woody species), as well as 14 leaf traits for each leaf. The results describe at which wavelengths in the infrared the leaves' spectra can differentiate most effectively between these plant species. A Quadratic Discrimination Analysis (QDA) shows that using five bands in the SWIR or the LWIR is enough to accurately differentiate these species (Kappa: 0.93, 0.94 respectively), while the MWIR has a lower classification accuracy (Kappa: 0.84). This study also shows that in the infrared spectra of fresh leaves, the identified species-specific features are correlated with leaf traits as well as changes in their values. Spectral features in the SWIR (1.66, 1.89 and 2.00 μm) are common to all species and match the main features of pure cellulose and lignin spectra. The depth of these features varies with changes of cellulose and leaf water content and can be used to differentiate species in this region. In the MWIR and LWIR, the absorption spectra of leaves are formed by key species-specific traits including lignin, cellulose, water, nitrogen and leaf thickness. The connection found in this study between leaf traits, features and spectral signatures are novel tools to assist when identifying plant species by spectroscopy and remote sensing.

  3. [Comparison of the antioxidant properties of selected parts of raspberry (Rubus idaeus) and blackberry (Rubus fruticosus)

    PubMed

    Zielonka-Brzezicka, Joanna; Nowak, Anna; Zielińska, Magdalena; Klimowicz, Adam

    Antioxidants contained in plant raw materials prevent oxidative stress, and reduce the degenerative effects of free radical reactions and damage caused by UV radiation. Antioxidant activity is exhibited, for example, in raspberry (Rubus idaeus), and blackberry (Rubus fruticosus), which have a high content of tannins, flavonoids, phenolic acids, vitamins and minerals. The raw plant material consisted of fresh and dried leaves and fruits of raspberries and blackberries harvested in 2014. This material was extracted using a Soxhlet apparatus and by an ultrasound-assisted technique. To evaluate antioxidant activity DPPH and FRAP methods were used in the first year and DPPH and ABTS methods in the second year of the study. The highest antioxidant activity, evaluated by the DPPH method in 2015, was demonstrated by both raspberry and blackberry fresh leaf extracts, whereas the highest reductive ability, assessed by FRAP, was demonstrated in fresh and dried blackberry leaves. In the next year of the study (2016), the activity of samples evaluated using the DPPH method did not differ significantly, with the exception of fresh raspberry leaf. In this year, the highest antioxidant properties, assessed using the ABTS method, were shown by extracts made of fresh blackberry leaves and fruits, and dried blackberry leaves. All the studied material, both ethanolic leaves and fruit extracts of raspberry and blackberry, reduced free radicals, which was examined using three methods of evaluation of the antioxidative properties (DPPH, FRAP and ABTS). The storage of ethanolic extracts at ambient temperature had no significant impact on the activity reduction of the evaluated plant material. The antioxidant activity of most examined extracts remained consistently high, which may be reflected in the use of the studied materials as sources of antioxidants in the cosmetics and pharmaceuticals industries.

  4. Influence of blue light on the leaf morphoanatomy of in vitro Kalanchoe pinnata (Lamarck) Persoon (Crassulaceae).

    PubMed

    Leal-Costa, Marcos Vinicius; Nascimento, Luana Beatriz dos Santos; Moreira, Nattacha dos Santos; Reinert, Fernanda; Costa, Sônia Soares; Lage, Celso Luiz Salgueiro; Tavares, Eliana Schwartz

    2010-10-01

    Kalanchoe pinnata (Lamarck) Persoon (Crassulaceae) (air plant, miracle leaf) is popularly used to treat gastrointestinal disorders and wounds. Recently, the species was tested to treat cutaneous leishmaniasis with successful results. This medicinal activity was associated with the phenolic fraction of the plant. Blue light induces biosynthesis of phenolic compounds and many changes in anatomical characteristics. We studied the effects of supplementary blue light on the leaf morphology of in vitro K. pinnata. Plants cultured under white light (W plants) only and white light plus blue light (WB plants) show petioles with plain-convex section, amphistomatic leaf blades with simple epidermis, homogeneous mesophyll with densely packed cells, and a single collateral vascular bundle in the midrib. W plants have longer branches, a larger number of nodes per branch, and smaller leaves, whereas WB plant leaves have a thicker upper epidermis and mesophyll. Leaf fresh weight and leaf dry weight were similar in both treatments. Phenolic idioblasts were observed in the plants supplemented with blue light, suggesting that blue light plays an important role in the biosynthesis of phenolic compounds in K. pinnata.

  5. [Effects of exogenous spermidine on lipid peroxidation and membrane proton pump activity of cucumber seedling leaves under high temperature stress].

    PubMed

    Tian, Jing; Guo, Shi-Rong; Sun, Jin; Wang, Li-Ping; Yang, Yan-Juan; Li, Bin

    2011-12-01

    Taking a relatively heat-resistant cucumber (Cucumis sativus) cultivar 'Jinchun No. 4' as test material, a sand culture experiment was conducted in growth chamber to investigate the effects of foliar spraying spermidine (Spd) on the lipid peroxidation, membrane proton pump activity, and corresponding gene expression of cucumber seedling leaves under high temperature stress. Compared with the control, foliar spraying Spd increased the plant height, stem diameter, dry and fresh mass, and leaf area significantly, and inhibited the increase of leaf relative conductivity, malondialdehyde (MDA) content, and lipoxygenase (LOX) activity effectively. Foliar spraying Spd also helped to the increase of leaf plasma membrane- and tonoplast H(+)-ATPase activity, but no significant difference was observed in the gene expression levels. These results suggested that exogenous Spd could significantly decrease the leaf lipid peroxidation and increase the proton pump activity, and thus, stabilize the leaf membrane structure and function, alleviate the damage induced by high temperature stress, and enhance the heat tolerance of cucumber seedlings.

  6. Biobanking of fresh frozen tissue from clinical surgical specimens: transport logistics, sample selection, and histologic characterization.

    PubMed

    Botling, Johan; Micke, Patrick

    2011-01-01

    Access to high-quality fresh frozen tissue is critical for translational cancer research and molecular -diagnostics. Here we describe a workflow for the collection of frozen solid tissue samples derived from fresh human patient specimens after surgery. The routines have been in operation at Uppsala University Hospital since 2001. We have integrated cryosection and histopathologic examination of each biobank sample into the biobank manual. In this way, even small, macroscopically ill-defined lesions can be -procured without a diagnostic hazard due to the removal of uncharacterized tissue from a clinical -specimen. Also, knowledge of the histomorphology of the frozen tissue sample - tumor cell content, stromal components, and presence of necrosis - is pivotal before entering a biobank case into costly molecular profiling studies.

  7. Observations on anatomical aspects of the fruit, leaf and stem tissues of four Citrullus spp.

    USDA-ARS?s Scientific Manuscript database

    Morphological characteristics of the fruit, stem and leaf tissues of four species of Citrullus (L.) Schrad. were examined using standard histological methods. Plant materials included the cultivated watermelon (C. lanatus (Thunb.) Matsum. & Nakai) and three of its related species; C. colocynthis (...

  8. Leaf tissue assay for lepidopteran pests of Bt cotton

    USDA-ARS?s Scientific Manuscript database

    Laboratory measurements of susceptibility to Bt toxins can be a poor indicator of the ability of an insect to survive on transgenic crops. We investigated the potential of using cotton leaf tissue for evaluating heliothine susceptibilities to two dual-gene Bt cottons. A preliminary study was conduct...

  9. Effect of Leaf Surface Chemical Properties on Efficacy of Sanitizer for Rotavirus Inactivation

    PubMed Central

    Fuzawa, Miyu; Ku, Kang-Mo; Palma-Salgado, Sindy Paola; Nagasaka, Kenya; Feng, Hao; Juvik, John A.; Sano, Daisuke; Shisler, Joanna L.

    2016-01-01

    ABSTRACT The use of sanitizers is essential for produce safety. However, little is known about how sanitizer efficacy varies with respect to the chemical surface properties of produce. To answer this question, the disinfection efficacies of an oxidant-based sanitizer and a new surfactant-based sanitizer for porcine rotavirus (PRV) strain OSU were examined. PRV was attached to the leaf surfaces of two kale cultivars with high epicuticular wax contents and one cultivar of endive with a low epicuticular wax content and then treated with each sanitizer. The efficacy of the oxidant-based sanitizer correlated with leaf wax content as evidenced by the 1-log10 PRV disinfection on endive surfaces (low wax content) and 3-log10 disinfection of the cultivars with higher wax contents. In contrast, the surfactant-based sanitizer showed similar PRV disinfection efficacies (up to 3 log10) that were independent of leaf wax content. A statistical difference was observed with the disinfection efficacies of the oxidant-based sanitizer for suspended and attached PRV, while the surfactant-based sanitizer showed similar PRV disinfection efficacies. Significant reductions in the entry and replication of PRV were observed after treatment with either disinfectant. Moreover, the oxidant-based-sanitizer-treated PRV showed sialic acid-specific binding to the host cells, whereas the surfactant-based sanitizer increased the nonspecific binding of PRV to the host cells. These findings suggest that the surface properties of fresh produce may affect the efficacy of virus disinfection, implying that food sanitizers should be carefully selected for the different surface characteristics of fresh produce. IMPORTANCE Food sanitizer efficacies are affected by the surface properties of vegetables. This study evaluated the disinfection efficacies of two food sanitizers, an oxidant-based sanitizer and a surfactant-based sanitizer, on porcine rotavirus strain OSU adhering to the leaf epicuticular surfaces of high- and low-wax-content cultivars. The disinfection efficacy of the oxidant-based sanitizer was affected by the surface properties of the vegetables, while the surfactant-based sanitizer was effective for both high- and low-wax leafy vegetable cultivars. This study suggests that the surface properties of vegetables may be an important factor that interacts with disinfection with food sanitizers of rotaviruses adhering to fresh produce. PMID:27520815

  10. Protective Effect of Polygonum orientale L. Extracts against Clavibater michiganense subsp. sepedonicum, the Causal Agent of Bacterial Ring Rot of Potato

    PubMed Central

    Cai, Jin; Xie, Shulian; Feng, Jia; Wang, Feipeng; Xu, Qiufeng

    2013-01-01

    The Polygonum orientale L. extracts were investigated for antibacterial activity against Clavibater michiganense subsp. sepedonicum (Spieckermann & Kotthoff) Davis et al., the causal agent of a serious disease called bacterial ring rot of potato. The results showed that the leaf extracts of P. orientale had significantly (p<0.05) greater antibacterial activity against C. michiganense subsp. sepedonicum than root, stem, flower extracts in vitro. According to the results of single factor experiments and L273(13) orthogonal experiments, optimum extraction conditions were A1B3C1, extraction time 6 h, temperature 80°C, solid to liquid ratio 1∶10 (g:mL). The highest (p<0.05) antibacterial activity was observed when pH was 5, excluding the effect of control. The extracts were stable under ultraviolet (UV). In vivo analysis revealed that 50 mg/mL of P. orientale leaf extracts was effective in controlling decay. Under field conditions, 50 mg/mL of P. orientale leaf extracts also improved growth parameters (whole plant length, shoot length, root length, plant fresh weight, shoot fresh weight, root fresh weight, dry weight, and number of leaves), in the 2010 and 2011 two growing seasons. Further solvent partition assays showed that the most active compounds were in the petroleum ether fractionation. Transmission electron microscopy (TEM) showed drastic ultrastructural changes caused by petroleum ether fractionation, including bacterial deformation, electron-dense particles, formation of vacuoles and lack of cytoplasmic materials. These results indicated that P. orientale extracts have strong antibacterial activity against C. michiganense subsp. sepedonicum and a promising effect in control of bacterial ring rot of potato disease. PMID:23861908

  11. Inhibition of α-amylase and α-glucosidase activities by ethanolic extract of Telfairia occidentalis (fluted pumpkin) leaf

    PubMed Central

    Oboh, G; Akinyemi, AJ; Ademiluyi, AO

    2012-01-01

    Objective To investigate the inhibitory effect of Telfairia occidentalis Hook f. (Curcubitaceae) (T. occidentalis) leaf on key enzyme linked to type-2 diabetes (α - amylase and α - glucosidase) as well as assess the effect of blanching (a commonly practiced food processing technique) of the vegetable on these key enzymes. Methods Fresh leaves of T. occidentalis were blanched in hot water for 10 minutes, and the extracts of both the fresh and blanched vegetables were prepared and used for subsequent analysis. The inhibitory effect of the extract on α - amylase and α - glucosidase activities as well as some antioxidant parameter was determined in vitro. Results The result revealed that unprocessed T. occidentalis leaf reduce Fe3+ to Fe2+ and also inhibited α - amylase and α - glucosidase activities in a dose dependent manner. However, blanching of the leafy vegetables caused a significant (P<0.05) increase in the antioxidant properties but decrease their ability to inhibit α - amylase and α - glucosidase activities. Conclusions This antioxidant properties and enzyme inhibition could be part of the mechanism by which they are used in the treatment/prevention of type-2 diabetes. However, the blanched vegetable reduces their ability to inhibit both α - amylase and α - glucosidase activity in vitro. PMID:23570004

  12. Estimating yields of salt- and water-stressed forages with remote sensing in the visible and near infrared.

    PubMed

    Poss, J A; Russell, W B; Grieve, C M

    2006-01-01

    In arid irrigated regions, the proportion of crop production under deficit irrigation with poorer quality water is increasing as demand for fresh water soars and efforts to prevent saline water table development occur. Remote sensing technology to quantify salinity and water stress effects on forage yield can be an important tool to address yield loss potential when deficit irrigating with poor water quality. Two important forages, alfalfa (Medicago sativa L.) and tall wheatgrass (Agropyron elongatum L.), were grown in a volumetric lysimeter facility where rootzone salinity and water content were varied and monitored. Ground-based hyperspectral canopy reflectance in the visible and near infrared (NIR) were related to forage yields from a broad range of salinity and water stress conditions. Canopy reflectance spectra were obtained in the 350- to 1000-nm region from two viewing angles (nadir view, 45 degrees from nadir). Nadir view vegetation indices (VI) were not as strongly correlated with leaf area index changes attributed to water and salinity stress treatments for both alfalfa and wheatgrass. From a list of 71 VIs, two were selected for a multiple linear-regression model that estimated yield under varying salinity and water stress conditions. With data obtained during the second harvest of a three-harvest 100-d growing period, regression coefficients for each crop were developed and then used with the model to estimate fresh weights for preceding and succeeding harvests during the same 100-d interval. The model accounted for 72% of the variation in yields in wheatgrass and 94% in yields of alfalfa within the same salinity and water stress treatment period. The model successfully predicted yield in three out of four cases when applied to the first and third harvest yields. Correlations between indices and yield increased as canopy development progressed. Growth reductions attributed to simultaneous salinity and water stress were well characterized, but the corrections for effects of varying tissue nitrogen (N) and very low leaf area index (LAI) are necessary.

  13. Impact of epidermal leaf mining by the aspen leaf miner (Phyllocnistis populiella) on the growth, physiology, and leaf longevity of quaking aspen.

    Treesearch

    Diane L. Wagner; Linda DeFoliart; Patricia Doak; Jenny Schneiderheinze

    2008-01-01

    The aspen leaf miner, Phyllocnistis populiella, feeds on the contents of epidermal cells on both top (adaxial) and bottom (abaxial) surfaces of quaking aspen leaves, leaving the photosynthetic tissue of the mesophyll intact. This type of feeding is taxonomically restricted to a small subset of leaf mining insects but can cause widespread plant...

  14. Brain tumor imaging of rat fresh tissue using terahertz spectroscopy

    NASA Astrophysics Data System (ADS)

    Yamaguchi, Sayuri; Fukushi, Yasuko; Kubota, Oichi; Itsuji, Takeaki; Ouchi, Toshihiko; Yamamoto, Seiji

    2016-07-01

    Tumor imaging by terahertz spectroscopy of fresh tissue without dye is demonstrated using samples from a rat glioma model. The complex refractive index spectrum obtained by a reflection terahertz time-domain spectroscopy system can discriminate between normal and tumor tissues. Both the refractive index and absorption coefficient of tumor tissues are higher than those of normal tissues and can be attributed to the higher cell density and water content of the tumor region. The results of this study indicate that terahertz technology is useful for detecting brain tumor tissue.

  15. Chemical compositions and antibacterial activity of the leaf and stem oils of Piper porphyrophyllum (Lindl.) N.E. Br.

    PubMed Central

    Salleh, Wan Mohd Nuzul Hakimi Wan; Ahmad, Farediah; Sirat, Hasnah Mohd; Yen, Khong Heng

    2012-01-01

    The essential oils obtained by hydrodistillation from the fresh leaf and stem of Piper porphyrophyllum N.E. Br. were analyzed by GC and GC/MS. Thirty four constituents were identified in the leaf oil, while thirty eight constituents were identified in the stems oil. The most abundant components in the leaf oil included bicyclogermacrene (14.7 %), α-copaene (13.2 %) and β-phellandrene (9.5 %) while sabinene (15.5 %), bicyclogermacrene (12.3 %) and α-copaene (8.1 %) were the main constituents in the stem oil. The evaluation of antibacterial activity by using micro-dilution method revealed that both oils were moderately active against all the Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis) and Gram-negative bacteria (Pseudomonas aeruginosa, Pseudomonas putida and Escherichia coli) with minimum inhibitory concentration (MIC) values in the range 125-1000 µg/ml. PMID:27418915

  16. Three-dimensional radiation transfer modeling in a dicotyledon leaf

    NASA Astrophysics Data System (ADS)

    Govaerts, Yves M.; Jacquemoud, Stéphane; Verstraete, Michel M.; Ustin, Susan L.

    1996-11-01

    The propagation of light in a typical dicotyledon leaf is investigated with a new Monte Carlo ray-tracing model. The three-dimensional internal cellular structure of the various leaf tissues, including the epidermis, the palisade parenchyma, and the spongy mesophyll, is explicitly described. Cells of different tissues are assigned appropriate morphologies and contain realistic amounts of water and chlorophyll. Each cell constituent is characterized by an index of refraction and an absorption coefficient. The objective of this study is to investigate how the internal three-dimensional structure of the tissues and the optical properties of cell constituents control the reflectance and transmittance of the leaf. Model results compare favorably with laboratory observations. The influence of the roughness of the epidermis on the reflection and absorption of light is investigated, and simulation results confirm that convex cells in the epidermis focus light on the palisade parenchyma and increase the absorption of radiation.

  17. Antimony (SbIII) reduces growth, declines photosynthesis, and modifies leaf tissue anatomy in sunflower (Helianthus annuus L.).

    PubMed

    Vaculík, Marek; Mrázová, Anna; Lux, Alexander

    2015-12-01

    The role of antimony (Sb)--a non-essential trace metalloid--in physiological processes running in crops is still poorly understood. Present paper describes the effect of Sb tartrate (SbIII) on growth, Sb uptake, photosynthesis, photosynthetic pigments, and leaf tissue organization in young sunflower plants grown in hydroponics. We found that growth of below- and aboveground part was reduced with increasing concentration of Sb in the medium. Although Sb was mostly taken up by sunflower roots and only small part (1-2%) was translocated to the shoots, decline in photosynthesis, transpiration, and decreased content of photosynthetic pigments were observed. This indicates that despite relatively low mobility of Sb in root-shoot system, Sb in shoot noticeably modifies physiological status and reduced plant growth. Additionally, leaf anatomical changes indicated that Sb reduced the size of intercellular spaces and made leaf tissue more compact.

  18. Microlensed dual-fiber probe for depth-resolved fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Choi, Hae Young; Ryu, Seon Young; Kim, Jae Young; Kim, Geon Hee; Park, Seong Jun; Lee, Byeong Ha; Chang, Ki Soo

    2011-07-01

    We propose and demonstrate a compact microlensed dual-fiber probe that has a good collection efficiency and a high depth-resolution ability for fluorescence measurements. The probe is formed with a conventional fusion splicer creating a common focusing lens on two fibers placed side by side. The collection efficiency of the fabricated probe was evaluated by measuring the fluorescence signal of a fresh ginkgo leaf. It was shown experimentally that the proposed probe could effectively collect the fluorescence signal with a six-fold increase compared to that of a general flat-tipped probe. The beam propagation method was used to design a probe with an optimized working distance and an improved resolving depth. It was found that the working distance depends mainly on the radius of curvature of the lens, whereas the resolving depth is determined by the core diameters of the illumination and collection fibers. The depth-resolved ability of probes with working distances of ~100 μm and 300 μm was validated by using a two-layer tissue phantom. The experimental results demonstrate that the microlensed dual-fiber probe has the potential to facilitate depth-resolved fluorescence detection of epithelial tissue.

  19. Changing environmental conditions and applying organic fertilizers in Origanum vulgare L.

    PubMed Central

    Murillo-Amador, Bernardo; Morales-Prado, Luis E.; Troyo-Diéguez, Enrique; Córdoba-Matson, Miguel V.; Hernández-Montiel, Luis G.; Rueda-Puente, Edgar O.; Nieto-Garibay, Alejandra

    2015-01-01

    Any improvement in agricultural systems that results in higher production should also reduce negative environmental impacts and enhance sustainability. The aim of this research was to investigate the effect of two different production systems, one open-field and the other shade-enclosure with four bocashi doses, in order to find the best environmental option in terms of yield, physiological and morphometric characteristics in one oregano (Origanum vulgare L.) cultivar. In this study a completely randomized block design was used with four replications and evaluated for photosynthetic and transpiration rate, stomatal conductance, chlorophyll, leaf area and temperature, aerial and roots fresh and dry biomass, fresh and dry yield. The results showed that oregano adapted best to the shade-enclosure with increase yield of fresh and dry leaf weight of 165% and 118%, respectively, when compared to open-field. Also, higher doses of bocashi improved yield in both environments but more so in shade-enclosure. Soil moisture retention was higher in shade-enclosure which was reflected in physiological variables for soil matric potential, transpiration, stomatal conductivity, photosynthesis being significantly higher in shade-enclosure compared to open-field, thus improving yield. It seems that oregano plants can be grown and perform better under shade-enclosure than open-field and bocashi is a suitable organic fertilizer. PMID:26257756

  20. Biological monitoring of an agricultural food chain: Soil cadmium and lead ruminant tissues. [Sorghum sudanense (Piper) Stapf-S. bicolor (L. ) Moench (Capra hircus)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Brams, E.; Anthony, W.; Weatherspoon, L.

    Low-level contamination of a sandy soil with toxicants Cd and Pb at 0.01 to 9.0 and 3.0 to 54.0 mg kg{sup {minus}1} soil induced a significant toxicant accumulation in sudan-sorghum hay (Sorghum sudanense (Piper) Stapf-S. bicolor (L.) Moench) (0.5-5.0 and 0.2-1.5 mg kg{sup {minus}1} dry biomass), respectively. Ingestion of 22 to 222 mg Cd kg{sup {minus}1} body wt. by pregnant dairy goats (Capra hircus) from the consumption of hay over 98 d resulted in a significant, but relatively diminutive accumulation of Cd in the doe livers (0.01-0.02 mg kg{sup {minus}1}) and brain cortex (0.002-0.007 mg kg{sup {minus}1}) fresh wt., butmore » not in doe kidneys and blood averaging 0.028 and 0.002 mg Cd kg{sup {minus}1} fresh tissues, respectively. Fetal blood and liver accumulated 2.0 and 4.0 mg Cd kg{sup {minus}1} fresh tissue, respectively, and fetal kidney exhibited a weak response (0.03-0.47 mg Cd kg{sup {minus}1} fresh tissue) to Cd ingested by the pregnant does. Consumption of 240 to 1230 {mu}g Pb kg{sup {minus}1} body wt. induced 1.0 to 43.0 mg Pb kg{sup {minus}1} fresh tissue in the doe brain cerebellum, but not in the doe liver and blood averaging 0.09 and 0.017 mg Pb kg{sup {minus}1} fresh tissue, respectively, Fetal liver and blood averaged 0.043 and 0.014 mg Pb kg{sup {minus}1} tissue. Only minuscule amounts of soil Cd and Pb were retained in the select animal tissues via the ingestion of this hay. Only one-ten-millionth of labile soil Cd and Pb, respectively, accumulated in the select tissues of the pregnant does via the hay pathway. All these amounts were comparable to the norm. If these select animal tissues were used as food, no deleterious effects to human health should be induced.« less

  1. Effectiveness of two-sided UV-C treatments in inhibiting natural microflora and extending the shelf-life of minimally processed 'Red Oak Leaf' lettuce.

    PubMed

    Allende, Ana; McEvoy, James L; Luo, Yaguang; Artes, Francisco; Wang, Chien Y

    2006-05-01

    The use of UV-C radiation treatments to inhibit the microbial growth and extend the shelf-life of minimally processed 'Red Oak Leaf' lettuce was investigated. Initially, UV-C resistance of 20 bacterial strains from different genera often associated with fresh produce (Enterobacter, Erwinia, Escherichia, Leuconostoc, Pantoea, Pseudomonas, Rahnela, Salmonella, Serratia and Yersinia) were tested in vitro. Most of the bacterial strains were inhibited with the minimum dose (30 J m(-2)). Erwinia carotovora, Leuconostoc carnosum, Salmonella typhimurium, and Yersinia aldovae were the most resistant strains requiring a UV-C dose of 85 J m(-2) to completely inhibit growth. An in vivo study consisted of treating minimally processed 'Red Oak Leaf' lettuce (Lactuca sativa) with UV-C at three radiation doses (1.18, 2.37 and 7.11 kJ m(-2)) on each side of the leaves and storing the product under passive MAP conditions at 5 degrees C for up to 10 days. The gas composition inside packages varied significantly among the treatments, with CO2 concentrations positively and O2 concentrations negatively correlating with the radiation dose. All the radiation doses were effective in reducing the natural microflora of the product, although the highest doses showed the greatest microbial inhibitions. Taking into account the microbial limit set by Spanish legislation [Boletín Oficial del Estado (BOE), 2001. Normas de higiene para la elaboración, distribución y comercio de comidas preparadas, Madrid, Spain, Real Decreto 3484/2000, pp. 1435-1441], all UV-C treatments extended the shelf-life of the product. However, the 7.11 kJ m(-2) dose induced tissue softening and browning after 7 days of storage at 5 degrees C. Therefore, the use of two sided UV-C radiation, at the proper dose, is effective in reducing the natural microflora and extending the shelf-life of minimally processed 'Red Oak Leaf' lettuce.

  2. Biosynthesis of silver nanoparticles using fresh extracts of Tridax procumbens linn.

    PubMed

    Bhati-Kushwaha, Himakshi

    2014-04-01

    A simple and eco-friendly method for the synthesis of biogenic nanoparticles (NP's) using an aqueous solution of T. procumbens fresh plant extract (leaf and stem) as a bioreductant is reported. The prepared biogenic nanoparticles were well characterized using U.V. visible spectroscopy, scanning electron microscopy, X-ray diffraction and Fourier-transform infrared spectroscopy. The particles were confirmed to be elemental crystal by X-ray diffraction. The potential applications of biosynthesized nanoparticles as antimicrobial (antibacterial and antifungal) against pathogens Escherichia coli, Vibrio cholerae, Aspergillus niger and Aspergillusflavus were demonstrated.

  3. Intra-plant variation in cyanogenesis and the continuum of foliar plant defense traits in the rainforest tree Ryparosa kurrangii (Achariaceae).

    PubMed

    Webber, Bruce L; Woodrow, Ian E

    2008-06-01

    At the intra-plant level, temporal and spatial variations in plant defense traits can be influenced by resource requirements, defensive priorities and storage opportunities. Across a leaf age gradient, cyanogenic glycoside concentrations in the rainforest understory tree Ryparosa kurrangii B.L. Webber were higher in young expanding leaves than in mature leaves (2.58 and 1.38 mg g(-1), respectively). Moreover, cyanogens, as an effective chemical defense against generalist herbivores, contributed to a defense continuum protecting foliar tissue during leaf development. Chemical (cyanogens and phenolic compounds) and phenological (delayed greening) defense traits protected young leaves, whereas mature leaves were largely protected by physical defense mechanisms (lamina toughness; explained primarily by leaf mass per area). Cyanogen concentration was considerably higher in floral tissue than in foliar tissue and decreased in floral tissue during development. Across contrasting tropical seasons, foliar cyanogenic concentration varied significantly, being highest in the late wet season and lowest during the pre-wet season, the latter coinciding with fruiting and leaf flushing. Cyanogens in R. kurrangii appear to be differentially allocated in a way that maximizes plant fitness but may also act as a store of reduced nitrogen that is remobilized during flowering and leaf flushing.

  4. Leaf water relations during summer water deficit: differential responses in turgor maintenance and variation in leaf structure among different plant communities in south-western Australia.

    PubMed

    Mitchell, Patrick J; Veneklaas, Erik J; Lambers, Hans; Burgess, Stephen S O

    2008-12-01

    We measured leaf water relations and leaf structural traits of 20 species from three communities growing along a topographical gradient. Our aim was to assess variation in seasonal responses in leaf water status and leaf tissue physiology between sites and among species in response to summer water deficit. Species from a ridge-top heath community showed the greatest reductions in pre-dawn leaf water potentials (Psi(leaf)) and stomatal conductance during summer; species from a valley-floor woodland and a midslope mallee community showed less reductions in these parameters. Heath species also displayed greater seasonal reduction in turgor-loss point (Psi(TLP)) than species from woodland or mallee communities. In general, species that had larger reductions in Psi(leaf) during summer showed significant shifts in either their osmotic potential at full turgor (Psi(pi 100); osmotic adjustment) or in tissue elasticity (epsilon(max)). Psi(pi 100) and epsilon(max) were negatively correlated, during both spring and summer, suggesting a trade-off between these different mechanisms to cope with water stress. Specific leaf area varied greatly among species, and was significantly correlated with seasonal changes in Psi(TLP) and pre-dawn Psi(leaf). These correlations suggest that leaf structure is a prerequisite for cellular mechanisms to be effective in adjusting to water deficit.

  5. [Comparison of productivity of different vitamin green technologies under the space station conditions].

    PubMed

    Levinskikh, M A

    2002-01-01

    At present, fresh plant products for nutrition of the International space station (ISS) crews are delivered from Earth in small quantities. Regular supply of additional fresh greens could be positive for improvement as of nutrition, so psychophysical state of ISS crews. Vitamin greens can be produced with the use of various technologies: planting leaf cultures in greenhouses, forcing the greens from onions and root vegetables (onion, garlic, chicory, beet, parsley etc.), and germinating seeds. Purpose of this study was to compare productivity of these technologies in order to specify inputs for designers of a vitamin greenhouse to be mounted in the space station and a Martian vehicle. Based on comparison of the productivity of various technologies, specific productivity of different greenhouses per a unit of power consumption, and a volume unit it will be maximal if used for germinating seeds and minimal if used for growing leaf vegetables in a greenhouse with a cylindrical crop surface.

  6. Gene expression profile analysis of Ligon lintless-1 (Li1) mutant reveals important genes and pathways in cotton leaf and fiber development.

    PubMed

    Ding, Mingquan; Jiang, Yurong; Cao, Yuefen; Lin, Lifeng; He, Shae; Zhou, Wei; Rong, Junkang

    2014-02-10

    Ligon lintless-1 (Li1) is a monogenic dominant mutant of Gossypium hirsutum (upland cotton) with a phenotype of impaired vegetative growth and short lint fibers. Despite years of research involving genetic mapping and gene expression profile analysis of Li1 mutant ovule tissues, the gene remains uncloned and the underlying pathway of cotton fiber elongation is still unclear. In this study, we report the whole genome-level deep-sequencing analysis of leaf tissues of the Li1 mutant. Differentially expressed genes in leaf tissues of mutant versus wild-type (WT) plants are identified, and the underlying pathways and potential genes that control leaf and fiber development are inferred. The results show that transcription factors AS2, YABBY5, and KANDI-like are significantly differentially expressed in mutant tissues compared with WT ones. Interestingly, several fiber development-related genes are found in the downregulated gene list of the mutant leaf transcriptome. These genes include heat shock protein family, cytoskeleton arrangement, cell wall synthesis, energy, H2O2 metabolism-related genes, and WRKY transcription factors. This finding suggests that the genes are involved in leaf morphology determination and fiber elongation. The expression data are also compared with the previously published microarray data of Li1 ovule tissues. Comparative analysis of the ovule transcriptomes of Li1 and WT reveals that a number of pathways important for fiber elongation are enriched in the downregulated gene list at different fiber development stages (0, 6, 9, 12, 15, 18dpa). Differentially expressed genes identified in both leaf and fiber samples are aligned with cotton whole genome sequences and combined with the genetic fine mapping results to identify a list of candidate genes for Li1. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Using fresh tissue dissection to teach human anatomy in the clinical years.

    PubMed

    Robinson, Alan G; Metten, Shaleen; Guiton, Gretchen; Berek, Jonathan

    2004-07-01

    Gross anatomy is taught in medical school with textbooks, cadaver dissection, plastic models, and multimedia illustration, but all lack the reality of color and texture that is possible with fresh tissue dissection. The authors studied the use of fresh tissue dissection of the thorax and abdomen of the rat to teach human anatomy. In a half-day exercise, 52 fourth-year medical students paired off and completed an exercise to dissect in less than three hours the thorax and abdomen of a euthanized rat. Observation of organs was augmented by active manipulation such as passing a tube down the esophagus, cannulating the trachea and inflating the lungs, injecting dye in the kidney to trace the ureter and bladder, and pulling the testis through the inguinal canal. Comparison of the rat and human was emphasized to enhance the education. The exercise ended with practice suturing fresh tissue. Students rated the exercise to teach anatomy as 4.9 positive on a 5.0 (high) scale. The significant positive structures (p <.05) for texture were heart, liver, lungs and trachea; for color they were lungs and spleen; for location and size they were adrenal gland and urinary bladder; and for function they were adrenal gland and esophagus. Fresh tissue dissection of the thorax and abdomen of the rat is a valuable tool for human anatomy education. The dissonances in human and rat anatomy enhance abstraction and transfer of knowledge. Active manipulation of organs promotes retention of knowledge, and suturing provides a "clinical" context. Fresh tissue dissection is an efficient innovative method to provide a global review of anatomy of the thorax and abdomen during the busy clinical years of medical education.

  8. Penetration of UV-A, UV-B, blue, and red light into leaf tissues of pecan measured by a fiber optic microprobe system

    NASA Astrophysics Data System (ADS)

    Qi, Yadong; Bai, Shuju; Vogelmann, Thomas C.; Heisler, Gordon M.

    2003-11-01

    The depth of light penetration from the adaxial surfaces of the mature leaves of pecan (Carya illinoensis) was measured using a fiber optic microprobe system at four wavelengths: UV-B (310nm), UV-A (360 nm), blue light (430nm), and red light (680nm). The average thickness of the leaf adaxial epidermal layer was 15um and the total leaf thickness was 219um. The patterns of the light attenuation by the leaf tissues exhibited strong wavelength dependence. The leaf adaxial epidermal layer was chiefly responsible for absorbing the UV-A UV-B radiation. About 98% of 310 nm light was steeply attenuated within the first 5 um of the adaxial epidermis; thus, very little UV-B radiation was transmitted to the mesophyll tissues where contain photosynthetically sensitive sites. The adaxial epidermis also attenuated 96% of the UV-A radiation. In contrast, the blue and red light penetrated much deeper and was gradually attenutated by the leaves. The mesophyll tissues attenuated 17% of the blue light and 42% of the red light, which were available for photosynthesis use. Since the epidermal layer absorbed nearly all UV-B light, it acted as an effective filter screening out the harmful radiation and protecting photosynthetically sensitive tissues from the UV-B damage. Therefore, the epidermal function of the UV-B screening effectiveness can be regarded as one of the UV-B protection mechanisms in pecan.

  9. An insect herbivore microbiome with high plant biomass-degrading capacity

    USDA-ARS?s Scientific Manuscript database

    Herbivores can gain indirect access to recalcitrant carbon present in plant cell walls through symbiotic associations with lignocellulolytic microbes. A paradigmatic example is the leaf-cutter ant (Tribe: Attini), which uses fresh leaves to cultivate a fungus for food in specialized gardens. Using a...

  10. A Vitis vinifera basic helix-loop-helix transcription factor enhances plant cell size, vegetative biomass and reproductive yield

    DOE PAGES

    Lim, Sung Don; Yim, Won Choel; Liu, Degao; ...

    2018-04-16

    Strategies for improving plant size are critical targets for plant biotechnology to increase vegetative biomass or reproductive yield. To improve biomass production, a codon-optimized helix–loop–helix transcription factor (VvCEB1 opt) from wine grape was overexpressed in Arabidopsis thaliana resulting in significantly increased leaf number, leaf and rosette area, fresh weight and dry weight. Cell size, but typically not cell number, was increased in all tissues resulting in increased vegetative biomass and reproductive organ size, number and seed yield. Ionomic analysis of leaves revealed the VvCEB1 opt-overexpressing plants had significantly elevated, K, S and Mo contents relative to control lines. Increased Kmore » content likely drives increased osmotic potential within cells leading to greater cellular growth and expansion. To understand the mechanistic basis of VvCEB1 opt action, one transgenic line was genotyped using RNA-Seq mRNA expression profiling and revealed a novel transcriptional reprogramming network with significant changes in mRNA abundance for genes with functions in delayed flowering, pathogen–defence responses, iron homeostasis, vesicle-mediated cell wall formation and auxin-mediated signalling and responses. Direct testing of VvCEB1 opt-overexpressing plants showed that they had significantly elevated auxin content and a significantly increased number of lateral leaf primordia within meristems relative to controls, confirming that cell expansion and organ number proliferation were likely an auxin-mediated process. VvCEB1 opt overexpression in Nicotiana sylvestris also showed larger cells, organ size and biomass demonstrating the potential applicability of this innovative strategy for improving plant biomass and reproductive yield in crops.« less

  11. A Vitis vinifera basic helix-loop-helix transcription factor enhances plant cell size, vegetative biomass and reproductive yield

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lim, Sung Don; Yim, Won Choel; Liu, Degao

    Strategies for improving plant size are critical targets for plant biotechnology to increase vegetative biomass or reproductive yield. To improve biomass production, a codon-optimized helix–loop–helix transcription factor (VvCEB1 opt) from wine grape was overexpressed in Arabidopsis thaliana resulting in significantly increased leaf number, leaf and rosette area, fresh weight and dry weight. Cell size, but typically not cell number, was increased in all tissues resulting in increased vegetative biomass and reproductive organ size, number and seed yield. Ionomic analysis of leaves revealed the VvCEB1 opt-overexpressing plants had significantly elevated, K, S and Mo contents relative to control lines. Increased Kmore » content likely drives increased osmotic potential within cells leading to greater cellular growth and expansion. To understand the mechanistic basis of VvCEB1 opt action, one transgenic line was genotyped using RNA-Seq mRNA expression profiling and revealed a novel transcriptional reprogramming network with significant changes in mRNA abundance for genes with functions in delayed flowering, pathogen–defence responses, iron homeostasis, vesicle-mediated cell wall formation and auxin-mediated signalling and responses. Direct testing of VvCEB1 opt-overexpressing plants showed that they had significantly elevated auxin content and a significantly increased number of lateral leaf primordia within meristems relative to controls, confirming that cell expansion and organ number proliferation were likely an auxin-mediated process. VvCEB1 opt overexpression in Nicotiana sylvestris also showed larger cells, organ size and biomass demonstrating the potential applicability of this innovative strategy for improving plant biomass and reproductive yield in crops.« less

  12. Genotype, development and tissue-derived variation of cell-wall properties in the lignocellulosic energy crop Miscanthus

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    da Costa, Ricardo M. F.; Lee, Scott J.; Allison, Gordon G.

    Species and hybrids of the genus Miscanthus contain attributes that make them front-runners among current selections of dedicated bioenergy crops. A key trait for plant biomass conversion to biofuels and biomaterials is cell-wall quality; however, knowledge of cell-wall composition and biology in Miscanthus species is limited. This study presents data on cell-wall compositional changes as a function of development and tissue type across selected genotypes, and considers implications for the development of miscanthus as a sustainable and renewable bioenergy feedstock. Cell-wall biomass was analysed for 25 genotypes, considering different developmental stages and stem vs. leaf compositional variability, by Fourier transformmore » mid-infrared spectroscopy and lignin determination. In addition, a Clostridium phytofermentans bioassay was used to assess cell-wall digestibility and conversion to ethanol. Important cell-wall compositional differences between miscanthus stem and leaf samples were found to be predominantly associated with structural carbohydrates. Lignin content increased as plants matured and was higher in stem tissues. Although stem lignin concentration correlated inversely with ethanol production, no such correlation was observed for leaves. Leaf tissue contributed significantly to total above-ground biomass at all stages, although the extent of this contribution was genotype-dependent. In conclusion, it is hypothesized that divergent carbohydrate compositions and modifications in stem and leaf tissues are major determinants for observed differences in cell-wall quality. The findings indicate that improvement of lignocellulosic feedstocks should encompass tissue-dependent variation as it affects amenability to biological conversion. For gene-trait associations relating to cell-wall quality, the data support the separate examination of leaf and stem composition, as tissue-specific traits may be masked by considering only total above-ground biomass samples, and sample variability could be mostly due to varying tissue contributions to total biomass.« less

  13. Genotype, development and tissue-derived variation of cell-wall properties in the lignocellulosic energy crop Miscanthus

    DOE PAGES

    da Costa, Ricardo M. F.; Lee, Scott J.; Allison, Gordon G.; ...

    2014-04-15

    Species and hybrids of the genus Miscanthus contain attributes that make them front-runners among current selections of dedicated bioenergy crops. A key trait for plant biomass conversion to biofuels and biomaterials is cell-wall quality; however, knowledge of cell-wall composition and biology in Miscanthus species is limited. This study presents data on cell-wall compositional changes as a function of development and tissue type across selected genotypes, and considers implications for the development of miscanthus as a sustainable and renewable bioenergy feedstock. Cell-wall biomass was analysed for 25 genotypes, considering different developmental stages and stem vs. leaf compositional variability, by Fourier transformmore » mid-infrared spectroscopy and lignin determination. In addition, a Clostridium phytofermentans bioassay was used to assess cell-wall digestibility and conversion to ethanol. Important cell-wall compositional differences between miscanthus stem and leaf samples were found to be predominantly associated with structural carbohydrates. Lignin content increased as plants matured and was higher in stem tissues. Although stem lignin concentration correlated inversely with ethanol production, no such correlation was observed for leaves. Leaf tissue contributed significantly to total above-ground biomass at all stages, although the extent of this contribution was genotype-dependent. In conclusion, it is hypothesized that divergent carbohydrate compositions and modifications in stem and leaf tissues are major determinants for observed differences in cell-wall quality. The findings indicate that improvement of lignocellulosic feedstocks should encompass tissue-dependent variation as it affects amenability to biological conversion. For gene-trait associations relating to cell-wall quality, the data support the separate examination of leaf and stem composition, as tissue-specific traits may be masked by considering only total above-ground biomass samples, and sample variability could be mostly due to varying tissue contributions to total biomass.« less

  14. Genotype, development and tissue-derived variation of cell-wall properties in the lignocellulosic energy crop Miscanthus.

    PubMed

    da Costa, Ricardo M F; Lee, Scott J; Allison, Gordon G; Hazen, Samuel P; Winters, Ana; Bosch, Maurice

    2014-10-01

    Species and hybrids of the genus Miscanthus contain attributes that make them front-runners among current selections of dedicated bioenergy crops. A key trait for plant biomass conversion to biofuels and biomaterials is cell-wall quality; however, knowledge of cell-wall composition and biology in Miscanthus species is limited. This study presents data on cell-wall compositional changes as a function of development and tissue type across selected genotypes, and considers implications for the development of miscanthus as a sustainable and renewable bioenergy feedstock. Cell-wall biomass was analysed for 25 genotypes, considering different developmental stages and stem vs. leaf compositional variability, by Fourier transform mid-infrared spectroscopy and lignin determination. In addition, a Clostridium phytofermentans bioassay was used to assess cell-wall digestibility and conversion to ethanol. Important cell-wall compositional differences between miscanthus stem and leaf samples were found to be predominantly associated with structural carbohydrates. Lignin content increased as plants matured and was higher in stem tissues. Although stem lignin concentration correlated inversely with ethanol production, no such correlation was observed for leaves. Leaf tissue contributed significantly to total above-ground biomass at all stages, although the extent of this contribution was genotype-dependent. It is hypothesized that divergent carbohydrate compositions and modifications in stem and leaf tissues are major determinants for observed differences in cell-wall quality. The findings indicate that improvement of lignocellulosic feedstocks should encompass tissue-dependent variation as it affects amenability to biological conversion. For gene-trait associations relating to cell-wall quality, the data support the separate examination of leaf and stem composition, as tissue-specific traits may be masked by considering only total above-ground biomass samples, and sample variability could be mostly due to varying tissue contributions to total biomass. © The Author 2014. Published by Oxford University Press on behalf of the Annals of Botany Company.

  15. Net primary production and phenology on a southern Appalachian watershed

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Day, F.P. Jr.; Monk, C.D.

    1977-01-01

    Net primary production (NPP) is an important function of plant communities which has not often been examined seasonally in a forested ecosystem. The major objective of the study was to measure above-ground NPP seasonally and relate it to phenological activity on a hardwood forest watershed at Coweeta Hydrologic Laboratory, North Carolina. NPP was estimated as the increase in biomass, estimated from regression equations on diameter. Diameter increases were measured by venier tree bands. Phenological observations were made on bud break, leaf emergence, flowering, mature fruit, leaf senescence, and leaf fall. The species studied intensively were Acer rubrum, Quercus prinus, Caryamore » glabra, Cornus florida, and Liriodendron tulipifera. Liriodendron was found to be the most productive species per individual, but Quercus prinus was the most productive per unit ground area. The total watershed estimate of aboveground NPP was 8,754 kg ha/sup -1/yr/sup -1/ and included 47.9 percent leaves, 33.2 percent wood, 7.8 percent bark, 4.8 percent reproductive tissues, 4.2 percent loss to consumers, and 2.1 percent twigs. Increases in leaf biomass were most rapid in the spring, but woody tissue production peaked in June and continued through August. Since leaf production peaked in the spring, the plants' photosynthetic machinery was activated early in the growing season to support woody tissue production, which followed the period of rapid leaf growth, and reproductive activity. Flowering occurred during the leaf expansion period except for Acer rubrum, which flowered before leaf emergence. Fruit maturation occurred during late summer to early fall, when there were no additional biomass increases. Acer rubrum was an exception as its fruit matured during the period of leaf expansion.« less

  16. Net primary production and phenology on a southern Appalachian watershed

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Day, F.P. Jr.; Monk, C.D.

    1977-10-01

    Net primary production (NPP) is an important function of plant communities which has not often been examined seasonally in a forested ecosystem. The major objective of the study was to measure above-ground NPP seasonally and relate it to phenological activity on a hardwood forest watershed at Coweeta Hydrologic Laboratory, North Carolina. NPP was estimated as the increase in biomass, estimated from regression equations on diameter. Diameter increases were measured by vernier tree bands. Phenological observations were made on bud break, leaf emergence, flowering, mature fruit, leaf senescence, and leaf fall. The species studied intensively were Acer rubrum, Quercus prinus, Caryamore » glabra, Cornus florida, and Liriodendron tulipifera. Liriodendron was found to be the most productive species per individual, but Quercus prinus was the most productive per unit ground area. The total watershed estimate of aboveground NPP was 8,754 kg ha/sup -1/ yr/sup -1/ and included 47.9% leaves, 33.2% wood, 7.8% bark, 4.8% reproductive tissues, 4.2% loss to consumers, and 2.1% twigs. Increases in leaf biomass were most rapid in the spring, but woody tissue production peaked in June and continued through August. Since leaf production peaked in the spring, the plants' photosynthetic machinery was activated early in the growing season to support woody tissue production, which followed the period of rapid leaf growth, and reproductive activity. Flowering ocurred during the leaf expansion period except for Acer rubrum, which flowered before leaf emergence. Fruit maturation occurred during late summer to early fall, when there were no additional biomass increases. Acer rubrum was an exception as its fruit matured during the period of leaf expansion.« less

  17. Detection of the multiphoton signals in stained tissue using nonlinear optical microscopy

    NASA Astrophysics Data System (ADS)

    Zeng, Yaping; Xu, Jian; Kang, Deyong; Lin, Jiangbo; Chen, Jianxin

    2016-10-01

    Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging, has become a powerful, important tool for tissue imaging at the molecular level. Recently, MPM is also used to image hematoxylin and eosin (H and E)-stained sections in cancer diagnostics. However, several studies have showed that the MPM images of tissue stained with H and E are significantly different from unstained tissue sections. Our aim was to detect of the multiphoton signals in stained tissue by using MPM. In this paper, MPM was used to image histological sections of esophageal invasive carcinoma tissues stained with H, E, H and E and fresh tissue. To detect of the multiphoton signals in stained tissue, the emission spectroscopic of tissue stained with H, E, H and E were obtained. For comparison, the fresh tissues were also investigated. Our results showed that the tissue stained with H, E, H and E could be detected by their TPEF signals. While the tissue stained with H and fresh tissue could be detected by their TPEF and SHG signals. In this work, we detect of the multiphoton signals in stained tissue. These findings will be useful for choosing suitable staining method so to improve the quality of MPM imaging in the future.

  18. Outside-Xylem Vulnerability, Not Xylem Embolism, Controls Leaf Hydraulic Decline during Dehydration1[CC-BY

    PubMed Central

    Townes, Shatara V.; Bartlett, Megan K.; Buckley, Thomas N.; McElrone, Andrew J.; Sack, Lawren

    2017-01-01

    Leaf hydraulic supply is crucial to maintaining open stomata for CO2 capture and plant growth. During drought-induced dehydration, the leaf hydraulic conductance (Kleaf) declines, which contributes to stomatal closure and, eventually, to leaf death. Previous studies have tended to attribute the decline of Kleaf to embolism in the leaf vein xylem. We visualized at high resolution and quantified experimentally the hydraulic vulnerability of xylem and outside-xylem pathways and modeled their respective influences on plant water transport. Evidence from all approaches indicated that the decline of Kleaf during dehydration arose first and foremost due to the vulnerability of outside-xylem tissues. In vivo x-ray microcomputed tomography of dehydrating leaves of four diverse angiosperm species showed that, at the turgor loss point, only small fractions of leaf vein xylem conduits were embolized, and substantial xylem embolism arose only under severe dehydration. Experiments on an expanded set of eight angiosperm species showed that outside-xylem hydraulic vulnerability explained 75% to 100% of Kleaf decline across the range of dehydration from mild water stress to beyond turgor loss point. Spatially explicit modeling of leaf water transport pointed to a role for reduced membrane conductivity consistent with published data for cells and tissues. Plant-scale modeling suggested that outside-xylem hydraulic vulnerability can protect the xylem from tensions that would induce embolism and disruption of water transport under mild to moderate soil and atmospheric droughts. These findings pinpoint outside-xylem tissues as a central locus for the control of leaf and plant water transport during progressive drought. PMID:28049739

  19. ELF Communications System Ecological Monitoring Program: A Summary Report for 1982-1992

    DTIC Science & Technology

    1993-10-01

    autumn- abscised alder leaves were used in the packs; since 1990, only fresh alder leaves have been used. Leaf packs were periodically retrieved and...red pine planting survival. Northern Journal of Applied Forestry, 5:14; 1988. 56. Connaughton, P. The effects of acid precipitation on nutrient levels

  20. Spinacia oleracea L. leaf stomata harboring Cryptosporidium parvum oocysts: A potential threat for food safety

    USDA-ARS?s Scientific Manuscript database

    Scientific literature documents the prevalence of Cryptosporidium oocysts in irrigation waters and on fresh produce. In the present study spinach leaves were experimentally exposed to Cryptosporidium oocysts which were subsequently irrigated with clean water daily for 5 days. As determined by confoc...

  1. Plant Histology: Clearing and the Optical Section.

    ERIC Educational Resources Information Center

    Freeman, H. E.

    1985-01-01

    Clearing is a simple and rapid technique in which 75 percent lactic acid is used to remove pigments and cytoplasmic contents of fresh leaves, enabling microscopic view of various internal leaf layers. Procedures for using the technique (which helps students gain a more thorough understanding of plant anatomy) are given. (DH)

  2. Indirect suppression of photosynthesis on individual leaves by arthropod herbivory

    PubMed Central

    Nabity, Paul D.; Zavala, Jorge A.; DeLucia, Evan H.

    2009-01-01

    Background Herbivory reduces leaf area, disrupts the function of leaves, and ultimately alters yield and productivity. Herbivore damage to foliage typically is assessed in the field by measuring the amount of leaf tissue removed and disrupted. This approach assumes the remaining tissues are unaltered, and plant photosynthesis and water balance function normally. However, recent application of thermal and fluorescent imaging technologies revealed that alterations to photosynthesis and transpiration propagate into remaining undamaged leaf tissue. Scope and Conclusions This review briefly examines the indirect effects of herbivory on photosynthesis, measured by gas exchange or chlorophyll fluorescence, and identifies four mechanisms contributing to the indirect suppression of photosynthesis in remaining leaf tissues: severed vasculature, altered sink demand, defence-induced autotoxicity, and defence-induced down-regulation of photosynthesis. We review the chlorophyll fluorescence and thermal imaging techniques used to gather layers of spatial data and discuss methods for compiling these layers to achieve greater insight into mechanisms contributing to the indirect suppression of photosynthesis. We also elaborate on a few herbivore-induced gene-regulating mechanisms which modulate photosynthesis and discuss the difficult nature of measuring spatial heterogeneity when combining fluorescence imaging and gas exchange technology. Although few studies have characterized herbivore-induced indirect effects on photosynthesis at the leaf level, an emerging literature suggests that the loss of photosynthetic capacity following herbivory may be greater than direct loss of photosynthetic tissues. Depending on the damage guild, ignoring the indirect suppression of photosynthesis by arthropods and other organisms may lead to an underestimate of their physiological and ecological impacts. PMID:18660492

  3. Carbon and nitrogen dynamics of the intertidal seagrass, Zostera japonica, on the southern coast of the Korean peninsula

    NASA Astrophysics Data System (ADS)

    Kim, Jong-Hyeob; Kim, Seung Hyeon; Kim, Young Kyun; Lee, Kun-Seop

    2016-12-01

    Seagrasses require a large amount of nutrient assimilation to support high levels of production, and thus nutrient limitation for growth often occurs in seagrass habitats. Seagrasses can take up nutrients from both the water column and sediments. However, since seagrasses inhabiting in the intertidal zones are exposed to the air during low tide, the intertidal species may exhibit significantly different carbon (C) and nitrogen (N) dynamics compared to the subtidal species. To examine C and N dynamics of the intertidal seagrass, Zostera japonica, C and N content and stable isotope ratios of above- and below-ground tissues were measured monthly at the three intertidal zones in Koje Bay on the southern coast of Korea. The C and N content and stable isotope (δ13C and δ15N) ratios of seagrass tissues exhibited significant seasonal variations. Both leaf and rhizome C content were not significantly correlated with productivity. Leaf δ13C values usually exhibited negative correlations with leaf productivity. These results of tissue C content and δ13C values suggest that photosynthesis of Z. japonica in the study site was not limited by inorganic C supply, and sufficient inorganic C was provided from the atmosphere. The tissue N content usually exhibited negative correlations with leaf productivity except at the upper intertidal zone, suggesting that Z. japonica growth was probably limited by N availability during high growing season. In the upper intertidal zone, no correlations between leaf productivity and tissue elemental content and stable isotope ratios were observed due to the severely suppressed growth caused by strong desiccation stress.

  4. The bionomics of the cottonwood leaf beetle, Chrysomela scripta Fab., on tissue culture hybrid poplars

    Treesearch

    T.R. Burkot; D.M. Benjamin

    1977-01-01

    Tissue culture methods are applied to poplars of the Aigeiros group in attempts to overcome premature decline thought to be associated with viral infections. Hybrid selections from such cultures outplanted in 1975 at the F. G. Wilson Nursery in Boscobel, Wisconsin subsequently were severely infested by the Cottonwood Leaf Beetle, Chrysomela scripta Fab. Beetle...

  5. A model using marginal efficiency of investment to analyse carbon and nitrogen interactions in forested ecosystems

    NASA Astrophysics Data System (ADS)

    Thomas, R. Q.; Williams, M.

    2014-12-01

    Carbon (C) and nitrogen (N) cycles are coupled in terrestrial ecosystems through multiple processes including photosynthesis, tissue allocation, respiration, N fixation, N uptake, and decomposition of litter and soil organic matter. Capturing the constraint of N on terrestrial C uptake and storage has been a focus of the Earth System modelling community. Here we explore the trade-offs and sensitivities of allocating C and N to different tissues in order to optimize the productivity of plants using a new, simple model of ecosystem C-N cycling and interactions (ACONITE). ACONITE builds on theory related to plant economics in order to predict key ecosystem properties (leaf area index, leaf C:N, N fixation, and plant C use efficiency) based on the optimization of the marginal change in net C or N uptake associated with a change in allocation of C or N to plant tissues. We simulated and evaluated steady-state and transient ecosystem stocks and fluxes in three different forest ecosystems types (tropical evergreen, temperate deciduous, and temperate evergreen). Leaf C:N differed among the three ecosystem types (temperate deciduous < tropical evergreen < temperature evergreen), a result that compared well to observations from a global database describing plant traits. Gross primary productivity (GPP) and net primary productivity (NPP) estimates compared well to observed fluxes at the simulation sites. A sensitivity analysis revealed that parameterization of the relationship between leaf N and leaf respiration had the largest influence on leaf area index and leaf C:N. Also, a widely used linear leaf N-respiration relationship did not yield a realistic leaf C:N, while a more recently reported non-linear relationship simulated leaf C:N that compared better to the global trait database than the linear relationship. Overall, our ability to constrain leaf area index and allow spatially and temporally variable leaf C:N can help address challenges simulating these properties in ecosystem and Earth System models. Furthermore, the simple approach with emergent properties based on coupled C-N dynamics has potential for use in research that uses data-assimilation methods to integrate data on both the C and N cycles to improve C flux forecasts.

  6. Pheophorbide a Content and Chlorophyllase Activity in Green Tea.

    PubMed

    Kohata, K; Hanada, K; Yamauchi, Y; Horie, H

    1998-01-01

    We investigated the total content of pheophorbide a (PB a), which is sum of the contents of newly produced PB a, including PB a initially present and that converted from chlorophyllide a (Chd a) by the chlorophyllase reaction during incubation, in green tea samples, and found that the total content of PB a markedly increased in both Sencha and Matcha, compared with the initially present PB a content in each. This result demonstrates that chlorophyllase activity still remains in green tea, even after processing fresh green leaves. A comparison of the total contents of PB a produced during the incubation of chlorophyll a (Chl a) with Sencha and fresh green leaf acetone powder indicates that the ratio of chlorophyllase activity in Sencha and in fresh green leaves was about 1:20.

  7. Production of human rotavirus and Salmonella antigens in plants and elicitation of fljB-specific humoral responses in mice.

    PubMed

    Bergeron-Sandoval, Louis-Philippe; Girard, Aurélie; Ouellet, François; Archambault, Denis; Sarhan, Fathey

    2011-02-01

    A Nicotiana benthamiana transient expression system was used to express single antigen and dimeric combinations of the human rotavirus (HRV) VP7 and a truncated VP4 (VP4Δ) proteins fused with Salmonella typhimurium's flagellin fljB subunit. Immunoblot analyses using rabbit antibodies generated against these proteins demonstrated that the constructs were successfully expressed with yields ranging from 0.85 to 31.97 μg of recombinant protein per gram of fresh leaf tissue. Expressing the single and dimeric antigens has no effect on plant growth and development except for VP7 and VP4Δ::VP7, which show mild necrotic lesions. Immunization of mice with proteins from leaves transformed with constructs bearing the fljB moiety elicited an fljB-specific humoral response. The Nicotiana benthamiana transient system is efficient to express multiple combinations of pathogen proteins and demonstrates the potential of generating a Salmonella typhimurium subunit vaccine in plants.

  8. Green synthesis of silver nanoparticles from Moringa oleifera leaf extracts and its antimicrobial potential

    NASA Astrophysics Data System (ADS)

    Moodley, Jerushka S.; Babu Naidu Krishna, Suresh; Pillay, Karen; Sershen; Govender, Patrick

    2018-03-01

    In this study we report on the synthesis of silver nanoparticles (AgNPs) from the leaf extracts of Moringa oleifera using sunlight irradiation as primary source of energy, and its antimicrobial potential. Silver nanoparticle formation was confirmed by surface plasmon resonance at 450 nm and 440 nm, respectively for both fresh and freeze-dried leaf samples. Crystanality of AgNPs was confirmed by transmission electron microscopy, scanning electron microscopy with energy dispersive x-ray spectroscopy and Fourier transform infrared (FTIR) spectroscopy analysis. FTIR spectroscopic analysis suggested that flavones, terpenoids and polysaccharides predominate and are primarily responsible for the reduction and subsequent capping of AgNPs. X-ray diffraction analysis also demonstrated that the size range of AgNPs from both samples exhibited average diameters of 9 and 11 nm, respectively. Silver nanoparticles showed antimicrobial activity on both bacterial and fungal strains. The biosynthesised nanoparticle preparations from M. oleifera leaf extracts exhibit potential for application as broad-spectrum antimicrobial agents.

  9. Growth and reflectance characteristics of winter wheat canopies

    NASA Technical Reports Server (NTRS)

    Hinzman, L. D.; Bauer, M. E.; Daughtry, C. S. T.

    1984-01-01

    A valuable input to crop growth and yield models would be estimates of current crop condition. If multispectral reflectance indicates crop condition, then remote sensing may provide an additional tool for crop assessment. The effects of nitrogen fertilization on the spectral reflectance and agronomic characteristics of winter wheat (Triticum aestivum L.) were determined through field experiments. Spectral reflectance was measured during the 1979 and 1980 growing seasons with a spectroradiometer. Agronomic data included total leaf N concentration, leaf chlorophyll concentration, stage of development, leaf area index (LAI), plant moisture, and fresh and dry phytomass. Nitrogen deficiency caused increased visible, reduced near infrared, and increased middle infrared reflectance. These changes were related to lower levels of chlorophyll and reduced leaf area in the N-deficient plots. Green LAI, an important descriptor of wheat canopies, could be reliably estimated with multispectral data. The potential of remote sensing in distinguishing stressed from healthy crops is demonstrated. Evidence suggests multispectral imagery may be useful for monitoring crop condition.

  10. Nondestructive and continuous monitoring of oxygen levels in modified atmosphere packaged ready-to-eat mixed salad products using optical oxygen sensors, and its effects on sensory and microbiological counts during storage.

    PubMed

    Hempel, A; O'Sullivan, M G; Papkovsky, D B; Kerry, J P

    2013-07-01

    The objective of this study was to determine the percentage oxygen consumption of fresh, respiring ready-to-eat (RTE) mixed leaf salad products (Iceberg salad leaf, Caesar salad leaf, and Italian salad leaf). These were held under different modified atmosphere packaging (MAP) conditions (5% O2 , 5% CO2 , 90% N2 (MAPC-commercial control), 21% O2 , 5% CO2 , 74% N2 (MAP 1), 45% O2 , 5% CO2 , 50% N2 (MAP 2), and 60% O2 , 5% CO2 , 35% N2 (MAP 3)) and 4 °C for up to 10 d. The quality and shelf-life stability of all packaged salad products were evaluated using sensory, physiochemical, and microbial assessment. Oxygen levels in all MAP packs were measured on each day of analysis using optical oxygen sensors allowing for nondestructive assessment of packs. Analysis showed that with the exception of control packs, oxygen levels for all MAP treatments decreased by approximately 10% after 7 d of storage. Oxygen levels in control packs were depleted after 7 d of storage. This appears to have had no detrimental effect on either the sensory quality or shelf-life stability of any of the salad products investigated. Additionally, the presence of higher levels of oxygen in modified atmosphere packs did not significantly improve product quality or shelf-life stability; however, these additional levels of oxygen were freely available to fresh respiring produce if required. This study shows that the application of optical sensors in MAP packs was successful in nondestructively monitoring oxygen level, or changes in oxygen level, during refrigerated storage of RTE salad products. © 2013 Institute of Food Technologists®

  11. Characteristics and DBP formation of dissolved organic matter from leachates of fresh and aged leaf litter.

    PubMed

    Jian, Qianyun; Boyer, Treavor H; Yang, Xiuhong; Xia, Beicheng; Yang, Xin

    2016-06-01

    Dissolved organic matter (DOM) was leached from leaves of two trees commonly grown in subtropical regions, Pinus elliottii (commonly known as slash pine) and Schima superba (S. superba), and its degradation pattern and potential for forming disinfection byproducts (DBPs) were evaluated. The leaves were exposed in the field for up to one year before leaching. The DOM leached from slash pine litter contained on average 10.4 mg of dissolved organic carbon (DOC) per gram of dry weight; for S. superba the average was 37.2 mg-DOC/g-dry weight. Ultraviolet and visible light absorbance, fluorescence, and molecular weight analysis indicated that more aromatic/humic and higher molecular weight compounds are formed as leaf litter ages. A 4-component parallel factor analysis of the fluorescence data showed that the intensity of peaks related with protein-like components decreased gradually during biodegradation, while that of peaks attributed to humic-acid-like components increased continuously. Fresh slash pine leachates formed on average 40.0 μg of trihalomethane (THM) per milligram of DOC, while S. superba leachates formed 45.6 μg. THM formation showed peak values of 55.7 μg/mg DOC for slash pine and 74.9 μg/mg DOC for S. superba after 8 months of aging. The formation of haloacetonitrile (HAN) and trichloronitromethane (TCNM) increased with increasing leaf age, while chloral hydrate (CH) formation did not show such a trend. Specific UV absorbance showed some positive correlation with DBPs, but humic-acid-like and protein-like absorbance peaks correlated with CH and TCNM yields in only some leaf samples. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Highly Decorated Lignins in Leaf Tissues of the Canary Island Date Palm Phoenix canariensis1[OPEN

    PubMed Central

    Bartuce, Allison; Free, Heather C.A.; Smith, Bronwen G.

    2017-01-01

    The cell walls of leaf base tissues of the Canary Island date palm (Phoenix canariensis) contain lignins with the most complex compositions described to date. The lignin composition varies by tissue region and is derived from traditional monolignols (ML) along with an unprecedented range of ML conjugates: ML-acetate, ML-benzoate, ML-p-hydroxybenzoate, ML-vanillate, ML-p-coumarate, and ML-ferulate. The specific functions of such complex lignin compositions are unknown. However, the distribution of the ML conjugates varies depending on the tissue region, indicating that they may play specific roles in the cell walls of these tissues and/or in the plant’s defense system. PMID:28894022

  13. Structural assessment of the impact of environmental constraints on Arabidopsis thaliana leaf growth: a 3D approach.

    PubMed

    Wuyts, Nathalie; Massonnet, Catherine; Dauzat, Myriam; Granier, Christine

    2012-09-01

    Light and soil water content affect leaf surface area expansion through modifications in epidermal cell numbers and area, while effects on leaf thickness and mesophyll cell volumes are far less documented. Here, three-dimensional imaging was applied in a study of Arabidopsis thaliana leaf growth to determine leaf thickness and the cellular organization of mesophyll tissues under moderate soil water deficit and two cumulative light conditions. In contrast to surface area, thickness was highly conserved in response to water deficit under both low and high cumulative light regimes. Unlike epidermal and palisade mesophyll tissues, no reductions in cell number were observed in the spongy mesophyll; cells had rather changed in volume and shape. Furthermore, leaf features of a selection of genotypes affected in leaf functioning were analysed. The low-starch mutant pgm had very thick leaves because of unusually large palisade mesophyll cells, together with high levels of photosynthesis and stomatal conductance. By means of an open stomata mutant and a 9-cis-epoxycarotenoid dioxygenase overexpressor, it was shown that stomatal conductance does not necessarily have a major impact on leaf dimensions and cellular organization, pointing to additional mechanisms for the control of CO(2) diffusion under high and low stomatal conductance, respectively. © 2012 Blackwell Publishing Ltd.

  14. The site of water stress governs the pattern of ABA synthesis and transport in peanut

    PubMed Central

    Hu, Bo; Cao, Jiajia; Ge, Kui; Li, Ling

    2016-01-01

    Abscisic acid (ABA) is one of the most important phytohormones involved in stress responses in plants. However, knowledge of the effect on ABA distribution and transport of water stress at different sites on the plant is limited. In this study, water stress imposed on peanut leaves or roots by treatment with PEG 6000 is termed “leaf stress” or “root stress”, respectively. Immunoenzyme localization technolony was first used to detect ABA distribution in peanut. Under root stress, ABA biosynthesis and distribution level were all more pronounced in root than in leaf. However, ABA transport and the ability to induce stomatal closure were still better in leaf than in root during root stress; However, ABA biosynthesis initially increased in leaf, then rapidly accumulated in the vascular cambium of leaves and induced stomatal closure under leaf stress; ABA produced in root tissues was also transported to leaf tissues to maintain stomatal closure. The vascular system was involved in the coordination and integration of this complex regulatory mechanism for ABA signal accumulation. Water stress subject to root or leaf results in different of ABA biosynthesis and transport ability that trigger stoma close in peanut. PMID:27694957

  15. Induction of direct and indirect plant responses by jasmonic acid, low spider mite densities, or a combination of jasmonic acid treatment and spider mite infestation.

    PubMed

    Gols, Rieta; Roosjen, Mara; Dijkman, Herman; Dicke, Marcel

    2003-12-01

    Jasmonic acid (JA) and the octadecanoid pathway are involved in both induced direct and induced indirect plant responses. In this study, the herbivorous mite, Tetranychus urticae, and its predator, Phytoseiulus persimilis, were given a choice between Lima bean plants induced by JA or spider mites and uninduced control plants. Infestation densities resulting in the induction of predator attractants were much lower than thus far assumed, i.e., predatory mites were significantly attracted to plants that were infested for 2 days with only one or four spider mites per plant. Phytoseiulus persimilis showed a density-dependent response to volatiles from plants that were infested with different numbers of spider mites. Similarly, treating plants with increasing concentrations of JA also led to increased attraction of P. persimilis. Moreover, the duration of spider mite infestation was positively correlated with the proportion of predators that were attracted to mite-infested plants. A pretreatment of the plants with JA followed by a spider mite infestation enhanced the attraction of P. persimilis to plant volatiles compared to attraction to volatiles from plants that were only infested with spider mites and did not receive a pretreatment with JA. The herbivore, T. urticae preferred leaf tissue that previously had been infested with conspecifics to uninfested leaf tissue. In the case of choice tests with JA-induced and control leaf tissue, spider mites slightly preferred control leaf tissue. When spider mites were given a choice between leaf discs induced by JA and leaf discs damaged by spider mite feeding, they preferred the latter. The presence of herbivore induced chemicals and/or spider mite products enhanced settlement of the mites, whereas treatment with JA seemed to impede settlement.

  16. Visualisation by high resolution synchrotron X-ray phase contrast micro-tomography of gas films on submerged superhydrophobic leaves.

    PubMed

    Lauridsen, Torsten; Glavina, Kyriaki; Colmer, Timothy David; Winkel, Anders; Irvine, Sarah; Lefmann, Kim; Feidenhans'l, Robert; Pedersen, Ole

    2014-10-01

    Floods can completely submerge terrestrial plants but some wetland species can sustain O2 and CO2 exchange with the environment via gas films forming on superhydrophobic leaf surfaces. We used high resolution synchrotron X-ray phase contrast micro-tomography in a novel approach to visualise gas films on submerged leaves of common cordgrass (Spartina anglica). 3D tomograms enabled a hitherto unmatched level of detail regarding the micro-topography of leaf gas films. Gas films formed only on the superhydrophobic adaxial leaf side (water droplet contact angle, Φ=162°) but not on the abaxial side (Φ=135°). The adaxial side of the leaves of common cordgrass is plicate with a longitudinal system of parallel grooves and ridges and the vast majority of the gas film volume was found in large ∼180μm deep elongated triangular volumes in the grooves and these volumes were connected to each neighbouring groove via a fine network of gas tubules (∼1.7μm diameter) across the ridges. In addition to the gas film retained on the leaf exterior, the X-ray phase contrast micro-tomography also successfully distinguished gas spaces internally in the leaf tissues, and the tissue porosity (gas volume per unit tissue volume) ranged from 6.3% to 20.3% in tip and base leaf segments, respectively. We conclude that X-ray phase contrast micro-tomography is a powerful tool to obtain quantitative data of exterior gas features on biological samples because of the significant difference in electron density between air, biological tissues and water. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Invasion of Solanum tuberosum L. by Aspergillus terreus: a microscopic and proteomics insight on pathogenicity.

    PubMed

    Louis, Bengyella; Waikhom, Sayanika Devi; Roy, Pranab; Bhardwaj, Pardeep Kumar; Singh, Mohendro Wakambam; Chandradev, Sharma K; Talukdar, Narayan Chandra

    2014-06-10

    Aspergillus terreus is one of the most harmful filamentous fungal pathogen of humans, animals and plants. Recently, researchers have discovered that A. terreus can cause foliar blight disease in potato (Solanum tuberosum L.). We used light and scanning electron microscopy, and performed proteomics analysis in an attempt to dissect the invasion process of A. terreus in this important crop. Microscopic study revealed that invasion of leaf tissue is marked by rapid germination of A. terreus phialidic conidia (PC) by 4 h after inoculation. By 8 h after inoculation, primary germ tubes from PC differentiated into irregular protuberance, often displayed stomata atropism, and failed to penetrate via the epidermal cells. Colonization of leaf tissues was associated with high rate of production of accessory conidia (AC). These analyses showed the occurrence of a unique opposing pattern of AC, tissue-specific and produced on melanized colonizing hyphae during the infection of leaf tissue. A significant proteome change hallmarked by differential expression of class I patatin, lipoxygenase, catalase-peroxidase complex, and cysteine proteinase inhibitor were observed during tuber colonization. These proteins are often involved in signal transduction pathways and crosstalk in pathogenic responses. A. terreus abundantly produced AC and multipolar germinating PC to invade potato leaf tissue. Additionally, A. terreus differentially induced enzymes in potato tuber during colonization which facilitates rapid disease development.

  18. Invasion of Solanum tuberosum L. by Aspergillus terreus: a microscopic and proteomics insight on pathogenicity

    PubMed Central

    2014-01-01

    Background Aspergillus terreus is one of the most harmful filamentous fungal pathogen of humans, animals and plants. Recently, researchers have discovered that A. terreus can cause foliar blight disease in potato (Solanum tuberosum L.). We used light and scanning electron microscopy, and performed proteomics analysis in an attempt to dissect the invasion process of A. terreus in this important crop. Results Microscopic study revealed that invasion of leaf tissue is marked by rapid germination of A. terreus phialidic conidia (PC) by 4 h after inoculation. By 8 h after inoculation, primary germ tubes from PC differentiated into irregular protuberance, often displayed stomata atropism, and failed to penetrate via the epidermal cells. Colonization of leaf tissues was associated with high rate of production of accessory conidia (AC). These analyses showed the occurrence of a unique opposing pattern of AC, tissue-specific and produced on melanized colonizing hyphae during the infection of leaf tissue. A significant proteome change hallmarked by differential expression of class I patatin, lipoxygenase, catalase-peroxidase complex, and cysteine proteinase inhibitor were observed during tuber colonization. These proteins are often involved in signal transduction pathways and crosstalk in pathogenic responses. Conclusion A. terreus abundantly produced AC and multipolar germinating PC to invade potato leaf tissue. Additionally, A. terreus differentially induced enzymes in potato tuber during colonization which facilitates rapid disease development. PMID:24917207

  19. Comparison of arabidopsis stomatal density mutants indicates variation in water stress responses and potential epistatic effects

    Treesearch

    Shaneka S. Lawson; Paula M. Pijut; Charles H. Michler

    2014-01-01

    Recent physiological analysis of Arabidopsis stomatal density (SD) mutants indicated that SD was not the major factor controlling aboveground biomass accumulation. Despite the general theory that plants with fewer stomata have limited biomass acquisition capabilities, epf1 and several other Arabidopsis mutants varied significantly in leaf fresh...

  20. Effect of spinach cultivar and strain variation on survival of Escherichia coli O157:H7 on spinach leaves

    USDA-ARS?s Scientific Manuscript database

    Introduction: Escherichia coli O157:H7 outbreaks of infections associated with the consumption of fresh produce have increased in recent years. Bacterial cell surface appendages such as curli and the spinach leaf structure topography influence pathogen attachment and subsequent survival on spinach ...

  1. Both phenolic and non-phenolic green tea fractions inhibit migration of cancer cells

    USDA-ARS?s Scientific Manuscript database

    Green tea consumption is associated with chemoprevention of many cancer types. Fresh tea leaves are rich in polyphenolic catechins, which can constitute up to 30% of the dry leaf weight. While the polyphenols of green tea have been well investigated, it is still largely unknown, whether or not non-p...

  2. Chemical composition and in vitro antioxidant activity of hydro-ethanolic extracts from Bauhinia forficata subsp. pruinosa and B. variegata.

    PubMed

    Sayago, Carla T M; Camargo, Vanessa B; Barbosa, F; Gularte, Cláudia; Pereira, Geovana; Miotto, Silvia; Cechinel Filho, V; Luiz Puntel, R; Folmer, V; Mendez, A

    2013-03-01

    Bauhinia species are known to have hypoglycemiant and antioxidant activities. Here, hydro-ethanolic leaf extracts from Bauhinia forficata subsp. pruinosa and Bauhinia variegata, collected in a Pampa biome region of Brazil, were investigated to characterize their chromatographic profile, flavonoid content and in vitro antioxidant activity (TBARS and DPH assays). The extracts were obtained from dried and fresh leaves. The total flavonoid content was assessed by spectrophotometric determination, and the results ranged between 572.08 and 1,102.99 μg mL-1. Moreover, flavonoids were more predominant in B. variegata than in B. forficata subsp. pruinosa. HPLC analysis detected a complex profile of phenolic compounds, being the flavonoid kaempferitrin founded B. forficata subsp. pruinosa; in addition, other kaempferol and quercetin derivatives were present. In vitro antioxidant assays demonstrated a different behavior depending on the species, leaf treatment and extract concentration. In general, B. variegata extracts obtained from fresh material presented higher antioxidant potential, which can be attributed to the predominance of flavonoids in their chemical composition.

  3. How Does Alkali Aid Protein Extraction in Green Tea Leaf Residue: A Basis for Integrated Biorefinery of Leaves.

    PubMed

    Zhang, Chen; Sanders, Johan P M; Xiao, Ting T; Bruins, Marieke E

    2015-01-01

    Leaf protein can be obtained cost-efficiently by alkaline extraction, but overuse of chemicals and low quality of (denatured) protein limits its application. The research objective was to investigate how alkali aids protein extraction of green tea leaf residue, and use these results for further improvements in alkaline protein biorefinery. Protein extraction yield was studied for correlation to morphology of leaf tissue structure, protein solubility and hydrolysis degree, and yields of non-protein components obtained at various conditions. Alkaline protein extraction was not facilitated by increased solubility or hydrolysis of protein, but positively correlated to leaf tissue disruption. HG pectin, RGII pectin, and organic acids were extracted before protein extraction, which was followed by the extraction of cellulose and hemi-cellulose. RGI pectin and lignin were both linear to protein yield. The yields of these two components were 80% and 25% respectively when 95% protein was extracted, which indicated that RGI pectin is more likely to be the key limitation to leaf protein extraction. An integrated biorefinery was designed based on these results.

  4. The hydrostatic gradient, not light availability, drives height-related variation in Sequoia sempervirens (Cupressaceae) leaf anatomy.

    PubMed

    Oldham, Alana R; Sillett, Stephen C; Tomescu, Alexandru M F; Koch, George W

    2010-07-01

    Leaves at the tops of most trees are smaller, thicker, and in many other ways different from leaves on the lowermost branches. This height-related variation in leaf structure has been explained as acclimation to differing light environments and, alternatively, as a consequence of hydrostatic, gravitational constraints on turgor pressure that reduce leaf expansion. • To separate hydrostatic effects from those of light availability, we used anatomical analysis of height-paired samples from the inner and outer tree crowns of tall redwoods (Sequoia sempervirens). • Height above the ground correlates much more strongly with leaf anatomy than does light availability. Leaf length, width, and mesophyll porosity all decrease linearly with height and help explain increases in leaf-mass-to-area ratio and decreases in both photosynthetic capacity and internal gas-phase conductance with increasing height. Two functional traits-leaf thickness and transfusion tissue-also increase with height and may improve water-stress tolerance. Transfusion tissue area increases enough that whole-leaf vascular volume does not change significantly with height in most trees. Transfusion tracheids become deformed with height, suggesting they may collapse under water stress and act as a hydraulic buffer that improves leaf water status and reduces the likelihood of xylem dysfunction. • That such variation in leaf structure may be caused more by gravity than by light calls into question use of the terms "sun" and "shade" to describe leaves at the tops and bottoms of tall tree crowns.

  5. Effect of ethanol treatment on physiological and quality attributes of fresh-cut eggplant.

    PubMed

    Hu, Wenzhong; Jiang, Aili; Tian, Mixia; Liu, Chenghui; Wang, Yanying

    2010-06-01

    Fresh-cut eggplants, as other vegetables, have relatively short shelf life because of the large amount of tissue disruption and increased metabolism. There is a very rapid onset of enzymatic browning and tissue softening with consequent decrease in sensorial and nutritional quality. To reduce respiration and maintain the quality, various treatments have been applied to find the optimum conditions that provide more fresh and natural fresh-cut produce after minimal processing. The objective of this study was to investigate the effects of ethanol vapour treatment on physiological and quality attributes of fresh-cut eggplant during the extension of shelf life. The fresh-cut eggplant treated with ethanol vapour showed that respiration rate and occurrence of enzymatic browning were reduced, and higher total phenol content was maintained during 8 days of storage at 10 degrees C. The polyphenol oxidase and peroxidase in fresh-cut eggplant were also inhibited significantly by ethanol treatment. The ethanol treatment reduced the weight loss and maintained the integrity of cell membranes, as confirmed by the low value of electrolyte leakage. The ethanol treatment applied for fresh-cut eggplant was a practical approach to reduce the activity of physiological metabolism and maintain the fresh quality of fresh-cut eggplant. The experimental results revealed that ethanol treatment was effective for extending the shelf life of fresh-cut eggplant as a cheap, environmentally acceptable method. Copyright (c) 2010 Society of Chemical Industry.

  6. Effects of elevated atmospheric CO2 concentrations, clipping regimen and differential day/night atmospheric warming on tissue nitrogen concentrations of a perennial pasture grass

    PubMed Central

    Volder, Astrid; Gifford, Roger M.; Evans, John R.

    2015-01-01

    Forecasting the effects of climate change on nitrogen (N) cycling in pastures requires an understanding of changes in tissue N. We examined the effects of elevated atmospheric CO2 concentration, atmospheric warming and simulated grazing (clipping frequency) on aboveground and belowground tissue N concentrations and C : N ratios of a C3 pasture grass. Phalaris aquatica L. cv. ‘Holdfast’ was grown in the field in six transparent temperature gradient tunnels (18 × 1.5 × 1.5 m each), three at ambient atmospheric CO2 and three at 759 p.p.m. CO2. Within each tunnel, there were three air temperature treatments: ambient control, +2.2/+4.0 °C above ambient day/night warming and +3.0 °C continuous warming. A frequent and an infrequent clipping treatment were applied to each warming × CO2 combination. Green leaf N concentrations were decreased by elevated CO2 and increased by more frequent clipping. Both warming treatments increased leaf N concentrations under ambient CO2 concentrations, but did not significantly alter leaf N concentrations under elevated CO2 concentrations. Nitrogen resorption from leaves was decreased under elevated CO2 conditions as well as by more frequent clipping. Fine root N concentrations decreased strongly with increasing soil depth and were further decreased at the 10–60 cm soil depths by elevated CO2 concentrations. The interaction between the CO2 and warming treatments showed that leaf N concentration was affected in a non-additive manner. Changes in leaf C : N ratios were driven by changes in N concentration. Overall, the effects of CO2, warming and clipping treatments on aboveground tissue N concentrations were much greater than on belowground tissue. PMID:26272874

  7. Long-Term Inhibition by Auxin of Leaf Blade Expansion in Bean and Arabidopsis1

    PubMed Central

    Keller, Christopher P.; Stahlberg, Rainer; Barkawi, Lana S.; Cohen, Jerry D.

    2004-01-01

    The role of auxin in controlling leaf expansion remains unclear. Experimental increases to normal auxin levels in expanding leaves have shown conflicting results, with both increases and decreases in leaf growth having been measured. Therefore, the effects of both auxin application and adjustment of endogenous leaf auxin levels on midrib elongation and final leaf size (fresh weight and area) were examined in attached primary monofoliate leaves of the common bean (Phaseolus vulgaris) and in early Arabidopsis rosette leaves. Aqueous auxin application inhibited long-term leaf blade elongation. Bean leaves, initially 40 to 50 mm in length, treated once with α-naphthalene acetic acid (1.0 mm), were, after 6 d, approximately 80% the length and weight of controls. When applied at 1.0 and 0.1 mm, α-naphthalene acetic acid significantly inhibited long-term leaf growth. The weak auxin, β-naphthalene acetic acid, was effective at 1.0 mm; and a weak acid control, benzoic acid, was ineffective. Indole-3-acetic acid (1 μm, 10 μm, 0.1 mm, and 1 mm) required daily application to be effective at any concentration. Application of the auxin transport inhibitor, 1-N-naphthylphthalamic acid (1% [w/w] in lanolin), to petioles also inhibited long-term leaf growth. This treatment also was found to lead to a sustained elevation of leaf free indole-3-acetic acid content relative to untreated control leaves. Auxin-induced inhibition of leaf growth appeared not to be mediated by auxin-induced ethylene synthesis because growth inhibition was not rescued by inhibition of ethylene synthesis. Also, petiole treatment of Arabidopsis with 1-N-naphthylphthalamic acid similarly inhibited leaf growth of both wild-type plants and ethylene-insensitive ein4 mutants. PMID:14988474

  8. Hyperforin accumulates in the translucent glands of Hypericum perforatum.

    PubMed

    Soelberg, Jens; Jørgensen, Lise Bolt; Jäger, Anna K

    2007-06-01

    Hypericum perforatum contains the therapeutically important compounds hypericin and hyperforin. Hypericin is known to accumulate in the dark glands. This investigation aimed to determine the accumulation site of hyperforin. Dark and translucent glands as well as non-secretory tissue in leaves were manually isolated under the microscope. Hyperforin content was quantified by UV HPLC. Secretory structures were surveyed anatomically. The hyperforin content of intact leaves was found to be about 3 mg g(-1) fresh tissue, whereas a content of about 7 mg g(-1) fresh material was found in isolated translucent glands. Hyperforin was found only to occur in minute amounts in dark glands (approx. 0.4 mg g(-1) fresh tissue). In non-secretory tissue no hyperforin was detected. The accumulation of hyperforin detected in the translucent glands supports the proposed hypothesis that hyperforin is synthesized by the same biosynthetic machinery as monoterpenes in the chloroplasts of cells delimiting the gland.

  9. Identification of Stevioside Using Tissue Culture-Derived Stevia (Stevia rebaudiana) Leaves

    PubMed Central

    Karim, Md. Ziaul; Uesugi, Daisuke; Nakayama, Noriyuki; Hossain, M. Monzur; Ishihara, Kohji; Hamada, Hiroki

    2015-01-01

    Stevioside is a natural sweetener from Stevia leaf, which is 300 times sweeter than sugar. It helps to reduce blood sugar levels dramatically and thus can be of benefit to diabetic people. Tissue culture is a very potential modern technology that can be used in large-scale disease-free stevia production throughout the year. We successfully produced stevia plant through in vitro culture for identification of stevioside in this experiment. The present study describes a potential method for identification of stevioside from tissue culture-derived stevia leaf. Stevioside in the sample was identified using HPLC by measuring the retention time. The percentage of stevioside content in the leaf samples was found to be 9.6%. This identification method can be used for commercial production and industrialization of stevia through in vitro culture across the world. PMID:28008268

  10. Wasabi leaf extracts attenuate adipocyte hypertrophy through PPARγ and AMPK.

    PubMed

    Oowatari, Yasuo; Ogawa, Tetsuro; Katsube, Takuya; Iinuma, Kiyohisa; Yoshitomi, Hisae; Gao, Ming

    2016-08-01

    Hypertrophy of adipocytes in obese adipose tissues causes metabolic abnormality by adipocytokine dysregulation, which promotes type 2 diabetes mellitus, hypertension, and dyslipidemia. We investigated the effects of wasabi (Wasabia japonica Matsum) leaf extracts on metabolic abnormalities in SHRSP.Z-Leprfa/IzmDmcr rats (SHRSP/ZF), which are a model of metabolic syndrome. Male SHRSP/ZF rats aged 7 weeks were divided into two groups: control and wasabi leaf extract (WLE) groups, which received water or oral treatment with 4 g/kg/day WLE for 6 weeks. WLE improved the body weight gain and high blood pressure in SHRSP/ZF rats, and the plasma triglyceride levels were significantly lower in the WLE group. Adipocyte hypertrophy was markedly prevented in adipose tissue. The expression of PPARγ and subsequent downstream genes was suppressed in the WLE group adipose tissues. Our data suggest that WLE inhibits adipose hypertrophy by suppressing PPARγ expression in adipose tissue and stimulating the AMPK activity by increased adiponectin.

  11. Reduction of Cr(VI) to Cr(III) by wetland plants: Potential for in situ heavy metal detoxification

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lytle, C.M.; Qian, J.H.; Hansen, D.

    1998-10-15

    Reduction of heavy metals in situ by plants may be a useful detoxification mechanism for phytoremediation. Using X-ray spectroscopy, the authors show that Eichhornia crassipes (water hyacinth), supplied with Cr(VI) in nutrient culture, accumulated nontoxic Cr(III) in root and shoot tissues. The reduction of Cr(VI) to Cr(III) appeared to occur in the fine lateral roots. The Cr(III) was subsequently translocated to leaf tissues. Extended X-ray absorption fine structure of Cr in leaf and petiole differed when compared to Cr in roots. In roots, Cr(III) was hydrated by water, but in petiole and more so in leaf, a portion of themore » Cr(III) may be bound to oxalate ligands. This suggests that E. crassipes detoxified Cr(VI) upon root uptake and transported a portion of the detoxified Cr to leaf tissues. Cr-rich crystalline structures were observed on the leaf surface. The chemical species of Cr in other plants, collected from wetlands that contained Cr(VI)-contaminated wastewater, was also found to be Cr(III). The authors propose that this plant-based reduction of Cr(VI) by E. crassipes has the potential to be used for the in situ detoxification of Cr(VI)-contaminated wastestreams.« less

  12. Photosynthesis and canopy characteristics in genetically defined families of silver birch (Betula pendula).

    PubMed

    Wang, T; Tigerstedt, P M; Viherä-Aarnio, A

    1995-10-01

    Net photosynthetic rates (A) of leaves in upper and lower crown layers (A(upper) and A(lower)), leaf area index (LAI), mean tilt angle (MTA), several leaf characteristics, and volume growth were observed in fast- and slow-growing families of a 14-year-old full-sib and half-sib family progeny test of Betula pendula Roth. Each measure of net photosynthetic rate was calculated after correcting measured net photosynthesis for the effects of environmental variables. The differences in A(upper) and LAI among families were significant. The proportions of the total variance assigned to family for A(upper), A(lower) and LAI were 33.64, 28.93 and 54.99%, respectively. The mean A(upper) and LAI of the fast-growing families were significantly higher than those of the slow-growing families, whereas the mean A(lower) of the fast-growing families was significantly lower than that of the slow-growing families. There were also significant differences among families in leaf size, leaf shape, and the ratios leaf fresh weight/area and leaf dry weight/area. Between 27.55 and 54.55% of the total variance in these characteristics could be assigned to the family effect. Volume growth was positively correlated with A(upper) and LAI, but it was most strongly correlated with A(upper) x LAI.

  13. Changes in vegetation spectra with leaf deterioration under two methods of preservation

    NASA Technical Reports Server (NTRS)

    Labovitz, M. L.; Masuoka, E. J.; Feldmann, S. G.

    1984-01-01

    Changes in leaf spectra caused by mineralization under different conditions of preservation are measured using a three-band portable radiometer which simulates three Thematic Mapper bands 3, 4, and 5. Daily spectral measurements of white oak (Quercus alba) leaves did not distinguish among the fresh, bottled, and bagged vegetation in the spectral bands 3 and 5 for up to four days after collection. The reflected energy of the preserved vegetation increased thereafter, reportedly due to the loss of chlorophyll and dehydration. It is concluded that the measurement procedure is sufficiently sensitive as to discern documented patterns of variation in reflectance measurements.

  14. Step changes in leaf oil accumulation via iterative metabolic engineering.

    PubMed

    Vanhercke, Thomas; Divi, Uday K; El Tahchy, Anna; Liu, Qing; Mitchell, Madeline; Taylor, Matthew C; Eastmond, Peter J; Bryant, Fiona; Mechanicos, Anna; Blundell, Cheryl; Zhi, Yao; Belide, Srinivas; Shrestha, Pushkar; Zhou, Xue-Rong; Ral, Jean-Philippe; White, Rosemary G; Green, Allan; Singh, Surinder P; Petrie, James R

    2017-01-01

    Synthesis and accumulation of plant oils in the entire vegetative biomass offers the potential to deliver yields surpassing those of oilseed crops. However, current levels still fall well short of those typically found in oilseeds. Here we show how transcriptome and biochemical analyses pointed to a futile cycle in a previously established Nicotiana tabacum line, accumulating up to 15% (dry weight) of the storage lipid triacylglycerol in leaf tissue. To overcome this metabolic bottleneck, we either silenced the SDP1 lipase or overexpressed the Arabidopsis thaliana LEC2 transcription factor in this transgenic background. Both strategies independently resulted in the accumulation of 30-33% triacylglycerol in leaf tissues. Our results demonstrate that the combined optimization of de novo fatty acid biosynthesis, storage lipid assembly and lipid turnover in leaf tissue results in a major overhaul of the plant central carbon allocation and lipid metabolism. The resulting further step changes in oil accumulation in the entire plant biomass offers the possibility of delivering yields that outperform current oilseed crops. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  15. Survival, dispersal, and potential soil-mediated suppression of Phytophthora ramorum in a California redwood-tanoak forest.

    PubMed

    Fichtner, E J; Lynch, S C; Rizzo, D M

    2009-05-01

    Because the role of soil inoculum of Phytophthora ramorum in the sudden oak death disease cycle is not well understood, this work addresses survival, chlamydospore production, pathogen suppression, and splash dispersal of the pathogen in infested forest soils. Colonized rhododendron and bay laurel leaf disks were placed in mesh sachets before transfer to the field in January 2005 and 2006. Sachets were placed under tanoak, bay laurel, and redwood at three vertical locations: leaf litter surface, litter-soil interface, and below the soil surface. Sachets were retrieved after 4, 8, 20, and 49 weeks. Pathogen survival was higher in rhododendron leaf tissue than in bay tissue, with >80% survival observed in rhododendron tissue after 49 weeks in the field. Chlamydospore production was determined by clearing infected tissue in KOH. Moist redwood-associated soils suppressed chlamydospore production. Rain events splashed inoculum as high as 30 cm from the soil surface, inciting aerial infection of bay laurel and tanoak. Leaf litter may provide an incomplete barrier to splash dispersal. This 2-year study illustrates annual P. ramorum survival in soil and the suppressive nature of redwood-associated soils to chlamydospore production. Infested soil may serve as primary inoculum for foliar infections by splash dispersal during rain events.

  16. Leaf area and photosynthesis of newly emerged trifoliolate leaves are regulated by mature leaves in soybean.

    PubMed

    Wu, Yushan; Gong, Wanzhuo; Wang, Yangmei; Yong, Taiwen; Yang, Feng; Liu, Weigui; Wu, Xiaoling; Du, Junbo; Shu, Kai; Liu, Jiang; Liu, Chunyan; Yang, Wenyu

    2018-03-29

    Leaf anatomy and the stomatal development of developing leaves of plants have been shown to be regulated by the same light environment as that of mature leaves, but no report has yet been written on whether such a long-distance signal from mature leaves regulates the total leaf area of newly emerged leaves. To explore this question, we created an investigation in which we collected data on the leaf area, leaf mass per area (LMA), leaf anatomy, cell size, cell number, gas exchange and soluble sugar content of leaves from three soybean varieties grown under full sunlight (NS), shaded mature leaves (MS) or whole plants grown in shade (WS). Our results show that MS or WS cause a marked decline both in leaf area and LMA in newly developing leaves. Leaf anatomy also showed characteristics of shade leaves with decreased leaf thickness, palisade tissue thickness, sponge tissue thickness, cell size and cell numbers. In addition, in the MS and WS treatments, newly developed leaves exhibited lower net photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (E), but higher carbon dioxide (CO 2 ) concentration in the intercellular space (Ci) than plants grown in full sunlight. Moreover, soluble sugar content was significantly decreased in newly developed leaves in MS and WS treatments. These results clearly indicate that (1) leaf area, leaf anatomical structure, and photosynthetic function of newly developing leaves are regulated by a systemic irradiance signal from mature leaves; (2) decreased cell size and cell number are the major cause of smaller and thinner leaves in shade; and (3) sugars could possibly act as candidate signal substances to regulate leaf area systemically.

  17. Red (anthocyanic) leaf margins do not correspond to increased phenolic content in New Zealand Veronica spp.

    PubMed Central

    Hughes, Nicole M.; Smith, William K.; Gould, Kevin S.

    2010-01-01

    Background and Aims Red or purple coloration of leaf margins is common in angiosperms, and is found in approx. 25 % of New Zealand Veronica species. However, the functional significance of margin coloration is unknown. We hypothesized that anthocyanins in leaf margins correspond with increased phenolic content in leaf margins and/or the leaf entire, signalling low palatability or leaf quality to edge-feeding insects. Methods Five species of Veronica with red leaf margins, and six species without, were examined in a common garden. Phenolic content in leaf margins and interior lamina regions of juvenile and fully expanded leaves was quantified using the Folin–Ciocalteu assay. Proportions of leaf margins eaten and average lengths of continuous bites were used as a proxy for palatability. Key Results Phenolic content was consistently higher in leaf margins compared with leaf interiors in all species; however, neither leaf margins nor more interior tissues differed significantly in phenolic content with respects to margin colour. Mean phenolic content was inversely correlated with the mean length of continuous bites, suggesting effective deterrence of grazing. However, there was no difference in herbivore consumption of red and green margins, and the plant species with the longest continuous grazing patterns were both red-margined. Conclusions Red margin coloration was not an accurate indicator of total phenolic content in leaf margins or interior lamina tissue in New Zealand Veronica. Red coloration was also ineffective in deterring herbivory on the leaf margin, though studies controlling for variations in leaf structure and biochemistry (e.g. intra-specific studies) are needed before more precise conclusions can be drawn. It is also recommended that future studies focus on the relationship between anthocyanin and specific defence compounds (rather than general phenolic pools), and evaluate possible alternative functions of red margins in leaves (e.g. antioxidants, osmotic adjustment). PMID:20145003

  18. Intraspecific Relationships among Wood Density, Leaf Structural Traits and Environment in Four Co-Occurring Species of Nothofagus in New Zealand

    PubMed Central

    Richardson, Sarah J.; Allen, Robert B.; Buxton, Rowan P.; Easdale, Tomás A.; Hurst, Jennifer M.; Morse, Christopher W.; Smissen, Rob D.; Peltzer, Duane A.

    2013-01-01

    Plant functional traits capture important variation in plant strategy and function. Recent literature has revealed that within-species variation in traits is greater than previously supposed. However, we still have a poor understanding of how intraspecific variation is coordinated among different traits, and how it is driven by environment. We quantified intraspecific variation in wood density and five leaf traits underpinning the leaf economics spectrum (leaf dry matter content, leaf mass per unit area, size, thickness and density) within and among four widespread Nothofagus tree species in southern New Zealand. We tested whether intraspecific relationships between wood density and leaf traits followed widely reported interspecific relationships, and whether variation in these traits was coordinated through shared responses to environmental factors. Sample sites varied widely in environmental variables, including soil fertility (25–900 mg kg–1 total P), precipitation (668–4875 mm yr–1), temperature (5.2–12.4 °C mean annual temperature) and latitude (41–46 °S). Leaf traits were strongly correlated with one another within species, but not with wood density. There was some evidence for a positive relationship between wood density and leaf tissue density and dry matter content, but no evidence that leaf mass or leaf size were correlated with wood density; this highlights that leaf mass per unit area cannot be used as a surrogate for component leaf traits such as tissue density. Trait variation was predicted by environmental factors, but not consistently among different traits; e.g., only leaf thickness and leaf density responded to the same environmental cues as wood density. We conclude that although intraspecific variation in wood density and leaf traits is strongly driven by environmental factors, these responses are not strongly coordinated among functional traits even across co-occurring, closely-related plant species. PMID:23527041

  19. Field performance of Populus expressing somaclonal variation in resistance to Septoria musiva

    Treesearch

    M. E. Ostry; K. T. Ward

    2003-01-01

    Over 1500 trees from two hybrid poplar clones regenerated from tissue culture and expressing somatic variation in leaf disease resistance in a laboratory leaf disk bioassay were field-tested for 5-11 years to examine their resistance to Septoria leaf spot and canker and to assess their growth characteristics compared with the source clones....

  20. A Technique to Perfuse Cadavers that Extends the Useful Life of Fresh Tissues: The Duke Experience

    ERIC Educational Resources Information Center

    Messmer, Caroline; Kellogg, Ryan T.; Zhang, Yixin; Baiak, Andresa; Leiweke, Clinton; Marcus, Jeffrey R.; Levin, L. Scott; Zenn, Michael R.; Erdmann, Detlev

    2010-01-01

    The demand for laboratory-based teaching and training is increasing worldwide as medical training and education confront the pressures of shorter training time and rising costs. This article presents a cost-effective perfusion technique that extends the useful life of fresh tissue. Refrigerated cadavers are preserved in their natural state for up…

  1. Chemical Composition and Activity of Essential Oils of Carissa macrocarpa (Eckl.) A.DC. Cultivated in Tunisia and its Anatomical Features.

    PubMed

    Souilem, Fedia; El Ayeb, Asma; Djlassi, Brahim; Ayari, Olfa; Chiboub, Wiem; Arbi, Faten; Ascrizzi, Roberta; Flamini, Guido; Harzallah-Skhiri, Fethia

    2018-06-15

    This is the first study investigating the chemical composition of essential oils (EOs) isolated from different tissues of Carissa macrocarpa (Eckl.)A.DC, their antimicrobial activity and the anatomical characters of the aerial organs and the fruits. The main EO components were pentadecanal and tetradecan-1-ol (31.9 and 16.5% in fresh leaf EO, resp.), (E)-nerolidol and caryophyllene oxide (27.3 and 15.0% in fruit EO, resp.), linalool and hexahydrofarnesyl acetone (30.9 and 24.9% in stem EO, resp.), benzyl benzoate (24.3% in flower EO). The fruit EO was more active against Candida albicans (MIC=0.46 mg/mL) compared to the reference antibiotic (17.66 mg/mL). Furthermore, at this concentration it inhibited all the Gram-positive bacteria. Concerning the anatomical features, it is noteworth the presence of a large cluster of Ca oxalate crystals inside some parenchymatous cells. Large ducts corresponding to non articulated laticifers were identified in the cortex of leaf, stem and fruit pericarp. The laticifers categories and their distribution are taxonomically important to discriminate this species from others acclimated in different countries. Considering the obtained results, EOs of C. macrocarpa can be a good source of antimicrobial compounds, contributing to solve the problem of microbial resistance to antibiotics. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  2. Leaf conductance and carbon gain under salt-stressed conditions

    NASA Astrophysics Data System (ADS)

    Volpe, V.; Manzoni, S.; Marani, M.; Katul, G.

    2011-12-01

    Exposure of plants to salt stress is often accompanied by reductions in leaf photosynthesis and in stomatal and mesophyll conductances. To separate the effects of salt stress on these quantities, a model based on the hypothesis that carbon gain is maximized subject to a water loss cost is proposed. The optimization problem of adjusting stomatal aperture for maximizing carbon gain at a given water loss is solved for both a non-linear and a linear biochemical demand function. A key novel theoretical outcome of the optimality hypothesis is an explicit relationship between the stomatal and mesophyll conductances that can be evaluated against published measurements. The approaches here successfully describe gas-exchange measurements reported for olive trees (Olea europea L.) and spinach (Spinacia oleraceaL.) in fresh water and in salt-stressed conditions. Salt stress affected both stomatal and mesophyll conductances and photosynthetic efficiency of both species. The fresh water/salt water comparisons show that the photosynthetic capacity is directly reduced by 30%-40%, indicating that reductions in photosynthetic rates under increased salt stress are not due only to a limitation of CO2diffusion. An increase in salt stress causes an increase in the cost of water parameter (or marginal water use efficiency) exceeding 100%, analogous in magnitude to findings from extreme drought stress studies. The proposed leaf-level approach can be incorporated into physically based models of the soil-plant-atmosphere system to assess how saline conditions and elevated atmospheric CO2 jointly impact transpiration and photosynthesis.

  3. Effects of agriculture production systems on nitrate and nitrite accumulation on baby-leaf salads

    PubMed Central

    Aires, Alfredo; Carvalho, Rosa; Rosa, Eduardo A S; Saavedra, Maria J

    2013-01-01

    Nitrate and nitrite are widespread contaminants of vegetables, fruits, and waters. The levels of these compounds are increased as a result of using organic wastes from chemical industries, domestic wastes, effluents, nitrogenous fertilizers, and herbicides in agriculture. Therefore, determining the nitrate and nitrite levels in biological, food, and environmental samples is important to protect human health and the environment. In this context, we set this study, in which we report the effect of production system (conventional and organic) on the accumulation of nitrates and nitrites in fresh baby-leaf samples. The average levels of the nitrate () and nitrite () contents in six different baby-leaf salads of a single species (green lettuce, red lettuce, watercress, rucola, chard, and corn salad) produced in organic and conventional agriculture system were evaluated. Spectrophotometric analytical method recently published was validated and used. Nitrates and nitrites were detected in all samples. The nitrates levels from organic production varied between 1.45 and 6.40 mg/kg fresh weight (FW), whereas those from conventional production ranged from 10.5 to 45.19 mg/kg FW. The nitrites content was lower than nitrates and ranged from 0.32 to 1.89 mg/kg FW in organic production system and between 0.14 and 1.41 mg/kg FW in conventional production system. Our results showed that the nitrate content was dependent on the agricultural production system, while for nitrites, this dependency was less pronounced. PMID:24804008

  4. Olive Leaf Extract Elevates Hepatic PPAR α mRNA Expression and Improves Serum Lipid Profiles in Ovariectomized Rats.

    PubMed

    Yoon, Leena; Liu, Ya-Nan; Park, Hyunjin; Kim, Hyun-Sook

    2015-07-01

    We hypothesized that olive leaf extract might alleviate dyslipidemia resulting from estrogen deficiency. Serum lipid profile and mRNA expression of the related genes in the liver and adipose tissue were analyzed after providing olive leaf extract (200 or 400 mg/kg body weight; n=7 for each group) to ovariectomized rats for 10 weeks. After 10 weeks' administration, the rats in the olive leaf extract-administered groups showed significantly lower levels of serum triglyceride and very-low-density lipoprotein (VLDL)-cholesterol compared with the rats in the control group, whereas the administration of olive leaf extract did not significantly change the elevated low-density lipoprotein cholesterol levels. In addition, administration of high dose of olive leaf extract significantly decreased the liver triglyceride and increased serum estradiol levels. mRNA expressions of peroxisome proliferator-activated receptor alpha (PPAR α) and acyl-CoA oxidase (ACO) were not affected by ovariectomy, however, administration of olive leaf extract significantly increased both PPAR α and ACO mRNA expression. Expression of adiponectin mRNA in adipose tissue was significantly decreased in the ovariectomized control group. Rats administered low-dose olive leaf extract showed significantly elevated adiponectin mRNA expression compared with rats in the ovariectomized control group. Even though dose-dependent effects were not observed in most of the measurements, these results suggest that genes involved in lipid metabolism may be regulated by olive leaf extract administration in ovariectomized rats.

  5. Global Climatic Controls On Leaf Size

    NASA Astrophysics Data System (ADS)

    Wright, I. J.; Prentice, I. C.; Dong, N.; Maire, V.

    2015-12-01

    Since the 1890s it's been known that the wet tropics harbour plants with exceptionally large leaves. Yet the observed latitudinal gradient of leaf size has never been fully explained: it is still unclear which aspects of climate are most important for understanding geographic trends in leaf size, a trait that varies many thousand-fold among species. The key is the leaf-to-air temperature difference, which depends on the balance of energy inputs (irradiance) and outputs (transpirational cooling, losses to the night sky). Smaller leaves track air temperatures more closely than larger leaves. Widely cited optimality-based theories predict an advantage for smaller leaves in dry environments, where transpiration is restricted, but are silent on the latitudinal gradient. We aimed to characterize and explain the worldwide pattern of leaf size. Across 7900 species from 651 sites, here we show that: large-leaved species predominate in wet, hot, sunny environments; smaller-leaved species typify hot, sunny environments only when arid; small leaves are required to avoid freezing in high latitudes and at high elevation, and to avoid overheating in dry environments. This simple pattern was unclear in earlier, more limited analyses. We present a simple but robust, fresh approach to energy-balance modelling for both day-time and night-time leaf-to-air temperature differences, and thus risk of overheating and of frost damage. Our analysis shows night-chilling is important as well as day-heating, and simplifies leaf temperature modelling. It provides both a framework for modelling leaf size constraints, and a solution to one of the oldest conundrums in ecology. Although the path forward is not yet fully clear, because of its role in controlling leaf temperatures we suggest that climate-related leaf size constraints could usefully feature in the next generation of land ecosystem models.

  6. Effect of Urtica dioica Leaf Alcoholic and Aqueous Extracts on the Number and the Diameter of the Islets in Diabetic Rats.

    PubMed

    Qujeq, Durdi; Tatar, Mohsen; Feizi, Farideh; Parsian, Hadi; Sohan Faraji, Alieh; Halalkhor, Sohrab

    2013-01-01

    Urtica dioica has been known as a plant that decreases blood glucose. Despite the importance of this plant in herbal medicine, relatively little research has been down on effects of this plant on islets yet. The objective of the current study was to evaluate the effect of dried Urtica dioica leaf alcoholic and aqueous extracts on the number and the diameter of the islets and histological parameters in streptozocin-induced diabetic rats. Six rats were used in each group. Group I: Normal rats were administered saline daily for 8 weeks. Group II: Diabetic rats were administered streptozotocin, 50 mg/kg of body weight; Group III: Diabetic rats were administered dried Urtica dioica leaf aqueous extracts for 8 weeks; Group IV: Diabetic rats were administered dried Urtica dioica leaf alcoholic extracts for 8 weeks. The animals, groups of diabetic and normal, were sacrificed by ether anaesthesia. Whole pancreas was dissected. The tissue samples were formalin fixed and paraffin embedded for microscopic examination. Histologic examination and grading were carried out on hematoxylin-eosin stained sections. The effects of administration of dried Urtica dioica leaf alcoholic and aqueous extracts to diabetic rats were determined by histopathologic examination. The pancreas from control rats showed normal pancreatic islets histoarchitecture. Our results also, indicate that the pancreas from diabetic rats show injury of pancreas tissue while the pancreas from diabetic rats treated with dried Urtica dioica leaf alcoholic and aqueous extracts show slight to moderate rearrangement of islets. According to our findings, dried Urtica dioica leaf alcoholic and aqueous extracts can cause a suitable repair of pancreatic tissue in streptozocin-induced diabetic experimental model.

  7. Effect of Urtica dioica Leaf Alcoholic and Aqueous Extracts on the Number and the Diameter of the Islets in Diabetic Rats

    PubMed Central

    Qujeq, Durdi; Tatar, Mohsen; Feizi, Farideh; Parsian, Hadi; Sohan Faraji, Alieh; Halalkhor, Sohrab

    2013-01-01

    Urtica dioica has been known as a plant that decreases blood glucose. Despite the importance of this plant in herbal medicine, relatively little research has been down on effects of this plant on islets yet. The objective of the current study was to evaluate the effect of dried Urtica dioica leaf alcoholic and aqueous extracts on the number and the diameter of the islets and histological parameters in streptozocin-induced diabetic rats. Six rats were used in each group. Group I: Normal rats were administered saline daily for 8 weeks. Group II: Diabetic rats were administered streptozotocin, 50 mg/kg of body weight; Group III: Diabetic rats were administered dried Urtica dioica leaf aqueous extracts for 8 weeks; Group IV: Diabetic rats were administered dried Urtica dioica leaf alcoholic extracts for 8 weeks. The animals, groups of diabetic and normal, were sacrificed by ether anaesthesia. Whole pancreas was dissected. The tissue samples were formalin fixed and paraffin embedded for microscopic examination. Histologic examination and grading were carried out on hematoxylin-eosin stained sections. The effects of administration of dried Urtica dioica leaf alcoholic and aqueous extracts to diabetic rats were determined by histopathologic examination. The pancreas from control rats showed normal pancreatic islets histoarchitecture. Our results also, indicate that the pancreas from diabetic rats show injury of pancreas tissue while the pancreas from diabetic rats treated with dried Urtica dioica leaf alcoholic and aqueous extracts show slight to moderate rearrangement of islets. According to our findings, dried Urtica dioica leaf alcoholic and aqueous extracts can cause a suitable repair of pancreatic tissue in streptozocin-induced diabetic experimental model. PMID:24551786

  8. The Distribution of Catalase Activity, Isozyme Protein, and Transcript in the Tissues of the Developing Maize Seedling 1

    PubMed Central

    Redinbaugh, Margaret G.; Sabre, Mara; Scandalios, John G.

    1990-01-01

    The catalase activity, CAT-2 and CAT-3 isozyme protein levels, and the steady-state mRNA levels for each of the three catalase genes were determined in the scutellum, root, epicotyl, and leaf of the developing maize (Zea mays L.) seedling. Catalase activity was highest in the scutellum, with 10-fold lower enzyme activity in the leaf and epicotyl. Very low levels of catalase activity were found in the root. The highest levels of CAT-2 protein were found in the scutellum, with about 10-fold lower levels in the green leaf. CAT-2 protein was present in trace amounts early in root development and no CAT-2 protein was detected in the epicotyl. Shortly after germination, CAT-3 protein was present at high levels in both the epicotyl and green leaf. With development, the amount of CAT-3 protein decreased slowly in the epicotyl and rapidly in the green leaf. Low levels of this isozyme were detected in the scutellum and root. The Cat1 transcript accumulated to low levels in all four tissues during the 14 day developmental period. High levels of the Cat2 transcript were found in the scutellum, with moderate levels of the mRNA in the green leaf. The Cat2 transcript levels were very low in the root and epicotyl. While the Cat3 mRNA level in the scutellum was low, high levels of the Cat3 transcript were detected in the root, epicotyl, and leaf. There was a positive correlation between the accumulation of a catalase isozyme and its transcript, indicating that the tissue specificity of maize catalase gene expression was regulated pretranslationally. Images Figure 3 Figure 4 PMID:16667285

  9. The HPLC-Fluorescence Detection of Coumarins in ‘Hamlin’ Sweet Orange and ‘Marsh’ Grapefruit Leaf Cankers

    USDA-ARS?s Scientific Manuscript database

    Canker is a devastating disease for the citrus fresh fruit market and is caused by the pathogenic bacterium Xanthomonas citri var. citri (Xcc). Infection occurs by bacterial penetration through physical damage of leaves, peel and stems, and also by bacterial entry through the stomates of these photo...

  10. Decomposition and nutrient dynamics of hardwood leaf litter in the Fernow Whole-Watershed Acidification Experiment

    Treesearch

    M.B. Adams; T.R. Angradi

    1996-01-01

    Two watersheds are part of an on-going long-term artificial acidification experiment: the treatment watershed (WS3) has received 60.5 kg S ha-1 year-1 and 54 kg N ha-1 year-1 via aerial applications of ammonium sulfate fertilizer since 1989. After 3 years of treatment, freshly...

  11. Host resistance to Botrytis bunch rot in Vitis spp. and its correlation with Botrytis leaf spot

    USDA-ARS?s Scientific Manuscript database

    Botrytis cinerea, the causal agent of Botrytis bunch rot, is the number one postharvest disease of fresh grapes in the U.S. Fungicide applications are used to manage the disease, but resistant isolates are common and postharvest losses occur annually. Host resistance is needed for long-term manageme...

  12. Evaluation of host resistance to Botrytis bunch rot in Vitis spp. and its correlation with Botrytis leaf spot

    USDA-ARS?s Scientific Manuscript database

    Botrytis cinerea, the causal agent of Botrytis bunch rot and gray mold, is the number one postharvest disease of fresh grapes in the United States. Fungicide applications are used to manage the disease, but fungicide-resistant isolates are common and postharvest losses occur annually. Host resistanc...

  13. Topical Olive Leaf Extract Improves Healing of Oral Mucositis in Golden Hamsters.

    PubMed

    Showraki, Najmeh; Mardani, Maryam; Emamghoreishi, Masoumeh; Andishe-Tadbir, Azadeh; Aram, Alireza; Mehriar, Peiman; Omidi, Mahmoud; Sepehrimanesh, Masood; Koohi-Hosseinabadi, Omid; Tanideh, Nader

    2016-12-01

    Oral mucositis (OM) is a common side effect of anti-cancer drugs and needs significant attention for its prevention. This study aimed to evaluate the healing effects of olive leaf extract on 5-fluorouracil-induced OM in golden hamster. OM was induced in 63 male golden hamsters by the combination of 5-fluorouracil injections (days 0, 5 and 10) and the abrasion of the cheek pouch (days 3 and 4). On day 12, hamsters were received topical olive leaf extract ointment, base of ointment, or no treatment (control) for 5 days. Histopathology evaluations, blood examinations, and tissue malondialdehyde level measurement were performed 1, 3 and 5 days after treatments. Histopathology score and tissue malondialdehyde level were significantly lower in olive leaf extract treated group in comparison with control and base groups ( p = 0.000). Significant decreases in white blood cell, hemoglobin, hematocrit , and mean corpuscular volume and an increase in mean corpuscular hemoglobin concentration were observed in olive leaf extract treated group in comparison with control and base groups ( p < 0.05). Our findings demonstrated that daily application of olive leaf extract ointment had healing effect on 5-fluorouracil induced OM in hamsters. Moreover, the beneficial effect of olive leaf extract on OM might be due to its antioxidant and anti-inflammatory properties.

  14. Effect of cryopreservation and transplantation on the expression of kit ligand and anti-Mullerian hormone in human ovarian tissue.

    PubMed

    David, Anu; Van Langendonckt, Anne; Gilliaux, Sébastien; Dolmans, Marie-Madeleine; Donnez, Jacques; Amorim, Christiani A

    2012-04-01

    Although cryopreservation and transplantation of ovarian tissue represent a promising alternative to safeguard fertility in cancer patients, low recovery rates of oocytes aspirated from antral follicles and a significant number of empty follicles have been observed in women with transplanted frozen-thawed ovarian tissue. In order to understand how freezing and/or grafting may affect follicular development, the follicular expression of kit ligand (KL) and anti-Müllerian hormone (AMH), two key factors activating and inhibiting follicle growth, were assessed after long-term grafting in severe combined immunodeficient (SCID) mice. Ovarian biopsies from eight patients were used for fresh and frozen-thawed tissue xenografting in 13 SCID mice for a period of 28 weeks, including 2 weeks of gonadotrophin stimulation. KL, AMH and proliferating cell nuclear antigen (PCNA) immunostaining were quantified before and after grafting in the two treatment groups (fresh and frozen-thawed grafted ovarian tissue). Lower expression of KL was found in primordial and primary follicles after grafting of both fresh and frozen-thawed tissue. Consistent expression of AMH was found in most growing follicles at a similar rate in both graft types. In fresh and frozen-thawed grafts, 13-14% of primordial follicles were PCNA-positive, indicating a similar maintenance of quiescent follicles despite follicle activation. Grafting and/or gonadotrophin stimulation appear to affect the follicular expression of KL, which may alter oocyte quality. AMH expression in growing follicles after ovarian tissue transplantation may be one of the factors contributing to the preservation of resting follicles in 28-week-old grafts.

  15. Fibre cables in the lacunae of Typha leaves contribute to a tensegrity structure.

    PubMed

    Witztum, Allan; Wayne, Randy

    2014-04-01

    Cables composed of long, non-lignified fibre cells enclosed in a cover of much shorter thin-walled, crystal-containing cells traverse the air chambers (lacunae) in leaves of the taller species of Typha. The non-lignified fibre cables are anchored in diaphragms composed of stellate cells of aerenchyma tissue that segment the long air chambers into smaller compartments. Although the fibre cables are easily observed and can be pulled free from the porous-to-air diaphragms, their structure and function have been ignored or misinterpreted. Leaves of various species of Typha were dissected and fibre cables were pulled free and observed with a microscope using bright-field and polarizing optics. Maximal tensile strength of freshly removed cables was measured by hanging weights from fibre cables, and Instron analysis was used to produce curves of load versus extension until cables broke. Polarized light microscopy revealed that the cellulose microfibrils that make up the walls of the cable fibres are oriented parallel to the long axis of the fibres. This orientation ensures that the fibre cables are mechanically stiff and strong under tension. Accordingly, the measured stiffness and tensile strength of the fibre cables were in the gigapascal range. In combination with the dorsal and ventral leaf surfaces and partitions that contain lignified fibre bundles and vascular strands that are strong in compression, the very fine fibre cables that are strong under tension form a tensegrity structure. The tensegrity structure creates multiple load paths through which stresses are redistributed throughout the 1-3 m tall upright leaves of Typha angustifolia, T. latifolia, T. × glauca, T. domingensis and T. shuttleworthii. The length of the fibre cables relative to the length of the leaf blades is reduced in the last-formed leaves of flowering individuals. Fibre cables are absent in the shorter leaves of Typha minima and, if present, only extend for a few centimetres from the sheath into the leaf blade of Typha laxmannii. The advantage of the structure of the Typha leaf blade, which enables stiffness to give way to flexibility under windy conditions, is discussed for both vegetative and flowering plants.

  16. Fibre cables in the lacunae of Typha leaves contribute to a tensegrity structure

    PubMed Central

    Witztum, Allan; Wayne, Randy

    2014-01-01

    Background and Aims Cables composed of long, non-lignified fibre cells enclosed in a cover of much shorter thin-walled, crystal-containing cells traverse the air chambers (lacunae) in leaves of the taller species of Typha. The non-lignified fibre cables are anchored in diaphragms composed of stellate cells of aerenchyma tissue that segment the long air chambers into smaller compartments. Although the fibre cables are easily observed and can be pulled free from the porous-to-air diaphragms, their structure and function have been ignored or misinterpreted. Methods Leaves of various species of Typha were dissected and fibre cables were pulled free and observed with a microscope using bright-field and polarizing optics. Maximal tensile strength of freshly removed cables was measured by hanging weights from fibre cables, and Instron analysis was used to produce curves of load versus extension until cables broke. Key Results and Conclusions Polarized light microscopy revealed that the cellulose microfibrils that make up the walls of the cable fibres are oriented parallel to the long axis of the fibres. This orientation ensures that the fibre cables are mechanically stiff and strong under tension. Accordingly, the measured stiffness and tensile strength of the fibre cables were in the gigapascal range. In combination with the dorsal and ventral leaf surfaces and partitions that contain lignified fibre bundles and vascular strands that are strong in compression, the very fine fibre cables that are strong under tension form a tensegrity structure. The tensegrity structure creates multiple load paths through which stresses are redistributed throughout the 1–3 m tall upright leaves of Typha angustifolia, T. latifolia, T. × glauca, T. domingensis and T. shuttleworthii. The length of the fibre cables relative to the length of the leaf blades is reduced in the last-formed leaves of flowering individuals. Fibre cables are absent in the shorter leaves of Typha minima and, if present, only extend for a few centimetres from the sheath into the leaf blade of Typha laxmannii. The advantage of the structure of the Typha leaf blade, which enables stiffness to give way to flexibility under windy conditions, is discussed for both vegetative and flowering plants. PMID:24532647

  17. Comparison of HeNe laser and sinusoidal non-uniform magnetic field seed pre-sowing treatment effect on Glycine max (Var 90-I) germination, growth and yield.

    PubMed

    Asghar, Tehseen; Iqbal, Munawar; Jamil, Yasir; Zia-Ul-Haq; Nisar, Jan; Shahid, Muhammad

    2017-01-01

    Recently, laser and magnetic field pre-sowing seed treatments attracted the attention of the scientific community in response to their positive effect on plant characteristics and the present study was exemplified for Glycine max Var 90-I. Seeds were exposed to laser (HeNe-wave length 632nm and density power of 1mW/cm 2 ) and magnetic field (sinusoidal non-uniform-50, 75 and 100mT for 3, 5min with exposure) and seed germination, seedling growth and yield attributes were compared. The germination (mean germination, germination percentage, emergence index, germination speed, relative germination coefficient, emergence coefficient of uniformity) growth (root dry weight, root length, shoot fresh weight and shoot dry weight, leaf dry & fresh weight, root fresh weight, leaf area, shoot length, plant total dry weight at different stages, stem diameter, number of leaves, vigor index I & II), biochemical (essential oil) and yield attributes (seed weight, count) were enhanced significantly in response to both laser and magnetic field treatments. However, magnetic field treatment furnished slightly higher response versus laser except relative water contents, whole plant weight and shoot length. Results revealed that both laser and magnetic field pre-sowing seed treatments affect the germination, seedling growth, and yield characteristics positively and could possibly be used to enhance Glycine max productivity. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Evidence of oleuropein degradation by olive leaf protein extract.

    PubMed

    De Leonardis, Antonella; Macciola, Vincenzo; Cuomo, Francesca; Lopez, Francesco

    2015-05-15

    The enzymatic activity of raw protein olive leaf extract has been investigated in vivo, on olive leaf homogenate and, in vitro with pure oleuropein and other phenolic substrates. At least two types of enzymes were found to be involved in the degradation of endogenous oleuropein in olive leaves. As for the in vitro experiments, the presence of active polyphenoloxidase and β-glucosidase was determined by HPLC and UV-Visible spectroscopy. Interestingly, both the enzymatic activities were found to change during the storage of olive leaves. Specifically, the protein extracts obtained from fresh leaves showed the presence of both the enzymatic activities, because oleuropein depletion occurred simultaneously with the formation of the oleuropein aglycon, 3,4-DHPEA-EA. In comparison leaves subjected to the drying process showed a polyphenoloxidase activity leading exclusively to the formation of oxidation products responsible for the typical brown coloration of the reaction solution. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. Staining paraffin embedded sections of scald of barley before paraffin removal.

    PubMed

    Xi, K; Burnett, P A

    1997-07-01

    Staining of paraffin embedded sections with periodic acid-Schiff reagent and fast green before paraffin removal resulted in differentiation of barley seed and leaf tissue from fungal structures of Rhynchosporium secalis. Crystal violet, toluidine blue O and antiline blue also successfully stained fungal structures of R. secalis in barley leaf tissues. Staining of embedded sections before paraffin removal allows simple processing of a series of sections, saves time and reduces solvent consumption.

  20. Infection and colonization of rice seedlings by the plant growth-promoting bacterium Herbaspirillum seropedicae Z67.

    PubMed

    James, Euan K; Gyaneshwar, Prasad; Mathan, Natarajan; Barraquio, Wilfredo L; Reddy, Pallavolu M; Iannetta, Pietro P M; Olivares, Fabio L; Ladha, Jagdish K

    2002-09-01

    A beta-glucoronidase (GUS)-marked strain of Herbaspirillum seropedicae Z67 was inoculated onto rice seedling cvs. IR42 and IR72. Internal populations peaked at over 10(6) log CFU per gram of fresh weight by 5 to 7 days after inoculation (DAI) but declined to 10(3) to 10(4) log CFU per gram of fresh weight by 28 DAI. GUS staining was most intense on coleoptiles, lateral roots, and at the junctions of some of the main and lateral roots. Bacteria entered the roots via cracks at the points of lateral root emergence, with cv. IR72 appearing to be more aggressively infected than cv. IR42. H. seropedicae subsequently colonized the root intercellular spaces, aerenchyma, and cortical cells, with a few penetrating the stele to enter the vascular tissue. Xylem vessels in leaves and stems were extensively colonized at 2 DAI but, in later harvests (7 and 13 DAI), a host defense reaction was often observed. Dense colonies of H. seropedicae with some bacteria expressing nitrogenase Fe-protein were seen within leaf and stem epidermal cells, intercellular spaces, and substomatal cavities up until 28 DAI. Epiphytic bacteria were also seen. Both varieties showed nitrogenase activity but only with added C, and the dry weights of the inoculated plants were significantly increased. Only cv. IR42 showed a significant (approximately 30%) increase in N content above that of the uninoculated controls, and it also incorporated a significant amount of 15N2.

  1. Climate Influences the Content and Chemical Composition of Foliar Tannins in Green and Senesced Tissues of Quercus rubra

    PubMed Central

    Top, Sara M.; Preston, Caroline M.; Dukes, Jeffrey S.; Tharayil, Nishanth

    2017-01-01

    Environmental stresses not only influence production of plant metabolites but could also modify their resorption during leaf senescence. The production-resorption dynamics of polyphenolic tannins, a class of defense compound whose ecological role extends beyond tissue senescence, could amplify the influence of climate on ecosystem processes. We studied the quantity, chemical composition, and tissue-association of tannins in green and freshly-senesced leaves of Quercus rubra exposed to different temperature (Warming and No Warming) and precipitation treatments (Dry, Ambient, Wet) at the Boston-Area Climate Experiment (BACE) in Massachusetts, USA. Climate influenced not only the quantity of tannins, but also their molecular composition and cell-wall associations. Irrespective of climatic treatments, tannin composition in Q. rubra was dominated by condensed tannins (CTs, proanthocyanidins). When exposed to Dry and Ambient*Warm conditions, Q. rubra produced higher quantities of tannins that were less polymerized. In contrast, under favorable conditions (Wet), tannins were produced in lower quantities, but the CTs were more polymerized. Further, even as the overall tissue tannin content declined, the content of hydrolysable tannins (HTs) increased under Wet treatments. The molecular composition of tannins influenced their content in senesced litter. Compared to the green leaves, the content of HTs decreased in senesced leaves across treatments, whereas the CT content was similar between green and senesced leaves in Wet treatments that produced more polymerized tannins. The content of total tannins in senesced leaves was higher in Warming treatments under both dry and ambient precipitation treatments. Our results suggest that, though climate directly influenced the production of tannins in green tissues (and similar patterns were observed in the senesced tissue), the influence of climate on tannin content of senesced tissue was partly mediated by the effect on the chemical composition of tannins. These different climatic impacts on leaves over the course of a growing season may alter forest dynamics, not only in decomposition and nutrient cycling dynamics, but also in herbivory dynamics. PMID:28559896

  2. Effects of nutrient addition on leaf chemistry, morphology, and photosynthetic capacity of three bog shrubs

    Treesearch

    Jill L. Bubier; Rose Smith; Sari Juutinen; Tim R. Moore; Rakesh Minocha; Stephanie Long; Subash Minocha

    2011-01-01

    Plants in nutrient-poor environments typically have low foliar nitrogen (N) concentrations, long-lived tissues with leaf traits designed to use nutrients efficiently, and low rates of photosynthesis. We postulated that increasing N availability due to atmospheric deposition would increase photosynthetic capacity, foliar N, and specific leaf area (SLA) of bog shrubs. We...

  3. Mangifera indica L. leaf extract alleviates doxorubicin induced cardiac stress

    PubMed Central

    Bhatt, Laxit; Joshi, Viraj

    2017-01-01

    Aim: The study was undertaken to evaluate the cardioprotective effect of the alcoholic leaf extract of Mangifera indica L. against cardiac stress caused by doxorubicin (DOX). Materials and Methods: Rats were treated with 100 mg/kg of M. indica leaf extract (MILE) in alone and interactive groups for 21 days. Apart from the normal and MILE control groups, all the groups were subjected to DOX (15 mg/kg, i.p.) toxicity for 21 days and effects of different treatments were analyzed by changes in serum biomarkers, tissue antioxidant levels, electrocardiographic parameters, lipid profile, and histopathological evaluation. Results: The MILE treated group showed decrease in serum biomarker enzyme levels and increase in tissue antioxidants levels. Compared to DOX control group, MILE treated animals showed improvement in lipid profile, electrocardiographic parameters, histological score, and mortality. Conclusion: These findings clearly suggest the protective role of alcoholic leaf extract of M. indica against oxidative stress induced by DOX. PMID:28894627

  4. De novo assembly and analysis of the Artemisia argyi transcriptome and identification of genes involved in terpenoid biosynthesis.

    PubMed

    Liu, Miaomiao; Zhu, Jinhang; Wu, Shengbing; Wang, Chenkai; Guo, Xingyi; Wu, Jiawen; Zhou, Meiqi

    2018-04-11

    Artemisia argyi Lev. et Vant. (A. argyi) is widely utilized for moxibustion in Chinese medicine, and the mechanism underlying terpenoid biosynthesis in its leaves is suggested to play an important role in its medicinal use. However, the A. argyi transcriptome has not been sequenced. Herein, we performed RNA sequencing for A. argyi leaf, root and stem tissues to identify as many as possible of the transcribed genes. In total, 99,807 unigenes were assembled by analysing the expression profiles generated from the three tissue types, and 67,446 of those unigenes were annotated in public databases. We further performed differential gene expression analysis to compare leaf tissue with the other two tissue types and identified numerous genes that were specifically expressed or up-regulated in leaf tissue. Specifically, we identified multiple genes encoding significant enzymes or transcription factors related to terpenoid synthesis. This study serves as a valuable resource for transcriptome information, as many transcribed genes related to terpenoid biosynthesis were identified in the A. argyi transcriptome, providing a functional genomic basis for additional studies on molecular mechanisms underlying the medicinal use of A. argyi.

  5. Prevention of pink-pigmented methylotrophic bacteria (Methylohacterium mesophilicum) contamination of plant tissue cultures.

    PubMed

    Chanprame, S; Todd, J J; Widholm, J M

    1996-12-01

    Pink-pigmented facultative methylotrophic bacteria (PPFMs) have been found on the surfaces of leaves of most plants tested. We found PPFMs on the leaf surfaces of all 40 plants (38 species) tested and on soybean pods by pressing onto AMS medium with methanol as the sole carbon source. The abundance ranged from 0.5 colony forming unit (cfu) /cm(2) to 69.4 cfu/cm(2) on the leaf surfaces. PPFMs were found in homogenized leaf tissues of only 4 of the species after surface disinfestation with 1.05% sodium hypochlorite and were rarely found in cultures initiated from surface disinfested Datura innoxia leaves or inside surface disinfested soybean pods. Of 20 antibiotics tested for PPFM growth inhibition, rifampicin was the most effective and of seven others which also inhibited PPFM growth, cefotaxime should be the most useful due to the expected low plant cell toxicity. These antibiotics could be used in concert with common surface sterilization procedures to prevent the introduction or to eliminate PPFM bacteria in tissue cultures. Thus, while PPFMs are present on the surfaces of most plant tissues, surface disinfestation alone can effectively remove them so that uncontaminated tissue cultures can be initiated in most cases.

  6. A surface wave elastography technique for measuring tissue viscoelastic properties.

    PubMed

    Zhang, Xiaoming

    2017-04-01

    A surface wave elastography method is proposed to study the viscoelastic properties of skin by measuring the surface wave speed and attenuation on the skin. Experiments were carried out on porcine skin tissues. The surface wave speed is measured by the change of phase with distance. The wave attenuation is measured by the decay of wave amplitude with distance. The change of viscoelastic properties with temperature was studied at room and body temperatures. The wave speed was 1.83m/s at 22°C but reduced to 1.52m/s at 33°C. The viscoelastic ratio was almost constant from 22°C to 33°C. Fresh and decayed tissues were studied. The wave speed of the decayed tissue increased from 1.83m/s of fresh state to 2.73m/s. The viscoelastic ratio was 0.412/mm at the decayed state compared to 0.215/mm at the fresh state. More tissue samples are needed to study these viscoelastic parameters according to specific applications. Copyright © 2017 IPEM. Published by Elsevier Ltd. All rights reserved.

  7. Developmental Role and Auxin Responsiveness of Class III Homeodomain Leucine Zipper Gene Family Members in Rice1[C][W][OA

    PubMed Central

    Itoh, Jun-Ichi; Hibara, Ken-Ichiro; Sato, Yutaka; Nagato, Yasuo

    2008-01-01

    Members of the Class III homeodomain leucine zipper (Class III HD-Zip) gene family are central regulators of crucial aspects of plant development. To better understand the roles of five Class III HD-Zip genes in rice (Oryza sativa) development, we investigated their expression patterns, ectopic expression phenotypes, and auxin responsiveness. Four genes, OSHB1 to OSHB4, were expressed in a localized domain of the shoot apical meristem (SAM), the adaxial cells of leaf primordia, the leaf margins, and the xylem tissue of vascular bundles. In contrast, expression of OSHB5 was observed only in phloem tissue. Plants ectopically expressing microRNA166-resistant versions of the OSHB3 gene exhibited severe defects, including the ectopic production of leaf margins, shoots, and radialized leaves. The treatment of seedlings with auxin quickly induced ectopic OSHB3 expression in the entire region of the SAM, but not in other tissues. Furthermore, this ectopic expression of OSHB3 was correlated with leaf initiation defects. Our findings suggest that rice Class III HD-Zip genes have conserved functions with their homologs in Arabidopsis (Arabidopsis thaliana), but have also acquired specific developmental roles in grasses or monocots. In addition, some Class III HD-Zip genes may regulate the leaf initiation process in the SAM in an auxin-dependent manner. PMID:18567825

  8. Freezing avoidance by supercooling in Olea europaea cultivars: the role of apoplastic water, solute content and cell wall rigidity.

    PubMed

    Arias, Nadia S; Bucci, Sandra J; Scholz, Fabian G; Goldstein, Guillermo

    2015-10-01

    Plants can avoid freezing damage by preventing extracellular ice formation below the equilibrium freezing temperature (supercooling). We used Olea europaea cultivars to assess which traits contribute to avoid ice nucleation at sub-zero temperatures. Seasonal leaf water relations, non-structural carbohydrates, nitrogen and tissue damage and ice nucleation temperatures in different plant parts were determined in five cultivars growing in the Patagonian cold desert. Ice seeding in roots occurred at higher temperatures than in stems and leaves. Leaves of cold acclimated cultivars supercooled down to -13 °C, substantially lower than the minimum air temperatures observed in the study site. During winter, leaf ice nucleation and leaf freezing damage (LT50 ) occurred at similar temperatures, typical of plant tissues that supercool. Higher leaf density and cell wall rigidity were observed during winter, consistent with a substantial acclimation to sub-zero temperatures. Larger supercooling capacity and lower LT50 were observed in cold-acclimated cultivars with higher osmotically active solute content, higher tissue elastic adjustments and lower apoplastic water. Irreversible leaf damage was only observed in laboratory experiments at very low temperatures, but not in the field. A comparative analysis of closely related plants avoids phylogenetic independence bias in a comparative study of adaptations to survive low temperatures. © 2015 John Wiley & Sons Ltd.

  9. Evaluation of the Xpert MTB/RIF Performance on Tissues: Potential Impact on Airborne Infection Isolation at a Tertiary Cancer Care Center.

    PubMed

    McMillen, Tracy; Usiak, Shauna C; Chen, Liang Hua; Gomez, Luz; Ntiamoah, Peter; Hameed, Meera R; Budvytiene, Indre; Banaei, Niaz; Kamboj, Mini; Babady, N Esther

    2018-04-01

    OBJECTIVES In this study, we sought to evaluate the performance of the Xpert MTB/RIF (Cepheid) assay for the detection of Mycobacterium tuberculosis (MTB) complex DNA on fresh and formalin-fixed, paraffin-embedded (FFPE) tissue specimens from oncology patients in an area with a low prevalence of tuberculosis. We also aimed to retrospectively assess the potential impact of Xpert MTB/RIF on the duration of airborne infection isolation (AII). SETTING A 473-bed, tertiary-care cancer center in New York City. DESIGN A total of 203 tissue samples (101 FFPE and 102 fresh) were tested using Xpert MTB/RIF, including 133 pulmonary tissue samples (65.5%) and 70 extrapulmonary tissue samples (34.5%). Acid-fast bacilli (AFB) culture was used as the diagnostic gold standard. The limit of detection (LOD) and reproducibility were also evaluated for both samples types using contrived specimens. The potential impact of the Xpert MTB PCR assay on tissue samples from AII patients on AII duration was retrospectively assessed. RESULTS Using the Xpert MTB/RIF for fresh tissue specimens, the sensitivity was 50% (95% CI, 1.3%-98.7%) and the specificity was 99% (95% CI, 94.5%-99.9%). For FFPE tissue specimens, the sensitivity was 100% (95% CI, 63.1%-100%) and the specificity was 98.3% (95% CI, 95.5%-100%. The LOD was 103 colony-forming units (CFU)/mL for both fresh and FFPE tissue specimens, and the Xpert MTB/RIF was 100% reproducible at concentrations 10 times that of the LOD. With an expected turnaround time of 24 hours, the Xpert MTB PCR could decrease the duration of AII from a median of 8 days to a median of 1 day. CONCLUSIONS The Xpert MTB/RIF assay offers a valid option for ruling out Mycobacterium tuberculosis complex (MTBC) on tissue samples from oncology patients and for minimizing AII resource utilization. Infect Control Hosp Epidemiol 2018;39:462-466.

  10. Fresh osteochondral allografts-procurement and tissue donation in Europe.

    PubMed

    Schmidt, S; Schulte, A; Schwarz, S; Hofmann, N; Tietz, S; Boergel, M; Sixt, S U

    2017-07-01

    Fresh osteochondral allografts are a well-established treatment for large, full-thickness cartilage defects. The clinical outcome for carefully selected patients is very favorable, especially for the young and active and graft survival up to 25 years has been described in the literature. Furthermore, a high patient satisfaction rate has been reported, but the biggest obstacle to overcome is the availability of tissue for transplantation. Large fresh bone allografts for cartilage damage repair only can be harvested from organ donors following organ removal or cadaveric donors, preferably in the setting of an operation room to minimize possible contamination of the tissue. Apart from the logistic challenges this entails, an experienced recovery team is needed. Furthermore, the public as well as medical staff is much less aware of the possibility and requirements of tissue donation than organ donation and families of deceased are rarely approached for bone and cartilage donation. This review aims to highlight the current situation of organ and tissue donation in Europe with special focus on the processing of bones and possible safety and quality concerns. We analyze what may prevent consent and what might be done to improve the situation of tissue donation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Spatially Different Tissue-Scale Diffusivity Shapes ANGUSTIFOLIA3 Gradient in Growing Leaves.

    PubMed

    Kawade, Kensuke; Tanimoto, Hirokazu; Horiguchi, Gorou; Tsukaya, Hirokazu

    2017-09-05

    The spatial gradient of signaling molecules is pivotal for establishing developmental patterns of multicellular organisms. It has long been proposed that these gradients could arise from the pure diffusion process of signaling molecules between cells, but whether this simplest mechanism establishes the formation of the tissue-scale gradient remains unclear. Plasmodesmata are unique channel structures in plants that connect neighboring cells for molecular transport. In this study, we measured cellular- and tissue-scale kinetics of molecular transport through plasmodesmata in Arabidopsis thaliana developing leaf primordia by fluorescence recovery assays. These trans-scale measurements revealed biophysical properties of diffusive molecular transport through plasmodesmata and revealed that the tissue-scale diffusivity, but not the cellular-scale diffusivity, is spatially different along the leaf proximal-to-distal axis. We found that the gradient in cell size along the developmental axis underlies this spatially different tissue-scale diffusivity. We then asked how this diffusion-based framework functions in establishing a signaling gradient of endogenous molecules. ANGUSTIFOLIA3 (AN3) is a transcriptional co-activator, and as we have shown here, it forms a long-range signaling gradient along the leaf proximal-to-distal axis to determine a cell-proliferation domain. By genetically engineering AN3 mobility, we assessed each contribution of cell-to-cell movement and tissue growth to the distribution of the AN3 gradient. We constructed a diffusion-based theoretical model using these quantitative data to analyze the AN3 gradient formation and demonstrated that it could be achieved solely by the diffusive molecular transport in a growing tissue. Our results indicate that the spatially different tissue-scale diffusivity is a core mechanism for AN3 gradient formation. This provides evidence that the pure diffusion process establishes the formation of the long-range signaling gradient in leaf development. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  12. Effects of different products of peach (Prunus persica L. Batsch) from a variety developed in southern Brazil on oxidative stress and inflammatory parameters in vitro and ex vivo

    PubMed Central

    Gasparotto, Juciano; Somensi, Nauana; Bortolin, Rafael Calixto; Moresco, Karla Suzana; Girardi, Carolina Saibro; Klafke, Karina; Rabelo, Thallita Kelly; Morrone, Maurilio Da Silva; Vizzotto, Márcia; Raseira, Maria do Carmo Bassols; Moreira, José Claudio Fonseca; Gelain, Daniel Pens

    2014-01-01

    Antioxidant, anti-glycation and anti-inflammatory activities of fresh and conserved peach fruits (Prunus persica L. Batsch) were compared. Fresh peach pulps, peels, preserve peach pulps and the preserve syrup were prepared at equal concentrations. Rat liver, kidney and brain cortex tissue slices were pre-incubated with peach samples, subjected to oxidative stress with FeSO4 and hydrogen peroxide. Fresh peach pulps and peel conferred higher protection against cytotoxicity and oxidative stress than preserve peach pulps in most tissues. Release of tumor necrosis factor-α and interleukin-1β was also significantly decreased by Fresh peach pulps and peel, followed by preserve peach pulps. Total phenolic determination and HPLC analysis of carotenoids showed that the content of secondary metabolites in Fresh peach pulps and peel is significantly higher than in preserve peach pulps, while the syrup had only small or trace amounts of these compounds. Fresh peach pulps and Peel demonstrated high antioxidant and anti-inflammatory effects preventing against induced damage. PMID:25320458

  13. Analyses of the leaf, fruit and seed of Thaumatococcus daniefii (Benth.): exploring potential uses.

    PubMed

    Chinedu, Shalom Nwodo; Oluwadamisi, Adetayo Y; Popoola, Samuel T; David, Bolaji J; Epelle, Tamunotonyesia

    2014-06-01

    Thaumatococcus daniellii is an economic plant with versatile uses in Southern Nigeria. The arils attached to the seeds contain thaumatin, a non-sugar sweetener and taste modifier. This study examined the chemical constituents of the leaf, fruit and seed of T. daniellii. The fresh fruit, on weight basis, consists of 4.8% aril, 22.8% seed and 72.4% fleshy part. The leaf contained (per 100 g): 10.67 g moisture, 8.95 g ash, 17.21 g fat, 21.06 g protein, 24.61 g crude fiber 17.50 g carbohydrate, 0.10 g calcium, 0.08 g magnesium, 0.01 g iron and 0.37 g phosphorus. The fruit (fleshy part) contained 10.04 g moisture, 21.08 g ash, 0.93 g fat, 11.53 g protein, 18.43 g crude fiber, 37.27 g carbohydrate, 0.34 g calcium, 0.30 g magnesium, 0.01 g iron and 0.21 g phosphorus. The seed contained 15.15 g moisture, 11.30 g ash, 0.21 g fat, 10.36 g protein, 20.52 g crude fiber and 42.46 g carbohydrate. Terpenoids, flavonoids, alkaloids and cardiac glycosides were significantly present in both the leaf and fruit whereas phlobatannin, saponin, steroids, anthraquinones and ascorbic acid were absent. Tannin was present only in the leaf. The leaf and fruit of T. daniellii have significant nutritional and medicinal benefits. The leaf is rich in protein and fat. The fruit is a good source of minerals, particularly, calcium and magnesium; the leaf is also rich in phosphorus.

  14. Allelopathic potential of leaf and seed of Mucuna bracteata DC. ex Kurz on Eleusine indica (L.) gaertn

    NASA Astrophysics Data System (ADS)

    Halimshah, Syamimi; Ismail B., S.; Ahmad, Wan Juliana Wan

    2015-09-01

    A study was conducted to determine the allelopathic potential of leaf and seed of Mucuna bracteata on the growth of E. indica through aqueous extract and debris (incorporated into the soil) experiment. Three concentrations of leaf and seed aqueous extract (16.7, 33.3 and 66.7 g/L) and debris (2.5, 5.0 and 10.0 g/500 g soil) of M. bracteata were used in the experiment. Complete randomized design (CRD) with three replications was applied in this experiment which was conducted twice. Results demonstrated that the leaf and seed extracts of M. bracteata exhibited higher suppression effect on the growth and germination of E. indica as the concentration increased. The leaf and seed extracts significantly reduced all measured parameters at all concentrations except for the shoot length and germination of E. indica by seed extract at 16.7 g/L which recorded insignificant reduction by 40.5% and 4% respectively. The leaf and seed debris significantly reduced the root length of E. indica at all treatments. Seed debris also showed significant reduction on the germination at all treatments and other seedling growth parameters (shoot length, fresh weight and dry weight) at 2.5 and 10.0 g/500 g soil. Meanwhile, the leaf debris demonstrated stimulation effect on the seedling growth parameters. As a whole, the leaf showed higher suppression effect in aqueous extract experiment while the seed recorded higher suppression effect in the debris experiment. Further studies need to be conducted to investigate the type of inhibition mechanism involved in both experiments.

  15. Dimensional Changes of Fresh Sockets With Reactive Soft Tissue Preservation: A Cone Beam CT Study.

    PubMed

    Crespi, Roberto; Capparé, Paolo; Crespi, Giovanni; Gastaldi, Giorgio; Gherlone, Enrico Felice

    2017-06-01

    The aim of this study was to assess dimensional changes of the fresh sockets grafted with collagen sheets and maintenance of reactive soft tissue, using cone beam computed tomography (CBCT). Tooth extractions were performed with maximum preservation of the alveolar housing, reactive soft tissue was left into the sockets and collagen sheets filled bone defects. Cone beam computed tomography were performed before and 3 months after extractions. One hundred forty-five teeth, 60 monoradiculars and 85 molars, were extracted. In total, 269 alveoli were evaluated. In Group A, not statistically significant differences were found between monoradiculars, whereas statistically significant differences (P < 0.05) were found between molars, both for mesial and distal alveoli. In Group B, not statistically significant differences were found between maxillary and mandibular bone changes values (P > 0.05) for all types of teeth. This study reported an atraumatic tooth extraction, reactive soft tissue left in situ, and grafted collagen sponge may be helpful to reduce fresh socket collapse after extraction procedures.

  16. Hyperforin Accumulates in the Translucent Glands of Hypericum perforatum

    PubMed Central

    Soelberg, Jens; Jørgensen, Lise Bolt; Jäger, Anna K.

    2007-01-01

    Background and Aims Hypericum perforatum contains the therapeutically important compounds hypericin and hyperforin. Hypericin is known to accumulate in the dark glands. This investigation aimed to determine the accumulation site of hyperforin. Methods Dark and translucent glands as well as non-secretory tissue in leaves were manually isolated under the microscope. Hyperforin content was quantified by UV HPLC. Secretory structures were surveyed anatomically. Key Results The hyperforin content of intact leaves was found to be about 3 mg g−1 fresh tissue, whereas a content of about 7 mg g−1 fresh material was found in isolated translucent glands. Hyperforin was found only to occur in minute amounts in dark glands (approx. 0·4 mg g−1 fresh tissue). In non-secretory tissue no hyperforin was detected. Conclusions The accumulation of hyperforin detected in the translucent glands supports the proposed hypothesis that hyperforin is synthesized by the same biosynthetic machinery as monoterpenes in the chloroplasts of cells delimiting the gland. PMID:17468113

  17. CO₂ processing and hydration of fruit and vegetable tissues by clathrate hydrate formation.

    PubMed

    Takeya, Satoshi; Nakano, Kohei; Thammawong, Manasikan; Umeda, Hiroki; Yoneyama, Akio; Takeda, Tohoru; Hyodo, Kazuyuki; Matsuo, Seiji

    2016-08-15

    CO2 hydrate can be used to preserve fresh fruits and vegetables, and its application could contribute to the processing of carbonated frozen food. We investigated water transformation in the frozen tissue of fresh grape samples upon CO2 treatment at 2-3 MPa and 3°C for up to 46 h. Frozen fresh bean, radish, eggplant and cucumber samples were also investigated for comparison. X-ray diffraction indicated that after undergoing CO2 treatment for several hours, structure I CO2 hydrate formed within the grape tissue. Phase-contrast X-ray imaging using the diffraction-enhanced imaging technique revealed the presence of CO2 hydrate within the intercellular spaces of these tissues. The carbonated produce became effervescent because of the dissociation of CO2 hydrate through the intercellular space, especially above the melting point of ice. In addition, suppressed metabolic activity resulting from CO2 hydrate formation, which inhibits water and nutrient transport through intercellular space, can be expected. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Possible Roles of Strigolactones during Leaf Senescence

    PubMed Central

    Yamada, Yusuke; Umehara, Mikihisa

    2015-01-01

    Leaf senescence is a complicated developmental process that involves degenerative changes and nutrient recycling. The progress of leaf senescence is controlled by various environmental cues and plant hormones, including ethylene, jasmonic acid, salicylic acid, abscisic acid, cytokinins, and strigolactones. The production of strigolactones is induced in response to nitrogen and phosphorous deficiency. Strigolactones also accelerate leaf senescence and regulate shoot branching and root architecture. Leaf senescence is actively promoted in a nutrient-poor soil environment, and nutrients are transported from old leaves to young tissues and seeds. Strigolactones might act as important signals in response to nutrient levels in the rhizosphere. In this review, we discuss the possible roles of strigolactones during leaf senescence. PMID:27135345

  19. Technetium-99 cycling in maple trees: characterization of changes in chemical form.

    PubMed

    Garten, C T; Lomax, R D

    1989-08-01

    Prior field studies near an old radioactive waste disposal site at Oak Ridge, TN, indicated that following root uptake, metabolism by deciduous trees rendered 99Tc less biogeochemically mobile than expected, based on chemistry of the pertechnetate (TcO-4) anion. Subsequently, the form of technetium (Tc) in maple tree (Acer sp.) sap, leaves, wood and forest leaf litter was characterized using one or more of the following methods: dialysis, physical fractionation, chemical extraction, gel permeation chromatography, enzymatic extraction, or thin layer chromatography (TLC) on silica gel. Chromatography (Sephadex G-25) of TcO-4 incubated in vitro with tree sap showed it to behave similar to TcO-4 anion. When labeled wood and leaf tissues were processed using a tissue homogenizer, 15% and 40%, respectively, of the Tc was solubilized into phosphate buffer. Most (65% to 80%) of the solubilized Tc passing a 0.45-micron filter also passed through an ultrafiltration membrane with a nominal molecular weight cutoff of 10,000 atomic mass units (amu). A majority (72% to 80%) of the Tc in wood could be chemically removed by successive extractions with ethanol, water and weak mineral acid. These same extractants removed only 23% to 31% of the Tc from maple leaves or forest floor leaf litter. Most of the Tc in leaves and leaf litter was removed only by strongly alkaline reagents typically used to release structural polysaccharides (hemicelluloses) from plant tissues. Chromatography (Sephadex G-25) of the ethanol-water extract from wood and the alkaline extract from leaves demonstrated that Tc in these extracts was not principally TcO-4 but was complexed with molecules greater than 1000 amu. Incubations of leaf and wood homogenates with protease approximately doubled the amount of Tc released from contaminated tissues. Ultrafiltration of protease-solubilized Tc from leaves and wood showed that 40% and 93%, respectively, of the Tc was less than 10,000 amu. TLC of the less than 10,000 amu fraction indicated the presence of TcO-4 in wood but not in leaves. In the leaf, TcO-4 is converted to less soluble forms apparently associated with structural components of leaf cell walls. This conversion explains why 99Tc is not easily leached by rainfall from tree foliage and why 99Tc appears to accumulate in forest floor leaf litter layers at the Oak Ridge study site.

  20. A noncoding RNA transcribed from the AGAMOUS (AG) second intron binds to CURLY LEAF and represses AG expression in leaves.

    PubMed

    Wu, Hui-Wen; Deng, Shulin; Xu, Haiying; Mao, Hui-Zhu; Liu, Jun; Niu, Qi-Wen; Wang, Huan; Chua, Nam-Hai

    2018-06-04

    Dispersed H3K27 trimethylation (H3K27me3) of the AGAMOUS (AG) genomic locus is mediated by CURLY LEAF (CLF), a component of the Polycomb Repressive Complex (PRC) 2. Previous reports have shown that the AG second intron, which confers AG tissue-specific expression, harbors sequences targeted by several positive and negative regulators. Using RACE reverse transcription polymerase chain reaction, we found that the AG intron 2 encodes several noncoding RNAs. RNAi experiment showed that incRNA4 is needed for CLF repressive activity. AG-incRNA4RNAi lines showed increased leaf AG mRNA levels associated with a decrease of H3K27me3 levels; these plants displayed AG overexpression phenotypes. Genetic and biochemical analyses demonstrated that the AG-incRNA4 can associate with CLF to repress AG expression in leaf tissues through H3K27me3-mediated repression and to autoregulate its own expression level. The mechanism of AG-incRNA4-mediated repression may be relevant to investigations on tissue-specific expression of Arabidopsis MADS-box genes. © 2018 The Authors New Phytologist © 2018 New Phytologist Trust.

  1. Analysis of terahertz dielectric properties of pork tissue

    NASA Astrophysics Data System (ADS)

    Huang, Yuqing; Xie, Qiaoling; Sun, Ping

    2017-10-01

    Seeing that about 70% component of fresh biological tissues is water, many scientists try to use water models to describe the dielectric properties of biological tissues. The classical water dielectric models are Debye model, Double Debye model and Cole-Cole model. This work aims to determine a suitable model by comparing three models above with experimental data. These models are applied to fresh pork tissue. By means of least square method, the parameters of different models are fitted with the experimental data. Comparing different models on both dielectric function, the Cole-Cole model is verified the best to describe the experiments of pork tissue. The correction factor α of the Cole-Cole model is an important modification for biological tissues. So Cole-Cole model is supposed to be a priority selection to describe the dielectric properties for biological tissues in the terahertz range.

  2. Allelopathic potential of Chromolaena odorata and Mikania micrantha on Brassica chinensis var. parachinensis

    NASA Astrophysics Data System (ADS)

    Yusoff, Nornasuha; Ismail B., S.

    2015-09-01

    Laboratory and greenhouse experiments were conducted to study the allelopathic potential of the aqueous leaf extract and leaf debris (incorporated into the soil) of Chromolaena odorata and Mikania micrantha on the germination indices and growth as well as the allelopathic effect response index of Brassica chinensis. Three concentrations each of the aqueous leaf extract (12.5, 25.0 and 50.0 g/L) and leaf debris (2.5, 5.0 and 10.0 g/500 g soil) were used in the experiments. The treatments were arranged in a Completely Randomized Design (CRD) with three replications, and the experiment was conducted twice. The aqueous leaf extracts of both species significantly inhibited all growth parameters of B. chinensis at 50.0 g/L concentration by more than 50% (compared to that by the control). In contrast, the leaf debris of both species at most of the concentrations showed significant stimulatory effects on all growth parameters of B. chinensis. However, M. micrantha leaf debris showed no significant effect on the fresh weight of B. chinensis at all concentrations. The total germination percentage of B. chinensis was significantly decreased as concentration of the aqueous leaf extracts of both species increased. The aqueous leaf extract of both species at concentrations higher than 25.0 g/L, significantly reduced the initial speed of germination as well as the cumulative speed of germination of B. chinensis. The allelopathic effect response index was negative for both species, indicating that the extracts of both species have inhibitory effects on the germination and seedling growth of B. chinensis. Results from the study suggested that the leaves of C. odorata and M. micrantha have phytotoxic properties and have potential for use directly or indirectly on susceptible weeds, and thereby reducing the use of chemical pesticides.

  3. Remote sensing of nutrient deficiency in Lactuca sativa using neural networks for terrestrial and advanced life support applications

    NASA Astrophysics Data System (ADS)

    Sears, Edie Seldon

    2000-12-01

    A remote sensing study using reflectance and fluorescence spectra of hydroponically grown Lactuca sativa (lettuce) canopies was conducted. An optical receiver was designed and constructed to interface with a commercial fiber optic spectrometer for data acquisition. Optical parameters were varied to determine effects of field of view and distance to target on vegetation stress assessment over the test plant growth cycle. Feedforward backpropagation neural networks (NN) were implemented to predict the presence of canopy stress. Effects of spatial and spectral resolutions on stress predictions of the neural network were also examined. Visual inspection and fresh mass values failed to differentiate among controls, plants cultivated with 25% of the recommended concentration of phosphorous (P), and those cultivated with 25% nitrogen (N) based on fresh mass and visual inspection. The NN's were trained on input vectors created using reflectance and test day, fluorescence and test day, and reflectance, fluorescence, and test day. Four networks were created representing four levels of spectral resolution: 100-nm NN, 10-nm NN, 1-nm NN, and 0.1-nm NN. The 10-nm resolution was found to be sufficient for classifying extreme nitrogen deficiency in freestanding hydroponic lettuce. As a result of leaf angle and canopy structure broadband scattering intensity in the 700-nm to 1000-nm range was found to be the most useful portion of the spectrum in this study. More subtle effects of "greenness" and fluorescence emission were believed to be obscured by canopy structure and leaf orientation. As field of view was not as found to be as significant as originally believed, systems implementing higher repetitions over more uniformly oriented, i.e. smaller, flatter, target areas would provide for more discernible neural network input vectors. It is believed that this technique holds considerable promise for early detection of extreme nitrogen deficiency. Further research is recommended using stereoscopic digital cameras to quantify leaf area index, leaf shape, and leaf orientation as well as reflectance. Given this additional information fluorescence emission may also prove a more useful biological assay of freestanding vegetation.

  4. Leaf economics and hydraulic traits are decoupled in five species-rich tropical-subtropical forests.

    PubMed

    Li, Le; McCormack, M Luke; Ma, Chengen; Kong, Deliang; Zhang, Qian; Chen, Xiaoyong; Zeng, Hui; Niinemets, Ülo; Guo, Dali

    2015-09-01

    Leaf economics and hydraulic traits are critical to leaf photosynthesis, yet it is debated whether these two sets of traits vary in a fully coordinated manner or there is room for independent variation. Here, we tested the relationship between leaf economics traits, including leaf nitrogen concentration and leaf dry mass per area, and leaf hydraulic traits including stomatal density and vein density in five tropical-subtropical forests. Surprisingly, these two suites of traits were statistically decoupled. This decoupling suggests that independent trait dimensions exist within a leaf, with leaf economics dimension corresponding to light capture and tissue longevity, and the hydraulic dimension to water-use and leaf temperature maintenance. Clearly, leaf economics and hydraulic traits can vary independently, thus allowing for more possible plant trait combinations. Compared with a single trait dimension, multiple trait dimensions may better enable species adaptations to multifarious niche dimensions, promote diverse plant strategies and facilitate species coexistence. © 2015 John Wiley & Sons Ltd/CNRS.

  5. V. THE IRON CONTENT OF BLOOD FREE TISSUES AND VISCERA

    PubMed Central

    Bogniard, Robert P.; Whipple, George H.

    1932-01-01

    When hemoglobin is set free in the circulation the kidney plays an important part in the conservation of iron. When the renal threshold is not exceeded by the hemoglobin in the blood there is little or no excess iron deposited in the kidney but when superthreshold doses of blood hemoglobin are given the epithelium of the convoluted tubules accumulates much iron and the iron analyses may show 5 times normal values. The normal dog (140 to 150 per cent hemoglobin) has a large reserve store of iron in the liver, spleen and marrow. Diets may modify this storage of iron in these tissues. To bring conclusive proof relating to the individual diet factors, the reserve store of iron should be depleted by an anemia period of 2 to 3 months. Complete removal of red cells from tissue capillaries is essential for accurate iron assays of fresh tissue. The method described accomplishes this without causing gross tissue edema. The lowest iron content is observed in the pancreas, stomach, jejunum, colon and urinary bladder. These figures average from 1 to 2 mg. iron per 100 gm. fresh tissue. This shows that smooth muscle and mucous membranes contain little iron. Striated muscle (heart, psoas) shows a relatively low iron content but uniform values close to 4 mg. per 100 gm. tissue. Lungs show a considerable fluctuation with low iron values in anemia (3.7 mg.) and higher values in health (6 to 7 mg.). The spleen shows maximal fluctuations and the highest reserve storage of iron per 100 gm. fresh tissue. The spleen iron analyses show low values in anemia (7 to 15 mg.) and wide differences in controls (25 to 50 mg.). With hemoglobin injections the iron storage is conspicuous and iron analyses may run as high as l50 to 175 mg. iron per 100 gm. fresh tissue. Bone marrow of the rib runs in parallel with the spleen as regards iron storage following hemoglobin injections and depletion following anemia periods. The liver because of its weight always contains the main bulk of the iron stored in the blood free tissues of the body. Its store is depleted by anemia even to levels of 4 to 5 mg. iron per 100 gm. fresh tissue. In the normal dog the iron store in the liver averages 25 mg. per 100 gm. tissue. Frequent hemoglobin injections may increase this level to 31 mg. iron per 100 gm. The liver is considered the most active clearing house for iron storage and utilization. PMID:19870020

  6. The effect of various media and hormones via suspension culture on secondary metabolic activities of (Cape Jasmine) Gardenia jasminoides Ellis.

    PubMed

    Farzinebrahimi, Reza; Mat Taha, Rosna; Rashid, Kamaludin; Syafawati Yaacob, Jamilah

    2014-01-01

    The leaf of Gardenia jasminoides Ellis was used as explants and was cultured on MS and WPM media supplemented with various concentrations of NAA, IAA, 2,4-D, IBA, TDZ, and Kn (0 to 5 mg L(-1) with 0.5 increment). After six months, the higher percentage of callus (100%) and the best dry and fresh weight of callus were formed on WPM medium supplemented with 2,4-D and NAA (2.0-3.0 mg L(-1)) and this amount was decreased from (84%) to (69%) when this media supplemented with Kinetin and TDZ (1 mg L(-1)) respectively were used. Leaf segments cultured on WPM media added with Kn (1 mg L(-1)) and TDZ (2 mg L(-1)) yielded the least amount of callus. It was found that WPM media added with IAA (4.5-5.0 mg L(-1)) were optimum for root induction from G. jasminoides plantlets. Antibacterial screening of leaf extracts (in vivo) showed no inhibitory effect against E. coli, P. aeruginosa, S. aureus, and B. cereus, in contrast to callus extracts from leaf cultures supplemented with NAA, which showed inhibition activity against E. coli and B. cereus. The callus extracts from leaf cultures grown on both MS and WPM media showed higher antioxidant and superoxide dismutase activities than leaf extracts.

  7. The Relationship between Anatomy and Photosynthetic Performance of Heterobaric Leaves1

    PubMed Central

    Nikolopoulos, Dimosthenis; Liakopoulos, Georgios; Drossopoulos, Ioannis; Karabourniotis, George

    2002-01-01

    Heterobaric leaves show heterogeneous pigmentation due to the occurrence of a network of transparent areas that are created from the bundle sheaths extensions (BSEs). Image analysis showed that the percentage of photosynthetically active leaf area (Ap) of the heterobaric leaves of 31 plant species was species dependent, ranging from 91% in Malva sylvestris to only 48% in Gynerium sp. Although a significant portion of the leaf surface does not correspond to photosynthetic tissue, the photosynthetic capacity of these leaves, expressed per unit of projected area (Pmax), was not considerably affected by the size of their transparent leaf area (At). This means that the photosynthetic capacity expressed per Ap (P*max) should increase with At. Moreover, the expression of P*max could be allowing the interpretation of the photosynthetic performance in relation to some critical anatomical traits. The P*max, irrespective of plant species, correlated with the specific leaf transparent volume (λt), as well as with the transparent leaf area complexity factor (CFAt), parameters indicating the volume per unit leaf area and length/density of the transparent tissues, respectively. Moreover, both parameters increased exponentially with leaf thickness, suggesting an essential functional role of BSEs mainly in thick leaves. The results of the present study suggest that although the Ap of an heterobaric leaf is reduced, the photosynthetic performance of each areole is increased, possibly due to the light transferring capacity of BSEs. This mechanism may allow a significant increase in leaf thickness and a consequent increase of the photosynthetic capacity per unit (projected) area, offering adaptive advantages in xerothermic environments. PMID:12011354

  8. FACS-based Isolation of Neural and Glioma Stem Cell Populations from Fresh Human Tissues Utilizing EGF Ligand

    PubMed Central

    Tome-Garcia, Jessica; Doetsch, Fiona; Tsankova, Nadejda M.

    2018-01-01

    Direct isolation of human neural and glioma stem cells from fresh tissues permits their biological study without prior culture and may capture novel aspects of their molecular phenotype in their native state. Recently, we demonstrated the ability to prospectively isolate stem cell populations from fresh human germinal matrix and glioblastoma samples, exploiting the ability of cells to bind the Epidermal Growth Factor (EGF) ligand in fluorescence-activated cell sorting (FACS). We demonstrated that FACS-isolated EGF-bound neural and glioblastoma populations encompass the sphere-forming colonies in vitro, and are capable of both self-renewal and multilineage differentiation. Here we describe in detail the purification methodology of EGF-bound (i.e., EGFR+) human neural and glioma cells with stem cell properties from fresh postmortem and surgical tissues. The ability to prospectively isolate stem cell populations using native ligand-binding ability opens new doors for understanding both normal and tumor cell biology in uncultured conditions, and is applicable for various downstream molecular sequencing studies at both population and single-cell resolution. PMID:29516026

  9. Herbivores alter plant-wind interactions by acting as a point mass on leaves and by removing leaf tissue.

    PubMed

    Kothari, Adit R; Burnett, Nicholas P

    2017-09-01

    In nature, plants regularly interact with herbivores and with wind. Herbivores can wound and alter the structure of plants, whereas wind can exert aerodynamic forces that cause the plants to flutter or sway. While herbivory has many negative consequences for plants, fluttering in wind can be beneficial for plants by facilitating gas exchange and loss of excess heat. Little is known about how herbivores affect plant motion in wind. We tested how the mass of an herbivore resting on a broad leaf of the tulip tree Liriodendron tulipifera , and the damage caused by herbivores, affected the motion of the leaf in wind. For this, we placed mimics of herbivores on the leaves, varying each herbivore's mass or position, and used high-speed video to measure how the herbivore mimics affected leaf movement and reconfiguration at two wind speeds inside a laboratory wind tunnel. In a similar setup, we tested how naturally occurring herbivore damage on the leaves affected leaf movement and reconfiguration. We found that the mass of an herbivore resting on a leaf can change that leaf's orientation relative to the wind and interfere with the ability of the leaf to reconfigure into a smaller, more streamlined shape. A large herbivore load slowed the leaf's fluttering frequency, while naturally occurring damage from herbivores increased the leaf's fluttering frequency. We conclude that herbivores can alter the physical interactions between wind and plants by two methods: (1) acting as a point mass on the plant while it is feeding and (2) removing tissue from the plant. Altering a plant's interaction with wind can have physical and physiological consequences for the plant. Thus, future studies of plants in nature should consider the effect of herbivory on plant-wind interactions, and vice versa.

  10. Inside out: efflux of carbon dioxide from leaves represents more than leaf metabolism.

    PubMed

    Stutz, Samantha S; Anderson, Jeremiah; Zulick, Rachael; Hanson, David T

    2017-05-17

    High concentrations of inorganic carbon in the xylem, produced from root, stem, and branch respiration, travel via the transpiration stream and eventually exit the plant through distant tissues as CO2. Unlike previous studies that focused on the efflux of CO2 from roots and woody tissues, we focus on efflux from leaves and the potential effect on leaf respiration measurements. We labeled transported inorganic carbon, spanning reported xylem concentrations, with 13C and then manipulated transpiration rates in the dark in order to vary the rates of inorganic carbon supply to cut leaves from Brassica napus and Populus deltoides. We used tunable diode laser absorbance spectroscopy to directly measure the rate of gross 13CO2 efflux, derived from inorganic carbon supplied from outside of the leaf, relative to gross 12CO2 efflux generated from leaf cells. These experiemnts showed that 13CO2 efflux was dependent upon the rate of inorganic carbon supply to the leaf and the rate of transpiration. Our data show that the gross leaf efflux of xylem-transported CO2 is likely small in the dark when rates of transpiration are low. However, gross leaf efflux of xylem-transported CO2 could approach half the rate of leaf respiration in the light when transpiration rates and branch inorganic carbon concentrations are high, irrespective of the grossly different petiole morphologies in our experiment. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  11. Ontogenetic changes in leaf traits of tropical rainforest trees differing in juvenile light requirement.

    PubMed

    Houter, Nico C; Pons, Thijs L

    2012-05-01

    Relationships between leaf traits and the gap dependence for regeneration, and ontogenetic changes therein, were investigated in juvenile and adult tropical rainforest tree species. The juveniles of the 17 species included in the study were grown in high light, similar to the exposed crowns of the adult trees. The traits were structural, biomechanical, chemical and photosynthetic. With increasing species gap dependence, leaf mass per area (LMA) decreased only slightly in juveniles and remained constant in adults, whereas punch strength together with tissue density decreased, and photosynthetic capacity and chlorophyll increased. Contrary to what has been mostly found in evergreen tropical rainforest, the trade-off between investment in longevity and in productivity was evident at an essentially constant LMA. Of the traits pertaining to the chloroplast level, photosynthetic capacity per unit chlorophyll increased with gap dependence, but the chlorophyll a/b ratio showed no relationship. Adults had a twofold higher LMA, but leaf strength was on average only about 50% larger. Leaf tissue density, and chlorophyll and leaf N per area were also higher, whereas chlorophyll and leaf N per unit dry mass were lower. Ranking of the species, relationships between traits and with the gap dependence of the species were similar for juveniles and adults. However, the magnitudes of most ontogenetic changes were not clearly related to a species' gap dependence. The adaptive value of the leaf traits for juveniles and adults is discussed.

  12. Relationship between fresh-packaged spinach leaves exposed to continuous light or dark and bioactive contents: Effects of cultivar, leaf size, and storage duration

    USDA-ARS?s Scientific Manuscript database

    Human-health benefits derived from consumption of fruits and vegetables are due to the many bioactive compounds found in produce. The concentrations of these bioactive compounds are heavily influenced by genetics (i.e. cultivar) and environment, especially the many pigments and vitamins that can ch...

  13. Quality of soluble organic C, N, and P produced by different types and species of litter: root litter versus leaf litter

    USDA-ARS?s Scientific Manuscript database

    In forested ecosystems, the quality of dissolved organic matter (DOM) produced by freshly senesced litter may differ by litter type and species, and these differences may influence the amount of DOM that is respired versus that which may either contribute to soil organic matter accumulation or be le...

  14. The role of forest humus in watershed management in New England

    Treesearch

    G. R., Jr. Trimble; Howard W. Lull

    1956-01-01

    Forest humus is one of the most interesting components of the forest environment. Its surface serves as a depository for leaf fall and needle fall, with successive depths marking stages of transmutation from the freshly fallen to the decomposed. And humus is responsive: humus type and depth are indicators of forest treatment and, to some extent, of site quality....

  15. Infrared sensor-based aerosol sanitization system for controlling Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on fresh produce.

    PubMed

    Kim, Sang-Oh; Ha, Jae-Won; Park, Ki-Hwan; Chung, Myung-Sub; Kang, Dong-Hyun

    2014-06-01

    An economical aerosol sanitization system was developed based on sensor technology for minimizing sanitizer usage, while maintaining bactericidal efficacy. Aerosol intensity in a system chamber was controlled by a position-sensitive device and its infrared value range. The effectiveness of the infrared sensor-based aerosolization (ISA) system to inactivate Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on spinach leaf surfaces was compared with conventional aerosolization (full-time aerosol treated), and the amount of sanitizer consumed was determined after operation. Three pathogens artificially inoculated onto spinach leaf surfaces were treated with aerosolized peracetic acid (400 ppm) for 15, 30, 45, and 60 min at room temperature (22 ± 2°C). Using the ISA system, inactivation levels of the three pathogens were equal or better than treatment with conventional full-time aerosolization. However, the amount of sanitizer consumed was reduced by ca. 40% using the ISA system. The results of this study suggest that an aerosol sanitization system combined with infrared sensor technology could be used for transportation and storage of fresh produce efficiently and economically as a practical commercial intervention.

  16. Endophytic colonization of date palm (Phoenix dactylifera L.) leaves by entomopathogenic fungi.

    PubMed

    Gómez-Vidal, S; Lopez-Llorca, L V; Jansson, H -B; Salinas, J

    2006-01-01

    Light and scanning electron microscopy together with fungal isolation techniques were used to detect entomopathogenic fungi within young and adult date palm (Phoenix dactylifera) petioles and to assess fungal survival in leaf tissues. The entomopathogenic fungi Beauveria bassiana, Lecanicillium dimorphum and Lecanicillium c.f. psalliotae survived inside leaf tissues at least 30 days after inoculation. Entomopathogenic fungi colonized inoculated petioles endophytically and were recovered up to 3cm from the inoculation site. Fungi were detected inside the parenchyma and sparsely within vascular tissue using microscopy techniques. Our results show that the entomopathogenic fungi used in this study survived and colonized date palm tissues in bioassays both under laboratory and field experimental conditions with no evidence of significant damage.

  17. L-Theanine Content and Related Gene Expression: Novel Insights into Theanine Biosynthesis and Hydrolysis among Different Tea Plant (Camellia sinensis L.) Tissues and Cultivars

    PubMed Central

    Liu, Zhi-Wei; Wu, Zhi-Jun; Li, Hui; Wang, Yong-Xin; Zhuang, Jing

    2017-01-01

    L-Theanine content has tissues and cultivars specificity in tea plant (Camellia sinensis L.), the correlations of theanine metabolic related genes expression profiles with theanine contents were explored in this study. L-theanine contents in the bud and 1st leaf, 2nd leaf, 3rd leaf, old leaf, stem, and lateral root were determined by HPLC from three C. sinensis cultivars, namely ‘Huangjinya’, ‘Anjibaicha’, and ‘Yingshuang’, respectively. The theanine contents in leaves and root of ‘Huangjinya’ were the highest, followed by ‘Anjibaicha’, and ‘Yingshuang’. The theanine contents in the leaves reduced as the leaf mature gradually, and in stem were the least. Seventeen genes encoding enzymes involved in theanine metabolism were identified from GenBank and our tea transcriptome database, including CsTS1, CsTS2, CsGS1, CsGS2, CsGOGAT-Fe, CsGOGAT-NAD(P)H, CsGDH1, CsGDH2, CsALT, CsSAMDC, CsADC, CsCuAO, CsPAO, CsNiR, CsNR, CsGGT1, and CsGGT3. The transcript profiles of those seventeen genes in the different tissues of three tea plant cultivars were analyzed comparatively. Among the different cultivars, the transcript levels of most selected genes in ‘Huangjinya’ were significantly higher than that in the ‘Anjibaicha’ and ‘Yingshuang’. Among the different tissues, the transcript levels of CsTS2, CsGS1, and CsGDH2 almost showed positive correlation with the theanine contents, while the other genes showed negative correlation with the theanine contents in most cases. The theanine contents showed correlations with related genes expression levels among cultivars and tissues of tea plant, and were determined by the integrated effect of the metabolic related genes. PMID:28439281

  18. Effects of Drought on Gene Expression in Maize Reproductive and Leaf Meristem Tissue Revealed by RNA-Seq1[W][OA

    PubMed Central

    Kakumanu, Akshay; Ambavaram, Madana M.R.; Klumas, Curtis; Krishnan, Arjun; Batlang, Utlwang; Myers, Elijah; Grene, Ruth; Pereira, Andy

    2012-01-01

    Drought stress affects cereals especially during the reproductive stage. The maize (Zea mays) drought transcriptome was studied using RNA-Seq analysis to compare drought-treated and well-watered fertilized ovary and basal leaf meristem tissue. More drought-responsive genes responded in the ovary compared with the leaf meristem. Gene Ontology enrichment analysis revealed a massive decrease in transcript abundance of cell division and cell cycle genes in the drought-stressed ovary only. Among Gene Ontology categories related to carbohydrate metabolism, changes in starch and Suc metabolism-related genes occurred in the ovary, consistent with a decrease in starch levels, and in Suc transporter function, with no comparable changes occurring in the leaf meristem. Abscisic acid (ABA)-related processes responded positively, but only in the ovaries. Related responses suggested the operation of low glucose sensing in drought-stressed ovaries. The data are discussed in the context of the susceptibility of maize kernel to drought stress leading to embryo abortion and the relative robustness of dividing vegetative tissue taken at the same time from the same plant subjected to the same conditions. Our working hypothesis involves signaling events associated with increased ABA levels, decreased glucose levels, disruption of ABA/sugar signaling, activation of programmed cell death/senescence through repression of a phospholipase C-mediated signaling pathway, and arrest of the cell cycle in the stressed ovary at 1 d after pollination. Increased invertase levels in the stressed leaf meristem, on the other hand, resulted in that tissue maintaining hexose levels at an “unstressed” level, and at lower ABA levels, which was correlated with successful resistance to drought stress. PMID:22837360

  19. Becoming less tolerant with age: sugar maple, shade, and ontogeny.

    PubMed

    Sendall, Kerrie M; Lusk, Christopher H; Reich, Peter B

    2015-12-01

    Although shade tolerance is often assumed to be a fixed trait, recent work suggests ontogenetic changes in the light requirements of tree species. We determined the influence of gas exchange, biomass distribution, and self-shading on ontogenetic variation in the instantaneous aboveground carbon balance of Acer saccharum. We quantified the aboveground biomass distributions of 18 juveniles varying in height and growing in low light in a temperate forest understory in Minnesota, USA. Gas exchange rates of leaf and stem tissues were measured, and the crown architecture of each individual was quantified. The YPLANT program was used to estimate the self-shaded fraction of each crown and to model net leaf-level carbon gain. Leaf respiration and photosynthesis per gram of leaf tissue increased with plant size. In contrast, stem respiration rates per gram of stem tissue declined, reflecting a shift in the distribution of stem diameter sizes from smaller (with higher respiration) to larger diameter classes. However, these trends were outweighed by ontogenetic increases in self-shading (which reduces the net photosynthesis realized) and stem mass fraction (which increases the proportion of purely respiratory tissue) in terms of influence on net carbon exchange. As a result, net carbon gain per gram of aboveground plant tissue declined with increasing plant size, and the instantaneous aboveground light compensation point increased. When estimates of root respiration were included to model whole-plant carbon gain and light compensation points, relationships with plant size were even more pronounced. Our findings show how an interplay of gas exchange, self-shading, and biomass distribution shapes ontogenetic changes in shade tolerance.

  20. Increased leaf angle1, a Raf-like MAPKKK that interacts with a nuclear protein family, regulates mechanical tissue formation in the Lamina joint of rice.

    PubMed

    Ning, Jing; Zhang, Baocai; Wang, Nili; Zhou, Yihua; Xiong, Lizhong

    2011-12-01

    Mitogen-activated protein kinase kinase kinases (MAPKKKs), which function at the top level of mitogen-activated protein kinase cascades, are clustered into three groups. However, no Group C Raf-like MAPKKKs have yet been functionally identified. We report here the characterization of a rice (Oryza sativa) mutant, increased leaf angle1 (ila1), resulting from a T-DNA insertion in a Group C MAPKKK gene. The increased leaf angle in ila1 is caused by abnormal vascular bundle formation and cell wall composition in the leaf lamina joint, as distinct from the mechanism observed in brassinosteroid-related mutants. Phosphorylation assays revealed that ILA1 is a functional kinase with Ser/Thr kinase activity. ILA1 is predominantly resident in the nucleus and expressed in the vascular bundles of leaf lamina joints. Yeast two-hybrid screening identified six closely related ILA1 interacting proteins (IIPs) of unknown function. Using representative IIPs, the interaction of ILA1 and IIPs was confirmed in vivo. IIPs were localized in the nucleus and showed transactivation activity. Furthermore, ILA1 could phosphorylate IIP4, indicating that IIPs may be the downstream substrates of ILA1. Microarray analyses of leaf lamina joints provided additional evidence for alterations in mechanical strength in ila1. ILA1 is thus a key factor regulating mechanical tissue formation at the leaf lamina joint.

  1. Preparation and Immunoaffinity Depletion of Fresh Frozen Tissue Homogenates for Mass Spectrometry-Based Proteomics in the Context of Drug Target/Biomarker Discovery.

    PubMed

    Prieto, DaRue A; Chan, King C; Johann, Donald J; Ye, Xiaoying; Whitely, Gordon; Blonder, Josip

    2017-01-01

    The discovery of novel drug targets and biomarkers via mass spectrometry (MS)-based proteomic analysis of clinical specimens has proven to be challenging. The wide dynamic range of protein concentration in clinical specimens and the high background/noise originating from highly abundant proteins in tissue homogenates and serum/plasma encompass two major analytical obstacles. Immunoaffinity depletion of highly abundant blood-derived proteins from serum/plasma is a well-established approach adopted by numerous researchers; however, the utilization of this technique for immunodepletion of tissue homogenates obtained from fresh frozen clinical specimens is lacking. We first developed immunoaffinity depletion of highly abundant blood-derived proteins from tissue homogenates, using renal cell carcinoma as a model disease, and followed this study by applying it to different tissue types. Tissue homogenate immunoaffinity depletion of highly abundant proteins may be equally important as is the recognized need for depletion of serum/plasma, enabling more sensitive MS-based discovery of novel drug targets, and/or clinical biomarkers from complex clinical samples. Provided is a detailed protocol designed to guide the researcher through the preparation and immunoaffinity depletion of fresh frozen tissue homogenates for two-dimensional liquid chromatography, tandem mass spectrometry (2D-LC-MS/MS)-based molecular profiling of tissue specimens in the context of drug target and/or biomarker discovery.

  2. DRAQ5 and Eosin (‘D&E’) as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues

    PubMed Central

    Elfer, Katherine N.; Sholl, Andrew B.; Wang, Mei; Tulman, David B.; Mandava, Sree H.; Lee, Benjamin R.; Brown, J. Quincy

    2016-01-01

    Real-time on-site histopathology review of biopsy tissues at the point-of-procedure has great potential for significant clinical value and improved patient care. For instance, on-site review can aid in rapid screening of diagnostic biopsies to reduce false-negative results, or in quantitative assessment of biospecimen quality to increase the efficacy of downstream laboratory and histopathology analysis. However, the only currently available rapid pathology method, frozen section analysis (FSA), is too time- and labor-intensive for use in screening large quantities of biopsy tissues and is too destructive for maximum tissue conservation in multiple small needle core biopsies. In this work we demonstrate the spectrally-compatible combination of the nuclear stain DRAQ5 and the anionic counterstain eosin as a dual-component fluorescent staining analog to hematoxylin and eosin intended for use on fresh, unsectioned tissues. Combined with optical sectioning fluorescence microscopy and pseudo-coloring algorithms, DRAQ5 and eosin (“D&E”) enables very fast, non-destructive psuedohistological imaging of tissues at the point-of-acquisition with minimal tissue handling and processing. D&E was validated against H&E on a one-to-one basis on formalin-fixed paraffin-embedded and frozen section tissues of various human organs using standard epi-fluorescence microscopy, demonstrating high fidelity of the staining mechanism as an H&E analog. The method was then applied to fresh, whole 18G renal needle core biopsies and large needle core prostate biospecimen biopsies using fluorescence structured illumination optical sectioning microscopy. We demonstrate the ability to obtain high-resolution histology-like images of unsectioned, fresh tissues similar to subsequent H&E staining of the tissue. The application of D&E does not interfere with subsequent standard-of-care H&E staining and imaging, preserving the integrity of the tissue for thorough downstream analysis. These results indicate that this dual-stain pseudocoloring method could provide a real-time histology-like image at the time of acquisition and valuable objective tissue analysis for the clinician at the time of service. PMID:27788264

  3. Susceptibility of field populations of the fall armyworm (Lepidoptera: Noctuidae) from Florida and Puerto Rico to purified Cry1F protein and corn leaf tissue containing single and pyramided Bt genes

    USDA-ARS?s Scientific Manuscript database

    Larval survival of Cry1F-susceptible (FL), -resistant (PR and Cry1F-RR), and -heterozygous (FL x PR and Cry1F-RS) populations of the fall armyworm, Spodoptera frugiperda (J.E. Smith) to purified Cry1F protein and corn leaf tissue of seven Bacillus thuringiensis (Bt) corn hybrids and five non-Bt corn...

  4. Live cell imaging reveals extensive intracellular cytoplasmic colonization of banana by normally non-cultivable endophytic bacteria.

    PubMed

    Thomas, Pious; Sekhar, Aparna Chandra

    2014-01-01

    It is generally believed that endophytic microorganisms are intercellular inhabitants present in either cultivable or non-cultivable form primarily as root colonizers. The objective of this study was to determine whether the actively mobile micro-particles observed in the intracellular matrix of fresh tissue sections of banana included endophytic bacteria. Tissue sections (50-100 µm) from apical leaf sheaths of surface-disinfected suckers (cv. Grand Naine) displayed 'Brownian motion'-reminiscent abundant motile micro-particles under bright-field and phase-contrast (×1000), which appeared similar in size and motility to the pure cultures of endophytes previously isolated from banana. Observations on callus, embryonic cells and protoplasts with intact cell wall/plasma membrane confirmed their cytoplasmic nature. The motility of these entities reduced or ceased upon tissue fixation or staining with safranin/crystal violet (0.5 % w/v), but continued uninterrupted following treatment with actin-disrupting drugs, ruling out the possibility of micro-organelles like peroxisomes. Staining with 2,3,5-triphenyl tetrazolium chloride (TTC) confirmed them to be live bacteria with similar observations after dilute safranin (0.005 %) treatment. Tissue staining with SYTO-9 coupled with epi-fluorescence or confocal laser scanning microscopy showed bacterial colonization along the peri-space between cell wall and plasma membrane initially. SYTO-9 counterstaining on TTC- or safranin-treated tissue and those subjected to enzymatic permeabilization revealed the cytoplasmic bacteria. These included organisms moving freely in the cytoplasm and those adhering to the nuclear envelope or vacuoles and the intravacuolar colonizers. The observations appeared ubiquitous to different genomes and genotypes of banana. Plating the tissue homogenate on nutrient media seldom yielded colony growth. This study, supported largely by live cell video-imaging, demonstrates enormous intracellular colonization in bananas by normally non-cultivable endophytic bacteria in two niches, namely cytoplasmic and periplasmic, designated as 'Cytobacts' and 'Peribacts', respectively. The integral intracellular association with their clonal perpetuation suggests a mutualistic relationship between endophytes and the host.

  5. Live cell imaging reveals extensive intracellular cytoplasmic colonization of banana by normally non-cultivable endophytic bacteria

    PubMed Central

    Thomas, Pious; Sekhar, Aparna Chandra

    2014-01-01

    It is generally believed that endophytic microorganisms are intercellular inhabitants present in either cultivable or non-cultivable form primarily as root colonizers. The objective of this study was to determine whether the actively mobile micro-particles observed in the intracellular matrix of fresh tissue sections of banana included endophytic bacteria. Tissue sections (50–100 µm) from apical leaf sheaths of surface-disinfected suckers (cv. Grand Naine) displayed ‘Brownian motion’-reminiscent abundant motile micro-particles under bright-field and phase-contrast (×1000), which appeared similar in size and motility to the pure cultures of endophytes previously isolated from banana. Observations on callus, embryonic cells and protoplasts with intact cell wall/plasma membrane confirmed their cytoplasmic nature. The motility of these entities reduced or ceased upon tissue fixation or staining with safranin/crystal violet (0.5 % w/v), but continued uninterrupted following treatment with actin-disrupting drugs, ruling out the possibility of micro-organelles like peroxisomes. Staining with 2,3,5-triphenyl tetrazolium chloride (TTC) confirmed them to be live bacteria with similar observations after dilute safranin (0.005 %) treatment. Tissue staining with SYTO-9 coupled with epi-fluorescence or confocal laser scanning microscopy showed bacterial colonization along the peri-space between cell wall and plasma membrane initially. SYTO-9 counterstaining on TTC- or safranin-treated tissue and those subjected to enzymatic permeabilization revealed the cytoplasmic bacteria. These included organisms moving freely in the cytoplasm and those adhering to the nuclear envelope or vacuoles and the intravacuolar colonizers. The observations appeared ubiquitous to different genomes and genotypes of banana. Plating the tissue homogenate on nutrient media seldom yielded colony growth. This study, supported largely by live cell video-imaging, demonstrates enormous intracellular colonization in bananas by normally non-cultivable endophytic bacteria in two niches, namely cytoplasmic and periplasmic, designated as ‘Cytobacts’ and ‘Peribacts’, respectively. The integral intracellular association with their clonal perpetuation suggests a mutualistic relationship between endophytes and the host. PMID:24790123

  6. A comparison of different radiative transfer model inversion methods for canopy water content retrieval

    NASA Astrophysics Data System (ADS)

    Boren, E. J.; Boschetti, L.; Johnson, D.

    2016-12-01

    With near-future droughts predicted to become both more frequent and more intense (Allen et al. 2015, Diffenbaugh et al. 2015), the estimation of satellite-derived vegetation water content would benefit a wide range of environmental applications including agricultural, vegetation, and fire risk monitoring. No vegetation water content thematic product is currently available (Yebra et al. 2013), but the successful launch of the Landsat 8 OLI and Sentinel 2A satellites, and the forthcoming Sentinel 2B, provide the opportunity for monitoring biophysical variables at a scale (10-30m) and temporal resolution (5 days) needed by most applications. Radiative transfer models (RTM) use a set of biophysical parameters to produce an estimated spectral response and - when used in inverse mode - provide a way to use satellite spectral data to estimate vegetation biophysical parameters, including water content (Zarco-Tejada et al. 2003). Using the coupled leaf and canopy level model PROSAIL5, and Landsat 8 OLI and Sentinel 2A MSI optical satellite data, the present research compares the results of three model inversion techniques: iterative optimization (OPT), look-up table (LUT), and artificial neural network (ANN) training. Ancillary biophysical data, needed for constraining the inversion process, were collected from various crop species grown in a controlled setting and under different water stress conditions. The measurements included fresh weight, dry weight, leaf area, and spectral leaf transmittance and reflectance in the 350-2500 nm range. Plot-level data, collected coincidently with satellite overpasses during three summer field campaigns in northern Idaho (2014 to 2016), are used to evaluate the results of the model inversion. Field measurements included fresh weight, dry weight, leaf area index, plant height, and top of canopy reflectance in the 350-2500 nm range. The results of the model inversion intercomparison exercised are used to characterize the uncertainties of vegetation water content estimation from Landsat 8 OLI and Sentinel 2A data.

  7. Inhibition of tomato shoot growth by over-irrigation is linked to nitrogen deficiency and ethylene.

    PubMed

    Fiebig, Antje; Dodd, Ian C

    2016-01-01

    Although physiological effects of acute flooding have been well studied, chronic effects of suboptimal soil aeration caused by over-irrigation of containerized plants have not, despite its likely commercial significance. By automatically scheduling irrigation according to soil moisture thresholds, effects of over-irrigation on soil properties (oxygen concentration, temperature and moisture), leaf growth, gas exchange, phytohormone [abscisic acid (ABA) and ethylene] relations and nutrient status of tomato (Solanum lycopersicum Mill. cv. Ailsa Craig) were studied. Over-irrigation slowly increased soil moisture and decreased soil oxygen concentration by 4%. Soil temperature was approximately 1°C lower in the over-irrigated substrate. Over-irrigating tomato plants for 2 weeks significantly reduced shoot height (by 25%) and fresh weight and total leaf area (by 60-70%) compared with well-drained plants. Over-irrigation did not alter stomatal conductance, leaf water potential or foliar ABA concentrations, suggesting that growth inhibition was not hydraulically regulated or dependent on stomatal closure or changes in ABA. However, over-irrigation significantly increased foliar ethylene emission. Ethylene seemed to inhibit growth, as the partially ethylene-insensitive genotype Never ripe (Nr) was much less sensitive to over-irrigation than the wild type. Over-irrigation induced significant foliar nitrogen deficiency and daily supplementation of small volumes of 10 mM Ca(NO3 )2 to over-irrigated soil restored foliar nitrogen concentrations, ethylene emission and shoot fresh weight of over-irrigated plants to control levels. Thus reduced nitrogen uptake plays an important role in inhibiting growth of over-irrigated plants, in part by stimulating foliar ethylene emission. © 2015 Scandinavian Plant Physiology Society.

  8. Distribution and uptake dynamics of mercury in leaves of common deciduous tree species in Minnesota, U.S.A.

    Treesearch

    Aicam Laacouri; Edward A. Nater; Randall K. Kolka

    2013-01-01

    A sequential extraction technique for compartmentalizing mercury (Hg) in leaves was developed based on a water extraction of Hg from the leaf surface followed by a solvent extraction of the cuticle. The bulk of leaf Hg was found in the tissue compartment (90-96%) with lesser amounts in the surface and cuticle compartments. Total leaf concentrations of Hg varied among...

  9. Elimination of two viruses which interact synergistically from sweetpotato by shoot tip culture and cryotherapy.

    PubMed

    Wang, Q C; Valkonen, J P T

    2008-12-01

    Sweet potato chlorotic stunt virus (SPCSV; Closteroviridae) and Sweet potato feathery mottle virus (SPFMV; Potyviridae) interact synergistically and cause severe diseases in co-infected sweetpotato plants (Ipomoea batatas). Sweetpotato is propagated vegetatively and virus-free planting materials are pivotal for sustainable production. Using cryotherapy, SPCSV and SPCSV were eliminated from all treated single-virus-infected and co-infected shoot tips irrespective of size (0.5-1.5mm including 2-4 leaf primordia). While shoot tip culture also eliminated SPCSV, elimination of SPFMV failed in 90-93% of the largest shoot tips (1.5mm) using this technique. Virus distribution to different leaf primordia and tissues within leaf primordia in the shoot apex and petioles was not altered by co-infection of the viruses in the fully virus-susceptible sweetpotato genotype used. SPFMV was immunolocalized to all types of tissues and up to the fourth-youngest leaf primordium. In contrast, SPCSV was detected only in the phloem and up to the fifth leaf primordium. Because only cells in the apical dome of the meristem and the two first leaf primordia survived cryotherapy, all data taken together could explain the results of virus elimination. The simple and efficient cryotherapy protocol developed for virus elimination can also be used for preparation of sweetpotato materials for long-term preservation.

  10. Space Pomology: Dwarf Plums for Fresh Food Production

    NASA Technical Reports Server (NTRS)

    Spencer, LaShelle; Graham, Thomas; Stutte, Gary; Massa, Gioia; Mickens, Matthew; Wheeler, Raymond

    2017-01-01

    Recently, USDA ARS researchers genetically modified plums for rapid breeding work and noticed the plants could flower and develop fruit rapidly on relatively small plants. We have tested several of these genetically modified (GM) plums in plant chambers to assess their potential as a space crop. We have been able to clone these genetic lines using cuttings that are rooted using growth regulating compounds. Results showed that the GM plums indeed flower and fruit on small plants in controlled environments similar to what might be used in space, but they require cross-pollination with pollen from a standard plum. Analysis of stomatal conductance and leaf transpiration showed that water use went up in the light period, as expected, and but that GM types typically showed higher conductance than a standard plum. Analysis of tissue showed that fruit could be a good source of potassium and phenolic compounds, which could be beneficial as a bone loss countermeasure (Smith et al., 2014). These findings are all promising for using dwarf GM plums as a supplemental food for space, but further horticultural testing is needed before they are ready.

  11. Differential distribution of metals in tree tissues growing on reclaimed coal mine overburden dumps, Jharia coal field (India).

    PubMed

    Rana, Vivek; Maiti, Subodh Kumar

    2018-04-01

    Opencast bituminous coal mining invariably generates huge amount of metal-polluted waste rocks (stored as overburden (OB) dumps) and reclaimed by planting fast growing hardy tree species which accumulate metals in their tissues. In the present study, reclaimed OB dumps located in Jharia coal field (Jharkhand, India) were selected to assess the accumulation of selected metals (Pb, Zn, Mn, Cu and Co) in tissues (leaf, stem bark, stem wood, root bark and root wood) of two commonly planted tree species (Acacia auriculiformis A.Cunn. ex Benth. and Melia azedarach L.). In reclaimed mine soil (RMS), the concentrations of pseudo-total and available metals (DTPA-extractable) were found 182-498 and 196-1877% higher, respectively, than control soil (CS). The positive Spearman's correlation coefficients between pseudo-total concentration of Pb and Cu (r = 0.717; p < 0.05), Pb and Co (r = 0.650; p < 0.05), Zn and Mn (0.359), Cu and Co (r = 0.896; p < 0.01) suggested similar sources for Pb-Cu-Co and Mn-Zn. Among the five tree tissues considered, Pb selectively accumulated in root bark, stem bark and leaves; Zn and Mn in leaves; and Cu in root wood and stem wood. These results suggested metal accumulation to be "tissue-specific". The biological indices (BCF, TF leaf , TF stem bark and TF stem wood ) indicated variation in metal uptake potential of different tree tissues. The study indicated that A. auriculiformis could be employed for Mn phytoextraction (BCF, TF leaf , TF stem bark and TF stem wood  > 1). The applicability of both the trees in Cu phytostabilization (BCF > 1; TF leaf , TF stem bark and TF stem wood  < 1) was suggested. The study enhanced knowledge about the selection of tree species for the phytoremediation of coal mine OB dumps and specific tree tissues for monitoring metal pollution.

  12. Successful slush nitrogen vitrification of human ovarian tissue.

    PubMed

    Talevi, Riccardo; Barbato, Vincenza; Fiorentino, Ilaria; Braun, Sabrina; De Stefano, Cristofaro; Ferraro, Raffaele; Sudhakaran, Sam; Gualtieri, Roberto

    2016-06-01

    To study whether slush nitrogen vitrification improves the preservation of human ovarian tissue. Control vs. treatment study. University research laboratory. Ovarian biopsies collected from nine women (aged 14-35 years) during laparoscopic surgery for benign gynecologic conditions. None. Ovarian cortical strips of 2 × 5 × 1 mm were vitrified with liquid or slush nitrogen. Fresh and vitrified cortical strips were analyzed for cryodamage and viability under light, confocal, and transmission electron microscopy. Compared with liquid nitrogen, vitrification with slush nitrogen preserves [1] follicle quality (grade 1 follicles: fresh control, 50%; liquid nitrogen, 27%; slush nitrogen, 48%); [2] granulosa cell ultrastructure (intact cells: fresh control, 92%; liquid nitrogen, 45%; slush nitrogen, 73%), stromal cell ultrastructure (intact cells: fresh control, 59.8%; liquid nitrogen, 24%; slush nitrogen, 48.7%), and DNA integrity (TUNEL-positive cells: fresh control, 0.5%; liquid nitrogen, 2.3%; slush nitrogen, 0.4%); and [3] oocyte, granulosa, and stromal cell viability (oocyte: fresh control, 90%; liquid nitrogen, 63%; slush nitrogen, 87%; granulosa cells: fresh control, 93%; liquid nitrogen, 53%; slush nitrogen, 81%; stromal cells: fresh control, 63%; liquid nitrogen, 30%; slush nitrogen, 52%). The histology, ultrastructure, and viability of follicles and stromal cells are better preserved after vitrification with slush nitrogen compared with liquid nitrogen. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  13. VirtualLeaf: an open-source framework for cell-based modeling of plant tissue growth and development.

    PubMed

    Merks, Roeland M H; Guravage, Michael; Inzé, Dirk; Beemster, Gerrit T S

    2011-02-01

    Plant organs, including leaves and roots, develop by means of a multilevel cross talk between gene regulation, patterned cell division and cell expansion, and tissue mechanics. The multilevel regulatory mechanisms complicate classic molecular genetics or functional genomics approaches to biological development, because these methodologies implicitly assume a direct relation between genes and traits at the level of the whole plant or organ. Instead, understanding gene function requires insight into the roles of gene products in regulatory networks, the conditions of gene expression, etc. This interplay is impossible to understand intuitively. Mathematical and computer modeling allows researchers to design new hypotheses and produce experimentally testable insights. However, the required mathematics and programming experience makes modeling poorly accessible to experimental biologists. Problem-solving environments provide biologically intuitive in silico objects ("cells", "regulation networks") required for setting up a simulation and present those to the user in terms of familiar, biological terminology. Here, we introduce the cell-based computer modeling framework VirtualLeaf for plant tissue morphogenesis. The current version defines a set of biologically intuitive C++ objects, including cells, cell walls, and diffusing and reacting chemicals, that provide useful abstractions for building biological simulations of developmental processes. We present a step-by-step introduction to building models with VirtualLeaf, providing basic example models of leaf venation and meristem development. VirtualLeaf-based models provide a means for plant researchers to analyze the function of developmental genes in the context of the biophysics of growth and patterning. VirtualLeaf is an ongoing open-source software project (http://virtualleaf.googlecode.com) that runs on Windows, Mac, and Linux.

  14. Low light and low ammonium are key factors for guayule leaf tissue shoot organogenesis and transformation.

    PubMed

    Dong, Niu; Montanez, Belen; Creelman, Robert A; Cornish, Katrina

    2006-02-01

    A new method has been developed for guayule tissue culture and transformation. Guayule leaf explants have a poor survival rate when placed on normal MS medium and under normal culture room light conditions. Low light and low ammonium treatment greatly improved shoot organogenesis and transformation from leaf tissues. Using this method, a 35S promoter driven BAR gene and an ubiquitin-3 promoter driven GUS gene (with intron) have been successfully introduced into guayule. These transgenic guayule plants were resistant to the herbicide ammonium-glufosinate and were positive to GUS staining. Molecular analysis showed the expected band and signal in all GUS positive transformants. The transformation efficiency with glufosinate selection ranged from 3 to 6%. Transformation with a pBIN19-based plasmid containing a NPTII gene and then selection with kanamycin also works well using this method. The ratio of kanamycin-resistant calli to total starting explants reached 50% in some experiments.

  15. Plasticity of vulnerability to leaf hydraulic dysfunction during acclimation to drought in grapevines: an osmotic-mediated process.

    PubMed

    Martorell, Sebastian; Medrano, Hipolito; Tomàs, Magdalena; Escalona, José M; Flexas, Jaume; Diaz-Espejo, Antonio

    2015-03-01

    Previous studies have reported correlation of leaf hydraulic vulnerability with pressure-volume parameters related to cell turgor. This link has been explained on the basis of the effects of turgor on connectivity among cells and tissue structural integrity, which affect leaf water transport. In this study, we tested the hypothesis that osmotic adjustment to water stress would shift the leaf vulnerability curve toward more negative water potential (Ψ leaf ) by increasing turgor at low Ψ leaf . We measured leaf hydraulic conductance (K leaf ), K leaf vulnerability [50 and 80% loss of K leaf (P50 and P80 ); |Ψ leaf | at 50 and 80% loss of K leaf , respectively), bulk leaf water relations, leaf gas exchange and sap flow in two Vitis vinifera cultivars (Tempranillo and Grenache), under two water treatments. We found that P50 , P80 and maximum K leaf decreased seasonally by more than 20% in both cultivars and watering treatments. However, K leaf at 2 MPa increased threefold, while osmotic potential at full turgor and turgor loss point decreased. Our results indicate that leaf resistance to hydraulic dysfunction is seasonally plastic, and this plasticity may be mediated by osmotic adjustment. © 2014 Scandinavian Plant Physiology Society.

  16. Comparative transcriptome analysis of different chemotypes elucidates withanolide biosynthesis pathway from medicinal plant Withania somnifera

    PubMed Central

    Gupta, Parul; Goel, Ridhi; Agarwal, Aditya Vikram; Asif, Mehar Hasan; Sangwan, Neelam Singh; Sangwan, Rajender Singh; Trivedi, Prabodh Kumar

    2015-01-01

    Withania somnifera is one of the most valuable medicinal plants synthesizing secondary metabolites known as withanolides. Despite pharmaceutical importance, limited information is available about the biosynthesis of withanolides. Chemo-profiling of leaf and root tissues of Withania suggest differences in the content and/or nature of withanolides in different chemotypes. To identify genes involved in chemotype and/or tissue-specific withanolide biosynthesis, we established transcriptomes of leaf and root tissues of distinct chemotypes. Genes encoding enzymes for intermediate steps of terpenoid backbone biosynthesis with their alternatively spliced forms and paralogous have been identified. Analysis suggests differential expression of large number genes among leaf and root tissues of different chemotypes. Study also identified differentially expressing transcripts encoding cytochrome P450s, glycosyltransferases, methyltransferases and transcription factors which might be involved in chemodiversity in Withania. Virus induced gene silencing of the sterol ∆7-reductase (WsDWF5) involved in the synthesis of 24-methylene cholesterol, withanolide backbone, suggests role of this enzyme in biosynthesis of withanolides. Information generated, in this study, provides a rich resource for functional analysis of withanolide-specific genes to elucidate chemotype- as well as tissue-specific withanolide biosynthesis. This genomic resource will also help in development of new tools for functional genomics and breeding in Withania. PMID:26688389

  17. Response of plant nutrient stoichiometry to fertilization varied with plant tissues in a tropical forest

    PubMed Central

    Mo, Qifeng; Zou, Bi; Li, Yingwen; Chen, Yao; Zhang, Weixin; Mao, Rong; Ding, Yongzhen; Wang, Jun; Lu, Xiankai; Li, Xiaobo; Tang, Jianwu; Li, Zhian; Wang, Faming

    2015-01-01

    Plant N:P ratios are widely used as indices of nutrient limitation in terrestrial ecosystems, but the response of these metrics in different plant tissues to altered N and P availability and their interactions remains largely unclear. We evaluated changes in N and P concentrations, N:P ratios of new leaves (<1 yr), older leaves (>1 yr), stems and mixed fine roots of seven species after 3-years of an N and P addition experiment in a tropical forest. Nitrogen addition only increased fine root N concentrations. P addition increased P concentrations among all tissues. The N × P interaction reduced leaf and stem P concentrations, suggesting a negative effect of N addition on P concentrations under P addition. The reliability of using nutrient ratios as indices of soil nutrient availability varied with tissues: the stoichiometric metrics of stems and older leaves were more responsive indicators of changed soil nutrient availability than those of new leaves and fine roots. However, leaf N:P ratios can be a useful indicator of inter-specific variation in plant response to nutrients availability. This study suggests that older leaf is a better choice than other tissues in the assessment of soil nutrient status and predicting plant response to altered nutrients using nutrients ratios. PMID:26416169

  18. Copper Oxide Nanoparticle Foliar Uptake, Phytotoxicity, and Consequences for Sustainable Urban Agriculture.

    PubMed

    Xiong, TianTian; Dumat, Camille; Dappe, Vincent; Vezin, Hervé; Schreck, Eva; Shahid, Muhammad; Pierart, Antoine; Sobanska, Sophie

    2017-05-02

    Throughout the world, urban agriculture supplies fresh local vegetables to city populations. However, the increasing anthropogenic uses of metal-containing nanoparticles (NPs) such as CuO-NPs in urban areas may contaminate vegetables through foliar uptake. This study focused on the CuO-NP transfer processes in leafy edible vegetables (i.e., lettuce and cabbage) to assess their potential phytotoxicity. Vegetables were exposed via leaves for 5, 10, or 15 days to various concentrations of CuO-NPs (0, 10, or 250 mg per plant). Biomass and gas exchange values were determined in relation to the Cu uptake rate, localization, and Cu speciation within the plant tissues. High foliar Cu uptake occurred after exposure for 15 days for lettuce [3773 mg (kg of dry weight) -1 ] and cabbage [4448 mg (kg of dry weight) -1 ], along with (i) decreased plant weight, net photosynthesis level, and water content and (ii) necrotic Cu-rich areas near deformed stomata containing CuO-NPs observed by scanning electron microscopy and energy dispersive X-ray microanalysis. Analysis of the CuO-NP transfer rate (7.8-242 μg day -1 ), translocation of Cu from leaves to roots and Cu speciation biotransformation in leaf tissues using electron paramagnetic resonance, suggests the involvement of plant Cu regulation processes. Finally, a potential health risk associated with consumption of vegetables contaminated with CuO-NPs was highlighted.

  19. Enterobacteriaceae resistant to third-generation cephalosporins and quinolones in fresh culinary herbs imported from Southeast Asia.

    PubMed

    Veldman, Kees; Kant, Arie; Dierikx, Cindy; van Essen-Zandbergen, Alieda; Wit, Ben; Mevius, Dik

    2014-05-02

    Since multidrug resistant bacteria are frequently reported from Southeast Asia, our study focused on the occurrence of ESBL-producing Enterobacteriaceae in fresh imported herbs from Thailand, Vietnam and Malaysia. Samples were collected from fresh culinary herbs imported from Southeast Asia in which ESBL-suspected isolates were obtained by selective culturing. Analysis included identification by MALDI-TOF mass spectrometry, susceptibility testing, XbaI-PFGE, microarray, PCR and sequencing of specific ESBL genes, PCR based replicon typing (PBRT) of plasmids and Southern blot hybridization. In addition, the quinolone resistance genotype was characterized by screening for plasmid mediated quinolone resistance (PMQR) genes and mutations in the quinolone resistance determining region (QRDR) of gyrA and parC. The study encompassed fifty samples of ten batches of culinary herbs (5 samples per batch) comprising nine different herb variants. The herbs originated from Thailand (Water morning glory, Acacia and Betel leaf), Vietnam (Parsley, Asian pennywort, Houttuynia leaf and Mint) and Malaysia (Holy basil and Parsley). By selective culturing 21 cefotaxime resistant Enterobacteriaceae were retrieved. Array analysis revealed 18 isolates with ESBL genes and one isolate with solely non-ESBL beta-lactamase genes. Mutations in the ampC promoter region were determined in two isolates with PCR and sequencing. The isolates were identified as Klebsiella pneumoniae (n=9), Escherichia coli (n=6), Enterobacter cloacae complex (n=5) and Enterobacter spp. (n=1). All isolates tested were multidrug resistant. Variants of CTX-M enzymes were predominantly found followed by SHV enzymes. PMQR genes (including aac(6')-1b-cr, qnrB and qnrS) were also frequently detected. In almost all cases ESBL and quinolone resistance genes were located on the same plasmid. Imported fresh culinary herbs from Southeast Asia are a potential source for contamination of food with multidrug resistant bacteria. Because these herbs are consumed without appropriate heating, transfer to human bacteria cannot be excluded. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Evaluation of Three Protein-Extraction Methods for Proteome Analysis of Maize Leaf Midrib, a Compound Tissue Rich in Sclerenchyma Cells.

    PubMed

    Wang, Ning; Wu, Xiaolin; Ku, Lixia; Chen, Yanhui; Wang, Wei

    2016-01-01

    Leaf morphology is closely related to the growth and development of maize (Zea mays L.) plants and final kernel production. As an important part of the maize leaf, the midrib holds leaf blades in the aerial position for maximum sunlight capture. Leaf midribs of adult plants contain substantial sclerenchyma cells with heavily thickened and lignified secondary walls and have a high amount of phenolics, making protein extraction and proteome analysis difficult in leaf midrib tissue. In the present study, three protein-extraction methods that are commonly used in plant proteomics, i.e., phenol extraction, TCA/acetone extraction, and TCA/acetone/phenol extraction, were qualitatively and quantitatively evaluated based on 2DE maps and MS/MS analysis using the midribs of the 10th newly expanded leaves of maize plants. Microscopy revealed the existence of substantial amounts of sclerenchyma underneath maize midrib epidermises (particularly abaxial epidermises). The spot-number order obtained via 2DE mapping was as follows: phenol extraction (655) > TCA/acetone extraction (589) > TCA/acetone/phenol extraction (545). MS/MS analysis identified a total of 17 spots that exhibited 2-fold changes in abundance among the three methods (using phenol extraction as a control). Sixteen of the proteins identified were hydrophilic, with GRAVY values ranging from -0.026 to -0.487. For all three methods, we were able to obtain high-quality protein samples and good 2DE maps for the maize leaf midrib. However, phenol extraction produced a better 2DE map with greater resolution between spots, and TCA/acetone extraction produced higher protein yields. Thus, this paper includes a discussion regarding the possible reasons for differential protein extraction among the three methods. This study provides useful information that can be used to select suitable protein extraction methods for the proteome analysis of recalcitrant plant tissues that are rich in sclerenchyma cells.

  1. Insights on the development, kinetics, and variation of photoinhibition using chlorophyll fluorescence imaging of a chilled, variegated leaf.

    PubMed

    Hogewoning, Sander W; Harbinson, Jeremy

    2007-01-01

    The effect of chilling on photosystem II (PSII) efficiency was studied in the variegated leaves of Calathea makoyana, in order to gain insight into the causes of chilling-induced photoinhibition. Additionally, a relationship was revealed between (chilling) stress and variation in photosynthesis. Chilling treatments (5 degrees C and 10 degrees C) were performed for different durations (1-7 d) under a moderate irradiance (120 micromol m-2 s-1). The individual leaves were divided into a shaded zone and two illuminated, chilled zones. The leaf tip and sometimes the leaf base were not chilled. Measurements of the dark-adapted Fv/Fm were made on the different leaf zones at the end of the chilling treatment, and then for several days thereafter to monitor recovery. Chilling up to 7 d in the dark did not affect PSII efficiency and visual appearance, whereas chilling in the light caused severe photoinhibition, sometimes followed by leaf necrosis. Photoinhibition increased with the duration of the chilling period, whereas, remarkably, chilling temperature had no effect. In the unchilled leaf tip, photoinhibition also occurred, whereas in the unchilled leaf base it did not. Whatever the leaf zone, photoinhibition became permanent if the mean value dropped below 0.4, although chlorosis and necrosis were associated solely with chilled illuminated tissue. Starch accumulated in the unchilled leaf tip, in contrast to the adjacent chilled irradiated zone. This suggests that photoinhibition was due to a secondary effect in the unchilled leaf tip (sink limitation), whereas it was a direct effect of chilling and irradiance in the chilled illuminated zones. The PSII efficiency and its coefficient of variation showed a unique negative linearity across all leaf zones and different tissue types. The slope of this curve was steeper for chilled leaves than it was for healthy, non-stressed leaves, suggesting that the coefficient of variation may be an important tool for assessing stress in leaves.

  2. Climatic signals registered as Carbon isotopic values in Metasequoia leaf tissues: A statistical analysis

    NASA Astrophysics Data System (ADS)

    Yang, H.; Blais, B.; Perez, G.; Pagani, M.

    2006-12-01

    To examine climatic signals registered as carbon isotopic values in leaf tissues of C3 plants, we collected mature leaf tissues from sun and shade leaves of Metasequoia trees germinated from the 1947 batch of seeds from China and planted along a latitudinal gradient of the United States. Samples from 40 individual trees, along with fossilized material from the early Tertiary of the Canadian Arctic, were analyzed for C and concentration and isotopic values using EA-IRMS after the removal of free lipids. The generated datasets were then merged with climate data compiled from each tree site recorded as average values over the past thirty years (1971-2002, NOAA database). When the isotope data were cross plotted against each geographic and climatic indicator, Latitude, Mean Annual Temperature (MAT), Average Summer Mean Temperature (ASMT)(June-August), Mean Annual Precipitation (MAP), and Average Summer Mean Precipitation (ASMP) respectively correlation patterns were revealed. The best correlating trend was obtained between temperature parameters and C isotopic values, and this correlation is stronger in the northern leaf samples than the southern samples. We discovered a strong positive correlation between latitude and the offset of C isotopic values between shade and sun leaves. This investigation represents a comprehensive examination on climatic signals registered as C isotopic values on a single species that is marked by single genetic source. The results bear implications on paleoclimatic interpretations of C isotopic signals obtained from fossil plant tissues.

  3. Effects of plant maturity and bacterial inoculum level on the colonization and internalization of escherichia coli 0157:H7 in growing spinach leaves.

    USDA-ARS?s Scientific Manuscript database

    The incidence of foodborne outbreaks linked to fresh produce has increased in the United States. Particularly noteworthy, was the 2006 Escherichia coli O157:H7 outbreak associated with pre-packaged baby spinach. The study aimed to determine whether E. coli O157:H7 would be present in the aerial leaf...

  4. Taxonomy, Traits, and Environment Determine Isoprenoid Emission from an Evergreen Tropical forest.

    NASA Astrophysics Data System (ADS)

    Taylor, T.; Alves, E. G.; Tota, J.; Oliveira Junior, R. C.; Camargo, P. B. D.; Saleska, S. R.

    2016-12-01

    Volatile isoprenoid emissions from the leaves of tropical forest trees significantly affects atmospheric chemistry, aerosols, and cloud dynamics, as well as the physiology of the emitting leaves. Emission is associated with plant tolerance to heat and drought stress. Despite a potentially central role of isoprenoid emissions in tropical forest-climate interactions, we have a poor understanding of the relationship between emissions and ecological axes of forest function. We used a custom instrument to quantify leaf isoprenoid emission rates from over 200 leaves and 80 trees at a site in the eastern Brazilian Amazon. We related standardized leaf emission capacity (EC: leaf emission rate at 1000 PAR) to tree taxonomy, height, light environment, wood traits, and leaf traits. Taxonomy was the strongest predictor of EC, and non-emitters could be found throughout the canopy. But we found that environment and leaf carbon economics constrained the upper bound of EC. For example, the relationship between EC and specific leaf area (SLA; fresh leaf area / dry mass) is described by an envelope with a decreasing upper bound on EC as SLA increases (quantile regression: 85th quantile, p<0.01). That result suggests a limitation on emissions related to leaf carbon investment strategies. EC was highest in the mid-canopy, and in leaves growing under less direct light. While inferences of ecosystem emissions focus on environmental conditions in the canopy, our results suggest that sub-canopy leaves are more responsive. This work is allowing us to develop an ecological understanding of isoprenoid emissions from forests, the basis for a predictive model of emissions that depends on both environmental factors and biological emission capacity that is grounded in plant traits and phylogeny.

  5. Secretory cavities and volatiles of Myrrhinium atropurpureum Schott var. atropurpureum (Myrtaceae): an endemic species collected in the restingas of Rio de Janeiro, Brazil.

    PubMed

    Victório, Cristiane Pimentel; Moreira, Claudio B; Souza, Marcelo da Costa; Sato, Alice; Arruda, Rosani do Carmo de Oliveira

    2011-07-01

    In this study, we investigated the leaf anatomy and the composition of volatiles in Myrrhinium atropurpureum var. atropurpureum endemic to Rio de Janeiro restingas. Particularly, leaf secretory structures were described using light microscopy, and histochemical tests were performed from fresh leaves to localize the secondary metabolites. To observe secretory cavities, fixed leaf samples were free-hand sectioned. To evaluate lipophilic compounds and terpenoids the following reagents were employed: Sudans III and IV, Red oil O and Nile blue. Leaf volatiles were characterized by gas chromatography after hydrodistillation (HD) or simultaneous distillation-extraction (SDE). Leaf analysis showed several cavities in mesophyll that are the main sites of lipophilic and terpenoid production. Monoterpenes, which represented more than 80% of the major volatiles, were characterized mainly by alpha- and beta-pinene and 1,8-cineole. In order to provide tools for M. atropurpureum identification, the following distinguishing characteristics were revealed by the following data: 1) adaxial face clear and densely punctuated by the presence of round or ellipsoidal secretory cavities randomly distributed in the mesophyll; 2) the presence of cells overlying the upper neck cells of secretory cavities; 3) the presence of numerous paracytic stomata distributed on the abaxial leaf surface, but absent in vein regions and leaf margin; and 4) non-glandular trichomes on both leaf surfaces. Our study of the compounds produced by the secretory cavities of M. atropurpureum led us to conclude that volatile terpenoid class are the main secretory compounds and that they consist of a high concentration of monoterpenes, which may indicate the phytotherapeutic importance of this plant.

  6. Effects of ultraviolet-B radiation on the growth, physiology and cannabinoid production of Cannabis sativa L

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lydon, J.

    The concentration of cannabinoids in Cannabis sativa L. is correlated with high ultraviolet-B (UV-B) radiation environments. ..delta../sup 9/-Tetrahydrocannabinolic acid and cannabidiolic acid, both major secondary products of C. sativa, absorb UV-B radiation and may function as solar screens. The object of this study was to test the effects of UV-B radiation on the physiology and cannabinoid production of C. sativa. Drug and fiber-type C. sativa were irradiated with three levels of UV-B radiation for 40 days in greenhouse experiments. Physiological measurements on leaf tissues were made by infra-red gas analysis. Drug and fiber-type control plants had similar CO/sub 2/ assimilationmore » rates from 26 to 32/sup 0/C. Drug-type control plant had higher dark respiration rates and stomatal conductances than fiber-type control plants. The concentration of ..delta../sup 9/-THC, but not of other cannabinoids) in both vegetative and reproductive tissues increased with UV-B dose in drug-type plants. None of the cannabinoids in fiber-type plants were affected by UV-B radiation. The increased level of ..delta../sup 9/-THC found in leaves after irradiation may account for the physiological and morphological insensitivity to UV-B radiation in the drug-type plants. However, fiber plants showed no comparable change in the level of cannabidoil (CBD). Resin stripped form fresh fiber-type floral tissue by sonication was spotted on filter paper and irradiated continuously for 7 days. Cannabidiol (CBD) gradually decreased when irradiated but ..delta../sup 9/-THC and cannabichromene did not.« less

  7. Foraging behavior of the mangrove sesarmid crab Neosarmatium trispinosum enhances food intake and nutrient retention in a low-quality food environment

    NASA Astrophysics Data System (ADS)

    Harada, Yota; Lee, S. Y.

    2016-06-01

    The large sesarmid crab Neosarmatium trispinosum has been reported to actively collect freshly fallen mangrove leaves and store them in its burrow where they are assumed to age prior to consumption. This leaf-catching behavior was hypothesized to improve the palatability and nutritional quality of leaves through leaching of feeding deterrent and microbial enrichment during storage. Earlier studies also hypothesized that N. trispinosum feeds on sediment or animal material to meet their N needs. A series of experiments was carried out to investigate the foraging behavior of N. trispinosum against these hypotheses. Study of foraging behavior using remotely operated cameras indicated that this crab spends the far majority of time (97.5 ± 2.5%, SD) underground and only a small percentage of time outside its burrow foraging (2.2 ± 2.3%). Collection of fresh mangrove litter was swift but no record of predation was evident over 31 h of video records. A field leaf tethering experiment showed that this crab started to consume the leaves immediately after collection rather than storing whole leaves, refuting the leaf-aging hypothesis. N. trispinosum also showed a preference for senescent yellow leaves over decaying brown leaves. This behavior may only aim to stock leaves (i.e. to ensure food availability) rather than conditioning them through decay (i.e. to improve food quality). Analysis of gut contents showed that vascular plant material was the dominant food item (83.3 ± 4.6%), followed by sediment (9.2 ± 4.6%) but no animal materials were recorded. N. trispinosum therefore relies minimally on animal food but are capable of removing 50% of the daily leaf litter production. Elemental C, N analysis shows that sediment inside the burrow is a sufficient potential food source (C/N = 13 to 15). While having a lower C/N ratio than fresh green or yellow leaves, the N content of sediment (∼0.1%) was significantly lower than those of mangrove leaves (0.3-0.9%), and may thus present as an inefficient source to meet N needs. Finally, stable isotope analysis indicated a dominant role of mangrove C in the diet of the crab but less important in terms of N. Behavioural coupled with physiological adaptations for a low-quality diet allows N. trispinosum to efficiently retain and recycle mangrove production in situ, with implications for local organic matter and microalgal distributions. Mechanisms by which N. trispinosum meets its N demand, however, remain obscure.

  8. Universal Algorithms for Plant Phenotyping: Are we there yet?

    NASA Astrophysics Data System (ADS)

    Kakani, V. G.; Kambham, R. R.; Zhao, D.; Foster, A. J.; Gowda, P. H.

    2017-12-01

    Hyperspectral remote sensing offers ability to capture spectral signatures of plant morpho-physio-biochemical traits at multiple scales (leaf to canopy to aerial). Experimental results on plant phenotype from pot, growth chamber and field studies at multiple location were used in this study. Pigment, leaf/plant water status, plant nutrient status, plant height, leaf area, fresh and dry weights of biomass and its components are correlated with hyperspectral reflectance signatures. Leaf reflectance was collected with spectroradiometer having a light source. Canopy hyperspectral reflectance was collected from 1.5 m above the canopy using a spectroradiometer, while multispectral images were acquired from aerial platforms ( 400m). Several statistical methods including simple ratios, principal component analysis, and partial least squares regression were used to identify hyperspectral reflectance bands that were tightly associated with plant phenotypic traits. Leaf level spectra best described the morpho-physio-biochemical traits (R2 = 0.6-0.9), while canopy reflectance best described plant height (R2 = 0.65), leaf area index (R2 = 0.67-0.74) and biomass (R2 = 0.69-0.78), while aerial spectra improved canopy level regression coefficients for plant height (R2 = 0.93) and leaf area index (R2 = 0.89). The comparison of multi-level spectra and resolution, clearly showed the advantage of hyperspectral reflectance data over the multispectral reflectance data, particularly for understanding the basis for spectral reflectance differences among species and traits. In conclusion, high resolution (1-2 cm) spectral imagery can help to bridge the gap across multiple levels of phenotype measurement.

  9. Responses of the two‐spotted oak buprestid, Agrilus biguttatus (Coleoptera: Buprestidae), to host tree volatiles

    PubMed Central

    Woodcock, Christine M; Sumner, Mary E; Caulfield, John C; Reed, Katy; Inward, Daegan JG; Leather, Simon R; Pickett, John A; Birkett, Michael A; Denman, Sandra

    2016-01-01

    Abstract BACKGROUND Agrilus bigutattus (Fabricius) is a forest pest of increasing importance in the United Kingdom. The larvae damage weakened native oaks and are thought to contribute to premature tree death. Suspected links with acute oak decline (AOD) are not yet confirmed, but AOD‐predisposed trees appear to become more susceptible to A. biguttatus attack. Thus, management may be necessary for control of this insect. To explore the possibility of monitoring beetle populations by baited traps, the host tree volatiles regulating A. biguttatus–oak interactions were studied. RESULTS Biologically active volatile organic compounds in dynamic headspace extracts of oak foliage and bark were identified initially by coupled gas chromatography–electroantennography (GC‐EAG) and GC–mass spectrometry (GC‐MS), and the structures were confirmed by GC coinjection with authentic compounds. Of two synthetic blends of these compounds comprising the active leaf volatiles, the simpler one containing three components evoked strongly positive behavioural responses in four‐arm olfactometer tests with virgin females and males, although fresh leaf material was more efficient than the blend. The other blend, comprising a five‐component mixture made up of bark volatiles, proved to be as behaviourally active for gravid females as bark tissue. CONCLUSIONS These initial results on A. biguttatus chemical ecology reveal aspects of the role of attractive tree volatiles in the host‐finding of beetles and underpin the development of semiochemically based surveillance strategies for this forest insect. © 2015 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. PMID:26663022

  10. [Relationship between leaf anatomical structure and heat resistance of 15 Rhododendron cultivars].

    PubMed

    Shen, Hui Fei; Zhao, Bing; Xu, Jing Jing

    2016-12-01

    In this study, 17 anatomical structure indexes of 15 Rhododendron cultivars were mea-sured by scanning electron microscope (SEM). Leaf anatomical structure indexes were screened via coefficient of variation, analysis of correlation and hierarchical cluster analysis, and comprehensive evaluation on heat resistance for each cultivar was conducted by the subordinate function. The results showed that the leaves of Rhododendron cultivars were typical bifacial leaf and the epidermal anticlinal walls showed slightly sinuate. The stomata only distributed in the lower epidermis and the shape was ruleless. The anatomical structure indexes all reached a significant level difference among 15 cultivars (P<0.01), except for lower epidermis thickness (P<0.05). Thickness of lamina corneum, stomatal density, stomatal width, the thickness palisade tissue and looseness of leaf spongy tissue were the main factors related to the hardness, while other indexes, such as stomatal length, stoma aperture, stomatal opening, length and thickness of upper epidermis, length and thickness of lower epidermis, thickness of spongy tissue, the ratio of the palisade tissue to spongy tissue, tightness of leaf palisade tissue, leaf thickness and media thickness didn't show much effect on heat resistance. There were some differences among 15 cultivars in heat resistance, and the order was Rhododendron 'Song Jiang Da Tao Hong' > Rhododendron 'Zhuang Yuan Hong' > Rhododendron 'Lv Se Guang Hui' > Rhododendron 'Fen Zhen Zhu' > Rhododendron 'Wai Guo Hong' > Rhododendron 'Lan Yin' > Rhododendron 'Bi Zhi' >Rhododendron 'Da He Zhi Chun' > Rhododendron 'Guo Qi Hong' > Rhododendron 'Yu Ling Long' > Rhododendron 'Hong Shan Hu' > Rhododendron 'Ning Bo Hong' > Rhododendron 'Tao Ban Zhu Sha' > Rhododendron 'Ai Ding Bao' > Rhododendron 'Liu Qiu Hong'. According to the heat hardiness, the cultivars could be divided into 4 groups: R. 'Song Jiang Da Tao Hong', R. 'Zhuang Yuan Hong' and R. 'Lv Se Guang Hui' with high heat resistance, R. 'Fen Zhen Zhu', R. 'Wai Guo Hong', R. 'Lan Yin', R. 'Bi Zhi', R. 'Da He Zhi Chun', R. 'Guo Qi Hong' and R. 'Yu Ling Long' with medium heat resistance, R. 'Hong Shan Hu', R. 'Ning Bo Hong', R. 'Tao Ban Zhu Sha' and R. 'Ai Ding Bao' with lower heat resistance, R. 'Liu Qiu Hong' without heat resistance. However, the accurate heat hardiness evaluation of Rhododendron still needs to consider other factors, including morphological structure, physiological and biochemical indicators and genetic factor of heat resistance, the harmfulness of Rhododendron, and the recovery state after being injured by high temperature.

  11. Dependence of Plant Uptake and Diffusion of Polycyclic Aromatic Hydrocarbons on the Leaf Surface Morphology and Micro-structures of Cuticular Waxes

    NASA Astrophysics Data System (ADS)

    Li, Qingqing; Li, Yungui; Zhu, Lizhong; Xing, Baoshan; Chen, Baoliang

    2017-04-01

    The uptake of organic chemicals by plants is considered of great significance as it impacts their environmental transport and fate and threatens crop growth and food safety. Herein, the dependence of the uptake, penetration, and distribution of sixteen polycyclic aromatic hydrocarbons (PAHs) on the morphology and micro-structures of cuticular waxes on leaf surfaces was investigated. Plant surface morphologies and wax micro-structures were examined by scanning emission microscopy, and hydrophobicities of plant surfaces were monitored through contact angle measurements. PAHs in the cuticles and inner tissues were distinguished by sequential extraction, and the cuticle was verified to be the dominant reservoir for the accumulation of lipophilic pollutants. The interspecies differences in PAH concentrations cannot be explained by normalizing them to the plant lipid content. PAHs in the inner tissues became concentrated with the increase of tissue lipid content, while a generally negative correlation between the PAH concentration in cuticles and the epicuticular wax content was found. PAHs on the adaxial and abaxial sides of a leaf were differentiated for the first time, and the divergence between these two sides can be ascribed to the variations in surface morphologies. The role of leaf lipids was redefined and differentiated.

  12. Nutritive value and haemolytic properties (in vitro) of the leaves of Vernonia amygdalina on human erythrocyte.

    PubMed

    Oboh, G

    2006-01-01

    Vernonia amygdalina (Bitter leaf) is popularly consumed in many homes in Nigeria as a vegetable in soup preparation, while the infusion is used in folk medicine in the treatment of malaria fever. In this study, fresh leaves of Vernonia amygdalina were subjected to abrasion (to remove the bitter taste) as is normally done in soup preparation, subsequently the nutrient, anti-nutrient and zinc bioavailability of the processed and unprocessed leaf were determined. The heamolytic effect of the leaf infusion on human erythrocytes (Genotype-SS, AS, AA) was also determined. The study revealed that Vernonia amygdalina leaf has high protein (33.3%), fat (10.1%), crude fibre (29.2%), ash (11.7%), mineral (Na, K, Ca, Mg, Zn & Fe), phytate (1015.4mg/100g) and tannin (0.6%) content, while it contain low cyanide (1.1mg/kg). Abrasion of the leaf caused a significant decrease (P<0.05) in both the nutrient and antinutrient content of the leaf except in carbohydrate where there was a significant increase (P<0.05) and Mg where there was no significant change (P>0.05). The result of the calculated [Ca][phytate]/ [Zn] molar ratio (the best index for the prediction of Zn bioavailability) indicated that the phytate content of both the processed and unprocessed Vernonia amygdalina leaf will not reduce the Zn content to a critical level (0.5mol./kg). Further studies on the leaf revealed that the infusion induced the haemolysis of human erythrocyte (In vitro). Human genotype-SS (1024) were highly susceptible to haemolysis induced by Vernonia amygdalina infusion, genotype-AS moderately susceptible (512) while genotype-AA (256) is highly resistant to haemolysis induced by the same infusion. Bitter leaf has high nutrient and antinutrient content, however, abrasion will bring about a significant decrease (P<0.05) in both. The infusion of the Vernonia amygdalina leaf induces the haemolysis of mammalian erythrocyte in vitro, with Human-SS having the highest susceptibility.

  13. Physiological, vascular and nanomechanical assessment of hybrid poplar leaf traits in micropropagated plants and plants propagated from root cuttings: A contribution to breeding programs.

    PubMed

    Ďurkovič, Jaroslav; Husárová, Hana; Javoříková, Lucia; Čaňová, Ingrid; Šuleková, Miriama; Kardošová, Monika; Lukáčik, Ivan; Mamoňová, Miroslava; Lagaňa, Rastislav

    2017-09-01

    Micropropagated plants experience significant stress from rapid water loss when they are transferred from an in vitro culture to either greenhouse or field conditions. This is caused both by inefficient stomatal control of transpiration and the change to a higher light intensity and lower humidity. Understanding the physiological, vascular and biomechanical processes that allow micropropagated plants to modify their phenotype in response to environmental conditions can help to improve both field performance and plant survival. To identify changes between the hybrid poplar [Populus tremula × (Populus × canescens)] plants propagated from in vitro tissue culture and those from root cuttings, we assessed leaf performance for any differences in leaf growth, photosynthetic and vascular traits, and also nanomechanical properties of the tracheary element cell walls. The micropropagated plants showed significantly higher values for leaf area, leaf length, leaf width and leaf dry mass. The greater leaf area and leaf size dimensions resulted from the higher transpiration rate recorded for this stock type. Also, the micropropagated plants reached higher values for chlorophyll a fluorescence parameters and for the nanomechanical dissipation energy of tracheary element cell walls which may indicate a higher damping capacity within the primary xylem tissue under abiotic stress conditions. The performance of the plants propagated from root cuttings was superior for instantaneous water-use efficiency which signifies a higher acclimation capacity to stressful conditions during a severe drought particularly for this stock type. Similarities were found among the majority of the examined leaf traits for both vegetative plant origins including leaf mass per area, stomatal conductance, net photosynthetic rate, hydraulic axial conductivity, indicators of leaf midrib vascular architecture, as well as for the majority of cell wall nanomechanical traits. This research revealed that there were no drawbacks in the leaf physiological performance which could be attributed to the micropropagated plants of fast growing hybrid poplar. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  14. Phytohormones signaling and crosstalk regulating leaf angle in rice.

    PubMed

    Luo, Xiangyu; Zheng, Jingsheng; Huang, Rongyu; Huang, Yumin; Wang, Houcong; Jiang, Liangrong; Fang, Xuanjun

    2016-12-01

    Leaf angle is an important agronomic trait in rice (Oryza sativa L.). It affects both the efficiency of sunlight capture and nitrogen reservoirs. The erect leaf phenotype is suited for high-density planting and thus increasing crop yields. Many genes regulate leaf angle by affecting leaf structure, such as the lamina joint, mechanical tissues, and the midrib. Signaling of brassinosteroids (BR), auxin (IAA), and gibberellins (GA) plays important roles in the regulation of lamina joint bending in rice. In addition, the biosynthesis and signaling of BR are known to have dominant effects on leaf angle development. In this review, we summarize the factors and genes associated with the development of leaf angle in rice, outline the regulatory mechanisms based on the signaling of BR, IAA, and GA, and discuss the contribution of crosstalk between BR and IAA or GA in the formation of leaf angle. Promising lines of research in the transgenic engineering of rice leaf angle to increase grain yield are proposed.

  15. Isochoric and isobaric freezing of fish muscle.

    PubMed

    Năstase, Gabriel; Lyu, Chenang; Ukpai, Gideon; Şerban, Alexandru; Rubinsky, Boris

    2017-04-01

    We have recently shown that, a living organism, which succumbs to freezing to -4 °C in an isobaric thermodynamic system (constant atmospheric pressure), can survive freezing to -4 °C in an isochoric thermodynamic system (constant volume). It is known that the mechanism of cell damage in an isobaric system is the freezing caused increase in extracellular osmolality, and, the consequent cell dehydration. An explanation for the observed survival during isochoric freezing is the thermodynamic modeling supported hypothesis that, in the isochoric frozen solution the extracellular osmolality is comparable to the cell intracellular osmolality. Therefore, cells in the isochoric frozen organism do not dehydrate, and the tissue maintains its morphological integrity. Comparing the histology of: a) fresh fish white muscle, b) fresh muscle frozen to -5 °C in an isobaric system and c) fresh muscle frozen to -5 °C I in an isochoric system, we find convincing evidence of the mechanism of cell dehydration during isobaric freezing. In contrast, the muscle tissue frozen to -5 °C in an isochoric system appears morphologically identical to fresh tissue, with no evidence of dehydration. This is the first experimental evidence in support of the hypothesis that in isochoric freezing there is no cellular dehydration and therefore the morphology of the frozen tissue remains intact. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Changes in water content and distribution in Quercus ilex leaves during progressive drought assessed by in vivo 1H magnetic resonance imaging

    PubMed Central

    2010-01-01

    Background Drought is a common stressor in many regions of the world and current climatic global circulation models predict further increases in warming and drought in the coming decades in several of these regions, such as the Mediterranean basin. The changes in leaf water content, distribution and dynamics in plant tissues under different soil water availabilities are not well known. In order to fill this gap, in the present report we describe our study withholding the irrigation of the seedlings of Quercus ilex, the dominant tree species in the evergreen forests of many areas of the Mediterranean Basin. We have monitored the gradual changes in water content in the different leaf areas, in vivo and non-invasively, by 1H magnetic resonance imaging (MRI) using proton density weighted (ρw) images and spin-spin relaxation time (T2) maps. Results ρw images showed that the distal leaf area lost water faster than the basal area and that after four weeks of similar losses, the water reduction was greater in leaf veins than in leaf parenchyma areas and also in distal than in basal leaf area. There was a similar tendency in all different areas and tissues, of increasing T2 values during the drought period. This indicates an increase in the dynamics of free water, suggesting a decrease of cell membranes permeability. Conclusions The results indicate a non homogeneous leaf response to stress with a differentiated capacity to mobilize water between its different parts and tissues. This study shows that the MRI technique can be a useful tool to follow non-intrusively the in vivo water content changes in the different parts of the leaves during drought stress. It opens up new possibilities to better characterize the associated physiological changes and provides important information about the different responses of the different leaf areas what should be taken into account when conducting physiological and metabolic drought stress studies in different parts of the leaves during drought stress. PMID:20735815

  17. Changes in water content and distribution in Quercus ilex leaves during progressive drought assessed by in vivo 1H magnetic resonance imaging.

    PubMed

    Sardans, Jordi; Peñuelas, Josep; Lope-Piedrafita, Silvia

    2010-08-24

    Drought is a common stressor in many regions of the world and current climatic global circulation models predict further increases in warming and drought in the coming decades in several of these regions, such as the Mediterranean basin. The changes in leaf water content, distribution and dynamics in plant tissues under different soil water availabilities are not well known. In order to fill this gap, in the present report we describe our study withholding the irrigation of the seedlings of Quercus ilex, the dominant tree species in the evergreen forests of many areas of the Mediterranean Basin. We have monitored the gradual changes in water content in the different leaf areas, in vivo and non-invasively, by 1H magnetic resonance imaging (MRI) using proton density weighted (rhow) images and spin-spin relaxation time (T2) maps. Rhow images showed that the distal leaf area lost water faster than the basal area and that after four weeks of similar losses, the water reduction was greater in leaf veins than in leaf parenchyma areas and also in distal than in basal leaf area. There was a similar tendency in all different areas and tissues, of increasing T2 values during the drought period. This indicates an increase in the dynamics of free water, suggesting a decrease of cell membranes permeability. The results indicate a non homogeneous leaf response to stress with a differentiated capacity to mobilize water between its different parts and tissues. This study shows that the MRI technique can be a useful tool to follow non-intrusively the in vivo water content changes in the different parts of the leaves during drought stress. It opens up new possibilities to better characterize the associated physiological changes and provides important information about the different responses of the different leaf areas what should be taken into account when conducting physiological and metabolic drought stress studies in different parts of the leaves during drought stress.

  18. Comparison of fluorescent and high-pressure sodium lamps on growth of leaf lettuce

    NASA Technical Reports Server (NTRS)

    Koontz, H. V.; Prince, R. P.; Koontz, R. F.; Knott, W. M. (Principal Investigator)

    1987-01-01

    Radiation from high-pressure sodium (HPS) lamps provided more than a 50% increased yield (fresh and dry weight of tops) of loose-leaf lettuce cultivars Grand Rapids Forcing and RubyConn, compared to that obtained by radiation from cool-white fluorescent (CWF) lamps at equal photosynthetic photon flux; yet, input wattage was approximately 36% less. It was postulated that the considerable output of 700 to 850 nm radiation from the HPS lamp was a significant factor of the increased yield. Under HPS lamps, the leaves of both cultivars were slightly less green with very little red pigmentation ('RubyConn') and slightly elongated, compared to CWF, but plant productivity per unit electrical energy input was vastly superior with HPS.

  19. Comparing five simple vascular storage protocols.

    PubMed

    van Doormaal, Tristan P C; Sluijs, Jurren H; Vink, Aryan; Tulleken, Cornelis A F; van der Zwan, Albert

    2014-11-01

    We aim to find a storage protocol for vessels that preserves their dimensional, histologic, and mechanical characteristics to facilitate reproducible anastomosis experiments and microsurgical training with constant quality. We compared stored rabbit aortas, harvested in a slaughterhouse, using five different protocols with fresh controls. Aortas were preserved for 125 d in (1) NaCl 0.9% at -18°C, (2) Roswell Park Memorial Institute 1640 90% with 10% dimethyl sulfoxide (RPMI/DMSO) at -18°C, (3) RPMI/DMSO at -70°C, (4) glycerol 85% at 4°C, and (5) glycerol in stepwise increased concentrations until 85% at 4°C. After preservation, we measured vessel diameter, wall thickness, and Young's Modulus indicating stiffness. Neurosurgeons compared stored vessels with fresh vessels, blinded for preservation subgroup. We performed histologic assessment blinded for preservation subgroup. Fresh rabbit aortas showed a mean diameter of 2.65 ± 0.14 mm, a mean wall thickness of 126 ± 22 μm, and a Young's Modulus of 11.4 ± 2.4 N/mm(2). NaCl 0.9%-preserved aortas showed a significantly increased vessel diameter and decreased stiffness. RPMI/DMSO-preserved aortas showed no significant differences from fresh aortas in dimensions and mechanical characteristics. Glycerol-preserved tissue showed a significant increase in wall thickness, a related significant decrease in diameter, and increase in stiffness. Neurosurgeons regarded RPMI/DMSO tissue as most comparable with fresh tissue. Histologic assessment revealed no differences between the different protocols and fresh control group. Storage of rabbit aortas in RPMI/DMSO most adequately preserves their dimensional and mechanical properties. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Evaluating relative contribution of osmotolerance and tissue tolerance mechanisms toward salinity stress tolerance in three Brassica species.

    PubMed

    Chakraborty, Koushik; Bose, Jayakumar; Shabala, Lana; Eyles, Alieta; Shabala, Sergey

    2016-10-01

    Three different species of Brassica, with differential salt sensitivity were used to understand physiological mechanisms of salt tolerance operating in these species and to evaluate the relative contribution of different strategies to cope with salt load. Brassica napus was the most tolerant species in terms of the overall performance, with Brassica juncea and Brassica oleracea being much more sensitive to salt stress with no obvious difference between them. While prominent reduction in net CO2 assimilation was observed in both sensitive species, physiological mechanisms beyond this reduction differed strongly. Brassica juncea plants possessed high osmotolerance and were able to maintain high transpiration rate but showed a significant reduction in leaf chlorophyll content and efficiency of leaf photochemistry. On the contrary, B. oleracea plants possessed the highest (among the three species) tissue tolerance but showed a very significant stomatal limitation of photosynthesis. Electrophysiological experiments revealed that the high tissue tolerance in B. oleracea was related to the ability of leaf mesophyll cells to maintain highly negative membrane potential in the presence of high apoplastic Na(+) . In addition to high osmotolerance, the most tolerant B. napus showed also lesser accumulation of toxic Na(+) and Cl(-) in the leaf, possessed moderate tissue tolerance and had a superior K(+) retention ability. Taken together, the results from this study indicate that the three Brassica species employ very different mechanisms to cope with salinity and, despite its overall sensitivity to salinity, B. oleracea could be recommended as a valuable 'donor' of tissue tolerance genes to confer this trait for marker-assisted breeding programs. © 2016 Scandinavian Plant Physiology Society.

  1. Double-filter identification of vascular-expressed genes using Arabidopsis plants with vascular hypertrophy and hypotrophy.

    PubMed

    Ckurshumova, Wenzislava; Scarpella, Enrico; Goldstein, Rochelle S; Berleth, Thomas

    2011-08-01

    Genes expressed in vascular tissues have been identified by several strategies, usually with a focus on mature vascular cells. In this study, we explored the possibility of using two opposite types of altered tissue compositions in combination with a double-filter selection to identify genes with a high probability of vascular expression in early organ primordia. Specifically, we generated full-transcriptome microarray profiles of plants with (a) genetically strongly reduced and (b) pharmacologically vastly increased vascular tissues and identified a reproducible cohort of 158 transcripts that fulfilled the dual requirement of being underrepresented in (a) and overrepresented in (b). In order to assess the predictive value of our identification scheme for vascular gene expression, we determined the expression patterns of genes in two unbiased subsamples. First, we assessed the expression patterns of all twenty annotated transcription factor genes from the cohort of 158 genes and found that seventeen of the twenty genes were preferentially expressed in leaf vascular cells. Remarkably, fifteen of these seventeen vascular genes were clearly expressed already very early in leaf vein development. Twelve genes with published leaf expression patterns served as a second subsample to monitor the representation of vascular genes in our cohort. Of those twelve genes, eleven were preferentially expressed in leaf vascular tissues. Based on these results we propose that our compendium of 158 genes represents a sample that is highly enriched for genes expressed in vascular tissues and that our approach is particularly suited to detect genes expressed in vascular cell lineages at early stages of their inception. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  2. The evaluation of long-term effects of cinnamon bark and olive leaf on toxicity induced by streptozotocin administration to rats.

    PubMed

    Onderoglu, S; Sozer, S; Erbil, K M; Ortac, R; Lermioglu, F

    1999-11-01

    The effects of cinnamon bark and olive leaf have been investigated on streptozotocin-induced tissue injury, and some biochemical and haematological changes in rats. The effects on glycaemia were also evaluated. Long-term administration of olive leaf caused significant improvement in tissue injury induced by streptozotocin treatment; the effect of cinnamon bark was less extent. No effects on blood glucose levels were detected. However, significant decreases in some increased biochemical and haematological parameters of streptozotocin-treated rats were observed. Aspartate aminotransferase, urea and cholesterol levels were significantly decreased by treatment with both plant materials, and alanine aminotransferase by treatment with olive leaf. Cinnamon bark also caused a significant decrease in platelet counts. In addition, any visible toxicity, except decrease in body weight gain, attributable to the long-term use of plant materials was not established in normal rats. The data indicate that long-term use of olive leaf and cinnamon bark may provide benefit against diabetic conditions. Determination of underlying mechanism(s) of beneficial effects, toxicity to other systems and clinical assessments of related plant materials are major topics requiring further studies.

  3. A Rice PECTATE LYASE-LIKE Gene Is Required for Plant Growth and Leaf Senescence1[OPEN

    PubMed Central

    Leng, Yujia; Yang, Yaolong; Ren, Deyong; Dai, Liping; Wang, Yuqiong; Chen, Long; Tu, Zhengjun; Gao, Yihong; Zhu, Li; Hu, Jiang; Gao, Zhenyu; Guo, Longbiao; Lin, Yongjun

    2017-01-01

    To better understand the molecular mechanisms behind plant growth and leaf senescence in monocot plants, we identified a mutant exhibiting dwarfism and an early-senescence leaf phenotype, termed dwarf and early-senescence leaf1 (del1). Histological analysis showed that the abnormal growth was caused by a reduction in cell number. Further investigation revealed that the decline in cell number in del1 was affected by the cell cycle. Physiological analysis, transmission electron microscopy, and TUNEL assays showed that leaf senescence was triggered by the accumulation of reactive oxygen species. The DEL1 gene was cloned using a map-based approach. It was shown to encode a pectate lyase (PEL) precursor that contains a PelC domain. DEL1 contains all the conserved residues of PEL and has strong similarity with plant PelC. DEL1 is expressed in all tissues but predominantly in elongating tissues. Functional analysis revealed that mutation of DEL1 decreased the total PEL enzymatic activity, increased the degree of methylesterified homogalacturonan, and altered the cell wall composition and structure. In addition, transcriptome assay revealed that a set of cell wall function- and senescence-related gene expression was altered in del1 plants. Our research indicates that DEL1 is involved in both the maintenance of normal cell division and the induction of leaf senescence. These findings reveal a new molecular mechanism for plant growth and leaf senescence mediated by PECTATE LYASE-LIKE genes. PMID:28455404

  4. Reversible Deformation of Transfusion Tracheids in Taxus baccata Is Associated with a Reversible Decrease in Leaf Hydraulic Conductance1[OPEN

    PubMed Central

    Zhang, Yong-Jiang; Rockwell, Fulton E.; Wheeler, James K.; Holbrook, N. Michele

    2014-01-01

    Declines in leaf hydraulic conductance (Kleaf) with increasing water stress have been attributed to cavitation of the leaf xylem. However, in the leaves of conifers, the reversible collapse of transfusion tracheids may provide an alternative explanation. Using Taxus baccata, a conifer species without resin, we developed a modified rehydration technique that allows the separation of declines in Kleaf into two components: one reversible and one irreversible upon relaxation of water potential to −1 MPa. We surveyed leaves at a range of water potentials for evidence of cavitation using cryo-scanning electron microscopy and quantified dehydration-induced structural changes in transfusion tracheids by cryo-fluorescence microscopy. Irreversible declines in Kleaf did not occur until leaf water potentials were more negative than −3 MPa. Declines in Kleaf between −2 and −3 MPa were reversible and accompanied by the collapse of transfusion tracheids, as evidenced by cryo-fluorescence microscopy. Based on cryo-scanning electron microscopy, cavitation of either transfusion or xylem tracheids did not contribute to declines in Kleaf in the reversible range. Moreover, the deformation of transfusion tracheids was quickly reversible, thus acting as a circuit breaker regulating the flux of water through the leaf vasculature. As transfusion tissue is present in all gymnosperms, the reversible collapse of transfusion tracheids may be a general mechanism in this group for the protection of leaf xylem from excessive loads generated in the living leaf tissue. PMID:24948828

  5. A Rice PECTATE LYASE-LIKE Gene Is Required for Plant Growth and Leaf Senescence.

    PubMed

    Leng, Yujia; Yang, Yaolong; Ren, Deyong; Huang, Lichao; Dai, Liping; Wang, Yuqiong; Chen, Long; Tu, Zhengjun; Gao, Yihong; Li, Xueyong; Zhu, Li; Hu, Jiang; Zhang, Guangheng; Gao, Zhenyu; Guo, Longbiao; Kong, Zhaosheng; Lin, Yongjun; Qian, Qian; Zeng, Dali

    2017-06-01

    To better understand the molecular mechanisms behind plant growth and leaf senescence in monocot plants, we identified a mutant exhibiting dwarfism and an early-senescence leaf phenotype, termed dwarf and early-senescence leaf1 ( del1 ). Histological analysis showed that the abnormal growth was caused by a reduction in cell number. Further investigation revealed that the decline in cell number in del1 was affected by the cell cycle. Physiological analysis, transmission electron microscopy, and TUNEL assays showed that leaf senescence was triggered by the accumulation of reactive oxygen species. The DEL1 gene was cloned using a map-based approach. It was shown to encode a pectate lyase (PEL) precursor that contains a PelC domain. DEL1 contains all the conserved residues of PEL and has strong similarity with plant PelC. DEL1 is expressed in all tissues but predominantly in elongating tissues. Functional analysis revealed that mutation of DEL1 decreased the total PEL enzymatic activity, increased the degree of methylesterified homogalacturonan, and altered the cell wall composition and structure. In addition, transcriptome assay revealed that a set of cell wall function- and senescence-related gene expression was altered in del1 plants. Our research indicates that DEL1 is involved in both the maintenance of normal cell division and the induction of leaf senescence. These findings reveal a new molecular mechanism for plant growth and leaf senescence mediated by PECTATE LYASE-LIKE genes. © 2017 American Society of Plant Biologists. All Rights Reserved.

  6. Olea europaea L. leaf extract and derivatives: antioxidant properties.

    PubMed

    Briante, Raffaella; Patumi, Maurizio; Terenziani, Stefano; Bismuto, Ettore; Febbraio, Ferdinando; Nucci, Roberto

    2002-08-14

    This paper reports a very simple and fast method to collect eluates with high amounts of hydroxytyrosol, biotransforming Olea europaea L. leaf extract by a thermophilic beta-glycosidase immobilized on chitosan. Some phenolic compounds in the leaf tissue and in the eluates obtained by biotransformation are identified. To propose the eluates as natural substances from a vegetal source, their antioxidant properties have been compared with those of the leaf extract from which they are originated. The eluates possess a higher concentration of simple phenols, characterized by a stronger antioxidant capacity, than those available in extra virgin olive oils and in many tablets of olive leaf extracts, commercially found as dietetic products and food integrators.

  7. Characterization anatomical leaf blade five species Nepenthes from Kerinci Seblat National Park, Kerinci regency, Jambi Province

    NASA Astrophysics Data System (ADS)

    Al Farishy, D. D.; Nisyawati, Metusala, D.

    2017-07-01

    Nepenthes is one of carnivorous plant genera which have key characters on leaf and pitcher as the modification. However, wide varieties of morphological features on pitcher intraspecies and between species could be tough for identification process. The objective was to provide alternative characters for identification process by anatomical features. Kerinci Seblat National Park was chosen because lack of update data on wild type of species there. Whole five species were collected at Lingkat Lake and Gunung Tujuh Lake as representative lowland and highland species. Leaves collected fresh, flawless, and has grown pitcher. Each leaf was separated into the paradermal and transversal section, dehydrated by series alcohol, and stained by safranin and fast green. Sections observed by light microscope. Result show there were specific differences between species that could be potential to be key characters. That features are stomatal density, stomatal length, sessile glands surface shaped, sessile glands density, trichome distribution, adaxial cuticle thickness, adaxial hypodermic thickness, and the number of layers of adaxial hypodermis

  8. Antioxidant and Antimicrobial Effects of Pistacia lentiscus L. Extracts in Pork Sausages

    PubMed Central

    Botsaris, George; Orphanides, Antia; Yiannakou, Evgenia; Gekas, Vassilis

    2015-01-01

    Summary Pistacia lentiscus fruits are ingredients of traditional Cypriot sausages. The objective of this study is to evaluate P. lentiscus extracts as natural additives to the sausages. First, the phenolic content and antioxidant activity of fruit and leaf extracts were determined. Results revealed that leaves are richer source of polyphenolic antioxidants than fruits, with methanol being the better extraction solvent. In the next step, the antioxidant and antimicrobial effects of methanolic extracts (300 mg/kg) in the pork sausage formulation were investigated. Peroxide, acid and thiobarbituric acid-reactive substance values demonstrated that both fruit and leaf extracts reduced the rate of lipid oxidation of sausages at 4 °C. Total viable count revealed significant differences on the fifth day of storage, with better microbial inhibition by leaf extract. No significant differences between the extracts were observed after the tenth day of storage. Overall, the extracts can be used to prevent lipid oxidation and reduce microbial spoilage during the first days of storage of fresh traditional pork sausages. PMID:27904382

  9. Antioxidant and Antimicrobial Effects of Pistacia lentiscus L. Extracts in Pork Sausages.

    PubMed

    Botsaris, George; Orphanides, Antia; Yiannakou, Evgenia; Gekas, Vassilis; Goulas, Vlasios

    2015-12-01

    Pistacia lentiscus fruits are ingredients of traditional Cypriot sausages. The objective of this study is to evaluate P. lentiscus extracts as natural additives to the sausages. First, the phenolic content and antioxidant activity of fruit and leaf extracts were determined. Results revealed that leaves are richer source of polyphenolic antioxidants than fruits, with methanol being the better extraction solvent. In the next step, the antioxidant and antimicrobial effects of methanolic extracts (300 mg/kg) in the pork sausage formulation were investigated. Peroxide, acid and thiobarbituric acid-reactive substance values demonstrated that both fruit and leaf extracts reduced the rate of lipid oxidation of sausages at 4 °C. Total viable count revealed significant differences on the fifth day of storage, with better microbial inhibition by leaf extract. No significant differences between the extracts were observed after the tenth day of storage. Overall, the extracts can be used to prevent lipid oxidation and reduce microbial spoilage during the first days of storage of fresh traditional pork sausages.

  10. A 115-year δ15N record of cumulative nitrogen pollution in California serpentine grasslands

    NASA Astrophysics Data System (ADS)

    Vallano, D.; Zavaleta, E. S.

    2010-12-01

    Until the 1980s, California’s biodiverse serpentine grasslands were threatened primarily by development and protected by reserve creation. However, nitrogen (N) fertilization due to increasing fossil fuel emissions in the expanding Bay Area is thought to be contributing to rapid, recent invasion of these ecosystems by exotic annual grasses that are displacing rare and endemic serpentine species. Documenting the cumulative effects of N deposition in this ecosystem can direct policy and management actions to mitigate the role of N deposition in its transformation. Natural abundance stable isotopes of N in vegetation have been increasingly used as bio-indicators of N deposition patterns and subsequent changes to plant N cycling and assimilation. However, the long-term record of atmospheric reactive N enrichment and the resulting changes in ecosystem N dynamics have yet to be adequately reconstructed in many ecosystems. Museum archives of vascular plant tissue are valuable sources of materials to reconstruct temporal and spatial isotopic patterns of N inputs to ecosystems. Here, we present N stable isotope data from archived and current specimens of an endemic California serpentine grassland species, leather oak (Quercus durata), since 1895 across the greater San Francisco Bay region. We measured spatial and temporal trends in stable isotope composition (δ15N and δ13C) and concentration (%N and %C) of historical and current samples of leather oak leaves from sites within the Bay Area, impacted by increasing development, and sites northeast of the Bay Area, with significantly lower rates of urbanization and industrialization. Specifically, we sampled dry museum and fresh leaf specimens from serpentine sites within Lake (n=27) and Santa Clara (n=30) counties dating from 1895 to 2010. Leaf δ15N values were stable from 1895 to the 1950s and then decreased strongly throughout the last 50 years as fossil fuel emissions rapidly increased in the Bay Area, indicating that N pollution is being retained in serpentine grassland ecosystems. Leaf δ15N values in the high-deposition region declined at a rate of -0.041‰ yr-1, while leaf δ15N values in the low-deposition region did not show a strong pattern. In both regions, leaf δ13C values declined through time as atmospheric CO2 concentrations increased in response to fossil fuel combustion (the Suess effect). Leaf %N and %C values did not present any clear patterns at sites within or outside of the Bay Area. We conclude that using natural abundance stable isotope values in leaves can indicate variation in N pollution inputs across wide spatial and temporal scales and that archived plant samples can provide valuable baselines against which to assess changes in regional N cycling and subsequent ecological impacts on vegetation.

  11. Evaluation of breast tissue with confocal strip-mosaicking microscopy: a test approach emulating pathology-like examination

    PubMed Central

    Abeytunge, Sanjee; Larson, Bjorg; Peterson, Gary; Morrow, Monica; Rajadhyaksha, Milind

    2017-01-01

    Abstract. Confocal microscopy is an emerging technology for rapid imaging of freshly excised tissue without the need for frozen- or fixed-section processing. Initial studies have described imaging of breast tissue using fluorescence confocal microscopy with small regions of interest, typically 750×750  μm2. We present exploration with a microscope, termed confocal strip-mosaicking microscope (CSM microscope), which images an area of 2×2  cm2 of tissue with cellular-level resolution in 10 min of excision. Using the CSM microscope, we imaged 34 fresh, human, large breast tissue specimens from 18 patients, blindly analyzed by a board-certified pathologist and subsequently correlated with the corresponding standard fixed histopathology. Invasive tumors and benign tissue were clearly identified in CSM strip-mosaic images. Thirty specimens were concordant for image-to-histopathology correlation while four were discordant. PMID:28327961

  12. A New Paradigm for Biospecimen Banking in the Personalized Medicine Era

    PubMed Central

    McDonald, Sandra A.; Watson, Mark A.; Rossi, Joan; Becker, Colleen M.; Jaques, David P.; Pfeifer, John D.

    2012-01-01

    Banking of high-quality, appropriately consented human tissue is crucial for the understanding of disease pathogenesis and translation of such knowledge into improvements in patient care. Traditionally, tissue banking has been thought of as primarily an academic research activity, but tissue and biospecimen banking is increasingly assuming clinical importance, especially with the advent of genetic and proteomic testing approaches that rely on fresh or fresh frozen tissue. These approaches are part of the revolution in personalized medicine. This revolution’s impact on biorepositories—their mission and day-to-day function—will be profound. Direct patient care will require structuring tissue procurement to become a routine part of patient care. Accordingly tissue banking will expand from its traditional research role in large academic medical centers into the everyday practice of surgical pathology. Successful implementation of this model will require consideration of several financial, medicolegal, and administrative issues. PMID:22031304

  13. Considerations for ex vivo thermal tissue testing exemplified using the fresh porcine longissimus muscle model for endometrial ablation

    NASA Astrophysics Data System (ADS)

    Fugett, James H.; Bennett, Haydon E.; Shrout, Joshua L.; Coad, James E.

    2017-02-01

    Expansions in minimally invasive medical devices and technologies with thermal mechanisms of action are continuing to advance the practice of medicine. These expansions have led to an increasing need for appropriate animal models to validate and quantify device performance. The planning of these studies should take into consideration a variety of parameters, including the appropriate animal model (test system - ex vivo or in vivo; species; tissue type), treatment conditions (test conditions), predicate device selection (as appropriate, control article), study timing (Day 0 acute to more than Day 90 chronic survival studies), and methods of tissue analysis (tissue dissection - staining methods). These considerations are discussed and illustrated using the fresh extirpated porcine longissimus muscle model for endometrial ablation.

  14. Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model.

    PubMed

    López, Luisa F; Muñoz, César O; Cáceres, Diego H; Tobón, Ángela M; Loparev, Vladimir; Clay, Oliver; Chiller, Tom; Litvintseva, Anastasia; Gade, Lalitha; González, Ángel; Gómez, Beatriz L

    2017-01-01

    Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis.

  15. Mechanical contribution of lamellar and interlamellar elastin along the mouse aorta.

    PubMed

    Clark, T E; Lillie, M A; Vogl, A W; Gosline, J M; Shadwick, R E

    2015-10-15

    The mechanical properties of aortic elastin vary regionally, but the microstructural basis for this variation is unknown. This study was designed to identify the relative contributions of lamellar and interlamellar elastin to circumferential load bearing in the mouse thoracic and abdominal aortas. Forces developed in uniaxial tests of samples of fresh and autoclaved aorta were correlated with elastin content and morphology obtained from histology and multiphoton laser scanning microscopy. Autoclaving should render much of the interlamellar elastin mechanically incompetent. In autoclaved tissue force per unit sample width correlated with lamellar elastin content (P≪0.001) but not total elastin content. In fresh tissue at low strain where elastin dominates the mechanical response, forces were higher than in the autoclaved tissue, but force did not correlate with total elastin content. Therefore although interlamellar elastin likely contributed to the stiffness in the fresh aorta, its contribution appeared not in proportion to its quantity. In both fresh and autoclaved tissue, elastin stiffness consistently decreased along the abdominal aorta, a key area for aneurysm development, and this difference could not be fully accounted for on the basis of either lamellar or total elastin content. These findings are relevant to the development of mathematical models of arterial mechanics, particularly for mouse models of arterial diseases involving elastic tissue. In microstructural based models the quantity of each mural constituent determines its contribution to the total response. This study shows elastin's mechanical response cannot necessarily be accounted for on the basis of fibre quantity, orientation, and modulus. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Multiplexed Molecular Imaging of Fresh Tissue Surfaces Enabled by Convection-Enhanced Topical Staining with SERS-Coded Nanoparticles.

    PubMed

    Wang, Yu W; Doerksen, Josh D; Kang, Soyoung; Walsh, Daniel; Yang, Qian; Hong, Daniel; Liu, Jonathan T C

    2016-10-01

    There is a need for intraoperative imaging technologies to guide breast-conserving surgeries and to reduce the high rates of re-excision for patients in which residual tumor is found at the surgical margins during postoperative pathology analyses. Feasibility studies have shown that utilizing topically applied surface-enhanced Raman scattering (SERS) nanoparticles (NPs), in conjunction with the ratiometric imaging of targeted versus untargeted NPs, enables the rapid visualization of multiple cell-surface biomarkers of cancer that are overexpressed at the surfaces of freshly excised breast tissues. In order to reliably and rapidly perform multiplexed Raman-encoded molecular imaging of large numbers of biomarkers (with five or more NP flavors), an enhanced staining method has been developed in which tissue surfaces are cyclically dipped into an NP-staining solution and subjected to high-frequency mechanical vibration. This dipping and mechanical vibration (DMV) method promotes the convection of the SERS NPs at fresh tissue surfaces, which accelerates their binding to their respective biomarker targets. By utilizing a custom-developed device for automated DMV staining, this study demonstrates the ability to simultaneously image four cell-surface biomarkers of cancer at the surfaces of fresh human breast tissues with a mixture of five flavors of SERS NPs (four targeted and one untargeted control) topically applied for 5 min and imaged at a spatial resolution of 0.5 mm and a raster-scanned imaging rate of >5 cm 2 min -1 . © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. The energetic and carbon economic origins of leaf thermoregulation.

    PubMed

    Michaletz, Sean T; Weiser, Michael D; McDowell, Nate G; Zhou, Jizhong; Kaspari, Michael; Helliker, Brent R; Enquist, Brian J

    2016-08-22

    Leaf thermoregulation has been documented in a handful of studies, but the generality and origins of this pattern are unclear. We suggest that leaf thermoregulation is widespread in both space and time, and originates from the optimization of leaf traits to maximize leaf carbon gain across and within variable environments. Here we use global data for leaf temperatures, traits and photosynthesis to evaluate predictions from a novel theory of thermoregulation that synthesizes energy budget and carbon economics theories. Our results reveal that variation in leaf temperatures and physiological performance are tightly linked to leaf traits and carbon economics. The theory, parameterized with global averaged leaf traits and microclimate, predicts a moderate level of leaf thermoregulation across a broad air temperature gradient. These predictions are supported by independent data for diverse taxa spanning a global air temperature range of ∼60 °C. Moreover, our theory predicts that net carbon assimilation can be maximized by means of a trade-off between leaf thermal stability and photosynthetic stability. This prediction is supported by globally distributed data for leaf thermal and photosynthetic traits. Our results demonstrate that the temperatures of plant tissues, and not just air, are vital to developing more accurate Earth system models.

  18. Endophytic fungi reduce leaf-cutting ant damage to seedlings

    PubMed Central

    Bittleston, L. S.; Brockmann, F.; Wcislo, W.; Van Bael, S. A.

    2011-01-01

    Our study examines how the mutualism between Atta colombica leaf-cutting ants and their cultivated fungus is influenced by the presence of diverse foliar endophytic fungi (endophytes) at high densities in tropical leaf tissues. We conducted laboratory choice trials in which ant colonies chose between Cordia alliodora seedlings with high (Ehigh) or low (Elow) densities of endophytes. The Ehigh seedlings contained 5.5 times higher endophyte content and a greater diversity of fungal morphospecies than the Elow treatment, and endophyte content was not correlated with leaf toughness or thickness. Leaf-cutting ants cut over 2.5 times the leaf area from Elow relative to Ehigh seedlings and had a tendency to recruit more ants to Elow plants. Our findings suggest that leaf-cutting ants may incur costs from cutting and processing leaves with high endophyte loads, which could impact Neotropical forests by causing variable damage rates within plant communities. PMID:20610420

  19. Isolation of guard cells from fresh epidermis using a piezo-power micro-dissection system with vibration-attenuated needles.

    PubMed

    Terpitz, Ulrich; Zimmermann, Dirk

    2010-01-01

    The Eppendorf Piezo-Power Microdissection (PPMD) system uses a tungsten needle (MicroChisel) oscillating in a forward-backward (vertical) mode to cut cells from surrounding tissue. This technology competes with laser-based dissection systems, which offer high accuracy and precision, but are more expensive and require fixed tissue. In contrast, PPMD systems can dissect freshly prepared tissue, but their accuracy and precision is lower due to unwanted lateral vibrations of the MicroChisel. Especially in tissues where elasticity is high, these vibrations can limit the cutting resolution or hamper the dissection. Here we describe a cost-efficient and simple glass capillary-encapsulation modification of MicroChisels for effective attenuation of lateral vibrations. The use of modified MicroChisels enables accurate and precise tissue dissection from highly elastic material.

  20. Strip mosaicing confocal microscopy for rapid imaging over large areas of excised tissue

    NASA Astrophysics Data System (ADS)

    Abeytunge, Sanjee; Li, Yongbiao; Larson, Bjorg; Peterson, Gary; Toledo-Crow, Ricardo; Rajadhyaksha, Milind

    2012-03-01

    Confocal mosaicing microscopy is a developing technology platform for imaging tumor margins directly in fresh tissue, without the processing that is required for conventional pathology. Previously, basal cell carcinoma margins were detected by mosaicing of confocal images of 12 x 12 mm2 of excised tissue from Mohs surgery. This mosaicing took 9 minutes. Recently we reported the initial feasibility of a faster approach called "strip mosaicing" on 10 x 10 mm2 of tissue that was demonstrated in 3 minutes. In this paper we report further advances in instrumentation and software. Rapid mosaicing of confocal images on large areas of fresh tissue potentially offers a means to perform pathology at the bedside. Thus, strip mosaicing confocal microscopy may serve as an adjunct to pathology for imaging tumor margins to guide surgery.

  1. Leaf gas films contribute to rice (Oryza sativa) submergence tolerance during saline floods.

    PubMed

    Herzog, Max; Konnerup, Dennis; Pedersen, Ole; Winkel, Anders; Colmer, Timothy David

    2018-05-01

    Floods and salinization of agricultural land adversely impact global rice production. We investigated whether gas films on leaves of submerged rice delay salt entry during saline submergence. Two-week-old plants with leaf gas films (+GF) or with gas films experimentally removed (-GF) were submerged in artificial floodwater with 0 or 50 mm NaCl for up to 16 d. Gas films were present >9 d on GF plants after which gas films were diminished. Tissue ion analysis (Na + , Cl - and K + ) showed that gas films caused some delay of Na + entry, as leaf Na + concentration was 36-42% higher in -GF leaves than +GF leaves on days 1-5. However, significant net uptakes of Na + and Cl - , and K + net loss, occurred despite the presence of gas films, indicating the likely presence of some leaf-to-floodwater contact, so that the gas layer must not have completely separated the leaf surfaces from the water. Natural loss and removal of gas films resulted in severe declines in growth, underwater photosynthesis, chlorophyll a and tissue porosity. Submergence was more detrimental to leaf P N and growth than the additional effect of 50 mm NaCl, as salt did not significantly affect underwater P N at 200 μm CO 2 nor growth. © 2016 John Wiley & Sons Ltd.

  2. Lotus leaf extract and L-carnitine influence different processes during the adipocyte life cycle.

    PubMed

    Siegner, Ralf; Heuser, Stefan; Holtzmann, Ursula; Söhle, Jörn; Schepky, Andreas; Raschke, Thomas; Stäb, Franz; Wenck, Horst; Winnefeld, Marc

    2010-08-05

    The cellular and molecular mechanisms of adipose tissue biology have been studied extensively over the last two decades. Adipose tissue growth involves both an increase in fat cell size and the formation of mature adipocytes from precursor cells. To investigate how natural substances influence these two processes, we examined the effects of lotus leaf extract (Nelumbo nucifera-extract solution obtained from Silab, France) and L-carnitine on human preadipocytes and adipocytes. For our in vitro studies, we used a lotus leaf extract solution alone or in combination with L-carnitine. Utilizing cultured human preadipocytes, we investigated lotus leaf extract solution-induced inhibition of triglyceride incorporation during adipogenesis and possible effects on cell viability. Studies on human adipocytes were performed aiming to elucidate the efficacy of lotus leaf extract solution to stimulate lipolytic activity. To further characterize lotus leaf extract solution-mediated effects, we determined the expression of the transcription factor adipocyte determination and differentiation factor 1 (ADD1/SREBP-1c) on the RNA- and protein level utilizing qRT-PCR and immunofluorescence analysis. Additionally, the effect of L-carnitine on beta-oxidation was analyzed using human preadipocytes and mature adipocytes. Finally, we investigated additive effects of a combination of lotus leaf extract solution and L-carnitine on triglyceride accumulation during preadipocyte/adipocyte differentiation. Our data showed that incubation of preadipocytes with lotus leaf extract solution significantly decreased triglyceride accumulation during adipogenesis without affecting cell viability. Compared to controls, adipocytes incubated with lotus leaf extract solution exhibited a significant increase in lipolysis-activity. Moreover, cell populations cultivated in the presence of lotus leaf extract solution showed a decrease in adipocyte differentiation capacity as indicated by a decrease in the ADD1/SREBP-1c signal. Importantly, our results demonstrated that a combination of lotus leaf extract solution and L-carnitine reduced triglyceride accumulation to a greater extent compared to incubation with either substance alone. Overall, our data demonstrate that a combination of lotus leaf extract and L-carnitine reduced triglyceride accumulation in human (pre)adipocytes by affecting different processes during the adipocyte life cycle. For this reason, this combination might represent a treatment option for obesity-related diseases.

  3. Responses of tropical legumes from the Brazilian Atlantic Rainforest to simulated acid rain.

    PubMed

    Andrade, Guilherme C; Silva, Luzimar C

    2017-07-01

    We investigated the morphological and anatomical effects of simulated acid rain on leaves of two species native to the Brazilian Atlantic Rainforest: Paubrasilia echinata and Libidibia ferrea var. leiostachya. Saplings were subjected to acid rain in a simulation chamber during 10 days for 15 min daily, using H 2 SO 4 solution pH 3.0 and, in the control, deionized water. At the end of the experiment, fragments from young and expanding leaves were anatomically analyzed. Although L. ferrea var. leiostachya leaves are more hydrophobic, rain droplets remained in contact with them for a longer time, as in the hydrophilic P. echinata leaves, droplets coalesce and rapidly run off. Visual symptomatology consisted in interveinal and marginal necrotic dots. Microscopic damage found included epicuticular wax flaking, turgor loss and epidermal cell shape alteration, hypertrophy of parenchymatous cells, and epidermal and mesophyll cell collapse. Formation of a wound tissue was observed in P. echinata, and it isolated the necrosis to the adaxial leaf surface. Acid rain increased thickness of all leaf tissues except spongy parenchyma in young leaves of L. ferrea var. leiostachya, and such thickness was maintained throughout leaf expansion. To our knowledge, this is the first report of acidity causing increase in leaf tissue thickness. This could represent the beginning of cell hypertrophy, which was seen in visually affected leaf regions. Paubrasilia echinata was more sensitive, showing earlier symptoms, but the anatomical damage in L. ferrea var. leiostachya was more severe, probably due to the higher time of contact with acid solution in this species.

  4. Leaf gas exchange performance and the lethal water potential of five European species during drought.

    PubMed

    Li, Shan; Feifel, Marion; Karimi, Zohreh; Schuldt, Bernhard; Choat, Brendan; Jansen, Steven

    2016-02-01

    Establishing physiological thresholds to drought-induced mortality in a range of plant species is crucial in understanding how plants respond to severe drought. Here, five common European tree species were selected (Acer campestre L., Acer pseudoplatanus L., Carpinus betulus L., Corylus avellana L. and Fraxinus excelsior L.) to study their hydraulic thresholds to mortality. Photosynthetic parameters during desiccation and the recovery of leaf gas exchange after rewatering were measured. Stem vulnerability curves and leaf pressure-volume curves were investigated to understand the hydraulic coordination of stem and leaf tissue traits. Stem and root samples from well-watered and severely drought-stressed plants of two species were observed using transmission electron microscopy to visualize mortality of cambial cells. The lethal water potential (ψlethal) correlated with stem P99 (i.e., the xylem water potential at 99% loss of hydraulic conductivity, PLC). However, several plants that were stressed beyond the water potential at 100% PLC showed complete recovery during the next spring, which suggests that the ψlethal values were underestimated. Moreover, we observed a 1 : 1 relationship between the xylem water potential at the onset of embolism and stomatal closure, confirming hydraulic coordination between leaf and stem tissues. Finally, ultrastructural changes in the cytoplasm of cambium tissue and mortality of cambial cells are proposed to provide an alternative approach to investigate the point of no return associated with plant death. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  5. Conversion of D-hamamelose into 2-carboxy-D-arabinitol and 2-carboxy-D-arabinitol 1-phosphate in leaves of Phaseolus vulgaris L.

    PubMed

    Andralojc, P J; Keys, A J; Martindale, W; Dawson, G W; Parry, M A

    1996-10-25

    [1-14C]Hamamelose (2-hydroxymethyl-D-ribose) was synthesized by reaction of ribulose 5-phosphate with potassium [14C]cyanide, catalytic hydrogenation of the resulting cyanohydrin, and dephosphorylation of the product. Its identity was established by a chromatographic comparison with hamamelose isolated from the bark of witch hazel (Hamamelis virginiana L.). Following vacuum infiltration of the [1-14C]hamamelose into leaf discs from Phaseolus vulgaris L., 14C-labeled 2carboxy-D-arabinitol (CA) and 2-carboxy-D-arabinitol 1-phosphate (CA1P) were formed, in the dark. Conversion of hamamelose to both CA and CA1P in the leaf discs was inhibited by dithiothreitol and sodium fluoride, although at high concentrations of these inhibitors conversion into CA was still evident when conversion into CA1P was totally inhibited. Wheat (Triticum aestivum L.) leaves converted hamamelose into CA without formation of CA1P. Leaves from P. vulgaris contained 68 nmol.g-1 fresh weight of hamamelose in the light and 35 nmol.g-1 fresh weight in the dark. A pathway for the biosynthesis of CA1P from Calvin cycle intermediates is proposed which includes the sequence: hamamelose --> CA --> CA1P.

  6. A linear spectral matching technique for retrieving equivalent water thickness and biochemical constituents of green vegetation

    NASA Technical Reports Server (NTRS)

    Gao, Bo-Cai; Goetz, Alexander F. H.

    1992-01-01

    Over the last decade, technological advances in airborne imaging spectrometers, having spectral resolution comparable with laboratory spectrometers, have made it possible to estimate biochemical constituents of vegetation canopies. Wessman estimated lignin concentration from data acquired with NASA's Airborne Imaging Spectrometer (AIS) over Blackhawk Island in Wisconsin. A stepwise linear regression technique was used to determine the single spectral channel or channels in the AIS data that best correlated with measured lignin contents using chemical methods. The regression technique does not take advantage of the spectral shape of the lignin reflectance feature as a diagnostic tool nor the increased discrimination among other leaf components with overlapping spectral features. A nonlinear least squares spectral matching technique was recently reported for deriving both the equivalent water thicknesses of surface vegetation and the amounts of water vapor in the atmosphere from contiguous spectra measured with the Airborne Visible/Infrared Imaging Spectrometer (AVIRIS). The same technique was applied to a laboratory reflectance spectrum of fresh, green leaves. The result demonstrates that the fresh leaf spectrum in the 1.0-2.5 microns region consists of spectral components of dry leaves and the spectral component of liquid water. A linear least squares spectral matching technique for retrieving equivalent water thickness and biochemical components of green vegetation is described.

  7. Metabolic dependence of green tea on plucking positions revisited: a metabolomic study.

    PubMed

    Lee, Jang-Eun; Lee, Bum-Jin; Hwang, Jeong-Ah; Ko, Kwang-Sup; Chung, Jin-Oh; Kim, Eun-Hee; Lee, Sang-Jun; Hong, Young-Shick

    2011-10-12

    The dependence of global green tea metabolome on plucking positions was investigated through (1)H nuclear magnetic resonance (NMR) analysis coupled with multivariate statistical data set. Pattern recognition methods, such as principal component analysis (PCA) and orthogonal projection on latent structure-discriminant analysis (OPLS-DA), were employed for a finding metabolic discrimination among fresh green tea leaves plucked at different positions from young to old leaves. In addition to clear metabolic discrimination among green tea leaves, elevations in theanine, caffeine, and gallic acid levels but reductions in catechins, such as epicatechin (EC), epigallocatechin (EGC), epicatechin-3-gallate (ECG), and epigallocatechin-3-gallate (EGCG), glucose, and sucrose levels were observed, as the green tea plant grows up. On the other hand, the younger the green tea leaf is, the more theanine, caffeine, and gallic acid but the lesser catechins accumlated in the green tea leaf, revealing a reverse assocation between theanine and catechins levels due to incorporaton of theanine into catechins with growing up green tea plant. Moreover, as compared to the tea leaf, the observation of marked high levels of theanine and low levels of catechins in green tea stems exhibited a distinct tea plant metabolism between the tea leaf and the stem. This metabolomic approach highlights taking insight to global metabolic dependence of green tea leaf on plucking position, thereby providing distinct information on green tea production with specific tea quality.

  8. Subtractive transcriptome analysis of leaf and rhizome reveals differentially expressed transcripts in Panax sokpayensis.

    PubMed

    Gurung, Bhusan; Bhardwaj, Pardeep K; Talukdar, Narayan C

    2016-11-01

    In the present study, suppression subtractive hybridization (SSH) strategy was used to identify rare and differentially expressed transcripts in leaf and rhizome tissues of Panax sokpayensis. Out of 1102 randomly picked clones, 513 and 374 high quality expressed sequenced tags (ESTs) were generated from leaf and rhizome subtractive libraries, respectively. Out of them, 64.92 % ESTs from leaf and 69.26 % ESTs from rhizome SSH libraries were assembled into different functional categories, while others were of unknown function. In particular, ESTs encoding galactinol synthase 2, ribosomal RNA processing Brix domain protein, and cell division cycle protein 20.1, which are involved in plant growth and development, were most abundant in the leaf SSH library. Other ESTs encoding protein KIAA0664 homologue, ubiquitin-activating enzyme e11, and major latex protein, which are involved in plant immunity and defense response, were most abundant in the rhizome SSH library. Subtractive ESTs also showed similarity with genes involved in ginsenoside biosynthetic pathway, namely farnesyl pyrophosphate synthase, squalene synthase, and dammarenediol synthase. Expression profiles of selected ESTs validated the quality of libraries and confirmed their differential expression in the leaf, stem, and rhizome tissues. In silico comparative analyses revealed that around 13.75 % of unigenes from the leaf SSH library were not represented in the available leaf transcriptome of Panax ginseng. Similarly, around 18.12, 23.75, 25, and 6.25 % of unigenes from the rhizome SSH library were not represented in available root/rhizome transcriptomes of P. ginseng, Panax notoginseng, Panax quinquefolius, and Panax vietnamensis, respectively, indicating a major fraction of novel ESTs. Therefore, these subtractive transcriptomes provide valuable resources for gene discovery in P. sokpayensis and would complement the available transcriptomes from other Panax species.

  9. Direct-to-PCR tissue preservation for DNA profiling.

    PubMed

    Sorensen, Amy; Berry, Clare; Bruce, David; Gahan, Michelle Elizabeth; Hughes-Stamm, Sheree; McNevin, Dennis

    2016-05-01

    Disaster victim identification (DVI) often occurs in remote locations with extremes of temperatures and humidities. Access to mortuary facilities and refrigeration are not always available. An effective and robust DNA sampling and preservation procedure would increase the probability of successful DNA profiling and allow faster repatriation of bodies and body parts. If the act of tissue preservation also released DNA into solution, ready for polymerase chain reaction (PCR), the DVI process could be further streamlined. In this study, we explored the possibility of obtaining DNA profiles without DNA extraction, by adding aliquots of preservative solutions surrounding fresh human muscle and decomposing human muscle and skin tissue samples directly to PCR. The preservatives consisted of two custom preparations and two proprietary solutions. The custom preparations were a salt-saturated solution of dimethyl sulfoxide (DMSO) with ethylenediaminetetraacetic (EDTA) and TENT buffer (Tris, EDTA, NaCl, Tween 20). The proprietary preservatives were DNAgard (Biomatrica(®)) and Tissue Stabilising Kit (DNA Genotek). We obtained full PowerPlex(®) 21 (Promega) and GlobalFiler(®) (Life Technologies) DNA profiles from fresh and decomposed tissue preserved at 35 °C for up to 28 days for all four preservatives. The preservative aliquots removed from the fresh muscle tissue samples had been stored at -80 °C for 4 years, indicating that long-term archival does not diminish the probability of successful DNA typing. Rather, storage at -80 °C seems to reduce PCR inhibition.

  10. Ontogeny of Daucus carota Infected with Meloidogyne hapla

    PubMed Central

    Slinger, L. A.; Bird, G. W.

    1978-01-01

    The ontogeny of carrots (Daucus carota cv. 'Spartan Premium') grown under greenhouse conditions in pots of organic soil infected with Meloidogyne hapla was influenced detrimentally as early as 4 days after seeding, as determined through analysis of plant surface area, dry weight, fresh weight, net assimilation rate, relative growth rate, and leaf-area ratio. Only 58% of the diseased carrots were suitable for fresh market, compared with 97% of those grown in nematode-free soil. Growth and development of the shoot system (height, surface area, dry weight, and fresh weight) were retarded by M. hapla as early as 12 days after seeding. During the first 12 days after seeding, root dry weight was greater for diseased plants than for controls. Root growth and development (surface area, dry weight, and fresh weight) associated with this nematode, however, were retarded as early as 16 days after seeding. M. hapla caused a delay in the occurrence of 2nd-, 4th-, and 5th-order roots, and an increase in the occurrence of 6th-order roots in infected plants. Parasitized plants had 44% fewer roots (primary through 6th-order) and 50% less total root length. PMID:19305837

  11. Potassium uptake and redistribution in Cabernet Sauvignon and Syrah grape tissues and its relationship with grape quality parameters.

    PubMed

    Ramos, María Concepción; Romero, María Paz

    2017-08-01

    The present study investigated the potassium (K) levels in petiole and other grape tissues during ripening in Vitis vinifera Shiraz and Cabernet Sauvignon, grown in areas with differences in vigour, as well as with and without leaf thinning. Potassium levels in petiole, seeds, skin and flesh were related to grape pH, acidity, berry weight and total soluble solids. Differences in K levels in petiole were in accordance with the differences in soil K. Leaf thinning gave rise to higher K levels in petiole but, in grape tissues, the differences were not significant in all samplings, with greater differences at the end of the growing cycle. Potassium levels per berry in grape tissues increased from veraison to harvest, with K mainly accumulated in skins and, to a lesser extent, in flesh. Potassium levels in flesh positively correlated with pH and total soluble solids, whereas the correlation with titratable acidity was negative. Grape juice pH and total soluble solids positively correlated with K, whereas titratable acidity correlated negatively. Leaf thinning increased K levels in petiole, although differences in K levels in grape tissues were not significant. This suggests the need to consider the K berry concentration when aiming to optimise K fertilisation programmes. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  12. Succession after fire: variation in \\delta13C of organic tissues and respired CO2 in boreal forests

    NASA Astrophysics Data System (ADS)

    Fessenden, J. E.; Li, H.; Mack, M.; Schuur, T.; Warren, S.; Randerson, J. T.

    2001-12-01

    Isotope ratios of carbon dioxide and leaf organic matter were measured in 5 neighboring forests of varying ages: 7, 14, 45, 140, and 160 years. These forests are composed primarily of black spruce (Picea Mariana) and quaking aspen (Populus tremuloides) with a shift in species dominance from aspen to spruce 50 years after fire disturbance. Research on the carbon isotope ratios of leaf material and CO2 was conducted to look for influences from species composition, forest age, and time after most recent burn. Samples of organic \\delta13C in whole leaf tissue were collected from the dominant species of each forest. Concurrent aboveground NPP measurements allowed us to estimate total ecosystem \\delta13C by providing a method for weighting \\delta13C of individual species and plant tissues. \\delta13CO2 and [CO2] were measured on canopy CO2 to determine the isotopic ratio of ecosystem respiration. The atmospheric results indicated that the \\delta13C of ecosystem respiration changes with successional stage. Specifically, the aspen dominating forests showed 13C depleted values relative to the spruce dominated forests. Organic results showed more 13C-enriched values with increased forest age and vegetation functional type. Specifically, oldest trees within the coniferous species had the most 13C-enriched values in leaf tissues. These results suggest that increases in the disturbance regime of northern boreal forests will lead to a decrease in the \\delta13C of ecosystem carbon with consequences for the atmospheric \\delta13C budget.

  13. Diurnal Cycling Transcription Factors of Pineapple Revealed by Genome-Wide Annotation and Global Transcriptomic Analysis

    PubMed Central

    Sharma, Anupma; Wai, Ching Man; Ming, Ray

    2017-01-01

    Abstract Circadian clock provides fitness advantage by coordinating internal metabolic and physiological processes to external cyclic environments. Core clock components exhibit daily rhythmic changes in gene expression, and the majority of them are transcription factors (TFs) and transcription coregulators (TCs). We annotated 1,398 TFs from 67 TF families and 80 TCs from 20 TC families in pineapple, and analyzed their tissue-specific and diurnal expression patterns. Approximately 42% of TFs and 45% of TCs displayed diel rhythmic expression, including 177 TF/TCs cycling only in the nonphotosynthetic leaf tissue, 247 cycling only in the photosynthetic leaf tissue, and 201 cycling in both. We identified 68 TF/TCs whose cycling expression was tightly coupled between the photosynthetic and nonphotosynthetic leaf tissues. These TF/TCs likely coordinate key biological processes in pineapple as we demonstrated that this group is enriched in homologous genes that form the core circadian clock in Arabidopsis and includes a STOP1 homolog. Two lines of evidence support the important role of the STOP1 homolog in regulating CAM photosynthesis in pineapple. First, STOP1 responds to acidic pH and regulates a malate channel in multiple plant species. Second, the cycling expression pattern of the pineapple STOP1 and the diurnal pattern of malate accumulation in pineapple leaf are correlated. We further examined duplicate-gene retention and loss in major known circadian genes and refined their evolutionary relationships between pineapple and other plants. Significant variations in duplicate-gene retention and loss were observed for most clock genes in both monocots and dicots. PMID:28922793

  14. Cryopreservation of Viable Human Lung Tissue for Versatile Post-thaw Analyses and Culture

    PubMed Central

    Baatz, John E.; Newton, Danforth A.; Riemer, Ellen C.; Denlinger, Chadrick E.; Jones, E. Ellen; Drake, Richard R.; Spyropoulos, Demetri D.

    2018-01-01

    Clinical trials are currently used to test therapeutic efficacies for lung cancer, infections and diseases. Animal models are also used as surrogates for human disease. Both approaches are expensive and time-consuming. The utility of human biospecimens as models is limited by specialized tissue processing methods that preserve subclasses of analytes (e.g. RNA, protein, morphology) at the expense of others. We present a rapid and reproducible method for the cryopreservation of viable lung tissue from patients undergoing lobectomy or transplant. This method involves the pseudo-diaphragmatic expansion of pieces of fresh lung tissue with cryoprotectant formulation (pseudo-diaphragmatic expansion-cryoprotectant perfusion or PDX-CP) followed by controlled-rate freezing in cryovials. Expansion-perfusion rates, volumes and cryoprotectant formulation were optimized to maintain tissue architecture, decrease crystal formation and increase long-term cell viability. Rates of expansion of 4 cc/min or less and volumes ranging from 0.8–1.2 × tissue volume were well-tolerated by lung tissue obtained from patients with chronic obstructive pulmonary disease or idiopathic pulmonary fibrosis, showing minimal differences compared to standard histopathology. Morphology was greatly improved by the PDX-CP procedure compared to simple fixation. Fresh versus post-thawed lung tissue showed minimal differences in histology, RNA integrity numbers and post-translational modified protein integrity (2-dimensional differential gel electrophoresis). It was possible to derive numerous cell types, including alveolar epithelial cells, fibroblasts and stem cells, from the tissue for at least three months after cryopreservation. This new method should provide a uniform, cost-effective approach to the banking of biospecimens, with versatility to be amenable to any post-acquisition process applicable to fresh tissue samples. PMID:24982205

  15. Synchrotron-Based Techniques Shed Light on Mechanisms of Plant Sensitivity and Tolerance to High Manganese in the Root Environment.

    PubMed

    Blamey, F Pax C; Hernandez-Soriano, Maria C; Cheng, Miaomiao; Tang, Caixian; Paterson, David J; Lombi, Enzo; Wang, Wei Hong; Scheckel, Kirk G; Kopittke, Peter M

    2015-11-01

    Plant species differ in response to high available manganese (Mn), but the mechanisms of sensitivity and tolerance are poorly understood. In solution culture, greater than or equal to 30 µm Mn decreased the growth of soybean (Glycine max), but white lupin (Lupinus albus), narrow-leafed lupin (Lupin angustifolius), and sunflower (Helianthus annuus) grew well at 100 µm Mn. Differences in species' tolerance to high Mn could not be explained simply by differences in root, stem, or leaf Mn status, being 8.6, 17.1, 6.8, and 9.5 mmol kg(-1) leaf fresh mass at 100 µm Mn. Furthermore, x-ray absorption near edge structure analyses identified the predominance of Mn(II), bound mostly to malate or citrate, in roots and stems of all four species. Rather, differences in tolerance were due to variations in Mn distribution and speciation within leaves. In Mn-sensitive soybean, in situ analysis of fresh leaves using x-ray fluorescence microscopy combined with x-ray absorption near edge structure showed high Mn in the veins, and manganite [Mn(III)] accumulated in necrotic lesions apparently through low Mn sequestration in vacuoles or other vesicles. In the two lupin species, most Mn accumulated in vacuoles as either soluble Mn(II) malate or citrate. In sunflower, Mn was sequestered as manganite at the base of nonglandular trichomes. Hence, tolerance to high Mn was ascribed to effective sinks for Mn in leaves, as Mn(II) within vacuoles or through oxidation of Mn(II) to Mn(III) in trichomes. These two mechanisms prevented Mn accumulation in the cytoplasm and apoplast, thereby ensuring tolerance to high Mn in the root environment. © 2015 American Society of Plant Biologists. All Rights Reserved.

  16. Synchrotron-Based Techniques Shed Light on Mechanisms of Plant Sensitivity and Tolerance to High Manganese in the Root Environment1[OPEN

    PubMed Central

    Blamey, F. Pax C.; Hernandez-Soriano, Maria C.; Cheng, Miaomiao; Tang, Caixian; Paterson, David J.; Lombi, Enzo; Wang, Wei Hong; Scheckel, Kirk G.; Kopittke, Peter M.

    2015-01-01

    Plant species differ in response to high available manganese (Mn), but the mechanisms of sensitivity and tolerance are poorly understood. In solution culture, greater than or equal to 30 µm Mn decreased the growth of soybean (Glycine max), but white lupin (Lupinus albus), narrow-leafed lupin (Lupin angustifolius), and sunflower (Helianthus annuus) grew well at 100 µm Mn. Differences in species’ tolerance to high Mn could not be explained simply by differences in root, stem, or leaf Mn status, being 8.6, 17.1, 6.8, and 9.5 mmol kg–1 leaf fresh mass at 100 µm Mn. Furthermore, x-ray absorption near edge structure analyses identified the predominance of Mn(II), bound mostly to malate or citrate, in roots and stems of all four species. Rather, differences in tolerance were due to variations in Mn distribution and speciation within leaves. In Mn-sensitive soybean, in situ analysis of fresh leaves using x-ray fluorescence microscopy combined with x-ray absorption near edge structure showed high Mn in the veins, and manganite [Mn(III)] accumulated in necrotic lesions apparently through low Mn sequestration in vacuoles or other vesicles. In the two lupin species, most Mn accumulated in vacuoles as either soluble Mn(II) malate or citrate. In sunflower, Mn was sequestered as manganite at the base of nonglandular trichomes. Hence, tolerance to high Mn was ascribed to effective sinks for Mn in leaves, as Mn(II) within vacuoles or through oxidation of Mn(II) to Mn(III) in trichomes. These two mechanisms prevented Mn accumulation in the cytoplasm and apoplast, thereby ensuring tolerance to high Mn in the root environment. PMID:26395840

  17. The effect of cutting origin and organic plant growth regulator on the growth of Daun Ungu (Graptophyllum pictum) through stem cutting method

    NASA Astrophysics Data System (ADS)

    Pratama, S. P.; Yunus, A.; Purwanto, E.; Widyastuti, Y.

    2018-03-01

    Graptophyllum pictum is one of medical plants which has important chemical content to treat diseases. Leaf, bark and flower can be used to facilitate menstruation, treat hemorrhoid, constipation, ulcers, ulcers, swelling, and earache. G. pictum is difficult to propagated by seedling due to the long duration of seed formation, thusvegetative propagation is done by stem cutting. The aims of this study are to obtain optimum combination of cutting origin and organic plant growth regulator in various consentration for the growth of Daun Ungu through stem cutting method. This research was conducted at Research center for Medicinal Plant and Traditional DrugTanjungsari, Tegal Gede, Karanganyar in June to August 2016. Origin of cuttings and organic plant growth regulator were used as treatments factor. A completely randomized design (RAL) is used and data were analyzed by F test (ANOVA) with a confidence level of 95%. Any significant differences among treatment followed with Duncan test at a = 5%. The research indicates that longest root was resulted from the treatment of 0,5 ml/l of organic plant growth regulator. The treatment of 1 ml/l is able to increase the fresh and dry weight of root, treatment of 1,5 ml/l of organic plant growth regulator was able to increase the percentage of growing shoots. Treatment of base part as origin of cuttings increases the length, fresh weight and and dry weight of shoot, increase the number of leaves. Interaction treatment between 1 ml/l consentration of organic plant growth regulator and central part origin of cuttings is capable of increasing the leaf area, whereas treatment without organic plant growth regulator and base part as planting material affects the smallest leaf area.

  18. Quality improvement by batch vacuum distillation and physicochemical characterization of clove leaf oil in Central Java, Indonesia

    NASA Astrophysics Data System (ADS)

    Alighiri, D.; Eden, W. T.; Cahyono, E.; Supardi, K. I.

    2018-03-01

    Clove leaf oil is one of the essential oils which have high economic value and produced in considerable amount in Indonesia. As many as 60% of clove oil produced by Indonesia has exported to many countries in the world. Musuk and Ringinlarik Village, Boyolali District, Central Java Province is one of the areas in Indonesia which became the primary supplier of Indonesian clove leaf oil. Although, the quality of the resulting yield is still low because it uses a simple distillation kettle in the form of the iron plate or used the drum. The clove leaf oil produced by steam distillation from the dry whole leaves of the clove tree. The color of the oil is brownish black and dirty with the odor are fresh distilled, spicy, warm, and terpenic. The specific gravity at 25 °C of 1.529, the refractive index at 20 °C of 1.030, and based on GC-MS analysis, eugenol levels only 68% and caryophyllene is too high, i.e., 20%. The quality of clove leaf oil produced does not meet the specifications of international market standards. This work aimed to improve the quality of Indonesian clove leaf oil. The purifications done in this research was used by batch vacuum distillation with mode operation at vacuum -76 cmHg and reflux ratios 5:1. Clove leaf oil produced by using this method has a better physicochemical characterization, i.e., the appearance that is yellow to pale color with the odor is spicy, woody, warm, and terpenic. The specific gravity at 25 °C of 1.533, the refractive index at 20 °C of 1.038, and eugenol and caryophyllene contents has yielded 80.58% and 10%, respectively. By The enhancement quality of clove leaf oil by batch vacuum distillation, these oil is already meet international standards and income of clove leaf oil grower in Musuk and Ringinlarik Village, Boyolali District, Central Java Province, Indonesia could be increased.

  19. A Multivariate Evaluation of Factors Affecting the Quality of Freshly Frozen Tissue Specimens.

    PubMed

    Wang, Tong-Hong; Chen, Chin-Chuan; Liang, Kung-Hao; Chen, Chi-Yuan; Chuang, Wen-Yu; Ueng, Shir-Hwa; Chu, Pao-Hsien; Huang, Chung-Guei; Chen, Tse-Ching; Hsueh, Chuen

    2017-08-01

    Well-prepared and preserved freshly frozen specimens are indispensable materials for clinical studies. To manage specimen quality and to understand the factors potentially affecting specimen quality during preservation processes, we analyzed the quality of RNA and genomic DNA of various tissues collected between 2002 and 2011 in Linkou Chang Gung Memorial Hospital, Taiwan. During this period, a total of 1059 freshly frozen specimens from eight major cancer categories were examined. It was found that preservation duration, organ origin, and tissue type could all influence the quality of RNA samples. The increased preservation period correlated with decreased RNA quality; the brain, breast, and stomach RNA specimens displayed faster degradation rates than those of other organs, and RNA specimens isolated from tumor tissues were apparently more stable than those of other tissues. These factors could all be used as quality predictors of RNA quality. In contrast, almost all analyses revealed that the genomic DNA samples had good quality, which was not influenced by the aforementioned factors. The results assisted us in determining preservation factors that affect specimen quality, which could provide evidence for improving processes of sample collection and preservation. Furthermore, the results are also useful for researchers to adopt as the evaluation criteria for choosing specimen collection and preservation strategies.

  20. A comparative study of vascular injection fluids in fresh-frozen and embalmed human cadaver forearms.

    PubMed

    Doomernik, D E; Kruse, R R; Reijnen, M M P J; Kozicz, T L; Kooloos, J G M

    2016-10-01

    Over the years, various vascular injection products have been developed to facilitate anatomical dissections. This study aimed to compare the most commonly used vascular injection products in fresh-frozen and formalin-embalmed cadaver specimens. An overview of the properties, advantages and limitations of each substance was given, and a comparison of vascular infusion procedures in both preservation methods was made. A literature search was performed in order to identify the most commonly used vascular injection products. Acrylic paint, latex, gelatin, silicone, Araldite F and Batson's No. 17 were selected for the study. One fresh-frozen and one embalmed cadaver forearm were infused with each injection product according to a uniform protocol. The curing time, skin- and subcutaneous tissue penetration, degree of filling of the arterial tree, extravasations, consistency of the injected vessels during dissection, and the costs of each injection fluid were noted. There was a large variation between the injection fluids in processing- and curing time, colour intensity, flexibility, fragility, elasticity, strength, toxicity and costs. All fluids were suitable for infusion. The penetration of injection fluid into the skin and subcutaneous tissue was significantly better in fresh-frozen specimens (P = 0.002 and P = 0.009, respectively), with significantly smaller branches casted (P = 0.004). Vascular infusion of fresh-frozen cadaver specimens results in a significantly better filled coloured arterial tree, enabling more detail to be achieved and smaller branches casted. The biomechanical properties of fresh-frozen soft tissues are less affected compared with formalin fixation. All the injection fluids studied are suitable for vascular infusion, but their different properties ensure that certain products and procedures are more suitable for specific study purposes. © 2016 Anatomical Society.

  1. Development of a rapid diagnostic assay for the detection of tomato chlorotic dwarf viroid based on isothermal reverse-transcription-recombinase polymerase amplification.

    PubMed

    Hammond, Rosemarie W; Zhang, Shulu

    2016-10-01

    A molecular diagnostic assay utilizing reverse transcription-recombinase polymerase amplification (RT-RPA) at an isothermal constant temperature of 39°C and target-specific primers and probe were developed for the rapid, sensitive, and specific detection of tomato chlorotic dwarf viroid (TCDVd) in infected leaf and seed tissues. The performance of the AmplifyRP(®) Acceler8™ RT-RPA diagnostic assay, utilizing a lateral flow strip contained within an amplicon detection chamber, was evaluated and the results were compared with a standard RT-PCR assay. The AmplifyRP(®) Acceler8™ assay was specific for TCDVd in leaf and seed tissues, its sensitivity was comparable to conventional RT-PCR in leaf tissues, and it does not require extensive sample purification, specialized equipment, or technical expertise. This is the first report utilizing an RT-RPA assay to detect viroids and the assay can be used both in the laboratory and in the field for TCDVd detection. Published by Elsevier B.V.

  2. Ex vivo applications of multiphoton microscopy in urology

    NASA Astrophysics Data System (ADS)

    Jain, Manu; Mukherjee, Sushmita

    2016-03-01

    Background: Routine urological surgery frequently requires rapid on-site histopathological tissue evaluation either during biopsy or intra-operative procedure. However, resected tissue needs to undergo processing, which is not only time consuming but may also create artifacts hindering real-time tissue assessment. Likewise, pathologist often relies on several ancillary methods, in addition to H&E to arrive at a definitive diagnosis. Although, helpful these techniques are tedious and time consuming and often show overlapping results. Therefore, there is a need for an imaging tool that can rapidly assess tissue in real-time at cellular level. Multiphoton microscopy (MPM) is one such technique that can generate histology-quality images from fresh and fixed tissue solely based on their intrinsic autofluorescence emission, without the need for tissue processing or staining. Design: Fresh tissue sections (neoplastic and non-neoplastic) from biopsy and surgical specimens of bladder and kidney were obtained. Unstained deparaffinized slides from biopsy of medical kidney disease and oncocytic renal neoplasms were also obtained. MPM images were acquired using with an Olympus FluoView FV1000MPE system. After imaging, fresh tissues were submitted for routine histopathology. Results: Based on the architectural and cellular details of the tissue, MPM could characterize normal components of bladder and kidney. Neoplastic tissue could be differentiated from non-neoplastic tissue and could be further classified as per histopathological convention. Some of the tumors had unique MPM signatures not otherwise seen on H&E sections. Various subtypes of glomerular lesions were identified as well as renal oncocytic neoplasms were differentiated on unstained deparaffinized slides. Conclusions: We envision MPM to become an integral part of regular diagnostic workflow for rapid assessment of tissue. MPM can be used to evaluate the adequacy of biopsies and triage tissues for ancillary studies. It can also be used as an adjunct to frozen section analysis for intra-operative margin assessment. Further, it can play an important role for pathologist for guiding specimen grossing, selecting tissue for tumor banking and as a rapid ancillary diagnostic tool.

  3. Interpretation of the fluorescence signatures from vegetation

    NASA Astrophysics Data System (ADS)

    Buschmann, C.

    Vegetation emits fluorescence as part of the energy taken up by absorption %of solar radiation from UV to the visible. This fluorescence consists of light with low intensity (only few percents of the reflected light) emitted from the leaves. The fluorescence emission of a green leaf is characterized by four bands with maxima in the blue (440 nm), green (520 nm), red (690 nm) and far red (740 nm) spectral region. The intensity of fluorescence in the maxima of the emission spectrum varies depending on the following six basic parameters which must be taken into account for the interpretation of fluorescence signatures from vegetation: (a) content of the fluorophores (ferulic acid, chlorophyll a), (b) temperature of the leaf, (c) penetration of excitation light into the leaf, (d) emission of fluorescence from the leaf (re-absorption inside the leaf tissue), (e) photosynthetic activity of the leaf, (f) non-radiative decay (heat production) parallel to the fluorescence The ratios between the intensities of the maxima (F440/F690, F440/F520, F690/F740) are used as characteristic fluorescence parameter. The wide range of changes of these ratios caused by differences in the leaf tissue (aerial interspaces, variegated/homogeneous green leaves), various types of stress (UV, photoinhibition, sun exposure, heat, water deficiency, N-deficiency) and chemicals (inhibitors, fertilizers) can be explained by changes of the six basic parameters. It will be shown that the interpretation of the fluorescence signatures, in most cases, must be based on a complex consideration of more than one of the basic parameters.

  4. The influence of elevated CO2 on non-structural carbohydrate distribution and fructan accumulation in wheat canopies

    NASA Technical Reports Server (NTRS)

    Smart, D. R.; Chatterton, N. J.; Bugbee, B.

    1994-01-01

    We grew 2.4 m2 wheat canopies in a large growth chamber under high photosynthetic photon flux (1000 micromoles m-2 s-1) and using two CO2 concentrations, 360 and 1200 micromoles mol-1. Photosynthetically active radiation (400-700 nm) was attenuated slightly faster through canopies grown in 360 micromoles mol-1 than through canopies grown in 1200 micromoles mol-1, even though high-CO2 canopies attained larger leaf area indices. Tissue fractions were sampled from each 5-cm layer of the canopies. Leaf tissue sampled from the tops of canopies grown in 1200 micromoles mol-1 accumulated significantly more total non-structural carbohydrate, starch, fructan, sucrose, and glucose (p < 0.05) than for canopies grown in 360 micromoles mol-1. Non-structural carbohydrate did not significantly increase in the lower canopy layers of the elevated CO2 treatment. Elevated CO2 induced fructan synthesis in all leaf tissue fractions, but fructan formation was greatest in the uppermost leaf area. A moderate temperature reduction of 10 degrees C over 5 d increased starch, fructan and glucose levels in canopies grown in 1200 micromoles mol-1, but concentrations of sucrose and fructose decreased slightly or remained unchanged. Those results may correspond with the use of fructosyl-residues and release of glucose when sucrose is consumed in fructan synthesis.

  5. Identification and Chacterization of new strains of Enterobacter spp. causing Mulberry (Morus alba) wilt disease in China

    USDA-ARS?s Scientific Manuscript database

    A new mulberry wilt disease (MWD) was recently identified in Hangzhou, Zhejiang province, China. Typical symptoms of the disease are dark brown discolorations in vascular tissues, leaf wilt, defoliation, and tree decline. Unlike the bacterial wilt disease caused by Ralstonia solanacearum, the leaf w...

  6. Leaf, woody, and root biomass of Populus irrigated with landfill leachate

    Treesearch

    Jill A. Zalesny; Ronald S., Jr. Zalesny; D.R. Coyle; R.B. Hall

    2007-01-01

    Poplar (Populus spp.) trees can be utilized for ecological leachate disposal when applied as an irrigation source for managed tree systems. Our objective was to evaluate differences in tree height, diameter, volume, and biomass of leaf, stem, branch, and root tissues of Populus trees after two seasons of irrigation with municipal...

  7. Theoretical and experimental errors for in situ measurements of plant water potential.

    PubMed

    Shackel, K A

    1984-07-01

    Errors in psychrometrically determined values of leaf water potential caused by tissue resistance to water vapor exchange and by lack of thermal equilibrium were evaluated using commercial in situ psychrometers (Wescor Inc., Logan, UT) on leaves of Tradescantia virginiana (L.). Theoretical errors in the dewpoint method of operation for these sensors were demonstrated. After correction for these errors, in situ measurements of leaf water potential indicated substantial errors caused by tissue resistance to water vapor exchange (4 to 6% reduction in apparent water potential per second of cooling time used) resulting from humidity depletions in the psychrometer chamber during the Peltier condensation process. These errors were avoided by use of a modified procedure for dewpoint measurement. Large changes in apparent water potential were caused by leaf and psychrometer exposure to moderate levels of irradiance. These changes were correlated with relatively small shifts in psychrometer zero offsets (-0.6 to -1.0 megapascals per microvolt), indicating substantial errors caused by nonisothermal conditions between the leaf and the psychrometer. Explicit correction for these errors is not possible with the current psychrometer design.

  8. Theoretical and Experimental Errors for In Situ Measurements of Plant Water Potential 1

    PubMed Central

    Shackel, Kenneth A.

    1984-01-01

    Errors in psychrometrically determined values of leaf water potential caused by tissue resistance to water vapor exchange and by lack of thermal equilibrium were evaluated using commercial in situ psychrometers (Wescor Inc., Logan, UT) on leaves of Tradescantia virginiana (L.). Theoretical errors in the dewpoint method of operation for these sensors were demonstrated. After correction for these errors, in situ measurements of leaf water potential indicated substantial errors caused by tissue resistance to water vapor exchange (4 to 6% reduction in apparent water potential per second of cooling time used) resulting from humidity depletions in the psychrometer chamber during the Peltier condensation process. These errors were avoided by use of a modified procedure for dewpoint measurement. Large changes in apparent water potential were caused by leaf and psychrometer exposure to moderate levels of irradiance. These changes were correlated with relatively small shifts in psychrometer zero offsets (−0.6 to −1.0 megapascals per microvolt), indicating substantial errors caused by nonisothermal conditions between the leaf and the psychrometer. Explicit correction for these errors is not possible with the current psychrometer design. PMID:16663701

  9. Physiology and quality of fresh-cut produce in CA/MA storage

    USDA-ARS?s Scientific Manuscript database

    Fresh-cut fruits and vegetables have exposed injured tissues due to the mechanical processes of peeling, slicing and/or cutting. Such processing consequently renders the produce highly susceptible to physiological breakdown and microbial spoilage. Product deterioration is usually accompanied with ph...

  10. Molecular Mechanisms of Phosphorus Metabolism and Transport during Leaf Senescence

    PubMed Central

    Stigter, Kyla A.; Plaxton, William C.

    2015-01-01

    Leaf senescence, being the final developmental stage of the leaf, signifies the transition from a mature, photosynthetically active organ to the attenuation of said function and eventual death of the leaf. During senescence, essential nutrients sequestered in the leaf, such as phosphorus (P), are mobilized and transported to sink tissues, particularly expanding leaves and developing seeds. Phosphorus recycling is crucial, as it helps to ensure that previously acquired P is not lost to the environment, particularly under the naturally occurring condition where most unfertilized soils contain low levels of soluble orthophosphate (Pi), the only form of P that roots can directly assimilate from the soil. Piecing together the molecular mechanisms that underpin the highly variable efficiencies of P remobilization from senescing leaves by different plant species may be critical for devising effective strategies for improving overall crop P-use efficiency. Maximizing Pi remobilization from senescing leaves using selective breeding and/or biotechnological strategies will help to generate P-efficient crops that would minimize the use of unsustainable and polluting Pi-containing fertilizers in agriculture. This review focuses on the molecular mechanisms whereby P is remobilized from senescing leaves and transported to sink tissues, which encompasses the action of hormones, transcription factors, Pi-scavenging enzymes, and Pi transporters. PMID:27135351

  11. An electron spin resonance study for real-time detection of ascorbyl free radicals after addition of dimethyl sulfoxide in murine hippocampus or plasma during kainic acid-induced seizures.

    PubMed

    Matsumoto, Shigekiyo; Shingu, Chihiro; Koga, Hironori; Hagiwara, Satoshi; Iwasaka, Hideo; Noguchi, Takayuki; Yokoi, Isao

    2010-07-01

    Electron spin resonance (ESR)-silent ascorbate solutions generate a detectable, likely concentration-dependent signal of ascorbyl free radicals (AFR) immediately upon addition of a molar excess of dimethyl sulfoxide (DMSO). We aimed to perform quantitative ESR analysis of AFR in real time after addition of DMSO (AFR/DMSO) to evaluate ascorbate concentrations in fresh hippocampus or plasma following systemic administration of kainate in mice. Use of a special tissue-type quartz cell allowed immediate detection of AFR/DMSO ESR spectra in fresh tissues from mice. AFR/DMSO content was increased significantly in fresh hippocampus or plasma obtained during kainate-induced seizures of mice, reaching maximum levels at 90 min after intraperitoneal administration of 50 mg/kg kainic acid. This suggests that oxidative injury of the hippocampus resulted from the accumulation of large amounts of ascorbic acid in the brain after kainic acid administration. AFR/DMSO content measured on an ESR spectrometer can be used for real-time evaluation of ascorbate content in fresh tissue. Due to the simplicity, good performance, low cost and real-time monitoring of ascorbate, this method may be applied to clinical research and treatment in the future.

  12. Isochoric and isobaric freezing of fish muscle

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Năstase, Gabriel; Department of Building Services, University of Transilvania, Braşov, Braşov, 500152; Lyu, Chenang

    We have recently shown that, a living organism, which succumbs to freezing to −4 °C in an isobaric thermodynamic system (constant atmospheric pressure), can survive freezing to −4 °C in an isochoric thermodynamic system (constant volume). It is known that the mechanism of cell damage in an isobaric system is the freezing caused increase in extracellular osmolality, and, the consequent cell dehydration. An explanation for the observed survival during isochoric freezing is the thermodynamic modeling supported hypothesis that, in the isochoric frozen solution the extracellular osmolality is comparable to the cell intracellular osmolality. Therefore, cells in the isochoric frozen organism do not dehydrate, andmore » the tissue maintains its morphological integrity. Comparing the histology of: a) fresh fish white muscle, b) fresh muscle frozen to −5 °C in an isobaric system and c) fresh muscle frozen to −5 °C I in an isochoric system, we find convincing evidence of the mechanism of cell dehydration during isobaric freezing. In contrast, the muscle tissue frozen to −5 °C in an isochoric system appears morphologically identical to fresh tissue, with no evidence of dehydration. This is the first experimental evidence in support of the hypothesis that in isochoric freezing there is no cellular dehydration and therefore the morphology of the frozen tissue remains intact. - Highlights: • Preservation of fish muscle at, subfreezing temperatures, in an isochoric system, is demonstrated. • Experiments were performed to an average pressure of 41.3 MPa and temperatures of −5 °C. • Isochoric subfreezing temperature is a new preservation method that does not require the.use of cryoprotectants. • No cellular dehydration and therefore the morphology of the frozen tissue remains intact.« less

  13. Simulation of plastic surgery and microvascular procedures using perfused fresh human cadavers.

    PubMed

    Carey, Joseph N; Rommer, Elizabeth; Sheckter, Clifford; Minneti, Michael; Talving, Peep; Wong, Alex K; Garner, Warren; Urata, Mark M

    2014-02-01

    Surgical simulation models are often limited by their lack of fidelity, which hinders their essential purpose, making a better surgeon. Fresh cadaveric tissue is a superior model of simulation owing to its approximation of live tissue. One major unresolved difference between dead and live tissue is perfusion. Here, we propose a means of enhancing the fidelity of cadaveric simulation through the development of a perfused cadaveric model whereby simulation is further able to approach life-like surgery and teach one of the more technically demanding skills of plastic surgery: microsurgery. Fresh tissue human cadavers were procured according to university protocol. Perfusion was performed via cannulation of large vessels, and arterial and venous pressure was maintained by centrifugal circulation. Skin perfusion was evaluated with incisions in the perfused regions and was evaluated using indocyanine green angiography. Surgical simulations were selected to broadly evaluate applicability to plastic surgical education. Surgical simulation of 38 procedures ranging in complexity from skin excisions to microsurgical cases was performed with high priority given to the accurate simulation of clinical procedures. Flap dissections included perforator flaps, muscle flaps, and fasciocutaneous flaps. Effective perfusion was noted with ICG angiography and notable bleeding vessels. Microsurgical flap transfer was successfully performed. We report the establishment of a high fidelity surgical simulation using a perfused fresh tissue model in a realistic environment akin to the operating room. We anticipate utilization of this model prior to entering the operating room will enhance surgical ability and offer a valuable resource in plastic surgical education. Copyright © 2013 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

  14. Protective mechanisms and acclimation to solar ultraviolet-B radiation in Oenothera stricta

    NASA Technical Reports Server (NTRS)

    Robberecht, R.; Caldwell, M. M.

    1981-01-01

    Plant adaptations ameliorating or repairing the damaging effects of ultraviolet-B (UV-B) radiation on plant tissue were investigated. The degree of phenotype plasticity in UV protective mechanisms and acclimation in relation to the natural solar UV-B radiation flux and in an enhanced UV-B irradiance environment was also examined. Mechanisms by which plants avoid radiation, adaptations altering the path of radiation incident on the leaf, and repair processes were considered. Attenuation of UV-B by tissues, UV-B irradiation into the leaf, and the effects of UV-B on photosynthesis were investigated.

  15. Hevea Linamarase—A Nonspecific β-Glycosidase 1

    PubMed Central

    Selmar, Dirk; Lieberei, Reinhard; Biehl, Böle; Voigt, Jürgen

    1987-01-01

    In the leaf tissue of the cyanogenic plant Hevea brasiliensis, which contains large amounts of linamarin, there is no specific linamarase. In Hevea leaves only one β-glucosidase is detectable. It is responsible for the cleavage of all β-glucosides and β-galactosides occurring in Hevea leaf tissue, including the cyanogenic glucoside linamarin. Therefore, the enzyme is referred to as a β-glycosidase instead of the term β-glucosidase. This β-glycosidase has a broad substrate spectrum and occurs in multiple forms. These homo-oligomeric forms are interconvertible by dissociation-association processes. The monomer is a single protein of 64 kilodaltons. PMID:16665288

  16. Morphological, ultrastructural and functional imaging of frozen/thawed and vitrified/warmed human ovarian tissue retrieved from oncological patients.

    PubMed

    Fabbri, R; Vicenti, R; Macciocca, M; Martino, N A; Dell'Aquila, M E; Pasquinelli, G; Morselli-Labate, A M; Seracchioli, R; Paradisi, R

    2016-08-01

    Which is the best method for human ovarian tissue cryopreservation: slow freezing/rapid thawing (SF/RT) or vitrification/warming (V/W)? The conventional SF/RT protocol used in this study seems to better preserve the morpho-functional status of human cryopreserved ovarian tissue than the used open carrier V/W protocol. Cryopreservation of human ovarian tissue is generally performed using the SF/RT method. However, reduction in the follicular pool and stroma damage are often observed. An emerging alternative procedure is represented by V/W which seems to allow the maintenance of the morphological integrity of the stroma. This is a retrospective cohort study including six patients affected by oncological diseases and enrolled from January to December 2014. Ovarian tissue was laparoscopically harvested from the right and left ovaries and was cryopreserved using a routinary SF/RT protocol or a V/W method, involving tissue incubation in two solutions (containing propylene glycol, ethylene glycol and sucrose at different concentrations) and vitrification in an open system. For each patient, three pieces from each ovary were collected at the time of laparoscopy (fresh tissue) and after storage (SF/RT or V/W) and processed for light microscopy (LM) and transmission electron microscopy (TEM), to assess the morphological and ultrastructural features of follicles and stroma, and for laser scanning confocal microscopy (LSCM), to determine the functional energetic/redox stroma status. The preservation status of SF/RT and V/W ovarian tissues was compared with that of fresh ones, as well as between them. By LM and TEM, SF/RT and V/W samples showed cryodamage of small entity. Interstitial oedema and increased stromal cell vacuolization and chromatin clumping were observed in SF/RT samples; in contrast, V/W samples showed oocyte nuclei with slightly thickened chromatin and irregular shapes. The functional imaging analysis by LSCM revealed that the mitochondrial activity and intracellular reactive oxygen species levels were reduced both in SF/RT and in V/W samples compared with fresh samples. The study also showed progressive dysfunction of the mitochondrial activity going from the outer to the inner serial section of the ovarian cortex. The reduction of mitochondrial activity of V/W samples compared with fresh samples was significantly higher in the inner section than in the outer section. The results report the bioenergetic and oxidative status assessment of fresh and cryopreserved human ovarian tissue by LSCM, a technique recently applied to tissue samples. The use of LSCM on human ovarian tissues after SF/RT or V/W is a new application that requires validation. The procedures for mitochondrial staining with functional probes and fixing are not yet standardized. Xenografting of the cryopreserved ovarian tissue in severe combined immunodeficient mice and in vitro culture have not yet been performed. The identification of a cryopreservation method able to maintain the morpho-functional integrity of the ovarian tissue and a number of follicles comparable with those observed in fresh tissue might optimize results in clinical practice, in terms of recovery, duration of ovarian function and increased delivery outcomes after replanting. The SF/RT protocol allowed better morpho-functional tissue integrity than the V/W procedure. Funding was provided by Fondazione del Monte di Bologna e Ravenna, Italy. Dr N.A.M. was granted by the project ONEV MIUR PONa3 00134-n.254/R&C 18 5 2011 and the project GR-2011-02351396 (Ministry of Health, Young Researchers Grant 2011/2012). There are no competing interests. Clinical trial 74/2001/0 (approved:13 2 2002): 'Pilot study on cryopreservation of human ovarian tissue: morphological and immunohistochemical analysis before and after cryopreservation'. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  17. DMSO‐ and Serum‐Free Cryopreservation of Wharton's Jelly Tissue Isolated From Human Umbilical Cord

    PubMed Central

    Shivakumar, Sharath Belame; Bharti, Dinesh; Subbarao, Raghavendra Baregundi; Jang, Si‐Jung; Park, Ji‐Sung; Ullah, Imran; Park, Ji‐Kwon; Byun, June‐Ho

    2016-01-01

    ABSTRACT The facile nature of mesenchymal stem cell (MSC) acquisition in relatively large numbers has made Wharton's jelly (WJ) tissue an alternative source of MSCs for regenerative medicine. However, freezing of such tissue using dimethyl sulfoxide (DMSO) for future use impedes its clinical utility. In this study, we compared the effect of two different cryoprotectants (DMSO and cocktail solution) on post‐thaw cell behavior upon freezing of WJ tissue following two different freezing protocols (Conventional [−1°C/min] and programmed). The programmed method showed higher cell survival rate compared to conventional method of freezing. Further, cocktail solution showed better cryoprotection than DMSO. Post‐thaw growth characteristics and stem cell behavior of Wharton's jelly mesenchymal stem cells (WJMSCs) from WJ tissue cryopreserved with a cocktail solution in conjunction with programmed method (Prog‐Cock) were comparable with WJMSCs from fresh WJ tissue. They preserved their expression of surface markers, pluripotent factors, and successfully differentiated in vitro into osteocytes, adipocytes, chondrocytes, and hepatocytes. They also produced lesser annexin‐V‐positive cells compared to cells from WJ tissue stored using cocktail solution in conjunction with the conventional method (Conv‐Cock). Real‐time PCR and Western blot analysis of post‐thaw WJMSCs from Conv‐Cock group showed significantly increased expression of pro‐apoptotic factors (BAX, p53, and p21) and reduced expression of anti‐apoptotic factor (BCL2) compared to WJMSCs from the fresh and Prog‐Cock group. Therefore, we conclude that freezing of fresh WJ tissue using cocktail solution in conjunction with programmed freezing method allows for an efficient WJ tissue banking for future MSC‐based regenerative therapies. J. Cell. Biochem. 117: 2397–2412, 2016. © 2016 The Authors. Journal of Cellular Biochemistry published by Wiley Periodicals, Inc. PMID:27038129

  18. Potential of Three Ethnomedicinal Plants as Antisickling Agents.

    PubMed

    Nurain, Ismaila O; Bewaji, Clement O; Johnson, Jarrett S; Davenport, Robertson D; Zhang, Yang

    2017-01-03

    Sickle cell disease (SCD) is a genetic blood disorder that affects the shape and transportation of red blood cells (RBCs) in blood vessels, leading to various clinical complications. Many drugs that are available for treating the disease are insufficiently effective, toxic, or too expensive. Therefore, there is a pressing need for safe, effective, and inexpensive therapeutic agents from indigenous plants used in ethnomedicines. The potential of aqueous extracts of Cajanus cajan leaf and seed, Zanthoxylum zanthoxyloides leaf, and Carica papaya leaf in sickle cell disease management was investigated in vitro using freshly prepared 2% sodium metabisulfite for sickling induction. The results indicated that the percentage of sickled cells, which was initially 91.6% in the control, was reduced to 29.3%, 41.7%, 32.8%, 38.2%, 47.6%, in the presence of hydroxyurea, C. cajan seed, C. cajan leaf, Z. zanthoxyloides leaf, and C. papaya leaf extracts, respectively, where the rate of polymerization inhibition was 6.5, 5.9, 8.0, 6.6, and 6.0 (×10 -2 ) accordingly. It was also found that the RBC resistance to hemolysis was increased in the presence of the tested agents as indicated by the reduction of the percentage of hemolyzed cells from 100% to 0%. The phytochemical screening results indicated the presence of important phytochemicals including tannins, saponins, alkaloids, flavonoids, and glycosides in all the plant extracts. Finally, gas chromatography-mass spectrometry analysis showed the presence of important secondary metabolites in the plants. These results suggest that the plant extracts have some potential to be used as alternative antisickling therapy to hydroxyurea in SCD management.

  19. Fresh-frozen Complete Extensor Mechanism Allograft versus Autograft Reconstruction in Rabbits

    PubMed Central

    Chen, Guanyin; Zhang, Hongtao; Ma, Qiong; Zhao, Jian; Zhang, Yinglong; Fan, Qingyu; Ma, Baoan

    2016-01-01

    Different clinical results have been reported in the repair of extensor mechanism disruption using fresh-frozen complete extensor mechanism (CEM) allograft, creating a need for a better understanding of fresh-frozen CME allograft reconstruction. Here, we perform histological and biomechanical analyses of fresh-frozen CEM allograft or autograft reconstruction in an in vivo rabbit model. Our histological results show complete incorporation of the quadriceps tendon into the host tissues, patellar survival and total integration of the allograft tibia, with relatively fewer osteocytes, into the host tibia. Vascularity and cellularity are reduced and delayed in the allograft but exhibit similar distributions to those in the autograft. The infrapatellar fat pad provides the main blood supply, and the lowest cellularity is observed in the patellar tendon close to the tibia in both the allograft and autograft. The biomechanical properties of the junction of quadriceps tendon and host tissues and those of the allograft patellar tendon are completely and considerably restored, respectively. Therefore, fresh-frozen CEM allograft reconstruction is viable, but the distal patellar tendon and the tibial block may be the weak links of the reconstruction. These findings provide new insight into the use of allograft in repairing disruption of the extensor mechanism. PMID:26911538

  20. Fresh-frozen Complete Extensor Mechanism Allograft versus Autograft Reconstruction in Rabbits.

    PubMed

    Chen, Guanyin; Zhang, Hongtao; Ma, Qiong; Zhao, Jian; Zhang, Yinglong; Fan, Qingyu; Ma, Baoan

    2016-02-25

    Different clinical results have been reported in the repair of extensor mechanism disruption using fresh-frozen complete extensor mechanism (CEM) allograft, creating a need for a better understanding of fresh-frozen CME allograft reconstruction. Here, we perform histological and biomechanical analyses of fresh-frozen CEM allograft or autograft reconstruction in an in vivo rabbit model. Our histological results show complete incorporation of the quadriceps tendon into the host tissues, patellar survival and total integration of the allograft tibia, with relatively fewer osteocytes, into the host tibia. Vascularity and cellularity are reduced and delayed in the allograft but exhibit similar distributions to those in the autograft. The infrapatellar fat pad provides the main blood supply, and the lowest cellularity is observed in the patellar tendon close to the tibia in both the allograft and autograft. The biomechanical properties of the junction of quadriceps tendon and host tissues and those of the allograft patellar tendon are completely and considerably restored, respectively. Therefore, fresh-frozen CEM allograft reconstruction is viable, but the distal patellar tendon and the tibial block may be the weak links of the reconstruction. These findings provide new insight into the use of allograft in repairing disruption of the extensor mechanism.

  1. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dai, Ziyu; Hooker, Brian S.; Anderson, Daniel B.

    Optimization of Acidothermus cellulolyticus endoglucanase (E1) gene expression in transgenic potato (Solanum tuberosum L.) was examined in this study, where the E1 coding sequence was transcribed under control of a leaf specific promoter (tomato RbcS-3C) or the Mac promoter (a hybrid promoter of mannopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region). Average E1 activity in leaf extracts of potato transformants, in which E1 protein was targeted by a chloroplast signal peptide and an apoplast signal peptide were much higher than those by an E1 native signal peptide and a vacuole signal peptide. E1 protein accumulated up tomore » 2.6% of total leaf soluble protein, where E1 gene was under control of the RbcS-3C promoter, alfalfa mosaic virus 5-untranslated leader, and RbcS-2A signal peptide. E1 protein production, based on average E1 activity and E1 protein accumulation in leaf extracts, is higher in potato than those measured previously in transgenic tobacco bearing the same transgene constructs. Comparisons of E1 activity, protein accumulation, and relative mRNA levels showed that E1 expression under control of tomato RbcS-3C promoter was specifically localized in leaf tissues, while E1 gene was expressed in both leaf and tuber tissues under control of Mac promoter. This suggests dual-crop applications in which potato vines serve as enzyme production `bioreactors' while tubers are preserved for culinary applications.« less

  2. Imaging of the interaction of low frequency electric fields with biological tissues by optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Peña, Adrian F.; Devine, Jack; Doronin, Alexander; Meglinski, Igor

    2014-03-01

    We report the use of conventional Optical Coherence Tomography (OCT) for visualization of propagation of low frequency electric field in soft biological tissues ex vivo. To increase the overall quality of the experimental images an adaptive Wiener filtering technique has been employed. Fourier domain correlation has been subsequently applied to enhance spatial resolution of images of biological tissues influenced by low frequency electric field. Image processing has been performed on Graphics Processing Units (GPUs) utilizing Compute Unified Device Architecture (CUDA) framework in the frequencydomain. The results show that variation in voltage and frequency of the applied electric field relates exponentially to the magnitude of its influence on biological tissue. The magnitude of influence is about twice more for fresh tissue samples in comparison to non-fresh ones. The obtained results suggest that OCT can be used for observation and quantitative evaluation of the electro-kinetic changes in biological tissues under different physiological conditions, functional electrical stimulation, and potentially can be used non-invasively for food quality control.

  3. PHANTASTICA regulates leaf polarity and petiole identity in Medicago truncatula

    PubMed Central

    Ge, Liangfa; Chen, Rujin

    2014-01-01

    Establishment of proper polarities along the adaxial-abaxial, proximodistal, and medial-lateral axes is a critical step for the expansion of leaves from leaf primordia. It has been shown that the MYB domain protein, ASYMMETRIC LEAVES1/ROUGH SHEATH2/PHANTASTICA (collectively named ARP) plays an important role in this process. Loss of function of ARP leads to severe leaf polarity defects, such as abaxialized or needle-like leaves. In addition to its role in leaf polarity establishment, we have recently shown that the Medicago truncatula ARP gene, MtPHAN, also plays a role in leaf petiole identity regulation. We show that a mutation of MtPHAN results in petioles acquiring characteristics of the motor organ, pulvinus, including small epidermal cells with extensive cell surface modifications and altered vascular tissue development. Taken together, our results reveal a previously unidentified function of ARP in leaf development. PMID:24603499

  4. Oriented cell division shapes carnivorous pitcher leaves of Sarracenia purpurea

    PubMed Central

    Fukushima, Kenji; Fujita, Hironori; Yamaguchi, Takahiro; Kawaguchi, Masayoshi; Tsukaya, Hirokazu; Hasebe, Mitsuyasu

    2015-01-01

    Complex morphology is an evolutionary outcome of phenotypic diversification. In some carnivorous plants, the ancestral planar leaf has been modified to form a pitcher shape. However, how leaf development was altered during evolution remains unknown. Here we show that the pitcher leaves of Sarracenia purpurea develop through cell division patterns of adaxial tissues that are distinct from those in bifacial and peltate leaves, subsequent to standard expression of adaxial and abaxial marker genes. Differences in the orientation of cell divisions in the adaxial domain cause bifacial growth in the distal region and adaxial ridge protrusion in the middle region. These different growth patterns establish pitcher morphology. A computer simulation suggests that the cell division plane is critical for the pitcher morphogenesis. Our results imply that tissue-specific changes in the orientation of cell division underlie the development of a morphologically complex leaf. PMID:25774486

  5. Oriented cell division shapes carnivorous pitcher leaves of Sarracenia purpurea.

    PubMed

    Fukushima, Kenji; Fujita, Hironori; Yamaguchi, Takahiro; Kawaguchi, Masayoshi; Tsukaya, Hirokazu; Hasebe, Mitsuyasu

    2015-03-16

    Complex morphology is an evolutionary outcome of phenotypic diversification. In some carnivorous plants, the ancestral planar leaf has been modified to form a pitcher shape. However, how leaf development was altered during evolution remains unknown. Here we show that the pitcher leaves of Sarracenia purpurea develop through cell division patterns of adaxial tissues that are distinct from those in bifacial and peltate leaves, subsequent to standard expression of adaxial and abaxial marker genes. Differences in the orientation of cell divisions in the adaxial domain cause bifacial growth in the distal region and adaxial ridge protrusion in the middle region. These different growth patterns establish pitcher morphology. A computer simulation suggests that the cell division plane is critical for the pitcher morphogenesis. Our results imply that tissue-specific changes in the orientation of cell division underlie the development of a morphologically complex leaf.

  6. Spatial Imaging, Speciation, and Quantification of Selenium in theHyperaccumulator Plants Astragalus bisulcatus and Stanleya pinnata

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Freeman, J.L.; Zhang, L.H.; Marcus, M.A.

    2006-09-01

    Astragalus bisulcatus and Stanleya pinnata hyperaccumulate selenium (Se) up to 1% of plant dry weight. In the field, Se was mostly present in the young leaves and reproductive tissues of both hyperaccumulators. Microfocused scanning x-ray fluorescence mapping revealed that Se was hyperaccumulated in trichomes in young leaves of A. bisulcatus. None of 10 other elements tested were accumulated in trichomes. Micro x-ray absorption spectroscopy and liquid chromatography-mass spectrometry showed that Se in trichomes was present in the organic forms methylselenocysteine (MeSeCys; 53%) and {gamma}-glutamyl-MeSeCys (47%). In the young leaf itself, there was 30% inorganic Se (selenate and selenite) in additionmore » to 70% MeSeCys. In young S. pinnata leaves, Se was highly concentrated near the leaf edge and surface in globular structures that were shown by energy-dispersive x-ray microanalysis to be mainly in epidermal cells. Liquid chromatography-mass spectrometry revealed both MeSeCys (88%) and selenocystathionine (12%) inside leaf edges. In contrast, both the Se accumulator Brassica juncea and the nonaccumulator Arabidopsis thaliana accumulated Se in their leaf vascular tissues and mesophyll cells. Se in hyperaccumulators appears to be mobile in both the xylem and phloem because Se-treated S. pinnata was found to be highly toxic to phloem-feeding aphids, and MeSeCys was present in the vascular tissues of a S. pinnata young leaf petiole as well as in guttation fluid. The compartmentation of organic selenocompounds in specific storage areas in the plant periphery appears to be a unique property of Se hyperaccumulators. The high concentration of Se in the plant periphery may contribute to Se tolerance and may also serve as an elemental plant defense mechanism.« less

  7. Molecular imaging of alkaloids in khat (Catha edulis) leaves with MeV-SIMS

    NASA Astrophysics Data System (ADS)

    Jenčič, Boštjan; Jeromel, Luka; Ogrinc Potočnik, Nina; Vogel-Mikuš, Katarina; Vavpetič, Primož; Rupnik, Zdravko; Bučar, Klemen; Vencelj, Matjaž; Kelemen, Mitja; Matsuo, Jiro; Kusakari, Masakazu; Siketić, Zdravko; Al-Jalali, Muhammad A.; Shaltout, Abdallah; Pelicon, Primož

    2017-08-01

    Imaging Mass Spectroscopy (IMS) is a unique research tool providing localization and identification of a wide range of biomolecules as essential data to understand biochemical processes in living organisms. Secondary Ion Mass Spectrometry with high-energy heavy ions (MeV-SIMS) is emerging as a promising IMS technique for chemical imaging of biological tissue. We measured the molecular mass spatial distributions in leaves of khat (Catha edulis). Khat is a natural drug plant, native to eastern Africa and the Arabian Peninsula. In these countries, fresh leaves are being chewed by significant part of population. It was reported that 80% of the adult population in Yemen chew the khat leaves. The main stimulating effects of khat are induced by a monoamine alkaloid called cathinone. During leaf ageing, cathinone is further metabolised to cathine and norephedrine. Earlier studies identified the alkaloids in khat, however little is known on their spatial distribution, reflecting the biosynthesis and accumulation in the tissue. Chemical mapping of alkaloids on cross-sections of khat leaves by MeV-SIMS was done at JSI by a pulsed 5.8 MeV 35Cl6+ beam, focused to a diameter of 15 μm, using a linear time-of-flight (TOF) spectrometer with a mass resolution of 500. In addition, measurements of MeV-SIMS mass spectra were performed at Kyoto University by a continuous broad beam of 6 MeV 63Cu4+ ions at an orthogonal TOF spectrometer with a high mass resolution of 11,000. Sections of leaves were analysed and mass spectra obtained at both MeV-SIMS setups were compared. Tissue-level distributions of detected alkaloids are presented and discussed.

  8. A model using marginal efficiency of investment to analyze carbon and nitrogen interactions in terrestrial ecosystems (ACONITE Version 1)

    NASA Astrophysics Data System (ADS)

    Thomas, R. Q.; Williams, M.

    2014-09-01

    Carbon (C) and nitrogen (N) cycles are coupled in terrestrial ecosystems through multiple processes including photosynthesis, tissue allocation, respiration, N fixation, N uptake, and decomposition of litter and soil organic matter. Capturing the constraint of N on terrestrial C uptake and storage has been a focus of the Earth System Modeling community. However, there is little understanding of the trade-offs and sensitivities of allocating C and N to different tissues in order to optimize the productivity of plants. Here we describe a new, simple model of ecosystem C-N cycling and interactions (ACONITE), that builds on theory related to plant economics in order to predict key ecosystem properties (leaf area index, leaf C : N, N fixation, and plant C use efficiency) based on the outcome of assessments of the marginal change in net C or N uptake associated with a change in allocation of C or N to plant tissues. We simulated and evaluated steady-state ecosystem stocks and fluxes in three different forest ecosystems types (tropical evergreen, temperate deciduous, and temperate evergreen). Leaf C : N differed among the three ecosystem types (temperate deciduous < tropical evergreen < temperature evergreen), a result that compared well to observations from a global database describing plant traits. Gross primary productivity (GPP) and net primary productivity (NPP) estimates compared well to observed fluxes at the simulation sites. Simulated N fixation at steady-state, calculated based on relative demand for N and the marginal return on C investment to acquire N, was an order of magnitude higher in the tropical forest than in the temperate forest, consistent with observations. A sensitivity analysis revealed that parameterization of the relationship between leaf N and leaf respiration had the largest influence on leaf area index and leaf C : N. A parameter governing how photosynthesis scales with day length had the largest influence on total vegetation C, GPP, and NPP. Multiple parameters associated with photosynthesis, respiration, and N uptake influenced the rate of N fixation. Overall, our ability to constrain leaf area index and allow spatially and temporally variable leaf C : N can help address challenges simulating these properties in ecosystem and Earth System models. Furthermore, the simple approach with emergent properties based on coupled C-N dynamics has potential for use in research that uses data-assimilation methods to integrate data on both the C and N cycles to improve C flux forecasts.

  9. Molecular characteristics of continuously released DOM during one year of root and leaf litter decomposition

    NASA Astrophysics Data System (ADS)

    Altmann, Jens; Jansen, Boris; Kalbitz, Karsten; Filley, Timothy

    2013-04-01

    Dissolved organic matter (DOM) is one of the most dynamic carbon pools linking the terrestrial with the aquatic carbon cycle. Besides the insecure contribution of terrestrial DOM to the greenhouse effect, DOM also plays an important role for the mobility and availability of heavy metals and organic pollutants in soils. These processes depend very much on the molecular characteristics of the DOM. Surprisingly the processes that determine the molecular composition of DOM are only poorly understood. DOM can originate from various sources, which influence its molecular composition. It has been recognized that DOM formation is not a static process and DOM characteristics vary not only between different carbon sources. However, molecular characteristics of DOM extracts have scarcely been studied continuously over a longer period of time. Due to constant molecular changes of the parent litter material or soil organic matter during microbial degradation, we assumed that also the molecular characteristics of litter derived DOM varies at different stages during root and needle decomposition. For this study we analyzed the chemical composition of root and leaf samples of 6 temperate tree species during one year of litter decomposition in a laboratory incubation. During this long-term experiment we measured continuously carbon and nitrogen contents of the water extracts and the remaining residues, C mineralization rates, and the chemical composition of water extracts and residues by Curie-point pyrolysis mass spectrometry with TMAH We focused on the following questions: (I) How mobile are molecules derived from plant polymers like tannin, lignin, suberin and cutin? (II) How does the composition of root and leaf derived DOM change over time in dependence on the stage of decomposition and species? Litter derived DOM was generally dominated by aromatic compounds. Substituded fatty acids as typically cutin or suberin derived were not detected in the water extracts. Fresh leaf and needle samples released a much higher amount of tannins than fresh root samples. At later litter decomposition stages the influence of tannins decreased and lignin derived phenols dominated the extracts. With ongoing litter degradation the degree of oxidation for the litter material increased, which was also reflected by the water extracted molecules.

  10. Antihypertensive potential of the aqueous extract which combine leaf of Persea americana Mill. (Lauraceae), stems and leaf of Cymbopogon citratus (D.C) Stapf. (Poaceae), fruits of Citrus medical L. (Rutaceae) as well as honey in ethanol and sucrose experimental model.

    PubMed

    Dzeufiet, Paul Désiré Djomeni; Mogueo, Amélie; Bilanda, Danielle Claude; Aboubakar, Bibi-Farouck Oumarou; Tédong, Léonard; Dimo, Théophile; Kamtchouing, Pierre

    2014-12-17

    The present study was designed to evaluate the effects of the aqueous extract obtained from the mixture of fresh leaf of Persea americana, stems and fresh leaf of Cymbopogon citratus, fruits of Citrus medica and honey on ethanol and sucrose induced hypertension in rats. Rats were divided into eight groups of 6 rats each and daily treated for 5 weeks. The control group received distilled water (1 mL/kg) while rats of groups 2, 3 and 4 received ethanol 40 degrees (3 g/kg/day), 10% sucrose as drinking water and the two substances respectively. The remaining groups received in addition to sucrose and ethanol, the aqueous extract (50, 100 and 150 mg/kg) or nifedipine (10 mg/kg) respectively. Many parameters including hemodynamic, biochemical and histopathological were assessed at the end of the study. The concomitant consumption of ethanol and sucrose significantly (p < 0.001) increased the blood pressure and the heart rate compared to distilled water treated-rats. The levels of total cholesterol, LDL-cholesterol, triglycerides, atherogenic index, glucose, proteins, AST, ALT, creatinin, potassium, sodium and albumin increased while the HDL-cholesterol decreased under ethanol and sucrose feeding. Chronic ethanol and sucrose intake significantly decreased the activities of superoxide dismutase (SOD) and catalase (CAT) as well as the contents of reduced glutathione (GSH) and nitrites whereas elevated the malondialdehyde (MDA) levels. Histological analysis revealed among other vascular congestion, inflammation, tubular clarification and thickening of the vessel wall in rats treated with alcohol and sucrose. Administration of the aqueous extract or nifedipine prevented the hemodynamic, biochemical, oxidative and histological impairments induced chronic ethanol and sucrose consumption. Current results suggest that the aqueous extract used in this study possess antihypertensive activity against ethanol and sucrose induced hypertension in rats by the improvement of biochemical and oxidative status, and by protecting liver, kidney and vascular endothelium against damages induced by chronic consumption of ethanol and sucrose.

  11. Biocontrol of Escherichia coli O157: H7 on fresh-cut leafy greens.

    PubMed

    Boyacioglu, Olcay; Sharma, Manan; Sulakvelidze, Alexander; Goktepe, Ipek

    2013-01-01

    The effect of a bacteriophage cocktail (EcoShield™) that is specific against Escherichia coli O157:H7 was evaluated against a nalidixic acid-resistant enterohemorrhagic E. coli O157:H7 RM4407 (EHEC) strain on leafy greens stored under either (1) ambient air or (2) modified atmosphere (MA; 5% O 2 /35% CO 2 /60% N 2 ). Pieces (~2 × 2 cm 2 ) of leafy greens (lettuce and spinach) inoculated with 4.5 log CFU/cm 2 EHEC were sprayed with EcoShield™ (6.5 log PFU/cm 2 ). Samples were stored at 4 or 10°C for up to 15 d. On spinach, the level of EHEC declined by 2.38 and 2.49 log CFU/cm 2 at 4 and 10°C, respectively, 30 min after phage application (p ≤ 0.05). EcoShield™ was also effective in reducing EHEC on the surface of green leaf lettuce stored at 4°C by 2.49 and 3.28 log units in 30 min and 2 h, respectively (p ≤ 0.05). At 4°C under atmospheric air, the phage cocktail significantly (p ≤ 0.05) lowered the EHEC counts in one day by 1.19, 3.21 and 3.25 log CFU/cm 2 on spinach, green leaf and romaine lettuce, respectively compared with control (no bacteriophage) treatments. When stored under MA at 4°C, phages reduced (p ≤ 0.05) EHEC populations by 2.18, 3.50 and 3.13 log CFU/cm 2 , on spinach, green leaf and romaine lettuce. At 10°C, EHEC reductions under atmospheric air storage were 1.99, 3.90 and 3.99 log CFU/cm 2 (p ≤ 0.05), while population reductions under MA were 3.08, 3.89 and 4.34 logs on spinach, green leaf and romaine lettuce, respectively, compared with controls (p ≤ 0.05). The results of this study showed that bacteriophages were effective in reducing the levels of E. coli O157:H7 on fresh leafy produce, and that the reduction was further improved when produce was stored under the MA conditions.

  12. Detection of triterpene acids distribution in loquat (Eriobotrya japonica) leaf using hyperspectral imaging

    NASA Astrophysics Data System (ADS)

    Shi, Jiyong; Chen, Wu; Zou, Xiaobo; Xu, Yiwei; Huang, Xiaowei; Zhu, Yaodi; Shen, Tingting

    2018-01-01

    Hyperspectral images (431-962 nm) and partial least squares (PLS) were used to detect the distribution of triterpene acids within loquat (Eriobotrya japonica) leaves. 72 fresh loquat leaves in the young group, mature group and old group were collected for hyperspectral imaging; and triterpene acids content of the loquat leaves was analyzed using high performance liquid chromatography (HPLC). Then the spectral data of loquat leaf hyperspectral images and the triterpene acids content were employed to build calibration models. After spectra pre-processing and wavelength selection, an optimum calibration model (Rp = 0.8473, RMSEP = 2.61 mg/g) for predicting triterpene acids was obtained by synergy interval partial least squares (siPLS). Finally, spectral data of each pixel in the loquat leaf hyperspectral image were extracted and substituted into the optimum calibration model to predict triterpene acids content of each pixel. Therefore, the distribution map of triterpene acids content was obtained. As shown in the distribution map, triterpene acids are accumulated mainly in the leaf mesophyll regions near the main veins, and triterpene acids concentration of young group is less than that of mature and old groups. This study showed that hyperspectral imaging is suitable to determine the distribution of active constituent content in medical herbs in a rapid and non-invasive manner.

  13. Histopathological investigations of the infection process and propagule development of Phytophthora ramorumon rhododendron leaves

    Treesearch

    Marko Riedel; Sabine Werres; Marianne Elliott; Katie McKeever; Simon Shamoun

    2012-01-01

    Studies on the relationship between rhododendron and Phytophthora ramorum include the influence of wounds on leaf infection and on the development of leaf necrosis (De Dobbelaere et al. 2010; Denman et al. 2005), the influence of the inoculum type (Widmer 2009), and tissue colonization by P. ramorum (Brown and Brasier 2007;...

  14. Somaclonal variation in hybrid poplars for resistance to Septoria leaf spot

    Treesearch

    M.E. Ostry; D. D. Skilling

    1987-01-01

    Tissue culture techniques have been used to obtain hybrid poplars with putative resistance to leaf spot caused by Septoria musiva from clones previously susceptible to the disease. Stem internode explants were used to obtain proliferating callus cultures. Adventitious bud formation and shoot proliferation were then induced. Elongated shoots were excised and rooted in a...

  15. Slow Freezing, but Not Vitrification Supports Complete Spermatogenesis in Cryopreserved, Neonatal Sheep Testicular Xenografts

    PubMed Central

    Pukazhenthi, Budhan S.; Nagashima, Jennifer; Travis, Alexander J.; Costa, Guilherme M.; Escobar, Enrique N.; França, Luiz R.; Wildt, David E.

    2015-01-01

    The ability to spur growth of early stage gametic cells recovered from neonates could lead to significant advances in rescuing the genomes of rare genotypes or endangered species that die unexpectedly. The purpose of this study was to determine, for the first time, the ability of two substantially different cryopreservation approaches, slow freezing versus vitrification, to preserve testicular tissue of the neonatal sheep and subsequently allow initiation of spermatogenesis post-xenografting. Testis tissue from four lambs (3-5 wk old) was processed and then untreated or subjected to slow freezing or vitrification. Tissue pieces (fresh, n = 214; slow freezing, then thawing, n = 196; vitrification, then warming, n = 139) were placed subcutaneously under the dorsal skin of SCID mice and then grafts recovered and evaluated 17 wk later. Grafts from fresh and slow frozen tissue contained the most advanced stages of spermatogenesis, including normal tubule architecture with elongating spermatids in ~1% (fresh) and ~10% (slow frozen) of tubules. Fewer than 2% of seminiferous tubules advanced to the primary spermatocyte stage in xenografts derived from vitrified tissue. Results demonstrate that slow freezing of neonatal lamb testes was far superior to vitrification in preserving cellular integrity and function after xenografting, including allowing ~10% of tubules to retain the capacity to resume spermatogenesis and yield mature spermatozoa. Although a first for any ruminant species, findings also illustrate the importance of preemptive studies that examine cryo-sensitivity of testicular tissue before attempting this type of male fertility preservation on a large scale. PMID:25923660

  16. Diurnal Cycling Transcription Factors of Pineapple Revealed by Genome-Wide Annotation and Global Transcriptomic Analysis.

    PubMed

    Sharma, Anupma; Wai, Ching Man; Ming, Ray; Yu, Qingyi

    2017-09-01

    Circadian clock provides fitness advantage by coordinating internal metabolic and physiological processes to external cyclic environments. Core clock components exhibit daily rhythmic changes in gene expression, and the majority of them are transcription factors (TFs) and transcription coregulators (TCs). We annotated 1,398 TFs from 67 TF families and 80 TCs from 20 TC families in pineapple, and analyzed their tissue-specific and diurnal expression patterns. Approximately 42% of TFs and 45% of TCs displayed diel rhythmic expression, including 177 TF/TCs cycling only in the nonphotosynthetic leaf tissue, 247 cycling only in the photosynthetic leaf tissue, and 201 cycling in both. We identified 68 TF/TCs whose cycling expression was tightly coupled between the photosynthetic and nonphotosynthetic leaf tissues. These TF/TCs likely coordinate key biological processes in pineapple as we demonstrated that this group is enriched in homologous genes that form the core circadian clock in Arabidopsis and includes a STOP1 homolog. Two lines of evidence support the important role of the STOP1 homolog in regulating CAM photosynthesis in pineapple. First, STOP1 responds to acidic pH and regulates a malate channel in multiple plant species. Second, the cycling expression pattern of the pineapple STOP1 and the diurnal pattern of malate accumulation in pineapple leaf are correlated. We further examined duplicate-gene retention and loss in major known circadian genes and refined their evolutionary relationships between pineapple and other plants. Significant variations in duplicate-gene retention and loss were observed for most clock genes in both monocots and dicots. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  17. Internalization of Escherichia coli O157:H7 gfp+ in rocket and Swiss chard baby leaves as affected by abiotic and biotic damage.

    PubMed

    Hartmann, R; Fricke, A; Stützel, H; Mansourian, S; Dekker, T; Wohanka, W; Alsanius, B

    2017-07-01

    Internalization of human pathogens in edible parts of vegetables eaten raw is a major concern, since once internalized they are protected from sanitizing treatments. In this study, we examined the invasion of gfp-labelled Escherichia coli O157:H7 into intact and biotically (infection with Xanthomonas campestris/Pseudomonas syringae) and abiotically (grating with silicon carbide) damaged leaves of wild rocket (Diplotaxis tenuifolia) and Swiss chard (Beta vulgaris subsp. cicla) using laser scanning confocal microscopy. Bacterial cells were found in internal locations of the tissue, irrespective of tissue health status. Contaminated leaf sections of biotically and abiotically damaged wild rocket leaves showed higher susceptibility to microbial invasion, while the pathogen was internalized in greater numbers into intact Swiss chard leaf sections when abiotically, but not biotically, damaged. The greatest differences were observed between the plant species; after surface sanitization, E. coli O157:H7 was still detected in wild rocket leaves, but not in Swiss chard leaves. Contamination of leafy vegetables with Escherichia coli O157:H7 is a growing problem, as reported outbreaks are increasing. However, establishment of this human pathogen in the phyllosphere is not completely understood. Using laser scanning confocal microscopy, we demonstrated that E. coli O157:H7gfp+ can invade plant tissue of Swiss chard and wild rocket leaves and that the bacterium is more sensitive to surface sanitization of Swiss chard leaves. Damage to leaf tissue promoted leaf invasion, but the nature of the damage (abiotic or biotic) and plant species had an impact. © 2017 The Society for Applied Microbiology.

  18. Whole organ, venation and epidermal cell morphological variations are correlated in the leaves of Arabidopsis mutants.

    PubMed

    Pérez-Pérez, José Manuel; Rubio-Díaz, Silvia; Dhondt, Stijn; Hernández-Romero, Diana; Sánchez-Soriano, Joaquín; Beemster, Gerrit T S; Ponce, María Rosa; Micol, José Luis

    2011-12-01

    Despite the large number of genes known to affect leaf shape or size, we still have a relatively poor understanding of how leaf morphology is established. For example, little is known about how cell division and cell expansion are controlled and coordinated within a growing leaf to eventually develop into a laminar organ of a definite size. To obtain a global perspective of the cellular basis of variations in leaf morphology at the organ, tissue and cell levels, we studied a collection of 111 non-allelic mutants with abnormally shaped and/or sized leaves, which broadly represent the mutational variations in Arabidopsis thaliana leaf morphology not associated with lethality. We used image-processing techniques on these mutants to quantify morphological parameters running the gamut from the palisade mesophyll and epidermal cells to the venation, whole leaf and rosette levels. We found positive correlations between epidermal cell size and leaf area, which is consistent with long-standing Avery's hypothesis that the epidermis drives leaf growth. In addition, venation parameters were positively correlated with leaf area, suggesting that leaf growth and vein patterning share some genetic controls. Positional cloning of the genes affected by the studied mutations will eventually establish functional links between genotypes, molecular functions, cellular parameters and leaf phenotypes. © 2011 Blackwell Publishing Ltd.

  19. Selective and cost-effective protocol to separate bioactive triterpene acids from plant matrices using alkalinized ethanol: Application to leaves of Myrtaceae species

    PubMed Central

    Lima, Adélia M. Belem; Siani, Antonio Carlos; Nakamura, Marcos Jun; D’Avila, Luiz Antonio

    2015-01-01

    Background: Triterpenes as betulinic (BA), oleanolic (OA) and ursolic acids (UA) have increasingly gained therapeutic relevance due to their wide scope of pharmacological activities. To fit large-scale demands, exploitable sources of these compounds have to be found and simple, cost-effective methods to extract them developed. Leaf material represents the best plant sustainable raw material. To obtain triterpene acid-rich extracts from leaves of Eugenia, Psidium and Syzygium species (Myrtaceae) by directly treating the dry plant material with alkalinized hydrated ethanol. This procedure was adapted from earlier methods to effect depolymerization of the leaf cutin. Materials and Methods: Extracts were prepared by shaking the milled dry leaves in freshly prepared 2% NaOH in 95% EtOH solution (1:4 w/v) at room temperature for 6 h. Working up the product in acidic aqueous medium led to clear precipitates in which BA, OA and UA were quantified by gas chromatography. Results: Pigment-free and low-polyphenol content extracts (1.2–2.8%) containing 6–50% of total triterpene acids were obtained for the six species assayed. UA (7–20%) predominated in most extracts, but BA preponderated in Eugenia florida (39%). Carried out in parallel, n-hexane defatted leaves led to up to 9% enhancement of total acids in the extracts. The hydroalcoholate treatment of Myrtaceae species dry leaves proved to be a cost-effective and environmentally friendly method to obtain triterpene acids, providing them be resistant to alkaline medium. These combined techniques might be applicable to other plant species and tissues. PMID:26246721

  20. Foliar trichome- and aquaporin-aided water uptake in a drought-resistant epiphyte Tillandsia ionantha Planchon.

    PubMed

    Ohrui, T; Nobira, H; Sakata, Y; Taji, T; Yamamoto, C; Nishida, K; Yamakawa, T; Sasuga, Y; Yaguchi, Y; Takenaga, H; Tanaka, Shigeo

    2007-12-01

    The atmospheric epiphyte Tillandsia ionantha is capable of surviving drought stress for 6 months or more without any exogenous water supply via an as of yet to be determined mechanism. When plants were soaked in water for 3 h, leaves absorbed a remarkably large amount of water (30-40% on the basis of fresh weight), exhibiting a bimodal absorption pattern. Radiolabeled water was taken up by the leaves by capillary action of the epidermal trichomes within 1 min (phase 1) and then transported intracellularly to leaf tissues over 3 h (phase 2). The removal of epidermal trichome wings from leaves as well as rinsing leaves with water significantly lowered the extracellular accumulation of water on leaf surfaces. The intracellular transport of water was inhibited by mercuric chloride, implicating the involvement of a water channel aquaporin in second-phase water absorption. Four cDNA clones (TiPIP1a, TiPIP1b, TiPIP1c, and TiPIP2a) homologous to PIP family aquaporins were isolated from the leaves, and RT-PCR showed that soaking plants in water stimulated the expression of TiPIP2a mRNA, suggesting the reinforcement in ability to rapidly absorb a large amount of water. The expression of TiPIP2a complementary RNA in Xenopus oocytes enhanced permeability, and treatment with inhibitors suggested that the water channel activity of TiPIP2a protein was regulated by phosphorylation. Thus, the high water uptake capability of T. ionantha leaves surviving drought is attributable to a bimodal trichome- and aquaporin-aided water uptake system based on rapid physical collection of water and subsequent, sustained chemical absorption.

  1. Overexpression of NAC gene from Lepidium latifolium L. enhances biomass, shortens life cycle and induces cold stress tolerance in tobacco: potential for engineering fourth generation biofuel crops.

    PubMed

    Grover, Atul; Singh, Sadhana; Pandey, Pankaj; Patade, Vikas Yadav; Gupta, Sanjay Mohan; Nasim, M

    2014-11-01

    We report elevated biomass and altered growth characteristics of tobacco plants up on transformation with a NAC (NAM, ATAF1/2,CUC2) gene (GenBank Accession FJ754254) isolated from Lepidium latifolium L. (LlaNAC). Transgenic plants showed significant differences in fresh weight, midrib length of longest leaf, leaf area, height of the plant, root and shoot weights, etc. during vegetative phase. On 100th day after sowing (DAS), plants of transgenic lines were 2-3 times taller than the wild type plants, though no significant difference was recorded in moisture contents of any of the plant tissues. Over-expression of NAC gene up to 2,000 fold was recorded in leaves of transgenic plants on 100th DAS. Interestingly, transgenic plants showed significantly shortened (P(t) = 0.02-0.04) life cycle, as they showed a completely altered growth behaviour. Transgenic plants entered reproductive phase earlier by 60 days, with lines NC2 and NC7b entering first, followed by line NC10. However, the time period spent in the reproductive phase by the plant was nearly twice in case of transgenic lines NC2, NC7b and NC10, as compared to the wild type plants. Despite that, these lines completed their life cycle in 45-60 days lesser than the time taken by wild-type tobacco plants. No difference was recorded in fruit and seed yield of transgenic or wild type plants. To the best of our knowledge, this is the first report on over-expression of NAC gene causing altered growth and biomass patterns. We expect this study to become an important reference towards future engineering of plants for fuel and fodder purposes.

  2. Cluster-root formation and carboxylate release in three Lupinus species as dependent on phosphorus supply, internal phosphorus concentration and relative growth rate

    PubMed Central

    Wang, Xing; Pearse, Stuart J.; Lambers, Hans

    2013-01-01

    Background and Aims Some Lupinus species produce cluster roots in response to low plant phosphorus (P) status. The cause of variation in cluster-root formation among cluster-root-forming Lupinus species is unknown. The aim of this study was to investigate if cluster-root formation is, in part, dependent on different relative growth rates (RGRs) among Lupinus species when they show similar shoot P status. Methods Three cluster-root-forming Lupinus species, L. albus, L. pilosus and L. atlanticus, were grown in washed river sand at 0, 7·5, 15 or 40 mg P kg−1 dry sand. Plants were harvested at 34, 42 or 62 d after sowing, and fresh and dry weight of leaves, stems, cluster roots and non-cluster roots of different ages were measured. The percentage of cluster roots, tissue P concentrations, root exudates and plant RGR were determined. Key Results Phosphorus treatments had major effects on cluster-root allocation, with a significant but incomplete suppression in L. albus and L. pilosus when P supply exceeded 15 mg P kg−1 sand. Complete suppression was found in L. atlanticus at the highest P supply; this species never invested more than 20 % of its root weight in cluster roots. For L. pilosus and L. atlanticus, cluster-root formation was decreased at high internal P concentration, irrespective of RGR. For L. albus, there was a trend in the same direction, but this was not significant. Conclusions Cluster-root formation in all three Lupinus species was suppressed at high leaf P concentration, irrespective of RGR. Variation in cluster-root formation among the three species cannot be explained by species-specific variation in RGR or leaf P concentration. PMID:24061491

  3. Cluster-root formation and carboxylate release in three Lupinus species as dependent on phosphorus supply, internal phosphorus concentration and relative growth rate.

    PubMed

    Wang, Xing; Pearse, Stuart J; Lambers, Hans

    2013-11-01

    Some Lupinus species produce cluster roots in response to low plant phosphorus (P) status. The cause of variation in cluster-root formation among cluster-root-forming Lupinus species is unknown. The aim of this study was to investigate if cluster-root formation is, in part, dependent on different relative growth rates (RGRs) among Lupinus species when they show similar shoot P status. Three cluster-root-forming Lupinus species, L. albus, L. pilosus and L. atlanticus, were grown in washed river sand at 0, 7·5, 15 or 40 mg P kg(-1) dry sand. Plants were harvested at 34, 42 or 62 d after sowing, and fresh and dry weight of leaves, stems, cluster roots and non-cluster roots of different ages were measured. The percentage of cluster roots, tissue P concentrations, root exudates and plant RGR were determined. Phosphorus treatments had major effects on cluster-root allocation, with a significant but incomplete suppression in L. albus and L. pilosus when P supply exceeded 15 mg P kg(-1) sand. Complete suppression was found in L. atlanticus at the highest P supply; this species never invested more than 20 % of its root weight in cluster roots. For L. pilosus and L. atlanticus, cluster-root formation was decreased at high internal P concentration, irrespective of RGR. For L. albus, there was a trend in the same direction, but this was not significant. Cluster-root formation in all three Lupinus species was suppressed at high leaf P concentration, irrespective of RGR. Variation in cluster-root formation among the three species cannot be explained by species-specific variation in RGR or leaf P concentration.

  4. Transgenic rice expressing the cry2AX1 gene confers resistance to multiple lepidopteran pests.

    PubMed

    Chakraborty, M; Reddy, P Sairam; Mustafa, G; Rajesh, G; Narasu, V M Laxmi; Udayasuriyan, V; Rana, Debashis

    2016-10-01

    A chimeric Bacillus thuringiensis toxin (Bt) gene, cry2AX1was cloned in a bi-selectable marker free binary vector construct. The cry2AX1 gene, driven by the Chrysanthemum rbcS1 promoter, was introduced into JK1044R, the restorer line (Oryza sativa L. ssp. Indica) of a notified commercially grown rice hybrid in India, by Agrobacterium-mediated transformation. Its effect against two major lepidopteran insect pests viz., yellow stem borer (YSB) Scirpophaga incertulas, rice leaf folder (RLF) Cnaphalocrocis medinalis and one minor insect pest, oriental army worm (OAW) Mythimna separata was demonstrated through bioassays of transgenic rice plants under laboratory and greenhouse conditions. The rbcS1 promoter with chloroplast signal peptide was used to avoid Cry2AX1 protein expression in rice seed endosperm tissue. A total of 37 independent transformants were generated, of which after preliminary molecular characterization and YSB bioassay screening, five events were selected for their protein expression and bioefficacy against all three rice insect. One elite transgenic rice line, BtE15, was identified with Cry2AX1 expression ranging from 0.68 to 1.34 µg g(-1) leaf fresh weight and with 80-92 % levels of resistance against rice pests at the vegetative and reproductive stages. Increase in Cry2AX1 protein concentration was also observed with crop maturity. The Cry2AX1protein concentration in the de-husked seeds was negligible (as low as 2.7-3.6 ng g(-1)). These results indicate the potential application of cry2AX1 gene in rice for protection against YSB, RLF and OAW.

  5. Influence of near null magnetic field on in vitro growth of potato and wild Solanum species.

    PubMed

    Rakosy-Tican, Lenuta; Aurori, C M; Morariu, V V

    2005-10-01

    The influence of near null magnetic field on in vitro growth of different cultures of potato and related Solanum species was investigated for various exposure times and dates. Potato (Solanum tuberosum L. cv. Désirée) in vitro cultures of shoot tips or nodal segments were used. Three different exposure periods revealed either stimulation or inhibition of root, stem, or leaf in vitro growth after 14 or 28 days of exposure. In one experiment the significant stimulation of leaf growth was also demonstrated at biochemical level, the quantity of chlorophyll a and b and carotenoids increasing more than two-fold. For the wild species Solanum chacoense, S. microdontum, and S. verrucosum, standardized in vitro cultures of nodal stem segments were used. Root and stem growth was either stimulated or slightly inhibited after 9 days exposure to near null magnetic field. Callus cultures obtained from potato dihaploid line 120/19 were maintained in near null magnetic field in 2 different months. For these experiments as well as for Solanum verrucosum, callus cultures recorded either slight inhibition or no effect on fresh weight. For all experiments significant growth variation was brought about only when geomagnetic activity (AP index) showed variations at the beginning of in vitro growth and when the explant had at least one meristematic tissue. Moreover longer maintenance in near null magnetic field, 28 days as compared to 14 days or the controls, can also make a difference in plant growth in response to geomagnetic field variations when static component was reduced to zero value. These results of in vitro plant growth stimulation by variable component of geomagnetic field also sustain the so-called seasonal "window" effect. (c) 2005 Wiley-Liss, Inc.

  6. Efficacy of chlorine, acidic electrolyzed water and aqueous chlorine dioxide solutions to decontaminate Escherichia coli O157:H7 from lettuce leaves.

    PubMed

    Keskinen, Lindsey A; Burke, Angela; Annous, Bassam A

    2009-06-30

    This study compared the efficacy of chlorine (20-200 ppm), acidic electrolyzed water (50 ppm chlorine, pH 2.6), acidified sodium chlorite (20-200 ppm chlorite ion concentration, Sanova), and aqueous chlorine dioxide (20-200 ppm chlorite ion concentration, TriNova) washes in reducing populations of Escherichia coli O157:H7 on artificially inoculated lettuce. Fresh-cut leaves of Romaine or Iceberg lettuce were inoculated by immersion in water containing E. coli O157:H7 (8 log CFU/ml) for 5 min and dried in a salad spinner. Leaves (25 g) were then washed for 2 min, immediately or following 24 h of storage at 4 degrees C. The washing treatments containing chlorite ion concentrations of 100 and 200 ppm were the most effective against E. coli O157:H7 populations on Iceberg lettuce, with log reductions as high as 1.25 log CFU/g and 1.05 log CFU/g for TriNova and Sanova wash treatments, respectively. All other wash treatments resulted in population reductions of less than 1 log CFU/g. Chlorine (200 ppm), TriNova, Sanova, and acidic electrolyzed water were all equally effective against E. coli O157:H7 on Romaine, with log reductions of approximately 1 log CFU/g. The 20 ppm chlorine wash was as effective as the deionized water wash in reducing populations of E. coli O157:H7 on Romaine and Iceberg lettuce. Scanning electron microscopy indicated that E. coli O157:H7 that was incorporated into biofilms or located in damage lettuce tissue remained on the lettuce leaf, while individual cells on undamaged leaf surfaces were more likely to be washed away.

  7. Detection of fresh bruises in apples by structured-illumination reflectance imaging

    USDA-ARS?s Scientific Manuscript database

    Detection of fresh bruises in apples remains a challenging task due to the absence of visual symptoms and significant chemical alterations of fruit tissues during the initial stage after the fruit have been bruised. This paper reports on a new structured-illumination reflectance imaging (SIRI) techn...

  8. Photoacoustic characterization of the left atrium wall: healthy and ablated tissue (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Iskander-Rizk, Sophinese; Kruizinga, Pieter; van der Steen, Antonius F. W.; van Soest, Gijs

    2017-03-01

    Radio-frequency ablation (RFA) creates a thermal lesion in the atrial wall, with clearly recognizable optical and structural changes to the tissue. This can be detected by photoacoustic (PA) imaging, and used for monitoring of lesion depth, lesion functionality, and limiting excessive ablation. Porcine left atrium tissue can be split into three visually distinguishable regions, a thick white endocardium, pinkish myocardium and a thin gelatinous epicardium. In this study, we characterize the layered left atrium tissue in terms of the relevant photoacoustic parameters (wavelength, frequency content, imaging depth, lesion contrast). Previous studies in the literature targeted the photoacoustic characterization of fresh and ablated ventricular myocardium in the range of 650nm to 900nm. In this study we target the characterization of fresh and ablated left atrial tissue from 410nm to 1000nm, including the endocardium and epicardium. We generate the photoacoustic signals using a tunable pulsed laser source, and record those signals using either a broadband 1 mm hydrophone or a L12-3v transducer connected to the Verasonics machine for more realistic conditions. Initial experiments on fresh porcine tissue show that the presence of the endocardium and epicardium layers do affect the photoacoustic signal received. The signal recorded is representative of the difference in optical and mechanical properties between the layers. Ablated and non-ablated tissue also present differences in spectra. The determined optical contrast could be used in the PA monitoring of RFA lesion to monitor the extension of the lesion to the edge of the myocardium-epicardium border avoiding complications related to over ablation.

  9. Does oolong tea (Camellia sinensis) made from a combination of leaf and stem smell more aromatic than leaf-only tea? Contribution of the stem to oolong tea aroma.

    PubMed

    Zeng, Lanting; Zhou, Ying; Fu, Xiumin; Mei, Xin; Cheng, Sihua; Gui, Jiadong; Dong, Fang; Tang, Jinchi; Ma, Shengzhou; Yang, Ziyin

    2017-12-15

    The raw materials used to make oolong tea (Camellia sinensis) are a combination of leaf and stem. Oolong tea made from leaf and stem is thought to have a more aromatic smell than leaf-only tea. However, there is no available evidence to support the viewpoint. In this study, sensory evaluation and detailed characterization of emitted and internal volatiles (not readily emitted, but stored in samples) of dry oolong teas and infusions indicated that the presence of stem did not significantly improve the total aroma characteristics. During the enzyme-active processes, volatile monoterpenes and theanine were accumulated more abundantly in stem than in leaf, while jasmine lactone, indole, and trans-nerolidol were lower in stem than in leaf. Tissue-specific aroma-related gene expression and availability of precursors of aroma compounds resulted in different aroma distributions in leaf and stem. This study presents the first determination of the contribution of stem to oolong tea aroma. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Salt tolerance, salt accumulation, and ionic homeostasis in an epidermal bladder-cell-less mutant of the common ice plant Mesembryanthemum crystallinum.

    PubMed

    Agarie, Sakae; Shimoda, Toshifumi; Shimizu, Yumi; Baumann, Kathleen; Sunagawa, Haruki; Kondo, Ayumu; Ueno, Osamu; Nakahara, Teruhisa; Nose, Akihiro; Cushman, John C

    2007-01-01

    The aerial surfaces of the common or crystalline ice plant Mesembryanthemum crystallinum L., a halophytic, facultative crassulacean acid metabolism species, are covered with specialized trichome cells called epidermal bladder cells (EBCs). EBCs are thought to serve as a peripheral salinity and/or water storage organ to improve survival under high salinity or water deficit stress conditions. However, the exact contribution of EBCs to salt tolerance in the ice plant remains poorly understood. An M. crystallinum mutant lacking EBCs was isolated from plant collections mutagenized by fast neutron irradiation. Light and electron microscopy revealed that mutant plants lacked EBCs on all surfaces of leaves and stems. Dry weight gain of aerial parts of the mutant was almost half that of wild-type plants after 3 weeks of growth at 400 mM NaCl. The EBC mutant also showed reduced leaf succulence and leaf and stem water contents compared with wild-type plants. Aerial tissues of wild-type plants had approximately 1.5-fold higher Na(+) and Cl(-) content than the mutant grown under 400 mM NaCl for 2 weeks. Na(+) and Cl(-) partitioning into EBCs of wild-type plants resulted in lower concentrations of these ions in photosynthetically active leaf tissues than in leaves of the EBC-less mutant, particularly under conditions of high salt stress. Potassium, nitrate, and phosphate ion content decreased with incorporation of NaCl into tissues in both the wild type and the mutant, but the ratios of Na(+)/K(+) and Cl(-)/NO(3)(-)content were maintained only in the leaf and stem tissues of wild-type plants. The EBC mutant showed significant impairment in plant productivity under salt stress as evaluated by seed pod and seed number and average seed weight. These results clearly show that EBCs contribute to succulence by serving as a water storage reservoir and to salt tolerance by maintaining ion sequestration and homeostasis within photosynthetically active tissues of M. crystallinum.

  11. Effects of perchlorate on growth of four wetland plants and its accumulation in plant tissues.

    PubMed

    He, Hongzhi; Gao, Haishuo; Chen, Guikui; Li, Huashou; Lin, Hai; Shu, Zhenzhen

    2013-10-01

    Perchlorate contamination in water is of concern because of uncertainties about toxicity and health effects, impact on ecosystems, and possible indirect exposure pathways to humans. Therefore, it is very important to investigate the ecotoxicology of perchlorate and to screen plant species for phytoremediation. Effects of perchlorate (20, 200, and 500 mg/L) on the growth of four wetland plants (Eichhornia crassipes, Acorus calamus L., Thalia dealbata, and Canna indica) as well as its accumulation in different plant tissues were investigated through water culture experiments. Twenty milligrams per liter of perchlorate had no significant effects on height, root length, aboveground part weight, root weight, and oxidizing power of roots of four plants, except A. calamus, and increasing concentrations of perchlorate showed that out of the four wetland plants, only A. calamus had a significant (p<0.05) dose-dependent decrease in these parameters. When treated with 500 mg/L perchlorate, these parameters and chlorophyll content in the leaf of plants showed significant decline contrasted to control groups, except the root length of E. crassipes and C. indica. The order of inhibition rates of perchlorate on root length, aboveground part weight and root weight, and oxidizing power of roots was: A. calamus > C. indica > T. dealbata > E. crassipes and on chlorophyll content in the leaf it was: A. calamus > T. dealbata > C. indica > E. crassipes. The higher the concentration of perchlorate used, the higher the amount of perchlorate accumulation in plants. Perchlorate accumulation in aboveground tissues was much higher than that in underground tissues and leaf was the main tissue for perchlorate accumulation. The order of perchlorate accumulation content and the bioconcentration factor in leaf of four plants was: E. crassipes > C. indica > T. dealbata > A. calamus. Therefore, E. crassipes might be an ideal plant with high tolerance ability and accumulation ability for constructing wetland to remediate high levels of perchlorate polluted water.

  12. Preservation of Plant Biomolecules and the Relevance to the Interpretation of Paleoenvironmental Signals: Tertiary Metasequoia Fossils as Examples

    NASA Astrophysics Data System (ADS)

    Yang, H.; Leng, Q.

    2004-12-01

    The degradation and preservation of biomolecules in plant tissues not only affects the inference on paleoecology of ancient plants but also bears significance in the interpretation of paleoenvironmental signals. Using a combined SEM and geochemical approach, we are able to show the source, liability, and preservation of structural biopolymers from morphologically well-preserved Metasequoia tissues from three Tertiary deposits. We detected a continuum of biomolecular preservation in this evolutionarily-conserved conifer. Pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) was applied to solvent-extracted residues from both fossil leaf and wood remains in comparison with tissues from their living counterparts. The late Paleocene-early Eocene leaves from Ellesmere Island, Canadian Arctic Archipelago, exhibit the best quality of biochemical preservation and show pyrolysis products derived from labile biomolecules characterized by large amounts of polysaccharides. These labile biomolecules are the oldest record of these kinds so far characterized by the pyrolysis technology. The middle Eocene leaf tissues from Axel Heiberg Island, Canadian Arctic Archipelago, yielded slightly lesser amounts of polysaccharide moieties, but the lignin products are similar to those identified from the Ellesmere Island fossils. Compared with these Arctic materials, the Metasequoia leaves from Miocene Clarkia, Idaho, USA, show the lowest quality of molecular preservation, characterized by a dramatic reduction of polysaccharides. This continuum of relative quality of biomolecular preservation is further confirmed by SEM observations of transverse sections of these fossil leaves. The investigation revealed tissue-specific degradation, and our data support the in-situ polymerization hypothesis for the origin of long-chain homologous pairs of aliphatic n-alk-1-enes/n-alkanes as leaf alteration products. The preferential degradation and selective removal of polysaccharides may be significant in estimating plant paleo-productivity whereas the addition of aliphatic components to the leaf wax lipid pool may potentially contribute to the accuracy of compound specific isotope analysis using these lipid markers.

  13. [PS II photochemical efficiency in flag leaf of wheat varieties and its adaptation to strong sun- light intensity on farmland of Xiangride in Qinghai Province, Northwest China].

    PubMed

    Shi, Sheng-Bo; Chen, Wen-Jie; Shi, Rui; Li, Miao; Zhang, Huai-Gang; Sun, Ya-Nan

    2014-09-01

    Taking four wheat varieties developed by Northwest Institute of Plateau Biology, Chinese Academy of Sciences, as test materials, with the measurement of content of photosynthetic pigments, leaf area, fresh and dry mass of flag leaf, the PS II photochemistry efficiency of abaxial and adaxial surface of flag leaf and its adaptation to strong solar radiation during the period of heading stage in Xiangride region were investigated with the pulse-modulated in-vivo chlorophyll fluorescence technique. The results indicated that flag leaf angle mainly grew in horizontal state in Gaoyuan 314, Gaoyuan 363 and Gaoyuan 584, and mainly in vertical state in Gaoyuan 913 because of its smaller leaf area and larger width. Photosynthetic pigments were different among the 4 varieties, and positively correlated with intrinsic PS II photochemistry efficiencies (Fv/Fm). In clear days, especially at noon, the photosynthetic photoinhibition was more serious in abaxial surface of flag leaf due to directly facing the solar radiation, but it could recover after reduction of sunlight intensity in the afternoon, which meant that no inactive damage happened in PS II reaction centers. There were significant differences of PS II actual and maximum photochemical efficiencies at the actinic light intensity (ΦPS II and Fv'/Fm') between abaxial and adaxial surface, and their relative variation trends were on the contrary. The photochemical and non-photochemical quenching coefficients (qP and NPQ) had a similar tendency in both abaxial and adaxial surfaces. Although ΦPS II and qP were lower in adaxial surface of flag leaf, the Fv'/Fm' was significantly higher, which indicated that the potential PS II capture efficiency of excited energy was higher. The results demonstrated that process of photochemical and non-photochemical quenching could effectively dissipate excited energy caused by strong solar radiation, and there were higher adaptation capacities in wheat varieties natively cultivated in Qinghai-Tibetan Plateau area.

  14. Lotus leaf extract and L-carnitine influence different processes during the adipocyte life cycle

    PubMed Central

    2010-01-01

    Background The cellular and molecular mechanisms of adipose tissue biology have been studied extensively over the last two decades. Adipose tissue growth involves both an increase in fat cell size and the formation of mature adipocytes from precursor cells. To investigate how natural substances influence these two processes, we examined the effects of lotus leaf extract (Nelumbo nucifera-extract solution obtained from Silab, France) and L-carnitine on human preadipocytes and adipocytes. Methods For our in vitro studies, we used a lotus leaf extract solution alone or in combination with L-carnitine. Utilizing cultured human preadipocytes, we investigated lotus leaf extract solution-induced inhibition of triglyceride incorporation during adipogenesis and possible effects on cell viability. Studies on human adipocytes were performed aiming to elucidate the efficacy of lotus leaf extract solution to stimulate lipolytic activity. To further characterize lotus leaf extract solution-mediated effects, we determined the expression of the transcription factor adipocyte determination and differentiation factor 1 (ADD1/SREBP-1c) on the RNA- and protein level utilizing qRT-PCR and immunofluorescence analysis. Additionally, the effect of L-carnitine on beta-oxidation was analyzed using human preadipocytes and mature adipocytes. Finally, we investigated additive effects of a combination of lotus leaf extract solution and L-carnitine on triglyceride accumulation during preadipocyte/adipocyte differentiation. Results Our data showed that incubation of preadipocytes with lotus leaf extract solution significantly decreased triglyceride accumulation during adipogenesis without affecting cell viability. Compared to controls, adipocytes incubated with lotus leaf extract solution exhibited a significant increase in lipolysis-activity. Moreover, cell populations cultivated in the presence of lotus leaf extract solution showed a decrease in adipocyte differentiation capacity as indicated by a decrease in the ADD1/SREBP-1c signal. Importantly, our results demonstrated that a combination of lotus leaf extract solution and L-carnitine reduced triglyceride accumulation to a greater extent compared to incubation with either substance alone. Conclusions Overall, our data demonstrate that a combination of lotus leaf extract and L-carnitine reduced triglyceride accumulation in human (pre)adipocytes by affecting different processes during the adipocyte life cycle. For this reason, this combination might represent a treatment option for obesity-related diseases. PMID:20687953

  15. Lyophilized platelet-rich fibrin (PRF) promotes craniofacial bone regeneration through Runx2.

    PubMed

    Li, Qi; Reed, David A; Min, Liu; Gopinathan, Gokul; Li, Steve; Dangaria, Smit J; Li, Leo; Geng, Yajun; Galang, Maria-Therese; Gajendrareddy, Praveen; Zhou, Yanmin; Luan, Xianghong; Diekwisch, Thomas G H

    2014-05-14

    Freeze-drying is an effective means to control scaffold pore size and preserve its composition. The purpose of the present study was to determine the applicability of lyophilized Platelet-rich fibrin (LPRF) as a scaffold for craniofacial tissue regeneration and to compare its biological effects with commonly used fresh Platelet-rich fibrin (PRF). LPRF caused a 4.8-fold±0.4-fold elevation in Runt-related transcription factor 2 (Runx2) expression in alveolar bone cells, compared to a 3.6-fold±0.2-fold increase when using fresh PRF, and a more than 10-fold rise of alkaline phosphatase levels and mineralization markers. LPRF-induced Runx2 expression only occurred in alveolar bone and not in periodontal or dental follicle cells. LPRF also caused a 1.6-fold increase in osteoblast proliferation (p<0.001) when compared to fresh PRF. When applied in a rat craniofacial defect model for six weeks, LPRF resulted in 97% bony coverage of the defect, compared to 84% for fresh PRF, 64% for fibrin, and 16% without scaffold. Moreover, LPRF thickened the trabecular diameter by 25% when compared to fresh PRF and fibrin, and only LPRF and fresh PRF resulted in the formation of interconnected trabeculae across the defect. Together, these studies support the application of lyophilized PRF as a biomimetic scaffold for craniofacial bone regeneration and mineralized tissue engineering.

  16. Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model

    PubMed Central

    López, Luisa F.; Muñoz, César O.; Cáceres, Diego H.; Tobón, Ángela M.; Loparev, Vladimir; Clay, Oliver; Chiller, Tom; Litvintseva, Anastasia; Gade, Lalitha; González, Ángel

    2017-01-01

    Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis. PMID:29287097

  17. Consumption and utilization of experimentally altered corn by southern armyworm: Iron, nitrogen, and cyclic hydroxamates.

    PubMed

    Manuwoto, S; Scriber, J M

    1985-11-01

    The effects of differential leaf water, leaf nitrogen and cyclic hydroxamate (DIMBOA) concentrations in corn seedlings were analyzed for a polyphagous insect, the southern armyworm (Spodoptera eridania Cram.). Six different combinations of nutrients and allelochemicals [DIMBOA = 2,4-dihydroxy-7-methoxy(2H)-benzoxazin-3(4H)-one] were generated using two corn genotypes (WF9 and CI3IA) and three fertility regimes (complete nutrient, Fe-deficient, and N-deficient solutions) in the University Biotron. Poorest larval growth was observed in the low-nitrogen treatments (1.2% and 1.7% leaf N) and was the result of both low consumption rates and high metabolic costs (low efficiency of conversion of digested food, ECD). Fastest growth rates were observed forthe larvae fed leaves from the high-nitrogen treatments (4.6% and 4.4% leaf N). It is noteworthy that these treatments also contained the highest concentration of cyclic hydroxamates, which are generally believed to be the primary defensive chemicals mediating resistance against the European corn borer,Ostrinia nubilalis (Hubner). If these hydroxamates do have any deleterious or costly effects (perhaps accounting for a large portion of metabolic expenditures), the high digestibility of the leaf tissue and the increased consumption rates more than compensate, resulting in rapid growth (growth rate = consumption rate × approximate digestibility × efficiency of conversion of the digested food). These studies illustrate that variation in key nutrients and allelochemicals within a single plant species (Zea mays L.) may have significantly different effects upon various potential leaf-chewing caterpillars, such as these armyworms versus corn borers (which cannot handle the cyclic hydroxamates, even if provided with young nutritious leaf tissues).

  18. Cryopreservation of testicular tissue before long-term testicular cell culture does not alter in vitro cell dynamics.

    PubMed

    Baert, Yoni; Braye, Aude; Struijk, Robin B; van Pelt, Ans M M; Goossens, Ellen

    2015-11-01

    To assess whether testicular cell dynamics are altered during long-term culture after testicular tissue cryopreservation. Experimental basic science study. Reproductive biology laboratory. Testicular tissue with normal spermatogenesis was obtained from six donors. None. Detection and comparison of testicular cells from fresh and frozen tissues during long-term culture. Human testicular cells derived from fresh (n = 3) and cryopreserved (n = 3) tissues were cultured for 2 months and analyzed with quantitative reverse-transcription polymerase chain reaction and immunofluorescence. Spermatogonia including spermatogonial stem cells (SSCs) were reliably detected by combining VASA, a germ cell marker, with UCHL1, a marker expressed by spermatogonia. The established markers STAR, ACTA2, and SOX9 were used to analyze the presence of Leydig cells, peritubular myoid cells, and Sertoli cells, respectively. No obvious differences were found between the cultures initiated from fresh or cryopreserved tissues. Single or small groups of SSCs (VASA(+)/UCHL1(+)) were detected in considerable amounts up to 1 month of culture, but infrequently after 2 months. SSCs were found attached to the feeder monolayer, which expressed markers for Sertoli cells, Leydig cells, and peritubular myoid cells. In addition, VASA(-)/UCHL1(+) cells, most likely originating from the interstitium, also contributed to this monolayer. Apart from Sertoli cells, all somatic cell types could be detected throughout the culture period. Testicular tissue can be cryopreserved before long-term culture without modifying its outcome, which encourages implementation of testicular tissue banking for fertility preservation. However, because of the limited numbers of SSCs available after 2 months, further exploration and optimization of the culture system is needed. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  19. Myocardial regeneration potential of adipose tissue-derived stem cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bai, Xiaowen, E-mail: baixw01@yahoo.com; Alt, Eckhard, E-mail: ealt@mdanderson.org

    Research highlights: {yields} Various tissue resident stem cells are receiving tremendous attention from basic scientists and clinicians and hold great promise for myocardial regeneration. {yields} For practical reasons, human adipose tissue-derived stem cells are attractive stem cells for future clinical application in repairing damaged myocardium. {yields} This review summarizes the characteristics of cultured and freshly isolated stem cells obtained from adipose tissue, their myocardial regeneration potential and the, underlying mechanisms, and safety issues. -- Abstract: Various tissue resident stem cells are receiving attention from basic scientists and clinicians as they hold promise for myocardial regeneration. For practical reasons, adipose tissue-derivedmore » stem cells (ASCs) are attractive cells for clinical application in repairing damaged myocardium based on the following advantages: abundant adipose tissue in most patients and easy accessibility with minimally invasive lipoaspiration procedure. Several recent studies have demonstrated that both cultured and freshly isolated ASCs could improve cardiac function in animal model of myocardial infarction. The mechanisms underlying the beneficial effect of ASCs on myocardial regeneration are not fully understood. Growing evidence indicates that transplantation of ASCs improve cardiac function via the differentiation into cardiomyocytes and vascular cells, and through paracrine pathways. Paracrine factors secreted by injected ASCs enhance angiogenesis, reduce cell apoptosis rates, and promote neuron sprouts in damaged myocardium. In addition, Injection of ASCs increases electrical stability of the injured heart. Furthermore, there are no reported cases of arrhythmia or tumorigenesis in any studies regarding myocardial regeneration with ASCs. This review summarizes the characteristics of both cultured and freshly isolated stem cells obtained from adipose tissue, their myocardial regeneration potential, and the underlying mechanisms for beneficial effect on cardiac function, and safety issues.« less

  20. Cryopreserved Dental Pulp Tissues of Exfoliated Deciduous Teeth Is a Feasible Stem Cell Resource for Regenerative Medicine

    PubMed Central

    Yamaza, Haruyoshi; Akiyama, Kentaro; Hoshino, Yoshihiro; Song, Guangtai; Kukita, Toshio; Nonaka, Kazuaki; Shi, Songtao; Yamaza, Takayoshi

    2012-01-01

    Human exfoliated deciduous teeth have been considered to be a promising source for regenerative therapy because they contain unique postnatal stem cells from human exfoliated deciduous teeth (SHED) with self-renewal capacity, multipotency and immunomodulatory function. However preservation technique of deciduous teeth has not been developed. This study aimed to evaluate that cryopreserved dental pulp tissues of human exfoliated deciduous teeth is a retrievable and practical SHED source for cell-based therapy. SHED isolated from the cryopreserved deciduous pulp tissues for over 2 years (25–30 months) (SHED-Cryo) owned similar stem cell properties including clonogenicity, self-renew, stem cell marker expression, multipotency, in vivo tissue regenerative capacity and in vitro immunomodulatory function to SHED isolated from the fresh tissues (SHED-Fresh). To examine the therapeutic efficacy of SHED-Cryo on immune diseases, SHED-Cryo were intravenously transplanted into systemic lupus erythematosus (SLE) model MRL/lpr mice. Systemic SHED-Cryo-transplantation improved SLE-like disorders including short lifespan, elevated autoantibody levels and nephritis-like renal dysfunction. SHED-Cryo amended increased interleukin 17-secreting helper T cells in MRL/lpr mice systemically and locally. SHED-Cryo-transplantation was also able to recover osteoporosis bone reduction in long bones of MRL/lpr mice. Furthermore, SHED-Cryo-mediated tissue engineering induced bone regeneration in critical calvarial bone-defect sites of immunocompromised mice. The therapeutic efficacy of SHED-Cryo transplantation on immune and skeletal disorders was similar to that of SHED-Fresh. These data suggest that cryopreservation of dental pulp tissues of deciduous teeth provide a suitable and desirable approach for stem cell-based immune therapy and tissue engineering in regenerative medicine. PMID:23251621

  1. The potential of epiphytic hydrocarbon-utilizing bacteria on legume leaves for attenuation of atmospheric hydrocarbon pollutants.

    PubMed

    Ali, Nida; Sorkhoh, Naser; Salamah, Samar; Eliyas, Mohamed; Radwan, Samir

    2012-01-01

    The leaves of two legumes, peas and beans, harbored on their surfaces up to 9×10⁷ cells g⁻¹ of oil-utilizing bacteria. Less numbers, up to 5×10⁵ cells g⁻¹ inhabited leaves of two nonlegume crops, namely tomato and sunflower. Older leaves accommodated more of such bacteria than younger ones. Plants raised in oily environments were colonized by much more oil-utilizing bacteria than those raised in pristine (oil-free) environments. Similar numbers were counted on the same media in which nitrogen salt was deleted, indicating that most phyllospheric bacteria were probably diazotrophic. Most dominant were Microbacterium spp. followed by Rhodococcus spp., Citrobacter freundii, in addition to several other minor species. The pure bacterial isolates could utilize leaf tissue hydrocarbons, and consume considerable proportions of crude oil, phenanthrene (an aromatic hydrocarbon) and n-octadecane (an alkane) in batch cultures. Bacterial consortia on fresh (but not on previously autoclaved) leaves of peas and beans could also consume substantial proportions of the surrounding volatile oil hydrocarbons in closed microcosms. It was concluded that phytoremediation through phyllosphere technology could be useful in remediating atmospheric hydrocarbon pollutants. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Norovirus Narita 104 Virus-Like Particles Expressed in Nicotiana benthamiana Induce Serum and Mucosal Immune Responses

    PubMed Central

    Mathew, Lolita George; Herbst-Kralovetz, Melissa M.; Mason, Hugh S.

    2014-01-01

    Narita 104 virus is a human pathogen belonging to the norovirus (family Caliciviridae) genogroup II. Noroviruses cause epidemic gastroenteritis worldwide. To explore the potential of developing a plant-based vaccine, a plant optimized gene encoding Narita 104 virus capsid protein (NaVCP) was expressed transiently in Nicotiana benthamiana using a tobacco mosaic virus expression system. NaVCP accumulated up to approximately 0.3 mg/g fresh weight of leaf at 4 days postinfection. Initiation of hypersensitive response-like symptoms followed by tissue necrosis necessitated a brief infection time and was a significant factor limiting expression. Transmission electron microscopy of plant-derived NaVCP confirmed the presence of fully assembled virus-like particles (VLPs). In this study, an optimized method to express and partially purify NaVCP is described. Further, partially purified NaVCP was used to immunize mice by intranasal delivery and generated significant mucosal and serum antibody responses. Thus, plant-derived Narita 104 VLPs have potential for use as a candidate subunit vaccine or as a component of a multivalent subunit vaccine, along with other genotype-specific plant-derived VLPs. PMID:24949472

  3. Niv versus dropping vitrification in cryopreservation of human ovarian tissue.

    PubMed

    Xiao, Z; Li, S W; Zhang, Y Y; Wang, Y; Li, L L; Fan, W

    2014-01-01

    The containers for vitrification of tissues include cryovials, copper grids, Pasteur pipettes, the solid-surface method and etc. Recently the acupuncture needle was used to achieve better result in vitrification of human ovarian tissue. To determine if the needle immersed vitrification method (NIV) is a promising approach to vitrify the human ovarian tissue. Human ovarian biopsies from five patients were vitrified using NIV and Dropping vitrification. After 14 days of in vitro culture, the incidence of apoptotic primordial follicles from fresh and vitrified groups was assessed by TUNEL assay. 17β-estradiol (E2) and progesterone (P4) were detected in the media after culturing of vitrified and fresh ovarian tissues. The incidence of apoptotic primordial follicles was significantly higher in the dropping vitrification group than in the NIV group (P < 0.05). E2 and P4 concentrations were significantly higher in NIV groups than in Dropping vitrification group (P < 0.05). NIV was an appropriate method to vitrify ovarian tissue by improving the growth potential of frozen-warmed ovarian tissue in vitro culture.

  4. Higher plant modelling for life support applications: first results of a simple mechanistic model

    NASA Astrophysics Data System (ADS)

    Hezard, Pauline; Dussap, Claude-Gilles; Sasidharan L, Swathy

    2012-07-01

    In the case of closed ecological life support systems, the air and water regeneration and food production are performed using microorganisms and higher plants. Wheat, rice, soybean, lettuce, tomato or other types of eatable annual plants produce fresh food while recycling CO2 into breathable oxygen. Additionally, they evaporate a large quantity of water, which can be condensed and used as potable water. This shows that recycling functions of air revitalization and food production are completely linked. Consequently, the control of a growth chamber for higher plant production has to be performed with efficient mechanistic models, in order to ensure a realistic prediction of plant behaviour, water and gas recycling whatever the environmental conditions. Purely mechanistic models of plant production in controlled environments are not available yet. This is the reason why new models must be developed and validated. This work concerns the design and test of a simplified version of a mathematical model coupling plant architecture and mass balance purposes in order to compare its results with available data of lettuce grown in closed and controlled chambers. The carbon exchange rate, water absorption and evaporation rate, biomass fresh weight as well as leaf surface are modelled and compared with available data. The model consists of four modules. The first one evaluates plant architecture, like total leaf surface, leaf area index and stem length data. The second one calculates the rate of matter and energy exchange depending on architectural and environmental data: light absorption in the canopy, CO2 uptake or release, water uptake and evapotranspiration. The third module evaluates which of the previous rates is limiting overall biomass growth; and the last one calculates biomass growth rate depending on matter exchange rates, using a global stoichiometric equation. All these rates are a set of differential equations, which are integrated with time in order to provide total biomass fresh weight during the full growth duration. The model predicts a growth with exponential rate at the beginning and then it becomes linear for the end of the growth; this follows rather accurately the experimental data. Even if this model is too simple to be realistic for more complex plants in changing environments, this is the first step for an integrated approach of plant growth accounting of architectural and mass transfer limitations.

  5. [Identification characters of leaf morphological and venation pattern of Houttuynia cordata with its confused herb Gymnotheca chinensis].

    PubMed

    Lu, Hai-Lin; Guo, Min; Liao, Yue-Kui; Huang, Ding-Ying; Huang, Chun-Ni; Wu, Xiao-Chen; He, Bao-Zuo

    2012-11-01

    To study the identification characters of Houttuynia cordata and its confused herb Gymnotheca chinensis and establish an identification method. LMVP (leaf morphological-venation pattern for identification Chinese herbs), and QAERM (quantitatively analyze and evaluate reliability for the method of identification Chinese herbs) were applied for the study. Both venations were brochidodromous-acrodromous and arising from the mid-petiole or the upper section of petiole. The main characteristic of the leaf of Houttuynia cordata: surface with small gray-white stoma protuberances; Ligulate process of stipule-petiole sheath were clear; Primary veins 7 or 5; The innermost pair of primary vein closed up the top of the sinus at blade base or above sinus, and the section of closed vein was straight; Emitted a smell of fish when fresh leaf was kneaded into pieces. The main feature of the leaf of Gymnotheca chinensis: no small gray-white stoma protuberances; Ligulate process of stipule-petiole sheath were not clear; Primary veins 5; The innermost pair of primary vein closed into the sinus at blade base, and the section of closed vein was slightly curve; No smell of fish. With the mentioned key differences, the both plants could be successfully identified from each other. The accuracy of identification results (AC) was 100%, the repeatability of identification results: agreement rate for observation (ARO) was 100% and Kappa value was 1.00. The established method is simple, rapid, economic and reliable.

  6. Leucoagaricus gongylophorus Produces Diverse Enzymes for the Degradation of Recalcitrant Plant Polymers in Leaf-Cutter Ant Fungus Gardens

    PubMed Central

    Aylward, Frank O.; Burnum-Johnson, Kristin E.; Tringe, Susannah G.; Teiling, Clotilde; Tremmel, Daniel M.; Moeller, Joseph A.; Scott, Jarrod J.; Barry, Kerrie W.; Piehowski, Paul D.; Nicora, Carrie D.; Malfatti, Stephanie A.; Monroe, Matthew E.; Purvine, Samuel O.; Goodwin, Lynne A.; Smith, Richard D.; Weinstock, George M.; Gerardo, Nicole M.; Suen, Garret; Lipton, Mary S.

    2013-01-01

    Plants represent a large reservoir of organic carbon comprised primarily of recalcitrant polymers that most metazoans are unable to deconstruct. Many herbivores gain access to nutrients in this material indirectly by associating with microbial symbionts, and leaf-cutter ants are a paradigmatic example. These ants use fresh foliar biomass as manure to cultivate gardens composed primarily of Leucoagaricus gongylophorus, a basidiomycetous fungus that produces specialized hyphal swellings that serve as a food source for the host ant colony. Although leaf-cutter ants are conspicuous herbivores that contribute substantially to carbon turnover in Neotropical ecosystems, the process through which plant biomass is degraded in their fungus gardens is not well understood. Here we present the first draft genome of L. gongylophorus, and, using genomic and metaproteomic tools, we investigate its role in lignocellulose degradation in the gardens of both Atta cephalotes and Acromyrmex echinatior leaf-cutter ants. We show that L. gongylophorus produces a diversity of lignocellulases in ant gardens and is likely the primary driver of plant biomass degradation in these ecosystems. We also show that this fungus produces distinct sets of lignocellulases throughout the different stages of biomass degradation, including numerous cellulases and laccases that likely play an important role in lignocellulose degradation. Our study provides a detailed analysis of plant biomass degradation in leaf-cutter ant fungus gardens and insight into the enzymes underlying the symbiosis between these dominant herbivores and their obligate fungal cultivar. PMID:23584789

  7. Growth Promoting Rhizospheric and Endophytic Bacteria from Curcuma longa L. as Biocontrol Agents against Rhizome Rot and Leaf Blight Diseases.

    PubMed

    Vinayarani, G; Prakash, H S

    2018-06-01

    Plant growth promoting rhizobacteria and endophytic bacteria were isolated from different varieties of turmeric ( Curcuma longa L.) from South India. Totally 50 strains representing, 30 PGPR and 20 endophytic bacteria were identified based on biochemical assays and 16S rDNA sequence analysis. The isolates were screened for antagonistic activity against Pythium aphanidermatum (Edson) Fitzp., and Rhizoctonia solani Kuhn., causing rhizome rot and leaf blight diseases in turmeric, by dual culture and liquid culture assays. Results revealed that only five isolates of PGPR and four endophytic bacteria showed more than 70% suppression of test pathogens in both assays. The SEM studies of interaction zone showed significant ultrastructural changes of the hyphae like shriveling, breakage and desication of the pathogens by PGPR B. cereus (RBac-DOB-S24) and endophyte P. aeruginosa (BacDOB-E19). Selected isolates showed multiple Plant growth promoting traits. The rhizome bacterization followed by soil application of B. cereus (RBacDOB-S24) showed lowest Percent Disease Incidence (PDI) of rhizome rot and leaf blight, 16.4% and 15.5% respectively. Similarly, P. aeruginosa (BacDOB-E19) recorded PDI of rhizome rot (17.5%) and leaf blight (17.7%). The treatment of these promising isolates exhibited significant increase in plant height and fresh rhizome yield/plant in comparison with untreated control under greenhouse condition. Thereby, these isolates can be exploited as a potential biocontrol agent for suppressing rhizome rot and leaf blight diseases in turmeric.

  8. Do Aphids Alter Leaf Surface Temperature Patterns During Early Infestation?

    PubMed Central

    Cahon, Thomas; Caillon, Robin

    2018-01-01

    Arthropods at the surface of plants live in particular microclimatic conditions that can differ from atmospheric conditions. The temperature of plant leaves can deviate from air temperature, and leaf temperature influences the eco-physiology of small insects. The activity of insects feeding on leaf tissues, may, however, induce changes in leaf surface temperatures, but this effect was only rarely demonstrated. Using thermography analysis of leaf surfaces under controlled environmental conditions, we quantified the impact of presence of apple green aphids on the temperature distribution of apple leaves during early infestation. Aphids induced a slight change in leaf surface temperature patterns after only three days of infestation, mostly due to the effect of aphids on the maximal temperature that can be found at the leaf surface. Aphids may induce stomatal closure, leading to a lower transpiration rate. This effect was local since aphids modified the configuration of the temperature distribution over leaf surfaces. Aphids were positioned at temperatures near the maximal leaf surface temperatures, thus potentially experiencing the thermal changes. The feedback effect of feeding activity by insects on their host plant can be important and should be quantified to better predict the response of phytophagous insects to environmental changes. PMID:29538342

  9. Leaf maximum photosynthetic rate and venation are linked by hydraulics.

    PubMed

    Brodribb, Tim J; Feild, Taylor S; Jordan, Gregory J

    2007-08-01

    Leaf veins are almost ubiquitous across the range of terrestrial plant diversity, yet their influence on leaf photosynthetic performance remains uncertain. We show here that specific physical attributes of the vascular plumbing network are key limiters of the hydraulic and photosynthetic proficiency of any leaf. Following the logic that leaf veins evolved to bypass inefficient water transport through living mesophyll tissue, we examined the hydraulic pathway beyond the distal ends of the vein system as a possible limiter of water transport in leaves. We tested a mechanistic hypothesis that the length of this final traverse, as water moves from veins across the mesophyll to where it evaporates from the leaf, governs the hydraulic efficiency and photosynthetic carbon assimilation of any leaf. Sampling 43 species across the breadth of plant diversity from mosses to flowering plants, we found that the post-vein traverse as determined by characters such as vein density, leaf thickness, and cell shape, was strongly correlated with the hydraulic conductivity and maximum photosynthetic rate of foliage. The shape of this correlation provided clear support for the a priori hypothesis that vein positioning limits photosynthesis via its influence on leaf hydraulic efficiency.

  10. Endogenous electromagnetic fields in plant leaves: a new hypothesis for vascular pattern formation.

    PubMed

    Pietak, Alexis Mari

    2011-06-01

    Electromagnetic (EM) phenomena have long been implicated in biological development, but few detailed, practical mechanisms have been put forth to connect electromagnetism with morphogenetic processes. This work describes a new hypothesis for plant leaf veination, whereby an endogenous electric field forming as a result of a coherent Frohlich process, and corresponding to an EM resonant mode of the developing leaf structure, is capable of instigating leaf vascularisation. In order to test the feasibility of this hypothesis, a three-dimensional, EM finite-element model (FEM) of a leaf primordium was constructed to determine if suitable resonant modes were physically possible for geometric and physical parameters similar to those of developing leaf tissue. Using the FEM model, resonant EM modes with patterns of relevance to developing leaf vein modalities were detected. On account of the existence of shared geometric signatures in a leaf's vascular pattern and the electric field component of EM resonant modes supported by a developing leaf structure, further theoretical and experimental investigations are warranted. Significantly, this hypothesis is not limited to leaf vascular patterning, but may be applicable to a variety of morphogenetic phenomena in a number of living systems.

  11. Potential of cultivar and crop management to affect phytochemical content in winter-grown sprouting broccoli (Brassica oleracea L. var. italica).

    PubMed

    Reilly, Kim; Valverde, Juan; Finn, Leo; Rai, Dilip K; Brunton, Nigel; Sorensen, Jens C; Sorensen, Hilmer; Gaffney, Michael

    2014-01-30

    Variety and crop management strategies affect the content of bioactive compounds (phenolics, flavonoids and glucosinolates) in green broccoli (calabrese) types, which are cultivated during summer and autumn in temperate European climates. Sprouting broccoli types are morphologically distinct and are grown over the winter season and harvested until early spring. Thus they show considerable potential for development as an import substitution crop for growers and consumers during the 'hungry gap' of early spring. The present study investigated the effect of variety and management practices on phytochemical content in a range of sprouting broccoli varieties. Yields were significantly higher in white sprouting broccoli varieties. Levels of phenolics and flavonoids were in the range 81.64-297.65 and 16.95-104.80 mg 100 g⁻¹ fresh weight, respectively, depending on year and cultivar, and were highest in variety 'TZ 5052' in both years. In-row spacing did not affect flavonoid content. Phenolic and flavonoid content generally increased with increasing floret maturity and levels were high in edible portions of the crop. Crop wastes (leaf and flower) contained 145.9-239.3 and 21.5-116.6 mg 100 g⁻¹ fresh weight total phenolics and flavonoids, respectively, depending on cultivar, tissue and year. Climatic factors had a significant effect on phenolic and flavonoid content. Levels of total and some individual glucosinolates were higher in sprouting broccoli than in the green broccoli variety 'Ironman'. Levels of total phenolics, flavonoids and glucosinolates are higher in sprouting than green broccoli types. Sprouting broccoli represents an excellent source of dietary bioactive compounds. © 2013 Society of Chemical Industry.

  12. Maize YABBY Genes drooping leaf1 and drooping leaf2 Regulate Plant Architecture[OPEN

    PubMed Central

    Briggs, Sarah; Bradbury, Peter J.

    2017-01-01

    Leaf architecture directly influences canopy structure, consequentially affecting yield. We discovered a maize (Zea mays) mutant with aberrant leaf architecture, which we named drooping leaf1 (drl1). Pleiotropic mutations in drl1 affect leaf length and width, leaf angle, and internode length and diameter. These phenotypes are enhanced by natural variation at the drl2 enhancer locus, including reduced expression of the drl2-Mo17 allele in the Mo17 inbred. A second drl2 allele, produced by transposon mutagenesis, interacted synergistically with drl1 mutants and reduced drl2 transcript levels. The drl genes are required for proper leaf patterning, development and cell proliferation of leaf support tissues, and for restricting auricle expansion at the midrib. The paralogous loci encode maize CRABS CLAW co-orthologs in the YABBY family of transcriptional regulators. The drl genes are coexpressed in incipient and emergent leaf primordia at the shoot apex, but not in the vegetative meristem or stem. Genome-wide association studies using maize NAM-RIL (nested association mapping-recombinant inbred line) populations indicated that the drl loci reside within quantitative trait locus regions for leaf angle, leaf width, and internode length and identified rare single nucleotide polymorphisms with large phenotypic effects for the latter two traits. This study demonstrates that drl genes control the development of key agronomic traits in maize. PMID:28698237

  13. Complex I-complex II ratio strongly differs in various organs of Arabidopsis thaliana.

    PubMed

    Peters, Katrin; Niessen, Markus; Peterhänsel, Christoph; Späth, Bettina; Hölzle, Angela; Binder, Stefan; Marchfelder, Anita; Braun, Hans-Peter

    2012-06-01

    In most studies, amounts of protein complexes of the oxidative phosphorylation (OXPHOS) system in different organs or tissues are quantified on the basis of isolated mitochondrial fractions. However, yield of mitochondrial isolations might differ with respect to tissue type due to varying efficiencies of cell disruption during organelle isolation procedures or due to tissue-specific properties of organelles. Here we report an immunological investigation on the ratio of the OXPHOS complexes in different tissues of Arabidopsis thaliana which is based on total protein fractions isolated from five Arabidopsis organs (leaves, stems, flowers, roots and seeds) and from callus. Antibodies were generated against one surface exposed subunit of each of the five OXPHOS complexes and used for systematic immunoblotting experiments. Amounts of all complexes are highest in flowers (likewise with respect to organ fresh weight or total protein content of the flower fraction). Relative amounts of protein complexes in all other fractions were determined with respect to their amounts in flowers. Our investigation reveals high relative amounts of complex I in green organs (leaves and stems) but much lower amounts in non-green organs (roots, callus tissue). In contrast, complex II only is represented by low relative amounts in green organs but by significantly higher amounts in non-green organs, especially in seeds. In fact, the complex I-complex II ratio differs by factor 37 between callus and leaf, indicating drastic differences in electron entry into the respiratory chain in these two fractions. Variation in amounts concerning complexes III, IV and V was less pronounced in different Arabidopsis tissues (quantification of complex V in leaves was not meaningful due to a cross-reaction of the antibody with the chloroplast form of this enzyme). Analyses were complemented by in gel activity measurements for the protein complexes of the OXPHOS system and comparative 2D blue native/SDS PAGE analyses using isolated mitochondria. We suggest that complex I has an especially important role in the context of photosynthesis which might be due to its indirect involvement in photorespiration and its numerous enzymatic side activities in plants.

  14. Seismomorphogenesis: a novel approach to acclimatization of tissue culture regenerated plants.

    PubMed

    Sarmast, Mostafa Khoshhal; Salehi, Hassan; Khosh-Khui, Morteza

    2014-12-01

    Plantlets under in vitro conditions transferred to ex vivo conditions are exposed to biotic and abiotic stresses. Furthermore, in vitro regenerated plants are typically frail and sometimes difficult to handle subsequently increasing their risk to damage and disease; hence acclimatization of these plantlets is the most important step in tissue culture techniques. An experiment was conducted under in vitro conditions to study the effects of shaking duration (twice daily at 6:00 a.m. and 9:00 p.m. for 2, 4, 8, and 16 min at 250 rpm for 14 days) on Sansevieria trifasciata L. as a model plant. Results showed that shaking improved handling, total plant height, and leaf characteristics of the model plant. Forty-eight hours after 14 days of shaking treatments with increasing shaking time, leaf length decreased but proline content of leaf increased. However, 6 months after starting the experiment different results were observed. In explants that received 16 min of shaking treatment, leaf length and area and photosynthesis rate were increased compared with control plantlets. Six months after starting the experiment, control plantlets had 12.5 % mortality; however, no mortality was observed in other treated explants. The results demonstrated that shaking improved the explants' root length and number and as a simple, cost-effective, and non-chemical novel approach may be substituted for other prevalent acclimatization techniques used for tissue culture regenerated plantlets. Further studies with sensitive plants are needed to establish this hypothesis.

  15. Bovine viral diarrhea virus 1b fetal infection with extensive hemorrhages

    USDA-ARS?s Scientific Manuscript database

    Bovine viral diarrhea virus (BVDV) subtype 1b was isolated from tissues of a term bovine fetus with hemorrhages in multiple tissues. At autopsy, multiple petechial hemorrhages were observed at gross examination throughout the body and placenta. Lung, kidney, thymus, and liver fresh tissues were exam...

  16. SU-E-T-331: Dosimetric Impact of Multileaf Collimator Leaf Width On Stereotactic Radiosurgery (SRS) RapidArc Treatment Plans for Single and Multiple Brain Metastases

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hossain, S; Keeling, V; Ahmad, S

    Purpose: To determine the effects of multileaf collimator (MLC) leaf width on normal-brain-tissue doses and dose conformity of SRS RapidArc treatment plans for brain tumors. Methods: Ten patients with 24 intracranial tumors (seven with 1–2 and three with 4–6 lesions) were planned using RapidArc for both Varian Millennium 120 MLC (5 mm leaf width) and high definition (HD) MLC (2.5 mm leaf width). Between 2 and 8 arcs were used with two full coplanar arcs and the rest non-coplanar half arcs. 6 MV beams were used and plans were optimized with a high priority to the Normal Tissue Objective (tomore » achieve dose conformity and sharp dose fall-off) and normal brain tissue. Calculation was done using AAA on a 1 mm grid size. The prescription dose ranged from 14–22 Gy. Plans were normalized such that 99% of the target received the prescription dose. Identical beam geometries, optimizations, calculations, and normalizations were used for both plans. Paddick Conformity Index (PCI), V4, V8 and V12 Gy for normal brain tissue and Integral Dose were used for analysis. Results: In all cases, HD MLC plans performed better in sparing normal brain tissue, achieving a higher PCI with a lower Integral Dose. The average PCI for all 24 targets was 0.75±0.23 and 0.70±0.23 (p ≤0.0015) for HD MLC and Millennium MLC plans, respectively. The average ratio of normal brain doses for Millennium MLC to HD MLC plans was 1.30±0.16, 1.27±0.15, and 1.31±0.18 for the V4, V8, and V12, respectively. The differences in normal brain dose for all criteria were statistically significant with p-value < 0.02. On average Millennium MLC plans had a 16% higher integral dose than HD MLC plans. Conclusion: Significantly better dose conformity with reduced volume of normal brain tissue and integral dose was achieved with HD MLC plans compared to Millennium MLC plans.« less

  17. Pathobiological investigation of naturally infected canine rabies cases from Sri Lanka.

    PubMed

    Beck, S; Gunawardena, P; Horton, D L; Hicks, D J; Marston, D A; Ortiz-Pelaez, A; Fooks, A R; Núñez, A

    2017-04-12

    The recommended screening of rabies in 'suspect' animal cases involves testing fresh brain tissue. The preservation of fresh tissue however can be difficult under field conditions and formalin fixation provides a simple alternative that may allow a confirmatory diagnosis. The occurrence and location of histopathological changes and immunohistochemical (IHC) labelling for rabies in formalin fixed paraffin embedded (FFPE) canine brain is described in samples from 57 rabies suspect cases from Sri-Lanka. The presence of Negri bodies and immunohistochemical detection of rabies virus antigen were evaluated in the cortex, hippocampus, cerebellum and brainstem. The effect of autolysis and artefactual degeneration of the tissue was also assessed. Rabies was confirmed in 53 of 57 (93%) cases by IHC. IHC labelling was statistically more abundant in the brainstem. Negri bodies were observed in 32 of 53 (60.4%) of the positive cases. Although tissue degradation had no effect on IHC diagnosis, it was associated with an inability to detect Negri bodies. In 13 cases, a confirmatory Polymerase chain reaction (PCR) testing for rabies virus RNA was undertaken by extracting RNA from fresh frozen tissue, and also attempted using FFPE samples. PCR detection using fresh frozen samples was in agreement with the IHC results. The PCR method from FFPE tissues was suitable for control material but unsuccessful in our field cases. Histopathological examination of the brain is essential to define the differential diagnoses of behaviour modifying conditions in rabies virus negative cases, but it is unreliable as the sole method for rabies diagnosis, particularly where artefactual change has occurred. Formalin fixation and paraffin embedding does not prevent detection of rabies virus via IHC labelling even where artefactual degeneration has occurred. This could represent a pragmatic secondary assay for rabies diagnosis in the field because formalin fixation can prevent sample degeneration. The brain stem was shown to be the site with most viral immunoreactivity; supporting recommended sampling protocols in favour of improved necropsy safety in the field. PCR testing of formalin fixed tissue may be successful in certain circumstances as an alternative test.

  18. Studies on the ultrastructure in Anacardium occidentale L. leaves from Amazon in northern Brazil by scanning microscopy.

    PubMed

    Ramos, Glenda Quaresma; Cotta, Eduardo Adriano; da Fonseca Filho, Henrique Duarte

    2016-07-01

    Leaves surfaces have various structures with specific functions and contribute to the relationship with the environment. On morphological studies are analyzed various parameters, ranging from macro scale through the micro scale to the nanometer scale, which contribute to the study of taxonomy, pharmacognosy, and ecology, among others. Functional structures found in leaves are responsible for the wide variety of surfaces and some behaviors are given in terms of cellular adaptation and the presence or absence of wax. This study reports the characterization of Anacardium occidentale L. leaf surface and the techniques used therein. A set of scanning electron microscope (SEM) and atomic force microscope (AFM) images performed on fresh leaf allowed observation of textured and heterogeneous profiles on both sides. SCANNING 38:329-335, 2016. © 2015 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

  19. Antimicrobial activity of essential oils of Xylopia aethiopica.

    PubMed

    Fleischer, T C; Mensah, M L K; Mensah, A Y; Komlaga, G; Gbedema, S Y; Skaltsa, H

    2008-06-18

    Xylopia aethiopica is a medicinal plant of great repute in West Africa which produces a variety of complex chemical compounds. The fresh and dried fruits, leaf, stem bark and root bark essential oils showed various degrees of activity against the gram positive bacteria, Bacillus subtilis and Staphylococcus aureus, the gram negative bacteria Pseudomonas aeruginosa and the yeast-like fungus Candida albicans, using the cup plate method. However, none of the oils showed activity against Escherichia coli.

  20. Anti-conjunctivitis effect of fresh juice of xGraptoveria (Crassulaceae): A phytochemical and ethnobotanical study.

    PubMed

    Markova, Nadezhda V; Batovska, Daniela I; Kozuharova, Ekaterina K; Enchev, Venelin G

    2015-01-01

    The parent of xGraptoveria, Graptopetalum paraguayense, is used in Chinese folk medicine for alleviating hepatic disorders, detumescence and detoxication, lowering of blood pressure, inhibition of cancer cells, exerting diuretic effects, relieving pain and infections. No data are available regarding its anti-conjunctivitis effect. The aim of this preliminary study is to test the anti-conjunctivitis properties of xGraptoveria (Crassulaceae) and to identify its bioactive constituents. Fresh watery juice of leaves of xGraptoveria was extracted with n-butanol and the extract was analyzed using gas chromatography-mass spectrometry (GC/MS). The ethnobotanical appraisal of the anti-conjunctivitis properties of xGraptoveria was based on 11 interviews about the symptoms against which this plant demonstrated positive effect. Fresh juice of xGraptoveria leaves applied directly to the irritated eye 2 times per day cured conjunctivitis in all reported cases. The main groups of organic compounds identified by GC/MS analysis in the fresh extracted leaf juice of xGraptoveria were: Alkylamines, hydroxycarboxylic acids, aliphatic and aromatic carboxylic acids, amino acids, alcohols, aromatic and aliphatic hydrocarbons. In this preliminary study, it is suggested that xGraptoveria exerts anti-conjunctivitis activity, through synergistic effect of different chemical compounds, most probably alkylamines and mainly hydroxycarboxylic, aliphatic, and aromatic carboxylic acids.

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