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Sample records for spinach leaf sucrose-phosphate

  1. Multisite phosphorylation of spinach leaf sucrose-phosphate synthase

    SciTech Connect

    Huber, J.L.; Huber, S.C. )

    1990-05-01

    Spinach leaf sucrose-phosphate synthase is phosphorylated both in vivo and in vitro on serine residues. Phosphorylation of SPS in vivo yields twelve major phosphopeptides after a tryptic digest and two dimensional mapping. The in vivo labeling of three of these SPS P-peptides is reduced in illuminated leaves where the extracted enzyme is activated relative to that of dark leaves. Two of these inhibitory sites are phosphorylated as well when SPS is inactivated in vitro using ({sup 32}P)ATP. In vivo phosphorylation of two other sites is enhanced during mannose feeding of the leaves (in light or dark) which produces the highest activation state of SPS. Overall, the results confirm that light-dark regulation of SPS activity occurs as a result of regulatory seryl-phosphorylation and involves a balance between phosphorylation of sites which inhibit or stimulate activity. Regulation of the SPS protein kinase that inhibits activity is relatively unaffected by phosphate but inhibited by G1c 6-P (IC{sub 50}{approx}5 mM), which may explain the control of SPS activation state by light-dark signals.

  2. Inactivation of highly activated spinach leaf sucrose-phosphate synthase by dephosphorylation. [Spinacia oleracea

    SciTech Connect

    Huber, J.L. ); Huber, S.C. North Carolina State Univ., Raleigh ); Hite, D.R.C.; Outlaw, W.H. Jr. )

    1991-01-01

    Spinach (Spinacia oleracea L.) leaf sucrose-phosphate synthase (SPS) can be phosphorylated and inactivated in vitro with ({gamma}-{sup 32}P)ATP. Thus, it was surprising to find that SPS, extracted from leaves fed mannose in the light to highly activate the enzyme, could be inactivated in an ATP-independent manner when desalted crude extracts were preincubated at 25{degrees}C before assay. The spontaneous inactivation involved a loss in activity measured with limiting substrate concentrations in the presence of the inhibitor, Pi, without affecting maximum catalytic activity. The spontaneous inactivation was unaffected by exogenous carrier proteins and protease inhibitors, but was inhibited by inorganic phosphate, fluoride, and molybdate, suggesting that a phosphatase may be involved. Okadaic acid, a potent inhibitor of mammalian type 1 and 2A protein phosphatases, had no effect up to 5 micromolar. Inactivation was stimulated about twofold by exogenous Mg{sup 2+} and was relatively insensitive to Ca{sup 2+} and to pH over the range pH 6.5 to 8.5. Radioactive phosphate incorporated into SPS during labeling of excised leaves with ({sup 32}P)Pi (initially in the dark and then in the light with mannose) was lost with time when desalted crude extracts were incubated at 25 C, and the loss in radiolabel was substantially reduced by fluoride. These results provide direct evidence for action of an endogenous phosphatase(s) using SPS as substrate.

  3. Protein phosphorylation as a mechanism for regulation of spinach leaf sucrose-phosphate synthase activity

    SciTech Connect

    Huber, J.L.A.; Huber, S.C. )

    1989-04-01

    Protein phosphorylation has been identified as a mechanism for the light-dark regulation of spinach sucrose-phosphate synthase (SPS) activity, previously shown to involve some type of covalent modification of the enzyme. The 120 kD subunit of SPS in extracts of light-treated leaves was labeled with {sup 32}P in the presence of ({gamma}-{sup 32}P) ATP. In this in vitro system, {sup 32}P incorporation into light-activated SPS was dependent upon ATP and magnesium concentrations as well as time, and was closely paralleled by inactivation of the enzyme. The soluble protein kinase involved in the interconversion of SPS between activated and deactivated forms may be specific for SPS as it co-purifies with SPS during partial purification of the enzyme. The kinase appears not to be calcium activated and no evidence has been obtained for metabolite control of SPS phosphorylation/inactivation.

  4. Site-directed mutagenesis of serine 158 demonstrates its role in spinach leaf sucrose-phosphate synthase modulation

    NASA Technical Reports Server (NTRS)

    Toroser, D.; McMichael, R. Jr; Krause, K. P.; Kurreck, J.; Sonnewald, U.; Stitt, M.; Huber, S. C.; Davies, E. (Principal Investigator)

    1999-01-01

    Site-directed mutagenesis of spinach sucrose-phosphate synthase (SPS) was performed to investigate the role of Ser158 in the modulation of spinach leaf SPS. Tobacco plants expressing the spinach wild-type (WT), S158A, S158T and S157F/S158E SPS transgenes were produced. Expression of transgenes appeared not to reduce expression of the tobacco host SPS. SPS activity in the WT and the S158T SPS transgenics showed light/dark modulation, whereas the S158A and S157F/S158E mutants were not similarly light/dark modulated: the S158A mutant enzyme was not inactivated in the dark, and the S157F/S158E was not activated in the light. The inability to modulate the activity of the S158A mutant enzyme by protein phosphorylation was demonstrated in vitro. The WT spinach enzyme immunopurified from dark transgenic tobacco leaves had a low initial activation state, and could be activated by PP2A and subsequently inactivated by SPS-kinase plus ATP. Rapid purification of the S158A mutant enzyme from dark leaves of transgenic plants using spinach-specific monoclonal antibodies yielded enzyme that had a high initial activation state, and pre-incubation with leaf PP2A or ATP plus SPS-kinase (the PKIII enzyme) caused little modulation of activity. The results demonstrate the regulatory significance of Ser158 as the major site responsible for dark inactivation of spinach SPS in vivo, and indicate that the significance of phosphorylation is the introduction of a negative charge at the Ser158 position.

  5. Site-specific regulatory interaction between spinach leaf sucrose-phosphate synthase and 14-3-3 proteins

    NASA Technical Reports Server (NTRS)

    Toroser, D.; Athwal, G. S.; Huber, S. C.; Davies, E. (Principal Investigator)

    1998-01-01

    We report an Mg2+-dependent interaction between spinach leaf sucrose-phosphate synthase (SPS) and endogenous 14-3-3 proteins, as evidenced by co-elution during gel filtration and co-immunoprecipitation. The content of 14-3-3s associated with an SPS immunoprecipitate was inversely related to activity, and was specifically reduced when tissue was pretreated with 5-aminoimidazole-4-carboxamide riboside, suggesting metabolite control in vivo. A synthetic phosphopeptide based on Ser-229 was shown by surface plasmon resonance to bind a recombinant plant 14-3-3, and addition of the phosphorylated SPS-229 peptide was found to stimulate the SPS activity of an SPS:14-3-3 complex. Taken together, the results suggest a regulatory interaction of 14-3-3 proteins with Ser-229 of SPS.

  6. Interaction between Silver Nanoparticles and Spinach Leaf

    NASA Astrophysics Data System (ADS)

    Tian, Y.; Li, H.; Zhang, Y.; Riser, E.; He, S.; Zhang, W.

    2013-12-01

    Interactions of engineered nanoparticles (ENPs) with plant surfaces are critical to assessing the bioavailability of ENPs to edible plants and to further evaluating impacts of ENPs on ecological health and food safety. Silver nanoparticles (i.e., nanoAg) could enter the agroecosystems either as an active ingredient in pesticides or from other industrial and consumer applications. Thus, in the events of pesticide application, rainfall, and irrigation, vegetable leaves could become in contact and then interact with nanoAg. The present study was to assess whether the interaction of nanoAg with spinach leaves can be described by classical sorption models and to what extent it depends on and varies with dispersion methods, environmental temperature, and ion release. We investigated the stability and ion release of nanoAg dispersed by sodium dodecyl sulfate (SDS, 1%) and humic acid (HA, 10 mg C/L) solutions, as well as sorption and desorption of nanoAg on and from the fresh spinach leaf. Results showed SDS-nanoAg released about 2%-8% more Ag ion than HA-nanoAg. The sorption of Ag ion, described by the Freundlich model in the initial concentration range of 0.6-50 mg/L, was 2-4 times higher than that of nanoAg. The sorption of nanoAg on spinach leaf can be fitted by the Langmuir model, and the maximum sorption amount of HA-nanoAg and SDS-nanoAg was 0.21 and 0.41 mg/g, respectively. The higher sorption of SDS-nanoAg relative to that of HA-nanoAg could be partially resulted from the higher release of Ag ion from the former. The maximum desorption amount of HA-nanoAg and SDS-nanoAg in 1% SDS solution was 0.08 and 0.10 mg/g, respectively. NanoAg attachment on and its penetration to the spinach leaf was visualized by the Scanning Electron Microscope equipped with an Energy Dispersive Spectrometer (SEM-EDS). It is equally important that the less sorption of nanoAg under low environmental temperature could be partially due to the closure of stomata, as verified by SEM-EDS. Cyto

  7. Association mapping of leaf traits in spinach (Spinacia oleracea L.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spinach (Spinacia oleracea L.) is an important leafy vegetable crop grown world-wide. Leaf traits, surface texture (smooth vs. savoy or semi-savoy), petiole color (green vs. purple), and edge shape (serrate vs. entire) are important for spinach. Association mapping of the three traits were conducted...

  8. Population structure and association analysis of bolting, plant height, and leaf erectness in spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spinach (Spinacia oleracea L.) is an important vegetable worldwide with high nutritional and health-promoting compounds. Bolting is an important trait to consider in order to grow spinach in different seasons and regions. Plant height and leaf erectness are important traits for machine-harvesting. B...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  10. Antioxidant capacity and phenolic content in leaf extracts of tree spinach (Cnidoscolus spp.).

    PubMed

    Kuti, Joseph O; Konuru, Hima B

    2004-01-14

    Total phenolic content and antioxidant capacity of two tree spinach species (Cnidoscolus chayamansa McVaugh and C. aconitifolius Miller.) were determined in raw and cooked leaf extracts. Antioxidant capacity was assessed by the oxygen radical absorbance capacity (ORAC) assay, and flavonoid glycoside composition was quantified by HPLC and identified by GC. Total phenolics and antioxidant capacity were higher in raw than in cooked leaf extracts. The ORAC values were strongly correlated with total phenolic content (r = 0.926) in all leaf extracts. The major flavonoids isolated from the leaf extracts were kaempferol-3-O-glycosides and quercetin-3-O-glycosides. C. aconitifolius leaves contained more varieties of the flavonoid glycosides than C. chayamansa. Cooking reduced antioxidant activity and phenolic content and resulted in losses of some kaempferol glycoside and quercetin glycoside residues in leaf extracts. The results of this study indicate that tree spinach leaves are a rich source of natural antioxidants for foods. PMID:14709023

  11. Abscisic acid accumulation in spinach leaf slices in the presence of penetrating and nonpenetrating solutes

    SciTech Connect

    Creelman, R.A.; Zeevaart, J.A.D.

    1985-01-01

    Abscisic acid (ABA) accumulated in detached, wilted leaves of spinach (Spinacia oleracea L. cv Savoy Hybrid 612) and reached a maximum level within 3 to 4 hours. The increase in ABA over that found in detached turgid leaves was approximately 10-fold. The effects of water stress could be mimicked by the use of thin slices of spinach leaves incubated in the presence of 0.6 molar mannitol, a compound which causes plasmolysis (loss of turgor). When spinach leaf slices were incubated with ethylene glycol, a compound which rapidly penetrates the cell membrane causing a decrease in the osmotic potential of the tissue and only transient loss of turgor, no ABA accumulated. Spinach leaf slices incubated in both ethylene glycol and mannitol had ABA levels similar to those found when slices were incubated with mannitol alone. Increases similar to those found with mannitol also occurred when Aquacide III, a highly purified form of polyethylene glycol, was used. When spinach leaf slices were incubated with solutes which are supposed to disturb membrane integrity no increase in ABA was observed. These data indicate that, with respect to the accumulation of ABA, mannitol caused a physical stress rather than a chemical stress.

  12. "Respirasome"-like supercomplexes in green leaf mitochondria of spinach.

    PubMed

    Krause, Frank; Reifschneider, Nicole H; Vocke, Dirk; Seelert, Holger; Rexroth, Sascha; Dencher, Norbert A

    2004-11-12

    Higher plant mitochondria have many unique features compared with their animal and fungal counterparts. This is to a large extent related to the close functional interdependence of mitochondria and chloroplasts, in which the two ATP-generating processes of oxidative phosphorylation and photosynthesis, respectively, take place. We show that digitonin treatment of mitochondria contaminated with chloroplasts from spinach (Spinacia oleracea) green leaves at two different buffer conditions, performed to solubilize oxidative phosphorylation supercomplexes, selectively extracts the mitochondrial membrane protein complexes and only low amounts of stroma thylakoid membrane proteins. By analysis of digitonin extracts from partially purified mitochondria of green leaves from spinach using blue and colorless native electrophoresis, we demonstrate for the first time that in green plant tissue a substantial proportion of the respiratory complex IV is assembled with complexes I and III into "respirasome"-like supercomplexes, previously observed in mammalian, fungal, and non-green plant mitochondria only. Thus, fundamental features of the supramolecular organization of the standard respiratory complexes I, III, and IV as a respirasome are conserved in all higher eukaryotes. Because the plant respiratory chain is highly branched possessing additional alternative enzymes, the functional implications of the occurrence of respiratory supercomplexes in plant mitochondria are discussed.

  13. The Path of Carbon in Photosynthesis XIX. The Identification of Sucrose Phosphate in Sugar Beet Leaves

    DOE R&D Accomplishments Database

    Buchanan, J. G.

    1952-09-01

    The recognition and characterization of a sucrose phosphate as an intermediate in sucrose by synthesis by green plants is described. A tentative structure for this phosphate is proposed and its mode of formation suggested.

  14. Spinach or amaranth may represent highest residue of thiophanate-methyl with open field application on six leaf vegetables.

    PubMed

    Fan, Sufang; Zhao, Pengyue; Zhang, Fengzu; Yu, Chuanshan; Pan, Canping

    2013-04-01

    To select representative crop among leaf vegetables which may contain the highest residue after fungicide uses, open field applications with thiophanate-methyl on six crops including pakchoi, rape, crown daisy, amaranth, spinach and lettuce were designed and conducted. In this study, a high-performance liquid chromatography and electrospray ionization-tandem mass spectrometry with selected reaction monitoring was used to simultaneously determine thiophanate methyl and its metabolite carbendazim residue in various samples. The limit of quantification for thiophanate methyl and carbendazim were established in the range of 0.005-0.01 mg kg(-1) for all samples. It was shown that recoveries ranged from 67.8 % to 102.3 % for thiophanate methyl, and 72.0 %-112.6 % for carbendazim at spiked levels of 0.01-0.1 mg kg(-1). It's found that thiophanate methyl converts to carbendazim very quickly. In supervised field trials, the half-lives of thiophanate methyl in six leaf vegetables were in the range of 1.26-2.65 days, and the half-lives of carbendazim were in the range of 2.53-4.28 days. It was also found that thiophanate methyl residue in spinach and amaranth was higher than others after application. It's recommended that spinach or amaranth can be selected as representative crop in leaf vegetables in study of systemic fungicides with similarity as thiophanate methyl.

  15. Photosynthesis-related infrared light transmission changes in spinach leaf segments

    SciTech Connect

    Akimoto, T.

    1985-10-01

    The time courses of infrared light transmission changes and fluorescence induced by light in spinach leaf segments were measured. The illumination by red light exhibited a complex wave pattern. The transmission approached the baseline after repeating decreases and increases. Illumination by far-red light decreased the transmission. One of the differences between the two responses was the difference between the two amplitudes of the first increasing component. The component in the red light response was larger than the component in the far-red light response. The transmission decrease by far-red light is supposed to correspond to ''red drop.'' The transmission decrease by far-red light was suppressed by red light. This is due to an activation of a transmission-increasing component. This probably corresponds to ''enhancement.'' A proportional correlation existed between the intensity of far-red light and the minimum intensity of red light that suppressed the transmission decrease induced by far-red light. The component which made Peak D in the time course of fluorescence yield and the first increasing component in the transmission changes were suppressed by intense light.

  16. Identification of Ser-543 as the major regulatory phosphorylation site in spinach leaf nitrate reductase

    NASA Technical Reports Server (NTRS)

    Bachmann, M.; Shiraishi, N.; Campbell, W. H.; Yoo, B. C.; Harmon, A. C.; Huber, S. C.; Davies, E. (Principal Investigator)

    1996-01-01

    Spinach leaf NADH:nitrate reductase (NR) responds to light/dark signals and photosynthetic activity in part as a result of rapid regulation by reversible protein phosphorylation. We have identified the major regulatory phosphorylation site as Ser-543, which is located in the hinge 1 region connecting the cytochrome b domain with the molybdenum-pterin cofactor binding domain of NR, using recombinant NR fragments containing or lacking the phosphorylation site sequence. Studies with NR partial reactions indicated that the block in electron flow caused by phosphorylation also could be localized to the hinge 1 region. A synthetic peptide (NR6) based on the phosphorylation site sequence was phosphorylated readily by NR kinase (NRk) in vitro. NR6 kinase activity tracked the ATP-dependent inactivation of NR during several chromatographic steps and completely inhibited inactivation/phosphorylation of native NR in vitro. Two forms of NRk were resolved by using anion exchange chromatography. Studies with synthetic peptide analogs indicated that both forms of NRk had similar specificity determinants, requiring a basic residue at P-3 (i.e., three amino acids N-terminal to the phosphorylated serine) and a hydrophobic residue at P-5. Both forms are strictly calcium dependent but belong to distinct families of protein kinases because they are distinct immunochemically.

  17. Expression, purification and preliminary crystallographic analysis of sucrose phosphate synthase (SPS) from Halothermothrix orenii

    SciTech Connect

    Huynh, Frederick; Tan, Tien-Chye; Swaminathan, Kunchithapadam; Patel, Bharat K. C.

    2005-01-01

    The first crystallographic study of a sucrose phosphate synthase from H. orenii, an organism that is both thermophilic and halophilic, is reported. The protein crystal diffracts X-rays to 3.01 Å. This is the first report of the crystallization of a sucrose phosphate synthase (SPS; EC 2.4.1.14). It also constitutes the first study of a sucrose phosphate synthase from a non-photosynthetic thermohalophilic anaerobic bacterium, Halothermothrix orenii. The purified recombinant spsA protein has been crystallized in the monoclinic space group C2, with unit-cell parameters a = 154.2, b = 47.9, c = 72.3 Å, β = 103.16°, using the hanging-drop vapour-diffusion method. The crystal diffracts X-rays to a resolution limit of 3.01 Å. Heavy-metal and halide-soaking trials are currently in progress to solve the structure.

  18. Antisense repression of sucrose phosphate synthase in transgenic muskmelon alters plant growth and fruit development

    SciTech Connect

    Tian, Hongmei; Ma, Leyuan; Zhao, Cong; Hao, Hui; Gong, Biao; Yu, Xiyan; Wang, Xiufeng

    2010-03-12

    To unravel the roles of sucrose phosphate synthase (SPS) in muskmelon (Cucumis melo L.), we reduced its activity in transgenic muskmelon plants by an antisense approach. For this purpose, an 830 bp cDNA fragment of muskmelon sucrose phosphate synthase was expressed in antisense orientation behind the 35S promoter of the cauliflower mosaic virus. The phenotype of the antisense plants clearly differed from that of control plants. The transgenic plant leaves were markedly smaller, and the plant height and stem diameter were obviously shorter and thinner. Transmission electron microscope observation revealed that the membrane degradation of chloroplast happened in transgenic leaves and the numbers of grana and grana lamella in the chloroplast were significantly less, suggesting that the slow growth and weaker phenotype of transgenic plants may be due to the damage of the chloroplast ultrastructure, which in turn results in the decrease of the net photosynthetic rate. The sucrose concentration and levels of sucrose phosphate synthase decreased in transgenic mature fruit, and the fruit size was smaller than the control fruit. Together, our results suggest that sucrose phosphate synthase may play an important role in regulating the muskmelon plant growth and fruit development.

  19. Physical and Kinetic Evidence for an Association between Sucrose-Phosphate Synthase and Sucrose-Phosphate Phosphatase.

    PubMed Central

    Echeverria, E.; Salvucci, M. E.; Gonzalez, P.; Paris, G.; Salerno, G.

    1997-01-01

    The possible formation of a multienzyme complex between sucrose (Suc)-phosphate synthase (SPS) and Suc-phosphate phosphatase (SPP) was examined by measuring the rates of Suc-6-phosphate (Suc-6-P) synthesis and hydrolysis in mixing experiments with partially purified enzymes from spinach (Spinacia oleracea) and rice (Oryza sativa) leaves. The addition of SPP to SPS stimulated the rate of Suc-6-P synthesis. SPS inhibited the hydrolysis of exogenous Suc-6-P by SPP when added in the absence of its substrate (i.e. UDP-glucose) but stimulated SPP activity when the SPS substrates were present and used to generate Suc-6-P directly in the reaction. Results from isotope-dilution experiments suggest that Suc-6-P was channeled between SPS and SPP. A portion of the SPS activity comigrated with SPP during native polyacrylamide gel electrophoresis, providing physical evidence for an enzyme-enzyme interaction. Taken together, these results strongly suggest that SPS and SPP associate to form a multienzyme complex. PMID:12223802

  20. Spinach leaf chloroplast CO sub 2 and NO sub 2 sup minus photoassimilations do not compete for photogenerated reductant

    SciTech Connect

    Robinson, J.M. )

    1988-12-01

    Potential competition between CO{sub 2} and NO{sub 2}{sup {minus}} photoassimilation for photogenerated reductant (e.g. reduced ferredoxin and NADPH) was examined employing isolates of mesophyll cells and intact chloroplasts derived from mature source spinach leaves. Variations in the magnitude of incident light energy were used to manipulate the supply of reductant in situ within chloroplasts. Leaf cell and plastid isolates were fed with saturating CO{sub 2} and/or NO{sub 2}{sup {minus}} to produce the highest demand for reductant by CO{sub 2} and/or NO{sub 2}{sup {minus}} assimilatory processes (enzymes). Even in the presence of CO{sub 2} fixation, NO{sub 2}{sup {minus}} reduction in intact leaf cell isolates as well as plastid isolates was maximal at light energies as low as 50 to 200 microeinsteins per second per square meter. Simultaneously, 500 to 800 microeinsteins per second per square meter were required to support maximal CO{sub 2} assimilation. Regardless of the magnitude of the incident light energy, CO{sub 2} assimilation did not repress NO{sub 2}{sup {minus}} reduction, nor were these two processes mutually repressed. These observations have been interpreted to mean that reduced ferredoxin levels in situ in the plastids of mature source leaf mesophyll cells were adequate to supply the concurrent maximal demands exerted by enzymes associated with CO{sub 2} as well as with inorganic nitrogen photoassimilation.

  1. Summer (subarctic) versus winter (subtropic) production affects spinach (Spinacia oleracea L.) leaf bionutrients: vitamins (C, E, Folate, K1, provitamin A), lutein, phenolics, and antioxidants.

    PubMed

    Lester, Gene E; Makus, Donald J; Hodges, D Mark; Jifon, John L

    2013-07-24

    Comparison of spinach (Spinacia oleracea L.) cultivars Lazio and Samish grown during the summer solstice in the subarctic versus the winter solstice in the subtropics provided insight into interactions between production environment (light intensity), cultivar, and leaf age/maturity/position affecting bionutrient concentrations of vitamins (C, E, folate, K1, provitamin A), lutein, phenolics, and antioxidants. Growing spinach during the winter solstice in the subtropics resulted in increased leaf dry matter %, oxidized (dehydro) ascorbic acid (AsA), α- and γ-tocopherol, and total phenols but lower reduced (free) AsA, α-carotene, folate, and antioxidant capacity compared to summer solstice-grown spinach in the subarctic. Both cultivars had similar bionutrients, except for higher dehydroAsA, and lower α- and γ-tocopherol in 'Samish' compared to 'Lazio'. For most bionutrients measured, there was a linear, and sometimes quadratic, increase in concentrations from bottom to top canopy leaves. However, total phenolics and antioxidant capacity increased basipetally. The current study has thus demonstrated that dehydroAsA, α-tocopherol, and γ-tocopherol were substantially lower in subarctic compared to subtropical-grown spinach, whereas the opposite relationship was found for antioxidant capacity, α-carotene, and folates (vitamin B9). The observations are consistent with previously reported isolated effects of growth environment on bionutrient status of crops. The current results clearly highlight the effect of production environment (predominantly radiation capture), interacting with genetics and plant phenology to alter the bionutrient status of crops. While reflecting the effects of changing growing conditions, these results also indicate potential alterations in the nutritive value of foods with anticipated shifts in global climatic conditions.

  2. 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.

  3. Activation of SPS from darkened spinach leaves by an endogenous protein phosphatase

    SciTech Connect

    Huber, S.C.; Huber, J.L. )

    1990-05-01

    Sucrose-phosphate synthase from darkened spinach leaves has a low activation state but can undergo a time-dependent activation in desalted leaf extracts that is inhibited by Pi, molybdate, okadaic acid and vanadate, but stimulated by fluoride. SPS labeled in vivo with ({sup 32}P)Pi in excised leaves in the dark loses incorporated {sup 32}P with time when extracts are incubated at 25{degree}C. This loss is largely prevented by vanadate, suggesting that an endogenous protein phosphatase can use SPS as substrate. Changes in phosphorylation state are closely paralleled by changes in SPS activation state. The spontaneous activation achieved in the extracts can be reversed by addition of 2 mM MgATP. Feeding okadaic acid to darkened leaves prevents light activation of SPS suggesting that the endogenous protein phosphatase is similar to the type-1 enzyme of animal tissues. Overall, the results are consistent with the notion that light activation of SPS involves dephosphorylation of inhibitory phosphorylation site(s). Regulation of the protein phosphatase by Pi may be of physiological significance.

  4. Nodule-enhanced expression of a sucrose phosphate synthase gene member (MsSPSA) has a role in carbon and nitrogen metabolism in the nodules of alfalfa (Medicago sativa L.).

    PubMed

    Aleman, Lorenzo; Ortega, Jose Luis; Martinez-Grimes, Martha; Seger, Mark; Holguin, Francisco Omar; Uribe, Diana J; Garcia-Ibilcieta, David; Sengupta-Gopalan, Champa

    2010-01-01

    Sucrose phosphate synthase (SPS) catalyzes the first step in the synthesis of sucrose in photosynthetic tissues. We characterized the expression of three different isoforms of SPS belonging to two different SPS gene families in alfalfa (Medicago sativa L.), a previously identified SPS (MsSPSA) and two novel isoforms belonging to class B (MsSPSB and MsSPSB3). While MsSPSA showed nodule-enhanced expression, both MsSPSB genes exhibited leaf-enhanced expression. Alfalfa leaf and nodule SPS enzymes showed differences in chromatographic and electrophoretic migration and differences in V (max) and allosteric regulation. The root nodules in legume plants are a strong sink for photosynthates with its need for ATP, reducing power and carbon skeletons for dinitrogen fixation and ammonia assimilation. The expression of genes encoding SPS and other key enzymes in sucrose metabolism, sucrose phosphate phosphatase and sucrose synthase, was analyzed in the leaves and nodules of plants inoculated with Sinorhizobium meliloti. Based on the expression pattern of these genes, the properties of the SPS isoforms and the concentration of starch and soluble sugars in nodules induced by a wild type and a nitrogen fixation deficient strain, we propose that SPS has an important role in the control of carbon flux into different metabolic pathways in the symbiotic nodules.

  5. Summer (sub-arctic) versus winter (sub-tropical) production affects on spinach leaf bio-nutrients: Vitamins (C, E, Folate, K1, provitamin A), lutein, phenolics, and antioxidants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Comparison of spinach (Spinacia oleracea L.) cultivars Lazio and Samish grown during the summer solstice in the sub-arctic versus the winter solstice in the sub-tropics provided insight into interactions between plant environment (day length, light intensity, ambient temperatures), cultivar and leaf...

  6. Nectar secretion requires sucrose phosphate synthases and the sugar transporter SWEET9.

    PubMed

    Lin, I Winnie; Sosso, Davide; Chen, Li-Qing; Gase, Klaus; Kim, Sang-Gyu; Kessler, Danny; Klinkenberg, Peter M; Gorder, Molly K; Hou, Bi-Huei; Qu, Xiao-Qing; Carter, Clay J; Baldwin, Ian T; Frommer, Wolf B

    2014-04-24

    Angiosperms developed floral nectaries that reward pollinating insects. Although nectar function and composition have been characterized, the mechanism of nectar secretion has remained unclear. Here we identify SWEET9 as a nectary-specific sugar transporter in three eudicot species: Arabidopsis thaliana, Brassica rapa (extrastaminal nectaries) and Nicotiana attenuata (gynoecial nectaries). We show that SWEET9 is essential for nectar production and can function as an efflux transporter. We also show that sucrose phosphate synthase genes, encoding key enzymes for sucrose biosynthesis, are highly expressed in nectaries and that their expression is also essential for nectar secretion. Together these data are consistent with a model in which sucrose is synthesized in the nectary parenchyma and subsequently secreted into the extracellular space via SWEET9, where sucrose is hydrolysed by an apoplasmic invertase to produce a mixture of sucrose, glucose and fructose. The recruitment of SWEET9 for sucrose export may have been a key innovation, and could have coincided with the evolution of core eudicots and contributed to the evolution of nectar secretion to reward pollinators.

  7. Functional analysis of sucrose phosphate synthase (SPS) and sucrose synthase (SS) in sugarcane (Saccharum) cultivars.

    PubMed

    Verma, A K; Upadhyay, S K; Verma, P C; Solomon, S; Singh, S B

    2011-03-01

    Sucrose phosphate synthase (SPS; EC 2.4.1.14) and sucrose synthase (SS; EC 2.4.1.13) are key enzymes in the synthesis and breakdown of sucrose in sugarcane. The activities of internodal SPS and SS, as well as transcript expression were determined using semi-quantitative RT-PCR at different developmental stages of high and low sucrose accumulating sugarcane cultivars. SPS activity and transcript expression was higher in mature internodes compared with immature internodes in all the studied cultivars. However, high sugar cultivars showed increased transcript expression and enzyme activity of SPS compared to low sugar cultivars at all developmental stages. SS activity was higher in immature internodes than in mature internodes in all cultivars; SS transcript expression showed a similar pattern. Our studies demonstrate that SPS activity was positively correlated with sucrose and negatively correlated with hexose sugars. However, SS activity was negatively correlated with sucrose and positively correlated with hexose sugars. The present study opens the possibility for improvement of sugarcane cultivars by increasing expression of the respective enzymes using transgene technology.

  8. Nectar secretion requires sucrose phosphate synthases and the sugar transporter SWEET9.

    PubMed

    Lin, I Winnie; Sosso, Davide; Chen, Li-Qing; Gase, Klaus; Kim, Sang-Gyu; Kessler, Danny; Klinkenberg, Peter M; Gorder, Molly K; Hou, Bi-Huei; Qu, Xiao-Qing; Carter, Clay J; Baldwin, Ian T; Frommer, Wolf B

    2014-04-24

    Angiosperms developed floral nectaries that reward pollinating insects. Although nectar function and composition have been characterized, the mechanism of nectar secretion has remained unclear. Here we identify SWEET9 as a nectary-specific sugar transporter in three eudicot species: Arabidopsis thaliana, Brassica rapa (extrastaminal nectaries) and Nicotiana attenuata (gynoecial nectaries). We show that SWEET9 is essential for nectar production and can function as an efflux transporter. We also show that sucrose phosphate synthase genes, encoding key enzymes for sucrose biosynthesis, are highly expressed in nectaries and that their expression is also essential for nectar secretion. Together these data are consistent with a model in which sucrose is synthesized in the nectary parenchyma and subsequently secreted into the extracellular space via SWEET9, where sucrose is hydrolysed by an apoplasmic invertase to produce a mixture of sucrose, glucose and fructose. The recruitment of SWEET9 for sucrose export may have been a key innovation, and could have coincided with the evolution of core eudicots and contributed to the evolution of nectar secretion to reward pollinators. PMID:24670640

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

  10. Extraction and characterization of mixed phase KNO2-KNO3 nanocrystals derived from flat-leaf green spinach

    NASA Astrophysics Data System (ADS)

    Hazarika, S.; Mohanta, D.

    2013-01-01

    Naturally available green spinach, which is a rich source of potassium, was used as the key ingredient to extract mixed-phase ferroelectric crystals of nitrite and nitrate derivatives (KNO2 + KNO3). The KNO3 phase was found to be dominant for higher pH values, as revealed by the x-ray diffraction patterns. The characteristic optical absorption spectra exhibited intra-band π-π* electronic transitions, whereas Fourier transform infrared spectra exhibited characteristic N-O stretching vibrations. Differential scanning calorimetry revealed a broad endothermic peak at ˜121.8 °C, highlighting a transition from phase II to I via phase III of KNO3. Obtaining nanoscale ferroelectrics via the adoption of green synthesis is economically viable for large-scale production and possible application in ferroelectric elements/devices.

  11. Testing models of fatty acid transfer and lipid synthesis in spinach leaf using in vivo oxygen-18 labeling

    SciTech Connect

    Pollard, M.; Ohlrogge, J.

    1999-12-01

    Oxygen-18 labeling has been applied to the study of plant lipid biosynthesis for the first time. [{sup 13}C{sub 2}{sup 18}O{sub 2}]Acetate was incubated with spinach (Spinacia oleracea) leaves and the {sup 18}O content in fatty acid methyl esters isolated from different lipid classes measured by gas chromatography-mass spectrometry. Fatty acids isolated from lipids synthesized within the plastid, such as monogalactosyldiacylglycerol, show an {sup 18}O content consistent with the exogenous acetate undergoing a single activation step and with the direct utilization of acyl-acyl carrier protein by the acyl transferases of the chloroplast. In contrast, fatty acids isolated from lipids assembled in the cytosol, such as phosphatidylcholine, show a 50% reduction in the {sup 18}O content. This is indicative of export of the fatty acyl groups from the plastid via a free carboxylate anion, and is consistent with the acyl-acyl carrier protein thioesterase:acyl-coenzyme A (CoA) synthetase mediated export mechanism. If this were not the case and the acyl group was transferred directly from acyl-acyl carrier protein to an acyl acceptor on the cytosolic side, there would be either complete retention of {sup 18}O or, less likely, complete loss of {sup 18}O, but not a 50% loss of {sup 18}O. Thus, existing models for fatty acid transfer from the plastid and for spatially separate synthesis of prokaryotic and eukaryotic lipids have both been confirmed.

  12. Activities of starch hydrolytic enzymes and sucrose-phosphate synthase in the stems of rice subjected to water stress during grain filling.

    PubMed

    Yang, J; Zhang, J; Wang, Z; Zhu, Q

    2001-11-01

    To understand the effect of water stress on the remobilization of prestored carbon reserves, the changes in the activities of starch hydrolytic enzymes and sucrose-phosphate synthase (SPS) in the stems of rice (Oryza sativa L.) during grain filling were investigated. Two rice cultivars, showing high lodging-resistance and slow remobilization, were grown in the field and subjected to well-watered (WW, psi(soil)=0) and water-stressed (WS, psi(soil)=-0.05 MPa) treatments 9 d after anthesis (DAA) till maturity. Leaf water potentials of both cultivars markedly decreased during the day as a result of WS treatment, but completely recovered by early morning. WS treatment accelerated the reduction of starch in the stems, promoted the reallocation of prefixed (14)C from the stems to grains, shortened the grain filling period, and increased the grain filling rate. More soluble sugars including sucrose were accumulated in the stems under WS than under WW treatments. Both alpha- and beta-amylase activities were enhanced by the WS, with the former enhanced more than the latter, and were significantly correlated with the concentrations of soluble sugars in the stems. The other two possible starch-breaking enzymes, alpha-glucosidase and starch phosphorylase, showed no significant differences in the activities between the WW and WS treatments. Water stress also increased the SPS activity that is responsible for sucrose production. Both V(limit) and V(max), the activities of the enzyme at limiting and saturating substrate concentrations, were enhanced and the activation state (V(limit)/V(max)) was also increased as a result of the more significant enhancement of V(limit). The enhanced SPS activity was closely correlated with an increase of sucrose accumulation in the stems. The results suggest that the fast hydrolysis of starch and increased carbon remobilization were attributed to the enhanced alpha-amylase activity and the high activation state of SPS when the rice was subjected

  13. Mild water stress of Phaseolus vulgaris plants leads to reduced starch synthesis and extractable sucrose phosphate synthase activity

    SciTech Connect

    Vassey, T.L.; Sharkey, T.D. )

    1989-04-01

    Mild water stress, on the order of {minus}1.0 megapascals xylem water potential, can reduce the rate of photosynthesis and eliminate the inhibition of photosynthesis caused by O{sub 2} in water-stress-sensitive plants such as Phaseolus vulgaris. To investigate the lack of O{sub 2} inhibition of photosynthesis, we measured stromal and cytosolic fructose-1,6-bisphosphatase, sucrose phosphate synthase, and partitioning of newly fixed carbon between starch and sucrose before, during, and after mild water stress. The extractable activity of the fructose bisphosphatases was unaffected by mild water stress. The extractable activity of SPS was inhibited by more than 60% in plants stressed to water potentials of {minus}0.9 megapascals. Water stress caused a decline in the starch/sucrose partitioning ratio indicating that starch synthesis was inhibited more than sucrose synthesis. We conclude that the reduced rate of photosynthesis during water stress is caused by stomatal closure, and that the restriction of CO{sub 2} supply caused by stomatal closure leads to a reduction in the capacity for both starch and sucrose synthesis. This causes the reduced O{sub 2} inhibition and abrupt CO{sub 2} saturation of photosynthesis.

  14. The Structure of Sucrose Phosphate Synthase from Halothermothrix orenii Reveals Its Mechanism of Action and Binding Mode

    SciTech Connect

    Chua,T.; Bujnicki, J.; Tan, T.; Huynh, F.; Patel, B.; Sivaraman, J.; Ogimoto, Y.; Miyano, K.; Sawa, H.

    2008-01-01

    Sucrose phosphate synthase (SPS) catalyzes the transfer of a glycosyl group from an activated donor sugar, such as uridine diphosphate glucose (UDP-Glc), to a saccharide acceptor D-fructose 6-phosphate (F6P), resulting in the formation of UDP and D-sucrose-6'-phosphate (S6P). This is a central regulatory process in the production of sucrose in plants, cyanobacteria, and proteobacteria. Here, we report the crystal structure of SPS from the nonphotosynthetic bacterium Halothermothrix orenii and its complexes with the substrate F6P and the product S6P. SPS has two distinct Rossmann-fold domains with a large substrate binding cleft at the interdomain interface. Structures of two complexes show that both the substrate F6P and the product S6P bind to the A-domain of SPS. Based on comparative analysis of the SPS structure with other related enzymes, the donor substrate, nucleotide diphosphate glucose, binds to the B-domain of SPS. Furthermore, we propose a mechanism of catalysis by H. orenii SPS. Our findings indicate that SPS from H. orenii may represent a valid model for the catalytic domain of plant SPSs and thus may provide useful insight into the reaction mechanism of the plant enzyme.

  15. Alternative soaking media for the FDA procedure in the detection of salmonella from tomatoes and spinach leaf using phage magnetoelastic biosensors

    NASA Astrophysics Data System (ADS)

    Chen, I.-Hsuan; Hu, Jiajia; Wang, Fengen; Horikawa, Shin; Barbaree, James M.; Chin, Bryan A.

    2016-05-01

    Efforts were made to incorporate the phage Magnetoelastic (ME) biosensor in FDA's Spinach Soaking procedures according to FDA 2015 BAM method. Three soaking materials (Lactose broth, LB broth, and Peptone water) and various soaking times were investigated. Using merely 100 Salmonella cells spiked on the produce surfaces, the phage biosensors detected Salmonella within 5 hours when soaking tomatoes in LB broth as opposed to taking up to 24 hours. Salmonella was detected on spinach leaves within 7 hours. These phage ME biosensors provide a promising rapid detection platform using LB broth in FDA's soaking procedures while shortening the incubation period.

  16. Sucrose Phosphate Synthase and Acid Invertase as Determinants of Sucrose Concentration in Developing Muskmelon (Cucumis melo L.) Fruits 1

    PubMed Central

    Hubbard, Natalie L.; Huber, Steven C.; Pharr, D. Mason

    1989-01-01

    Fruits of orange-fleshed and green-fleshed muskmelon (Cucumis melo L.) were harvested at different times throughout development to evaluate changes in metabolism which lead to sucrose accumulation, and to determine the basis of differences in fruit sucrose accumulation among genotypes. Concentrations of sucrose, raffinose saccharides, hexoses and starch, as well as activities of the sucrose metabolizing enzymes sucrose phosphate synthase (SPS) (EC 2.4.1.14), sucrose synthase (EC 2.4.1.13), and acid and neutral invertases (EC 3.2.1.26) were measured. Sucrose synthase and neutral invertase activities were relatively low (1.7 ± 0.3 micromole per hour per gram fresh weight and 2.2 ± 0.2, respectively) and changed little throughout fruit development. Acid invertase activity decreased during fruit development, (from as high as 40 micromoles per hour per gram fresh weight) in unripe fruit, to undetectable activity in mature, ripened fruits, while SPS activity in the fruit increased (from 7 micromoles per hour per gram fresh weight) to as high as 32 micromoles per hour per gram fresh weight. Genotypes which accumulated different amounts of sucrose had similar acid invertase activity but differed in SPS activity. Our results indicate that both acid invertase and SPS are determinants of sucrose accumulation in melon fruit. However, the decline in acid invertase appears to be a normal function of fruit maturation, and is not the primary factor which determines sucrose accumulation. Rather, the capacity for sucrose synthesis, reflected in the activity of SPS, appears to determine sucrose accumulation, which is an important component of fruit quality. PMID:16667212

  17. Conversion of L-sorbosone to L-ascorbic acid by a NADP-dependent dehydrogenase in bean and spinach leaf. [Phaseolus vulgaris L. ; Spinacia oleracea L

    SciTech Connect

    Loewus, M.W.; Bedgar, D.L.; Saito, Kazumi; Loewus, F.A. )

    1990-11-01

    An NADP-dependent dehydrogenase catalyzing the conversion of L-sorbosone to L-ascorbic acid has been isolated from Phaseolus vulgaris L. and Spinacia oleracea L. and partially purified. It is stable at {minus}20{degree}C for up to 8 months. Molecular masses, as determined by gel filtration, were 21 and 29 kilodaltons for bean and spinach enzymes, respectively. K{sub m} for sorbosone were 12 {plus minus} 2 and 18 {plus minus} 2 millimolar and for NADP{sup +}, 0.14 {plus minus} 0.05 and 1.2 {plus minus} 0.5 millimolar, for bean and spinach, respectively. Lycorine, a purported inhibitor of L-ascorbic acid biosynthesis, had no effect on the reaction.

  18. Characterization of technetium species induced in spinach

    SciTech Connect

    Harms, A.V.; Krijger, G.C.; Elteren, J.T. van; Goeij, J.J.M. de

    1999-08-01

    Plants have the ability to accumulate the long-lived fission product {sup 99}Tc. In this work, an attempt was made to separate and characterize technetium species induced by spinach plants (Spinacia oleracea L.) grown on a TcO{sub 4}{sup {minus}} containing nutrient solution. Combination of data obtained with selective extraction and chromatography gave us insight into Tc speciation in spinach plants. The following classes of Tc species in spinach leaf homogenate were found after an incubation period of 11 d: TcO{sub 4}{sup {minus}} (ca. 7%), Tc{sup V}-cysteine (ca. 25%), Tc bound to insoluble cell-wall polysaccharides (ca. 17%), Tc bound to proteins (ca. 26%), and hydrophilic non-protein Tc species (ca. 25%). These results may yield a new insight into the metabolic pathways of Tc in plants.

  19. Lettuce and spinach breeding

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lettuce and spinach production is beset by numerous biotic an abiotic challenges. This report to the California Leafy Greens Research Program annual meeting provides an update by the ‘Genetic Enhancement of Lettuce, Spinach, Melon, and Related Species’ project at Salinas on the genetics and breeding...

  20. Analysis of gene-disruption mutants of a sucrose phosphate synthase gene in rice, OsSPS1, shows the importance of sucrose synthesis in pollen germination.

    PubMed

    Hirose, Tatsuro; Hashida, Yoichi; Aoki, Naohiro; Okamura, Masaki; Yonekura, Madoka; Ohto, Chikara; Terao, Tomio; Ohsugi, Ryu

    2014-08-01

    The molecular function of an isoform of sucrose phosphate synthase (SPS) in rice, OsSPS1, was investigated using gene-disruption mutant lines generated by retrotransposon insertion. The progeny of the heterozygote of disrupted OsSPS1 (SPS1(+/-)) segregated into SPS1(+/+), SPS1(+/-), and SPS1(-/-) at a ratio of 1:1:0. This distorted segregation ratio, together with the expression of OsSPS1 in the developing pollen revealed by quantitative RT-PCR analysis and promoter-beta-glucuronidase (GUS) fusion assay, suggested that the disruption of OsSPS1 results in sterile pollen. This hypothesis was reinforced by reciprocal crosses of SPS1(+/-) plants with wild-type plants in which the disrupted OsSPS1 was not paternally transmitted to the progeny. While the pollen grains of SPS(+/-) plants normally accumulated starch during their development, pollen germination on the artificial media was reduced to half of that observed in the wild-type control. Overall, our data suggests that sucrose synthesis via OsSPS1 is essential in pollen germination in rice. PMID:25017165

  1. International collaborative study of the endogenous reference gene, sucrose phosphate synthase (SPS), used for qualitative and quantitative analysis of genetically modified rice.

    PubMed

    Jiang, Lingxi; Yang, Litao; Zhang, Haibo; Guo, Jinchao; Mazzara, Marco; Van den Eede, Guy; Zhang, Dabing

    2009-05-13

    One rice ( Oryza sativa ) gene, sucrose phosphate synthase (SPS), has been proven to be a suitable endogenous reference gene for genetically modified (GM) rice detection in a previous study. Herein are the reported results of an international collaborative ring trial for validation of the SPS gene as an endogenous reference gene and its optimized qualitative and quantitative polymerase chain reaction (PCR) systems. A total of 12 genetically modified organism (GMO) detection laboratories from seven countries participated in the ring trial and returned their results. The validated results confirmed the species specificity of the method through testing 10 plant genomic DNAs, low heterogeneity, and a stable single-copy number of the rice SPS gene among 7 indica varieties and 5 japonica varieties. The SPS qualitative PCR assay was validated with a limit of detection (LOD) of 0.1%, which corresponded to about 230 copies of haploid rice genomic DNA, while the limit of quantification (LOQ) for the quantitative PCR system was about 23 copies of haploid rice genomic DNA, with acceptable PCR efficiency and linearity. Furthermore, the bias between the test and true values of eight blind samples ranged from 5.22 to 26.53%. Thus, we believe that the SPS gene is suitable for use as an endogenous reference gene for the identification and quantification of GM rice and its derivates.

  2. Expression patterns, activities and carbohydrate-metabolizing regulation of sucrose phosphate synthase, sucrose synthase and neutral invertase in pineapple fruit during development and ripening.

    PubMed

    Zhang, Xiu-Mei; Wang, Wei; Du, Li-Qing; Xie, Jiang-Hui; Yao, Yan-Li; Sun, Guang-Ming

    2012-01-01

    Differences in carbohydrate contents and metabolizing-enzyme activities were monitored in apical, medial, basal and core sections of pineapple (Ananas comosus cv. Comte de paris) during fruit development and ripening. Fructose and glucose of various sections in nearly equal amounts were the predominant sugars in the fruitlets, and had obvious differences until the fruit matured. The large rise of sucrose/hexose was accompanied by dramatic changes in sucrose phosphate synthase (SPS) and sucrose synthase (SuSy) activities. By contrast, neutral invertase (NI) activity may provide a mechanism to increase fruit sink strength by increasing hexose concentrations. Furthermore, two cDNAs of Ac-sps (accession no. GQ996582) and Ac-ni (accession no. GQ996581) were first isolated from pineapple fruits utilizing conserved amino-acid sequences. Homology alignment reveals that the amino acid sequences contain some conserved function domains. Transcription expression analysis of Ac-sps, Ac-susy and Ac-ni also indicated distinct patterns related to sugar accumulation and composition of pineapple fruits. It suggests that differential expressions of multiple gene families are necessary for sugar metabolism in various parts and developmental stages of pineapple fruit. A cycle of sucrose breakdown in the cytosol of sink tissues could be mediated through both Ac-SuSy and Ac-NI, and Ac-NI could be involved in regulating crucial steps by generating sugar signals to the cells in a temporally and spatially restricted fashion.

  3. Effect of spinach cultivar and bacterial adherence factors on survival of Escherichia coli O157:H7 on spinach leaves.

    PubMed

    Macarisin, Dumitru; Patel, Jitendra; Bauchan, Gary; Giron, Jorge A; Ravishankar, Sadhana

    2013-11-01

    Similar to phytopathogens, human bacterial pathogens have been shown to colonize the plant phylloplane. In addition to environmental factors, such as temperature, UV, relative humidity, etc., the plant cultivar and, specifically, the leaf blade morphological characteristics may affect the persistence of enteropathogens on leafy greens. This study was conducted to evaluate the effect of cultivar-dependent leaf topography and the role of strain phenotypic characteristics on Escherichia coli O157:H7 persistence on organic spinach. Spinach cultivars Emilia, Lazio, Space, and Waitiki were experimentally inoculated with the foodborne E. coli O157:H7 isolate EDL933 and its isogenic mutants deficient in cellulose, curli, or both curli and cellulose production. Leaves of 6-week-old plants were inoculated with 6.5 log CFU per leaf in a biosafety level 2 growth chamber. At 0, 1, 7, and 14 days, E. coli O157:H7 populations were determined by plating on selective medium and verified by laser scanning confocal microscopy. Leaf morphology (blade roughness and stoma density) was evaluated by low-temperature and variable-pressure scanning electron microscopy. E. coli O157:H7 persistence on spinach was significantly affected by cultivar and strain phenotypic characteristics, specifically, the expression of curli. Leaf blade roughness and stoma density influenced the persistence of E. coli O157:H7 on spinach. Cultivar Waitiki, which had the greatest leaf roughness, supported significantly higher E. coli O157:H7 populations than the other cultivars. These two morphological characteristics of spinach cultivars should be taken into consideration in developing intervention strategies to enhance the microbial safety of leafy greens.

  4. Nitrogen Uptake in Spinach

    NASA Astrophysics Data System (ADS)

    Ramirez, J.; VanBenthem, P.

    2013-12-01

    A plant's absorption of nitrogen can be encouraged by a variety of environmental factors, especially the application of fertilizers. As a common limiting factor in plant growth, not up taking enough nitrogen can be a result of an unhealthy plant. Moreover, as farmers seek out methods to increase growth of plants, fertilizers are used as a solution to the issue of nitrogen deficiency to incorporate additional nitrogen from chemical or organic sources, by not using the right fertilizer can greatly affect the plats. The point of this research project is to determine the effect of various fertilizers on the plant growth, and to correlate the measured nitrogen, water and chlorophyll content in spinach leaves. Spinach leaves were used because they are known to quickly uptake chemicals in the environment. The spinach plants were exposed to four different growing parameters, which are referred to as control, ammonium nitrate, MiracleGro , and organic. The spinach was originally placed in nitrogen deficient soil with only 2.2x10 4 weight percent (wt. %) nitrogen. The leaves in the control group were grown in this nitrogen deficient soil without any fertilizer added. Ammomium nitrate and MiracleGro were added to the spinach in the A and MG groups, respectively, and organic chicken stool was used for the O group. By using a spectral imaging system and flame combustion techniques, the chlorophyll content can be related to the nitrogen content in the spinach leaves. In these spinach leaves, nitrogen and chlorophyll content were measured, chlorophyll is a green pigment that plays a crucial role in producing nutrients for green plants. The lack of chlorophyll will allow the plant to become susceptible to diseases, so it is extremely important that the plants have a high content of chlorophyll. The role of nitrogen in chlorophyll is very important and helps in the creation of chlorophyll; therefore it is necessary that an appropriate amount of nitrogen is added for optimal growth

  5. Physical and mechanical properties of spinach for whole-surface online imaging inspection

    NASA Astrophysics Data System (ADS)

    Tang, Xiuying; Mo, Chang Y.; Chan, Diane E.; Peng, Yankun; Qin, Jianwei; Yang, Chun-Chieh; Kim, Moon S.; Chao, Kuanglin

    2011-06-01

    The physical and mechanical properties of baby spinach were investigated, including density, Young's modulus, fracture strength, and friction coefficient. The average apparent density of baby spinach leaves was 0.5666 g/mm3. The tensile tests were performed using parallel, perpendicular, and diagonal directions with respect to the midrib of each leaf. The test results showed that the mechanical properties of spinach are anisotropic. For the parallel, diagonal, and perpendicular test directions, the average values for the Young's modulus values were found to be 2.137MPa, 1.0841 MPa, and 0.3914 MPa, respectively, and the average fracture strength values were 0.2429 MPa, 0.1396 MPa, and 0.1113 MPa, respectively. The static and kinetic friction coefficient between the baby spinach and conveyor belt were researched, whose test results showed that the average coefficients of kinetic and maximum static friction between the adaxial (front side) spinach leaf surface and conveyor belt were 1.2737 and 1.3635, respectively, and between the abaxial (back side) spinach leaf surface and conveyor belt were 1.1780 and 1.2451 respectively. These works provide the basis for future development of a whole-surface online imaging inspection system that can be used by the commercial vegetable processing industry to reduce food safety risks.

  6. Betaine aldehyde dehydrogenase isozymes of spinach

    SciTech Connect

    Hanson, A.D.; Weretilnyk, E.A.; Weigel, P.

    1986-04-01

    Betaine is synthesized in spinach chloroplasts via the pathway Choline ..-->.. Betaine Aldehyde ..-->.. Betaine; the second step is catalyzed by betaine aldehyde dehydrogenase (BADH). The subcellular distribution of BADH was determined in leaf protoplast lysates; BADH isozymes were separated by 6-9% native PAGE. The chloroplast stromal fraction contains a single BADH isozyme (number1) that accounts for > 80% of the total protoplast activity; the extrachloroplastic fraction has a minor isozyme (number2) which migrates more slowly than number1. Both isozymes appear specific for betaine aldehyde, are more active with NAD than NADP, and show a ca. 3-fold activity increase in salinized leaves. The phenotype of a natural variant of isozyme number1 suggests that the enzyme is a dimer.

  7. Association analysis for oxalate concentration in spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Screening and breeding low-oxalate germplasm is a major objective in spinach breeding. This research aims to conduct association analysis and identify SNP markers associated with oxalate concentration in spinach germplasm. A total of 310 spinach genotypes including 300 USDA germplasm accessions and ...

  8. Understanding the molecular mechanism of transcriptional regulation of banana Sucrose phosphate synthase (SPS) gene during fruit ripening: an insight into the functions of various cis-acting regulatory elements.

    PubMed

    Choudhury, Swarup Roy; Roy, Sujit; Singh, Sanjay Kumar; Sengupta, Dibyendu N

    2010-05-01

    Recently, we have reported the characterization of promoter region of Sucrose phosphate synthase (SPS) gene in banana and investigated the role of some cis-elements/motifs, present in the promoter of SPS, in the transcriptional regulation of the gene. DNA-protein interaction studies have demonstrated the presence of specific trans-acting factors which showed specific interactions with ethylene, auxin, low temperature and light responsive elements in regulating SPS transcription. Transient expression analyses have demonstrated the functional significance of the various cis-acting regulatory elements present in banana SPS promoter in regulating SPS expression during ripening. (1) Here, we have further discussed the possible role of these regulatory sequences in the regulation of transcriptional network and comment on their function in relation to sucrose metabolism during banana fruit ripening. PMID:20139735

  9. Ammonium reduces oxalate accumulation in different spinach (Spinacia oleracea L.) genotypes by inhibiting root uptake of nitrate.

    PubMed

    Liu, Xiaoxia; Lu, Lingli; Chen, Qiuhui; Ding, Wenya; Dai, Peibin; Hu, Yan; Yu, Yan; Jin, Chongwei; Lin, Xianyong

    2015-11-01

    Excessive accumulation of oxalate negatively affects nutritional value of many vegetables, such as spinach (Spinacia oleracea L.). Mixed solution of ammonium and nitrate could effectively reduce oxalate accumulation, while the mechanism involved remains unknown. High (Heizhenzhu) and low (Weilv) oxalate-accumulated spinach genotypes were used in this study to investigate the association of oxalate accumulation and root uptake of nitrogen. Exposure of increasing nitrate or mixed-nitrogen (nitrate:ammonium = 1:1) significantly increased leaf total and soluble oxalate contents. In contrast, increasing ammonium did not result in elevation of leaf oxalate. Correlation analysis confirmed that leaf oxalate accumulation was positively associated with root uptake of nitrate but not ammonium. Moreover, addition of ammonium significantly reduced nitrate uptake rate, and subsequently decreased leaf oxalate accumulation. The results suggest that oxalate synthesis in spinach leaves is associated with its root uptake of nitrate, and ammonium is able to reduce oxalate accumulation by inhibiting uptake of nitrate.

  10. Genetic diversity and association analysis of leafminer (Liriomyza langei) resistance in spinach (Spinacia oleracea).

    PubMed

    Shi, Ainong; Mou, Beiquan

    2016-08-01

    Leafminer (Liriomyza langei) is a major insect pest of many important agricultural crops, including spinach (Spinacia oleracea). Use of genetic resistance is an efficient, economic, and environment-friendly method to control this pest. The objective of this research was to conduct association analysis and identify single nucleotide polymorphism (SNP) markers associated with leafminer resistance in spinach germplasm. A total of 300 USDA spinach germplasm accessions were used for the association analysis of leafminer resistance. Genotyping by sequencing (GBS) was used for genotyping and 783 SNPs from GBS were used for association analysis. The leafminer resistance showed a near normal distribution with a wide range from 1.1 to 11.7 stings per square centimeter leaf area, suggesting that the leafminer resistance in spinach is a complex trait controlled by multiple genes with minor effect in this spinach panel. Association analysis indicated that five SNP markers, AYZV02040968_7171, AYZV02076752_412, AYZV02098618_4615, AYZV02147304_383, and AYZV02271373_398, were associated with the leafminer resistance with LOD 2.5 or higher. The SNP markers may be useful for breeders to select plants and lines for leafminer resistance in spinach breeding programs through marker-assisted selection. PMID:27490441

  11. Genetic diversity and association analysis of leafminer (Liriomyza langei) resistance in spinach (Spinacia oleracea).

    PubMed

    Shi, Ainong; Mou, Beiquan

    2016-08-01

    Leafminer (Liriomyza langei) is a major insect pest of many important agricultural crops, including spinach (Spinacia oleracea). Use of genetic resistance is an efficient, economic, and environment-friendly method to control this pest. The objective of this research was to conduct association analysis and identify single nucleotide polymorphism (SNP) markers associated with leafminer resistance in spinach germplasm. A total of 300 USDA spinach germplasm accessions were used for the association analysis of leafminer resistance. Genotyping by sequencing (GBS) was used for genotyping and 783 SNPs from GBS were used for association analysis. The leafminer resistance showed a near normal distribution with a wide range from 1.1 to 11.7 stings per square centimeter leaf area, suggesting that the leafminer resistance in spinach is a complex trait controlled by multiple genes with minor effect in this spinach panel. Association analysis indicated that five SNP markers, AYZV02040968_7171, AYZV02076752_412, AYZV02098618_4615, AYZV02147304_383, and AYZV02271373_398, were associated with the leafminer resistance with LOD 2.5 or higher. The SNP markers may be useful for breeders to select plants and lines for leafminer resistance in spinach breeding programs through marker-assisted selection.

  12. Spinach and mustard greens response to soil type, sulfur addition and lithium level

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A greenhouse experiment was conducted near Weslaco, Texas (Lat. 26o 8' N, Long. 97o 57' W) between Dec. 2006 and Feb 2007 to evaluate the effect of soil type, added sulfur and lithium level on the growth and leaf nutrients, particularly biofortified levels of Li and S, in spinach and mustard gree...

  13. [Fluorescence used to investigate the sensitivity of spinach chloroplast membrane to low intensity electromagnetic radiation].

    PubMed

    Xi, Gang; Yang, Yun-Jing; Lu, Hong

    2009-07-01

    A system for studying biological effect of radio frequency electromagnetic field was developed. The system can form an area where electromagnetic wave with large frequency range is well distributed. The strength of electromagnetic wave was measured easily. Electromagnetic wave in the system did not have effect on environment. The sensitivity of spinach chloroplast membrane to low intensity electromagnetic radiation of 300 MHz under power density of 5 mW x cm(-2) was studied by the spectral analysis method of fluorescence of 8-anilino-1-naphthalene-sulfonic acid (ANS) and the changes in chlorophyll a (Chla) fluorescence parameters of spinach chloroplast membrane. The result showed that the position of spectrum of ANS fluorescence of spinach chloroplast membrane did not change, but the intensity of ANS fluorescence was obviously increased under the action of electromagnetic radiation with power density of 1-5 mW x cm(-2). There was an increase in the intensity of ANS fluorescence with the increase in electromagnetic radiation. The increase of ANS fluorescence of spinach chloroplast membrane showed that low level electromagnetic field induced the decrease in fluidity of chloroplast membrane compared with control experiment. The cause of the change in the fluidity could be related to the polarization of chloroplast membrane under the electromagnetic field. The analysis of Chla fluorescence parameters of spinach chloroplast membrane indicated that low level electromagnetic field of 300 MHz made the fluorescence parameters F0 and F(VI/)F(V) decrease, and F(V)/Fo, Fv/F(m) and deltaF(V)/T increase. It was showed that low level electromagnetic field caused the change of non-active center of photosystem II of spinach chloroplast membrane to active center and the increase in potential active and photochemical efficiency of PSII, and promoted the transmit process of electron in photosynthesis of chloroplast membrane of photosynthesis cell in spinach leaf. The study confirmed

  14. Characterization of multiple SPS knockout mutants reveals redundant functions of the four Arabidopsis sucrose phosphate synthase isoforms in plant viability, and strongly indicates that enhanced respiration and accelerated starch turnover can alleviate the blockage of sucrose biosynthesis.

    PubMed

    Bahaji, Abdellatif; Baroja-Fernández, Edurne; Ricarte-Bermejo, Adriana; Sánchez-López, Ángela María; Muñoz, Francisco José; Romero, Jose M; Ruiz, María Teresa; Baslam, Marouane; Almagro, Goizeder; Sesma, María Teresa; Pozueta-Romero, Javier

    2015-09-01

    We characterized multiple knock-out mutants of the four Arabidopsis sucrose phosphate synthase (SPSA1, SPSA2, SPSB and SPSC) isoforms. Despite their reduced SPS activity, spsa1/spsa2, spsa1/spsb, spsa2/spsb, spsa2/spsc, spsb/spsc, spsa1/spsa2/spsb and spsa2/spsb/spsc mutants displayed wild type (WT) vegetative and reproductive morphology, and showed WT photosynthetic capacity and respiration. In contrast, growth of rosettes, flowers and siliques of the spsa1/spsc and spsa1/spsa2/spsc mutants was reduced compared with WT plants. Furthermore, these plants displayed a high dark respiration phenotype. spsa1/spsb/spsc and spsa1/spsa2/spsb/spsc seeds poorly germinated and produced aberrant and sterile plants. Leaves of all viable sps mutants, except spsa1/spsc and spsa1/spsa2/spsc, accumulated WT levels of nonstructural carbohydrates. spsa1/spsc leaves possessed high levels of metabolic intermediates and activities of enzymes of the glycolytic and tricarboxylic acid cycle pathways, and accumulated high levels of metabolic intermediates of the nocturnal starch-to-sucrose conversion process, even under continuous light conditions. Results presented in this work show that SPS is essential for plant viability, reveal redundant functions of the four SPS isoforms in processes that are important for plant growth and nonstructural carbohydrate metabolism, and strongly indicate that accelerated starch turnover and enhanced respiration can alleviate the blockage of sucrose biosynthesis in spsa1/spsc leaves.

  15. Purification and characterization of recombinant sugarcane sucrose phosphate synthase expressed in E. coli and insect Sf9 cells: an importance of the N-terminal domain for an allosteric regulatory property.

    PubMed

    Sawitri, Widhi Dyah; Narita, Hirotaka; Ishizaka-Ikeda, Etsuko; Sugiharto, Bambang; Hase, Toshiharu; Nakagawa, Atsushi

    2016-06-01

    Sucrose phosphate synthase (SPS) catalyses the transfer of glycosyl group of uridine diphosphate glucose to fructose-6-phosphate to form sucrose-6-phosphate. Plant SPS plays a key role in photosynthetic carbon metabolisms, which activity is modulated by an allosteric activator glucose-6-phosphate (G6P). We produced recombinant sugarcane SPS using Escherichia coli and Sf9 insect cells to investigate its structure-function relationship. When expressed in E. coli, two forms of SPS with different sizes appeared; the larger was comparable in size with the authentic plant enzyme and the shorter was trimmed the N-terminal 20 kDa region off. In the insect cells, only enzyme with the authentic size was produced. We purified the trimmed SPS and the full size enzyme from insect cells and found their enzymatic properties differed significantly; the full size enzyme was activated allosterically by G6P, while the trimmed one showed a high activity even without G6P. We further introduced a series of N-terminal truncations up to 171 residue and found G6P-independent activity was enhanced by the truncation. These combined results indicated that the N-terminal region of sugarcane SPS is crucial for the allosteric regulation by G6P and may function like a suppressor domain for the enzyme activity. PMID:26826371

  16. Characterization of multiple SPS knockout mutants reveals redundant functions of the four Arabidopsis sucrose phosphate synthase isoforms in plant viability, and strongly indicates that enhanced respiration and accelerated starch turnover can alleviate the blockage of sucrose biosynthesis.

    PubMed

    Bahaji, Abdellatif; Baroja-Fernández, Edurne; Ricarte-Bermejo, Adriana; Sánchez-López, Ángela María; Muñoz, Francisco José; Romero, Jose M; Ruiz, María Teresa; Baslam, Marouane; Almagro, Goizeder; Sesma, María Teresa; Pozueta-Romero, Javier

    2015-09-01

    We characterized multiple knock-out mutants of the four Arabidopsis sucrose phosphate synthase (SPSA1, SPSA2, SPSB and SPSC) isoforms. Despite their reduced SPS activity, spsa1/spsa2, spsa1/spsb, spsa2/spsb, spsa2/spsc, spsb/spsc, spsa1/spsa2/spsb and spsa2/spsb/spsc mutants displayed wild type (WT) vegetative and reproductive morphology, and showed WT photosynthetic capacity and respiration. In contrast, growth of rosettes, flowers and siliques of the spsa1/spsc and spsa1/spsa2/spsc mutants was reduced compared with WT plants. Furthermore, these plants displayed a high dark respiration phenotype. spsa1/spsb/spsc and spsa1/spsa2/spsb/spsc seeds poorly germinated and produced aberrant and sterile plants. Leaves of all viable sps mutants, except spsa1/spsc and spsa1/spsa2/spsc, accumulated WT levels of nonstructural carbohydrates. spsa1/spsc leaves possessed high levels of metabolic intermediates and activities of enzymes of the glycolytic and tricarboxylic acid cycle pathways, and accumulated high levels of metabolic intermediates of the nocturnal starch-to-sucrose conversion process, even under continuous light conditions. Results presented in this work show that SPS is essential for plant viability, reveal redundant functions of the four SPS isoforms in processes that are important for plant growth and nonstructural carbohydrate metabolism, and strongly indicate that accelerated starch turnover and enhanced respiration can alleviate the blockage of sucrose biosynthesis in spsa1/spsc leaves. PMID:26259182

  17. Investigations on heat resistance of spinach leaves.

    PubMed

    Santarius, K A; Müller, M

    1979-10-01

    Exposure of spinach plants to high temperature (35° C) increased the heat resistance of the leaves by about 3° C. This hardening process occurred within 4 to 6 h, whereas dehardening at 20°/15° C required 1 to 2 days. At 5° C dehardening did not take place. Hardening and dehardening occurred in both the dark and the light. The hardiness was tested by exposure of the leaves to heat stress and subsequent measurements of chlorophyll fluorescence induction and light-induced absorbance changes at 535 nm on the leaves and of the photosynthetic electron transport in thylakoids isolated after heat treatment. Heat-induced damage to both heat-hardened and non-hardened leaves seemed to consist primarily in a breakdown of the membrane potential of the thylakoids accompanied by partial inactivation of electron transport through photosystem II. The increase in heat resistance was not due to temperature-induced changes in lipid content and fatty acid composition of the thylakoids, and no conspicuous changes in the polypeptide composition of the membranes were observed. Prolonged heat treatment at 35° C up to 3 days significantly decreased the total lipid content and the degree of unsaturation of the fatty acids of membrane lipids without further increase in the thermostability of the leaves. Intact chloroplasts isolated from heat-hardened leaves retained increased heat resistance. When the stroma of the chloroplasts was removed, the thermostability of the thylakoids was decreased and was comparable to the heat resistance of chloroplast membranes obtained from non-hardened control plants. Compartmentation studies demonstrated that the content of soluble sugars within the chloroplasts and the whole leaf tissue decreased as heat hardiness increased. This indicated that in spinach leaves, sugars play no protective role in heat hardiness. The results suggest that changes in the ultrastructure of thylakoids in connection with a stabilizing effect of soluble non-sugar stroma

  18. Comparison of Survival of Campylobacter jejuni in the Phyllosphere with That in the Rhizosphere of Spinach and Radish Plants

    PubMed Central

    Brandl, Maria T.; Haxo, Aileen F.; Bates, Anna H.; Mandrell, Robert E.

    2004-01-01

    Campylobacter jejuni has been isolated previously from market produce and has caused gastroenteritis outbreaks linked to produce. We have tested the ability of this human pathogen to utilize organic compounds that are present in leaf and root exudates and to survive in the plant environment under various conditions. Carbon utilization profiles revealed that C. jejuni can utilize many organic acids and amino acids available on leaves and roots. Despite the presence of suitable substrates in the phyllosphere and the rhizosphere, C. jejuni was unable to grow on lettuce and spinach leaves and on spinach and radish roots of plants incubated at 33°C, a temperature that is conducive to its growth in vitro. However, C. jejuni was cultured from radish roots and from the spinach rhizosphere for at least 23 and 28 days, respectively, at 10°C. This enteric pathogen also persisted in the rhizosphere of spinach for prolonged periods of time at 16°C, a temperature at which many cool-season crops are grown. The decline rate constants of C. jejuni populations in the spinach and radish rhizosphere were 10- and 6-fold lower, respectively, than on healthy spinach leaves at 10°C. The enhanced survival of C. jejuni in soil and in the rhizosphere may be a significant factor in its contamination cycle in the environment and may be associated with the sporadic C. jejuni incidence and campylobacteriosis outbreaks linked to produce. PMID:14766604

  19. Functional metagenomics of Escherichia coli O157:H7 interactions with spinach indigenous microorganisms during biofilm formation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The increase in foodborne outbreaks worldwide attributed to fresh fruit and vegetables suggests that produce may serve as an ecological niche for enteric pathogens. Here we examined the interaction of E. coli O157:H7 (EcO157) with spinach leaf microflora during co-colonization and establishment of a...

  20. Differential transcriptional regulation of banana sucrose phosphate synthase gene in response to ethylene, auxin, wounding, low temperature and different photoperiods during fruit ripening and functional analysis of banana SPS gene promoter.

    PubMed

    Roy Choudhury, Swarup; Roy, Sujit; Das, Ranjan; Sengupta, Dibyendu N

    2008-12-01

    Sucrose phosphate synthase (SPS) (EC 2.3.1.14) is the key regulatory component in sucrose formation in banana (Musa acuminata subgroup Cavendish, cv Giant governor) fruit during ripening. This report illustrates differential transcriptional responses of banana SPS gene following ethylene, auxin, wounding, low temperature and different photoperiods during ripening in banana fruit. Whereas ethylene strongly stimulated SPS transcript accumulation, auxin and cold treatment only marginally increased the abundance of SPS mRNA level, while wounding negatively regulated SPS gene expression. Conversely, SPS transcript level was distinctly increased by constant exposure to white light. Protein level, enzymatic activity of SPS and sucrose synthesis were substantially increased by ethylene and increased exposure to white light conditions as compared to other treatments. To further study the transcriptional regulation of SPS in banana fruit, the promoter region of SPS gene was cloned and some cis-acting regulatory elements such as a reverse GCC-box ERE, two ARE motifs (TGTCTC), one LTRE (CCGAA), a GAGA-box (GAGA...) and a GATA-box LRE (GATAAG) were identified along with the TATA and CAAT-box. DNA-protein interaction studies using these cis-elements indicated a highly specific cis-trans interaction in the banana nuclear extract. Furthermore, we specifically studied the light responsive characteristics of GATA-box containing synthetic as well as native banana SPS promoter. Transient expression assays using banana SPS promoter have also indicated the functional importance of the SPS promoter in regulating gene expression. Together, these results provide insights into the transcriptional regulation of banana SPS gene in response to phytohormones and other environmental factors during fruit ripening. PMID:18830708

  1. Analysis of energy utilization in spinach processing

    SciTech Connect

    Chhinnan, M.S.; Singh, R.P.; Pedersen, L.D.; Carroad, P.A.; Rose, W.W.; Jacob, N.L.

    1980-03-01

    The equipment and methods used to monitor the electrical and thermal energy consumed in various unit operations in a spinach processing plant are described and the results of a processing plant energy audit are presented. It is concluded that it requires 6.5 MJ of natural gas and fuel oil and 0.072 MJ of electric power to process one kg of new spinach; the energy intensive operations in spinach processing are associated with exhaust boxes, blanchers, and retorts; uniform product flow through the canning line is essential to energy conservation; and design improvements are needed for the blancher, exhaust box, and retort. (LCL)

  2. Choline oxidation by intact chloroplasts isolated directly from spinach leaves

    SciTech Connect

    Weigel, P.; Hanson, A.D.

    1986-04-01

    Illuminated chloroplasts derived from spinach leaf protoplasts synthesize betaine from choline via the intermediate betaine aldehyde (BAL) (PNAS 82:3678). Photosynthetically active chloroplasts isolated directly from spinach leaves oxidized (/sup 14/C)choline in the light at rates 10 times higher (25-80 nmol/mg chl b) than protoplast-derived chloroplasts. Up to 20% of the (/sup 14/C)choline supplied during a 30 min incubation was oxidized in the light; the main product was (/sup 14/C)BAL. Rates of (/sup 14/C)choline oxidation in darkness were only 5-30% of rates in light. Light-dependent (/sup 14/C)choline oxidation was abolished by DCMU and 5 mM DTT. Pre-illumination of the chloroplasts did not promote (/sup 14/C)choline oxidation in darkness. The uncouplers nigericin and CCCP at concentrations which eliminated CO/sub 2/-dependent O/sub 2/ evolution did not affect (/sup 14/C)choline oxidation in the light. They hypothesize that (/sup 14/C)choline oxidation is not dependent upon light activation of an enzymatic system or upon the electrochemical proton gradient but requires an oxidant generated in the light.

  3. Diversity of the spinach (Spinacia oleracea) spermosphere and phyllosphere bacterial communities.

    PubMed

    Lopez-Velasco, Gabriela; Carder, Phyllis A; Welbaum, Gregory E; Ponder, Monica A

    2013-09-01

    The bacterial diversity of seeds, transmission of bacteria from seed to phyllosphere, and fate of seed-transmitted bacteria on mature plants are poorly characterized. Understanding the dynamics of microbial communities is important for finding bio-control or mitigation strategies for human and plant pathogens. Bacterial populations colonizing spermosphere and phyllosphere of spinach (Spinacia oleracea) seedlings and plants were characterized using pyrosequencing of 16S rRNA gene amplicons. Spinach seed microbiota was composed of three bacterial phyla: Proteobacteria, Firmicutes and Actinobacteria, belonging to > 250 different operational taxonomic units (OTUs). Seed and cotyledon bacterial communities were similar in richness and diversity. Richness of 3-4 leaf-stage of development plants increased markedly to > 850 OTUs classified within 11 phyla. Although some bacterial OTUs were detected on seeds, cotyledons and plants, the breadth of new sequences indicates the importance of multiple sources outside the seed in shaping phyllosphere community. Most classified sequences were from previously undescribed taxa, highlighting the benefits of pyrosequencing in describing seed diversity and phyllosphere bacterial communities. Bacterial community richness increased from 250 different OTUs for spinach seeds and cotyledons, to 800 OTUs for seedlings. To our knowledge this is the first comprehensive characterization of the spinach microbiome, complementing previous culture-based and clone library studies. PMID:23859062

  4. Diversity of the spinach (Spinacia oleracea) spermosphere and phyllosphere bacterial communities.

    PubMed

    Lopez-Velasco, Gabriela; Carder, Phyllis A; Welbaum, Gregory E; Ponder, Monica A

    2013-09-01

    The bacterial diversity of seeds, transmission of bacteria from seed to phyllosphere, and fate of seed-transmitted bacteria on mature plants are poorly characterized. Understanding the dynamics of microbial communities is important for finding bio-control or mitigation strategies for human and plant pathogens. Bacterial populations colonizing spermosphere and phyllosphere of spinach (Spinacia oleracea) seedlings and plants were characterized using pyrosequencing of 16S rRNA gene amplicons. Spinach seed microbiota was composed of three bacterial phyla: Proteobacteria, Firmicutes and Actinobacteria, belonging to > 250 different operational taxonomic units (OTUs). Seed and cotyledon bacterial communities were similar in richness and diversity. Richness of 3-4 leaf-stage of development plants increased markedly to > 850 OTUs classified within 11 phyla. Although some bacterial OTUs were detected on seeds, cotyledons and plants, the breadth of new sequences indicates the importance of multiple sources outside the seed in shaping phyllosphere community. Most classified sequences were from previously undescribed taxa, highlighting the benefits of pyrosequencing in describing seed diversity and phyllosphere bacterial communities. Bacterial community richness increased from 250 different OTUs for spinach seeds and cotyledons, to 800 OTUs for seedlings. To our knowledge this is the first comprehensive characterization of the spinach microbiome, complementing previous culture-based and clone library studies.

  5. Degradation kinetics and pathways of spirotetramat in different parts of spinach plant and in the soil.

    PubMed

    Chen, Xiaojun; Meng, Zhiyuan; Zhang, Yanyan; Gu, Haotian; Ren, Yajun; Lu, Chunliang

    2016-08-01

    Spirotetramat is a new pesticide against a broad spectrum of sucking insects and exhibits a unique property with a two-way systemicity. In order to formulate a scientific rationale for a reasonable spray dose and the safe interval period of 22.4 % spirotetramat suspension concentrate on controlling vegetable pests, we analyzed degradation dynamics and pathways of spirotetramat in different parts of spinach plant (leaf, stalk, and root) and in the soil. We conducted experimental trials under field conditions and adopted a simple and reliable method (dispersive solid phase extraction) combined with liquid chromatography-triple quadrupole tandem mass spectrometry to evaluate the dissipation rates of spirotetramat residue and its metabolites. The results showed that the spirotetramat was degraded into different metabolite residues in different parts of spinach plant (leaf, stalk, and root) and in the soil. Specifically, spirotetramat was degraded into B-keto, B-glu, and B-enol in the leaf; B-glu and B-enol in the stalk; and only B-enol in the root. In the soil where the plants grew, spirotetramat followed a completely different pathway compared to the plant and degraded into B-keto and B-mono. Regardless of different degradation pathways, the dissipation dynamic equations of spirotetramat in different parts of spinach plant and in the soil were all based on the first-order reaction dynamic equations. This work provides guidelines for the safe use of spirotetramat in spinach fields, which would help prevent potential health threats to consumers. PMID:27083908

  6. Spinach downy mildew – Threat, prevention and control

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Downy mildew disease is a widespread and destructive disease of spinach in California and elsewhere where spinach is grown, and is caused by Peronospora effusa, an obligate, plant pathogenic oomycete. As in the case with most other crops damaged by downy mildews, spinach downy mildew culminates in ...

  7. Persistence of poultry associated Salmonella spp. on spinach plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Pre-harvest spinach contamination can occur via irrigation water and can influence the persistence of Salmonella on spinach leaves. Salmonella persistence on spinach plants should be evaluated as nearby poultry farms can be a critical source of contaminated water run-off. Purpose: The...

  8. Comparison of two possible routes of pathogen contamination of spinach leaves in a hydroponic cultivation system.

    PubMed

    Koseki, Shigenobu; Mizuno, Yasuko; Yamamoto, Kazutaka

    2011-09-01

    The route of pathogen contamination (from roots versus from leaves) of spinach leaves was investigated with a hydroponic cultivation system. Three major bacterial pathogens, Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes, were inoculated into the hydroponic solution, in which the spinach was grown to give concentrations of 10⁶ and 10³ CFU/ml. In parallel, the pathogens were inoculated onto the growing leaf surface by pipetting, to give concentrations of 10⁶ and 10³ CFU per leaf. Although contamination was observed at a high rate through the root system by the higher inoculum (10⁶ CFU) for all the pathogens tested, the contamination was rare when the lower inoculum (10³ CFU) was applied. In contrast, contamination through the leaf occurred at a very low rate, even when the inoculum level was high. For all the pathogens tested in the present study, the probability of contamination was promoted through the roots and with higher inoculum levels. The probability of contamination was analyzed with logistic regression. The logistic regression model showed that the odds ratio of contamination from the roots versus from the leaves was 6.93, which suggested that the risk of contamination from the roots was 6.93 times higher than the risk of contamination from the leaves. In addition, the risk of contamination by L. monocytogenes was about 0.3 times that of Salmonella enterica subsp. enterica serovars Typhimurium and Enteritidis and E. coli O157:H7. The results of the present study indicate that the principal route of pathogen contamination of growing spinach leaves in a hydroponic system is from the plant's roots, rather than from leaf contamination itself.

  9. Microtubules in Mesophyll Cells of Nonacclimated and Cold-Acclimated Spinach 1

    PubMed Central

    Bartolo, Michael E.; Carter, John V.

    1991-01-01

    Responses of cortical microtubules in spinach (Spinacia oleracea L. cv Bloomsdale) mesophyll cells to freezing, thawing, supercooling, and dehydration were assessed. Microtubules were visualized using a modified procedure for indirect immunofluorescence microscopy. Leaf sections of nonacclimated and cold-acclimated spinach were slowly frozen to various temperatures, fixed while frozen, and microtubules immunolabelled. Both nonacclimated and cold-acclimated cells exhibited nearly complete microtubule depolymerization after ice formation. After 1 hour thawing at 23°C, microtubules in both nonacclimated and cold-acclimated cells repolymerized. With time, however, microtubules in nonacclimated cells again depolymerized. Since microtubules in cells of leaf tissue frozen slowly are subjected to dehydration as well as subzero temperatures, these stresses were applied separately and their effects on microtubules noted. Supercooling induced microtubule depolymerization in both nonacclimated and cold-acclimated cells, but to a smaller extent than did freezing. Exposing leaf sections to solutions of sorbitol (a cell wall-penetrating osmoticum) or polyethylene glycol 10,000 (a nonpenetrating osmoticum) at room temperature caused microtubule depolymerization. The effects of low temperature and dehydration are roughly additive in producing the observed microtubule responses during freezing. Only small differences in microtubule stability were resolved between nonacclimated and cold-acclimated cells. ImagesFigure 2 PMID:16668366

  10. Proximate composition and mineral content of two edible species of Cnidoscolus (tree spinach).

    PubMed

    Kuti, J O; Kuti, H O

    1999-01-01

    Proximate composition and mineral content of raw and cooked leaves of two edible tree spinach species (Cnidoscolus chayamansa and C. aconitifolius), known locally as 'chaya', were determined and compared with that of a traditional green vegetable, spinach (Spinicia oleraceae). Results of the study indicated that the edible leafy parts of the two chaya species contained significantly (p<0.05) greater amounts of crude protein, crude fiber, Ca, K, Fe, ascorbic acid and beta-carotene than the spinach leaf. However, no significant (p>0.05) differences were found in nutritional composition and mineral content between the chaya species, except minor differences in the relative composition of fatty acids, protein and amino acids. Cooking of chaya leaves slightly reduced nutritional composition of both chaya species. Cooking is essential prior to consumption to inactivate the toxic hydrocyanic glycosides present in chaya leaves. Based on the results of this study, the edible chaya leaves may be good dietary sources of minerals (Ca, K and Fe) and vitamins (ascorbic acid and beta-carotene). PMID:10540979

  11. Rapid detection of Salmonella typhimurium on fresh spinach leaves using phage-immobilized magnetoelastic biosensors

    NASA Astrophysics Data System (ADS)

    Horikawa, Shin; Li, Suiqiong; Chai, Yating; Park, Mi-Kyung; Shen, Wen; Barbaree, James M.; Vodyanoy, Vitaly J.; Chin, Bryan A.

    2011-06-01

    This paper presents an investigation into the use of magnetoelastic biosensors for the rapid detection of Salmonella typhimurium on fresh spinach leaves. The biosensors used in this investigation were comprised of a strip-shaped, goldcoated sensor platform (2 mm-long) diced from a ferromagnetic, amorphous alloy and a filamentous fd-tet phage which specifically binds with S. typhimurium. After surface blocking with bovine serum albumin, these biosensors were, without any preceding sample preparation, directly placed on wet spinach leaves inoculated with various concentrations of S. typhimurium. Upon contact with cells, the phage binds S. typhimurium to the sensor thereby increasing the total mass of the sensor. This change in mass causes a corresponding decrease in the sensor's resonant frequency. After 25 min, the sensors were collected from the leaf surface and measurements of the resonant frequency were performed immediately. The total assay time was less than 30 min. The frequency changes for measurement sensors (i.e., phageimmobilized) were found to be statistically different from those for control sensors (sensors without phage), down to 5 × 106 cells/ml. The detection limit may be improved by using smaller, micron-sized sensors that will have a higher probability of contacting Salmonella on the rough surfaces of spinach leaves.

  12. A Spinach molecular beacon triggered by strand displacement.

    PubMed

    Bhadra, Sanchita; Ellington, Andrew D

    2014-08-01

    We have re-engineered the fluorescent RNA aptamer Spinach to be activated in a sequence-dependent manner. The original Spinach aptamer was extended at its 5'- and 3'-ends to create Spinach.ST, which is predicted to fold into an inactive conformation and thus prevent association with the small molecule fluorophore DFHBI. Hybridization of a specific trigger oligonucleotide to a designed toehold leads to toehold-initiated strand displacement and refolds Spinach into the active, fluorophore-binding conformation. Spinach.ST not only specifically detects its target oligonucleotide but can discriminate readily against single-nucleotide mismatches. RNA amplicons produced during nucleic acid sequence-based amplification (NASBA) of DNA or RNA targets could be specifically detected and reported in real-time by conformational activation of Spinach.ST generated by in vitro transcription. In order to adapt any target sequence to detection by a Spinach reporter we used a primer design technique that brings together otherwise distal toehold sequences via hairpin formation. The same techniques could potentially be used to adapt common Spinach reporters to non-nucleic acid analytes, rather than by making fusions between aptamers and Spinach.

  13. A Spinach molecular beacon triggered by strand displacement

    PubMed Central

    Bhadra, Sanchita; Ellington, Andrew D.

    2014-01-01

    We have re-engineered the fluorescent RNA aptamer Spinach to be activated in a sequence-dependent manner. The original Spinach aptamer was extended at its 5′- and 3′-ends to create Spinach.ST, which is predicted to fold into an inactive conformation and thus prevent association with the small molecule fluorophore DFHBI. Hybridization of a specific trigger oligonucleotide to a designed toehold leads to toehold-initiated strand displacement and refolds Spinach into the active, fluorophore-binding conformation. Spinach.ST not only specifically detects its target oligonucleotide but can discriminate readily against single-nucleotide mismatches. RNA amplicons produced during nucleic acid sequence-based amplification (NASBA) of DNA or RNA targets could be specifically detected and reported in real-time by conformational activation of Spinach.ST generated by in vitro transcription. In order to adapt any target sequence to detection by a Spinach reporter we used a primer design technique that brings together otherwise distal toehold sequences via hairpin formation. The same techniques could potentially be used to adapt common Spinach reporters to non-nucleic acid analytes, rather than by making fusions between aptamers and Spinach. PMID:24942625

  14. Inhibition of Escherichia coli O157:H7 and Salmonella enterica on spinach and identification of antimicrobial substances produced by a commercial Lactic Acid Bacteria food safety intervention.

    PubMed

    Cálix-Lara, Thelma F; Rajendran, Mahitha; Talcott, Stephen T; Smith, Stephen B; Miller, Rhonda K; Castillo, Alejandro; Sturino, Joseph M; Taylor, T Matthew

    2014-04-01

    The microbiological safety of fresh produce is of concern for the U.S. food supply. Members of the Lactic Acid Bacteria (LAB) have been reported to antagonize pathogens by competing for nutrients and by secretion of substances with antimicrobial activity, including organic acids, peroxides, and antimicrobial polypeptides. The objectives of this research were to: (i) determine the capacity of a commercial LAB food antimicrobial to inhibit Escherichia coli O157:H7 and Salmonella enterica on spinach leaf surfaces, and (ii) identify antimicrobial substances produced in vitro by the LAB comprising the food antimicrobial. Pathogens were inoculated on freshly harvested spinach, followed by application of the LAB antimicrobial. Treated spinach was aerobically incubated up to 12 days at 7 °C and surviving pathogens enumerated via selective/differential plating. l-Lactic acid and a bacteriocin-like inhibitory substance (BLIS) were detected and quantified from cell-free fermentates obtained from LAB-inoculated liquid microbiological medium. Application of 8.0 log10 CFU/g LAB produced significant (p < 0.05) reductions in E. coli O157:H7 and Salmonella populations on spinach of 1.6 and 1.9 log10 CFU/g, respectively. It was concluded the LAB antimicrobial inhibited foodborne pathogens on spinach during refrigerated storage, likely the result of the production of metabolites with antimicrobial activity.

  15. Frequency of Verticillium Species in Commercial Spinach Fields and Transmission of V. dahliae from Spinach to Subsequent Lettuce Crops.

    PubMed

    Short, D P G; Gurung, S; Koike, S T; Klosterman, S J; Subbarao, K V

    2015-01-01

    Verticillium wilt caused by V. dahliae is a devastating disease of lettuce in California (CA). The disease is currently restricted to a small geographic area in central coastal CA, even though cropping patterns in other coastal lettuce production regions in the state are similar. Infested spinach seed has been implicated in the introduction of V. dahliae into lettuce fields but direct evidence linking this inoculum to wilt epidemics in lettuce is lacking. In this study, 100 commercial spinach fields in four coastal CA counties were surveyed to evaluate the frequency of Verticillium species recovered from spinach seedlings and the area under spinach production in each county was assessed. Regardless of the county, V. isaacii was the most frequently isolated species from spinach followed by V. dahliae and, less frequently, V. klebahnii. The frequency of recovery of Verticillium species was unrelated to the occurrence of Verticillium wilt on lettuce in the four counties but was related to the area under spinach production in individual counties. The transmission of V. dahliae from infested spinach seeds to lettuce was investigated in microplots. Verticillium wilt developed on lettuce following two or three plantings of Verticillium-infested spinach, in independent experiments. The pathogen recovered from the infected lettuce from microplots was confirmed as V. dahliae by polymerase chain reaction assays. In a greenhouse study, transmission of a green fluorescence protein-tagged mutant strain of V. dahliae from spinach to lettuce roots was demonstrated, after two cycles of incorporation of infected spinach residue into the soil. This study presents conclusive evidence that V. dahliae introduced via spinach seed can cause Verticillium wilt in lettuce.

  16. SpinachDB: A Well-Characterized Genomic Database for Gene Family Classification and SNP Information of Spinach.

    PubMed

    Yang, Xue-Dong; Tan, Hua-Wei; Zhu, Wei-Min

    2016-01-01

    Spinach (Spinacia oleracea L.), which originated in central and western Asia, belongs to the family Amaranthaceae. Spinach is one of most important leafy vegetables with a high nutritional value as well as being a perfect research material for plant sex chromosome models. As the completion of genome assembly and gene prediction of spinach, we developed SpinachDB (http://222.73.98.124/spinachdb) to store, annotate, mine and analyze genomics and genetics datasets efficiently. In this study, all of 21702 spinach genes were annotated. A total of 15741 spinach genes were catalogued into 4351 families, including identification of a substantial number of transcription factors. To construct a high-density genetic map, a total of 131592 SSRs and 1125743 potential SNPs located in 548801 loci of spinach genome were identified in 11 cultivated and wild spinach cultivars. The expression profiles were also performed with RNA-seq data using the FPKM method, which could be used to compare the genes. Paralogs in spinach and the orthologous genes in Arabidopsis, grape, sugar beet and rice were identified for comparative genome analysis. Finally, the SpinachDB website contains seven main sections, including the homepage; the GBrowse map that integrates genome, genes, SSR and SNP marker information; the Blast alignment service; the gene family classification search tool; the orthologous and paralogous gene pairs search tool; and the download and useful contact information. SpinachDB will be continually expanded to include newly generated robust genomics and genetics data sets along with the associated data mining and analysis tools. PMID:27148975

  17. Frequency of Verticillium Species in Commercial Spinach Fields and Transmission of V. dahliae from Spinach to Subsequent Lettuce Crops.

    PubMed

    Short, D P G; Gurung, S; Koike, S T; Klosterman, S J; Subbarao, K V

    2015-01-01

    Verticillium wilt caused by V. dahliae is a devastating disease of lettuce in California (CA). The disease is currently restricted to a small geographic area in central coastal CA, even though cropping patterns in other coastal lettuce production regions in the state are similar. Infested spinach seed has been implicated in the introduction of V. dahliae into lettuce fields but direct evidence linking this inoculum to wilt epidemics in lettuce is lacking. In this study, 100 commercial spinach fields in four coastal CA counties were surveyed to evaluate the frequency of Verticillium species recovered from spinach seedlings and the area under spinach production in each county was assessed. Regardless of the county, V. isaacii was the most frequently isolated species from spinach followed by V. dahliae and, less frequently, V. klebahnii. The frequency of recovery of Verticillium species was unrelated to the occurrence of Verticillium wilt on lettuce in the four counties but was related to the area under spinach production in individual counties. The transmission of V. dahliae from infested spinach seeds to lettuce was investigated in microplots. Verticillium wilt developed on lettuce following two or three plantings of Verticillium-infested spinach, in independent experiments. The pathogen recovered from the infected lettuce from microplots was confirmed as V. dahliae by polymerase chain reaction assays. In a greenhouse study, transmission of a green fluorescence protein-tagged mutant strain of V. dahliae from spinach to lettuce roots was demonstrated, after two cycles of incorporation of infected spinach residue into the soil. This study presents conclusive evidence that V. dahliae introduced via spinach seed can cause Verticillium wilt in lettuce. PMID:25098494

  18. SpinachDB: A Well-Characterized Genomic Database for Gene Family Classification and SNP Information of Spinach

    PubMed Central

    Zhu, Wei-Min

    2016-01-01

    Spinach (Spinacia oleracea L.), which originated in central and western Asia, belongs to the family Amaranthaceae. Spinach is one of most important leafy vegetables with a high nutritional value as well as being a perfect research material for plant sex chromosome models. As the completion of genome assembly and gene prediction of spinach, we developed SpinachDB (http://222.73.98.124/spinachdb) to store, annotate, mine and analyze genomics and genetics datasets efficiently. In this study, all of 21702 spinach genes were annotated. A total of 15741 spinach genes were catalogued into 4351 families, including identification of a substantial number of transcription factors. To construct a high-density genetic map, a total of 131592 SSRs and 1125743 potential SNPs located in 548801 loci of spinach genome were identified in 11 cultivated and wild spinach cultivars. The expression profiles were also performed with RNA-seq data using the FPKM method, which could be used to compare the genes. Paralogs in spinach and the orthologous genes in Arabidopsis, grape, sugar beet and rice were identified for comparative genome analysis. Finally, the SpinachDB website contains seven main sections, including the homepage; the GBrowse map that integrates genome, genes, SSR and SNP marker information; the Blast alignment service; the gene family classification search tool; the orthologous and paralogous gene pairs search tool; and the download and useful contact information. SpinachDB will be continually expanded to include newly generated robust genomics and genetics data sets along with the associated data mining and analysis tools. PMID:27148975

  19. Detection and quantification of Verticillium dahliae in spinach seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Verticillium dahliae is a soilborne fungus that causes Verticillium wilt on multiple crops in central coastal California. Although spinach crops grown in this region for fresh and processing commercial production do not display Verticillium wilt symptoms, spinach seeds produced in the United States ...

  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. PMID:19681281

  1. cDNA sequence and heterologous expression of monomeric spinach pullulanase: multiple isomeric forms arise from the same polypeptide.

    PubMed Central

    Renz, A; Schikora, S; Schmid, R; Kossmann, J; Beck, E

    1998-01-01

    The spinach pullulanase gene was cloned and sequenced using peptide sequences of the purified enzyme as a starting point and employing PCR techniques and cDNA library screening. Its open reading frame codes for a protein of 964 amino acids which represents a precursor of the pullulanase. The N-terminal transit peptide consists of 65 amino acids, and the mature protein, comprising 899 amino acids, has a calculated molecular mass of 99kDa. Pullulanase is a member of the alpha-amylase family. In addition to a characteristic catalytic (beta/alpha)8-barrel domain, it contains a domain, F, that is specific for branching and debranching enzymes. Pullulanase cDNA was expressed in Escherichia coli, and the purified protein was compared with the enzyme from spinach leaves. Identity of the two proteins was confirmed in terms of catalytic properties, N-terminal amino acid sequences and molecular masses. The pullulanase produced by E. coli showed the same microheterogeneity as the spinach leaf enzyme: it could be resolved into two substrate-induced forms by electrophoresis in amylopectin-containing polyacrylamide gels, and, in the absence of substrate, into several free forms (charge isomers) by isoelectric focusing or chromatofocusing. Rechromatofocusing of single free forms resulted in the originally observed pattern of molecular forms. However, heterogeneity of the protein disappeared on isoelectric focusing under completely denaturing conditions when only one protein band was observed. Post-translational modifications such as glycosylation and phosphorylation could be excluded as potential explanations for the protein heterogeneity. Therefore the microheterogeneity of spinach leaf pullulanase results from neither genetic variation nor post-translational modifications, but is a property of the single unmodified gene product. The different interconvertible forms of the pullulanase represent protein populations of different tertiary structure of the same polypeptide. PMID

  2. Choline Oxidation by Intact Spinach Chloroplasts 1

    PubMed Central

    Weigel, Pierre; Lerma, Claudia; Hanson, Andrew D.

    1988-01-01

    Plants synthesize betaine by a two-step oxidation of choline (choline → betaine aldehyde → betaine). Protoplast-derived chloroplasts of spinach (Spinacia oleracea L.) carry out both reactions, more rapidly in light than in darkness (AD Hanson et al. 1985 Proc Natl Acad Sci USA 82: 3678-3682). We investigated the light-stimulated oxidation of choline, using spinach chloroplasts isolated directly from leaves. The rates of choline oxidation obtained (dark and light rates: 10-50 and 100-300 nanomoles per hour per milligram chlorophyll, respectively) were approximately 20-fold higher than for protoplast-derived chloroplasts. Betaine aldehyde was the main product. Choline oxidation in darkness and light was suppressed by hypoxia. Neither uncouplers nor the Calvin cycle inhibitor glyceraldehyde greatly affected choline oxidation in the light, and maximal choline oxidation was attained far below light saturation of CO2 fixation. The light stimulation of choline oxidation was abolished by the PSII inhibitors DCMU and dibromothymoquinone, and was partially restored by adding reduced diaminodurene, an electron donor to PSI. Both methyl viologen and phenazine methosulfate prevented choline oxidation. Adding dihydroxyacetone phosphate, which can generate NADPH in organello, doubled the dark rate of choline oxidation. These results indicate that choline oxidation in chloroplasts requires oxygen, and reducing power generated from PSI. Enzymic reactions consistent with these requirements are discussed. Images Fig. 1 PMID:16665893

  3. Impact of pigeon pea biochar on cadmium mobility in soil and transfer rate to leafy vegetable spinach.

    PubMed

    Coumar, M Vassanda; Parihar, R S; Dwivedi, A K; Saha, J K; Rajendiran, S; Dotaniya, M L; Kundu, S

    2016-01-01

    Introduction of heavy metals in the environment by various anthropogenic activities has become a potential treat to life. Among the heavy metals, cadmium (Cd) shows relatively high soil mobility and has high phyto-mammalian toxicity. Integration of soil remediation and ecosystem services, such as carbon sequestration in soils through organic amendments, may provide an attractive land management option for contaminated sites. The application of biochar in agriculture has recently received much attention globally due to its associated multiple benefits, particularly, long-term carbon storage in soil. However, the application of biochar from softwood crop residue for heavy metal immobilization, as an alternative to direct field application, has not received much attention. Hence, a pot experiment was conducted to study the effect of pigeon pea biochar on cadmium mobility in a soil-plant system in cadmium-spiked sandy loam soil. The biochar was prepared from pigeon pea stalk through a slow pyrolysis method at 300 °C. The experiment was designed with three levels of Cd (0, 5, and 10 mg Cd kg(-1) soil) and three levels of biochar (0, 2.5, and 5 g kg(-1) soil) using spinach as a test crop. The results indicate that with increasing levels of applied cadmium at 5 and 10 mg kg(-1) soil, the dry matter yield (DMY) of spinach leaf decreased by 9.84 and 18.29 %, respectively. However, application of biochar (at 2.5 and 5 g kg(-1) soil) significantly increased the dry matter yield of spinach leaf by 5.07 and 15.02 %, respectively, and root by 14.0 and 24.0 %, respectively, over the control. Organic carbon content in the post-harvest soil increased to 34.9 and 60.5 % due to the application of biochar 2.5 and 5 g kg(-1) soil, respectively. Further, there was a reduction in the diethylene triamine pentaacetic acid (DTPA)-extractable cadmium in the soil and in transfer coefficient values (soil to plant), as well as its concentrations in spinach leaf and root, indicating that

  4. Impact of pigeon pea biochar on cadmium mobility in soil and transfer rate to leafy vegetable spinach.

    PubMed

    Coumar, M Vassanda; Parihar, R S; Dwivedi, A K; Saha, J K; Rajendiran, S; Dotaniya, M L; Kundu, S

    2016-01-01

    Introduction of heavy metals in the environment by various anthropogenic activities has become a potential treat to life. Among the heavy metals, cadmium (Cd) shows relatively high soil mobility and has high phyto-mammalian toxicity. Integration of soil remediation and ecosystem services, such as carbon sequestration in soils through organic amendments, may provide an attractive land management option for contaminated sites. The application of biochar in agriculture has recently received much attention globally due to its associated multiple benefits, particularly, long-term carbon storage in soil. However, the application of biochar from softwood crop residue for heavy metal immobilization, as an alternative to direct field application, has not received much attention. Hence, a pot experiment was conducted to study the effect of pigeon pea biochar on cadmium mobility in a soil-plant system in cadmium-spiked sandy loam soil. The biochar was prepared from pigeon pea stalk through a slow pyrolysis method at 300 °C. The experiment was designed with three levels of Cd (0, 5, and 10 mg Cd kg(-1) soil) and three levels of biochar (0, 2.5, and 5 g kg(-1) soil) using spinach as a test crop. The results indicate that with increasing levels of applied cadmium at 5 and 10 mg kg(-1) soil, the dry matter yield (DMY) of spinach leaf decreased by 9.84 and 18.29 %, respectively. However, application of biochar (at 2.5 and 5 g kg(-1) soil) significantly increased the dry matter yield of spinach leaf by 5.07 and 15.02 %, respectively, and root by 14.0 and 24.0 %, respectively, over the control. Organic carbon content in the post-harvest soil increased to 34.9 and 60.5 % due to the application of biochar 2.5 and 5 g kg(-1) soil, respectively. Further, there was a reduction in the diethylene triamine pentaacetic acid (DTPA)-extractable cadmium in the soil and in transfer coefficient values (soil to plant), as well as its concentrations in spinach leaf and root, indicating that

  5. Response of spinach and komatsuna to biogas effluent made from source-separated kitchen garbage.

    PubMed

    Furukawa, Yuichiro; Hasegawa, Hiroshi

    2006-01-01

    Recycling of kitchen garbage is an urgent task for reducing public spending and environmental burdens by incineration and/or landfill. There is an interesting regional effort in Ogawa, Saitama prefecture, Japan, in which source-separated kitchen garbage is anaerobically fermented with a biogas plant and the resultant effluent is used as a quick-release organic fertilizer by surrounding farmers. However, scientific assessments of fertilizer values and risks in the use of the effluent were lacking. Thus, a field experiment was conducted from 2003 to 2004 in Tohoku National Agricultural Research Center to grow spinach (Spinacia oleracea L.) and komatsuna (Brassica rapa var. perviridis L. H. Bailey) for evaluating the fertilizer value of the kitchen garbage effluent (KGE), nitrate, coliform group (CG), Escherichia coli, fecal streptococci (FS), and Vibrio parahaemolyticus concentrations of KGE and in the soil and the plant leaves. A cattle manure effluent (CME) and chemical fertilizers (NPK) were used as controls. Total nitrogen (N) and ammonium N concentrations of the KGE were 1.47 and 1.46 g kg(-1), respectively. The bacteria tested were detected in both biogas effluents in the order of 2 to 3 log CFU g(-1), but there was little evidence that the biogas effluents increased these bacteria in the soil and the plant leaves. At the rate of 22 g N m(-2), yield, total N uptake, apparent N recovery rate, and leaf nitrate ion concentration at harvest of spinach and komatsuna in the KGE plot were mostly comparable to those in the NPK and CME plots. We conclude that the KGE is a quick-release N fertilizer comparable to chemical fertilizers and does not cause contamination of CG, E. coli, FS, or V. parahaemolyticus in the soil and spinach and komatsuna leaves.

  6. Response of spinach and komatsuna to biogas effluent made from source-separated kitchen garbage.

    PubMed

    Furukawa, Yuichiro; Hasegawa, Hiroshi

    2006-01-01

    Recycling of kitchen garbage is an urgent task for reducing public spending and environmental burdens by incineration and/or landfill. There is an interesting regional effort in Ogawa, Saitama prefecture, Japan, in which source-separated kitchen garbage is anaerobically fermented with a biogas plant and the resultant effluent is used as a quick-release organic fertilizer by surrounding farmers. However, scientific assessments of fertilizer values and risks in the use of the effluent were lacking. Thus, a field experiment was conducted from 2003 to 2004 in Tohoku National Agricultural Research Center to grow spinach (Spinacia oleracea L.) and komatsuna (Brassica rapa var. perviridis L. H. Bailey) for evaluating the fertilizer value of the kitchen garbage effluent (KGE), nitrate, coliform group (CG), Escherichia coli, fecal streptococci (FS), and Vibrio parahaemolyticus concentrations of KGE and in the soil and the plant leaves. A cattle manure effluent (CME) and chemical fertilizers (NPK) were used as controls. Total nitrogen (N) and ammonium N concentrations of the KGE were 1.47 and 1.46 g kg(-1), respectively. The bacteria tested were detected in both biogas effluents in the order of 2 to 3 log CFU g(-1), but there was little evidence that the biogas effluents increased these bacteria in the soil and the plant leaves. At the rate of 22 g N m(-2), yield, total N uptake, apparent N recovery rate, and leaf nitrate ion concentration at harvest of spinach and komatsuna in the KGE plot were mostly comparable to those in the NPK and CME plots. We conclude that the KGE is a quick-release N fertilizer comparable to chemical fertilizers and does not cause contamination of CG, E. coli, FS, or V. parahaemolyticus in the soil and spinach and komatsuna leaves. PMID:16973635

  7. Glycinebetaine biosynthesis and its control in detached secondary leaves of spinach.

    PubMed

    Coughlan, S J; Wyn Jones, R G

    1982-03-01

    In secondary leaves from spinach plants pretreated in vermiculite for 24 h with 300 mM NaCl, glycinebetaine accumulated at a rate of circa 0.16 μmol 100 μg(-1) Chl d(-1) (2 μmol g(-1) FW d(-1)), about three times the rate of control plants. The soluble carbohydrate and free amino acid contents did not increase significantly following salinisation until after 4 d when the relative growth rate also decreased. Leaf proline levels remained very low throughout the experimental period. K(+) on a tissue water basis remained constant at 200 mM while Cl(-) and Na(+) levels increased linearly to reach 175 and 100 mM respectively after 5 d of saline treatment. The osmotic pressure of leaf tissue also increased from 300 to 500 mosmol kg(-1). These experimental conditions were considered suitable to study glycinebetaine biosynthesis and its induction by salinity in the absence of marked growth inhibition or metabolic disturbance. Radioactive labelled [(14)C]serine, ethanolamine and choline (all 1 μmol, 13.3 MBq in 10 μl) were fed to detached secondary leaves via the petiole 24 h after the exposure of plants to salt. The rate of isotope incorporation into water soluble products, lipids and residue was measured over a further 24 h. The major metabolic fate of exogenous [(14)C]choline and [(14)C]ethanolamine was incorporation into glycinebetaine while less (14)C-label was found in phosphatidyl choline and phosphatidyl ethanolamine. Incorporation rates were identical in control and salinised leaves and were adequate to account for observed values of glycinebetaine accumulation previously reported in spinach. In contrast the labelling of glycinebetaine from [(14)C]serine was twice as great in salinated plants as in the controls. These results, together with short term labelling experiment with [(14)C]ethanolamine using leaf slices, were consistent with the formation of glycinebetaine via serine, ethanolamine and its methylated derivatives to choline with some control being

  8. Growth Conditions To Reduce Oxalic Acid Content of Spinach

    NASA Technical Reports Server (NTRS)

    Johnson-Rutzke, Corinne

    2003-01-01

    A controlled-environment agricultural (CEA) technique to increase the nutritive value of spinach has been developed. This technique makes it possible to reduce the concentration of oxalic acid in spinach leaves. It is desirable to reduce the oxalic acid content because oxalic acid acts as an anti-nutritive calcium-binding component. More than 30 years ago, an enzyme (an oxidase) that breaks down oxalic acid into CO2 and H2O2 was discovered and found to be naturally present in spinach leaves. However, nitrate, which can also be present because of the use of common nitratebased fertilizers, inactivates the enzyme. In the CEA technique, one cuts off the supply of nitrate and keeps the spinach plants cool while providing sufficient oxygen. This technique provides the precise environment that enables the enzyme to naturally break down oxalate. The result of application of this technique is that the oxalate content is reduced by 2/3 in one week.

  9. Functional metagenomics of Escherichia coli O157:H7 interactions with spinach indigenous microorganisms during biofilm formation.

    PubMed

    Carter, Michelle Q; Xue, Kai; Brandl, Maria T; Liu, Feifei; Wu, Liyou; Louie, Jacqueline W; Mandrell, Robert E; Zhou, Jizhong

    2012-01-01

    The increase in foodborne outbreaks worldwide attributed to fresh fruit and vegetables suggests that produce may serve as an ecological niche for enteric pathogens. Here we examined the interaction of E. coli O157:H7 (EcO157) with spinach leaf indigenous microorganisms during co-colonization and establishment of a mixed biofilm on a stainless steel surface. Stainless steel surface was selected to mimic the surface of produce-processing equipment, where retention of foodborne pathogens such as EcO157 could serve as a potential source for transmission. We observed a positive effect of spinach-associated microbes on the initial attachment of EcO157, but an antagonistic effect on the EcO157 population at the later stage of biofilm formation. Metagenomic analyses of the biofilm community with the GeoChip revealed an extremely diverse community (gene richness, 23409; Shannon-Weiner index H, 9.55). Presence of EcO157 in the mixed biofilm resulted in a significant decrease in the community α-diversity (t test, P<0.05), indicating a putative competition between the pathogen and indigenous spinach microbes. The decrease in the β-diversity of the EcO157-inoculated biofilm at 48 h (ANOVA, P<0.05) suggested a convergent shift in functional composition in response to EcO157 invasion. The success of EcO157 in the mixed biofilm is likely associated with its metabolic potential in utilizing spinach nutrients: the generation time of EcO157 in spinach lysates at 28°C is ~ 38 min, which is comparable to that in rich broth. The significant decrease in the abundance of many genes involved in carbon, nitrogen, and phosphorus cycling in the EcO157-inoculated biofilms (t test, P<0.05) further support our conclusion that competition for essential macronutrients is likely the primary interaction between the EcO157 and indigenous spinach-biofilm species.

  10. Evidence that spinach leaves express calreticulin but not calsequestrin.

    PubMed Central

    Navazio, L; Baldan, B; Dainese, P; James, P; Damiani, E; Margreth, A; Mariani, P

    1995-01-01

    The presence of either calreticulin (CR) or calsequestrin (CS-like proteins in spinach (Spinacia oleracea L.) leaves has been previously described. Here we report the purification from spinach leaves of two highly acidic (isoelectric point 5.2) Ca(2+)-binding proteins of 56 and 54 kD by means of DEAE-cellulose chromatography followed by phenyl-Sepharose chromatography in the presence of Zn(2+) (i.e., under experimental conditions that allowed the purification of CR from human liver). On the other hand, we failed to identify any protein sharing with animal CS the ability to bind to phenyl-Sepharose in the absence of Ca(2+). Based on the N-terminal amino acid sequence, the 56- and 54-kD spinach Ca(2+)-binding proteins were identified as two distinct isoforms of CR. Therefore, we conclude that CR, and not CS, is expressed in spinach leaves. The 56-kD spinach CR isoform was found to be glycosylated, as judged by ligand blot techniques with concanavalin A and affinity chromatography with concanavalin A-Sepharose. Furthermore, the 56-kD CR was found to differ from rabbit liver CR in amino acid sequence, peptide mapping after partial digestion with Staphylococcus aureus V8 protease, pH-dependent shift of electrophoretic mobility, and immunological cross-reactivity with an antiserum raised to spinach CR, indicating a low degree of structural homology with animal CRs. PMID:8552722

  11. Functional properties of spinach (Spinacia oleracea L.) phytochemicals and bioactives.

    PubMed

    Roberts, Joseph L; Moreau, Régis

    2016-08-10

    Overwhelming evidence indicates that diets rich in fruits and vegetables are protective against common chronic diseases, such as cancer, obesity and cardiovascular disease. Leafy green vegetables, in particular, are recognized as having substantial health-promoting activities that are attributed to the functional properties of their nutrients and non-essential chemical compounds. Spinach (Spinacia oleracea L.) is widely regarded as a functional food due to its diverse nutritional composition, which includes vitamins and minerals, and to its phytochemicals and bioactives that promote health beyond basic nutrition. Spinach-derived phytochemicals and bioactives are able to (i) scavenge reactive oxygen species and prevent macromolecular oxidative damage, (ii) modulate expression and activity of genes involved in metabolism, proliferation, inflammation, and antioxidant defence, and (iii) curb food intake by inducing secretion of satiety hormones. These biological activities contribute to the anti-cancer, anti-obesity, hypoglycemic, and hypolipidemic properties of spinach. Despite these valuable attributes, spinach consumption remains low in comparison to other leafy green vegetables. This review examines the functional properties of spinach in cell culture, animals and humans with a focus on the molecular mechanisms by which spinach-derived non-essential phytochemicals and bioactives, such as glycolipids and thylakoids, impart their health benefits.

  12. Functional properties of spinach (Spinacia oleracea L.) phytochemicals and bioactives.

    PubMed

    Roberts, Joseph L; Moreau, Régis

    2016-08-10

    Overwhelming evidence indicates that diets rich in fruits and vegetables are protective against common chronic diseases, such as cancer, obesity and cardiovascular disease. Leafy green vegetables, in particular, are recognized as having substantial health-promoting activities that are attributed to the functional properties of their nutrients and non-essential chemical compounds. Spinach (Spinacia oleracea L.) is widely regarded as a functional food due to its diverse nutritional composition, which includes vitamins and minerals, and to its phytochemicals and bioactives that promote health beyond basic nutrition. Spinach-derived phytochemicals and bioactives are able to (i) scavenge reactive oxygen species and prevent macromolecular oxidative damage, (ii) modulate expression and activity of genes involved in metabolism, proliferation, inflammation, and antioxidant defence, and (iii) curb food intake by inducing secretion of satiety hormones. These biological activities contribute to the anti-cancer, anti-obesity, hypoglycemic, and hypolipidemic properties of spinach. Despite these valuable attributes, spinach consumption remains low in comparison to other leafy green vegetables. This review examines the functional properties of spinach in cell culture, animals and humans with a focus on the molecular mechanisms by which spinach-derived non-essential phytochemicals and bioactives, such as glycolipids and thylakoids, impart their health benefits. PMID:27353735

  13. Thermal inactivation kinetics of hepatitis A virus in spinach.

    PubMed

    Bozkurt, Hayriye; Ye, Xiaofei; Harte, Federico; D'Souza, Doris H; Davidson, P Michael

    2015-01-16

    Leafy vegetables have been recognized as important vehicles for the transmission of foodborne viral pathogens. To control hepatitis A viral foodborne illness outbreaks associated with mildly heated (e.g., blanched) leafy vegetables such as spinach, generation of adequate thermal processes is important both for consumers and the food industry. Therefore, the objectives of this study were to determine the thermal inactivation behavior of hepatitis A virus (HAV) in spinach, and provide insights on HAV inactivation in spinach for future studies and industrial applications. The D-values calculated from the first-order model (50-72 °C) ranged from 34.40 ± 4.08 to 0.91 ± 0.12 min with a z-value of 13.92 ± 0.87 °C. The calculated activation energy value was 162 ± 11 kJ/mol. Using the information generated in the present study and the thermal parameters of industrial blanching conditions for spinach as a basis (100 °C for 120-180 s), the blanching of spinach in water at 100 °C for 120-180 s under atmospheric conditions will provide greater than 6 log reduction of HAV. The results of this study may be useful to the frozen food industry in designing blanching conditions for spinach to inactivate or control hepatitis A virus outbreaks.

  14. Role of curli and plant cultivation conditions on Escherichia coli O157:H7 internalization into spinach grown on hydroponics and in soil.

    PubMed

    Macarisin, Dumitru; Patel, Jitendra; Sharma, Vijay K

    2014-03-01

    Contamination of fresh produce could represent a public health concern because no terminal kill step is applied during harvest or at the processing facility to kill pathogens. In addition, once contaminated, pathogens may internalize into produce and be protected from disinfectants during the postharvest processing step. The objective of the current study was to determine the potential internalization of Escherichia coli O157:H7 into spinach roots and subsequent transfer to the edible parts. Because curli are involved in biofilm formation, we investigated whether their presence influence the internalization of E. coli O157:H7 into spinach. Further, the effect of the spinach cultivar on E. coli O157:H7 internalization was evaluated. Spinach plants were grown in contaminated soil as well as hydroponically to prevent mechanical wounding of the roots and inadvertent transfer of pathogens from the contamination source to the non-exposed plant surfaces. Results showed that E. coli O157:H7 could internalize into hydroponically grown intact spinach plants through the root system and move to the stem and leaf level. The incidence of internalization was significantly higher in hydroponically grown plants when roots were exposed to 7 log CFU/mL compared to those exposed to 5 log CFU/mL. The effect of cultivar on E. coli O157:H7 internalization was not significant (P>0.05) for the analyzed spinach varieties, internalization incidences showing almost equal distribution between Space and Waitiki, 49.06% and 50.94% respectively. Wounding of the root system in hydroponically grown spinach increased the incidence of E. coli O157:H7 internalization and translocation to the edible portions of the plant. Experimental contamination of the plants grown in soil resulted in a greater number of internalization events then in those grown hydroponically, suggesting that E. coli O157:H7 internalization is dependent on root damage, which is more likely to occur when plants are grown in soil

  15. Light intensity is the main factor affecting fresh market spinach tolerance for Phenmedipham

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The few available herbicides for fresh market spinach do not provide adequate weed control, and there is need for additional herbicide tools. Phenmedipham is registered for use in processing spinach but not in fresh spinach due to its crop injury potential and short time window from application to h...

  16. Colonization of spinach (Spinacia oleracea L.) by GFP-tagged verticillium dahliae.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The soilborne fungus, Verticillium dahliae, causes wilt in a wide range of hosts, including spinach (Spinacia oleracea L.). The interaction between a green fluorescent protein (GFP)-tagged V. dahliae strain and spinach was studied by confocal laser scanning microscopy. The roots of spinach seedlings...

  17. Biosynthesis of Sucrose and Mannitol as a Function of Leaf Age in Celery (Apium graveolens L.).

    PubMed

    Davis, J M; Fellman, J K; Loescher, W H

    1988-01-01

    In celery (Apium graveolens L.), the two major translocated carbohydrates are sucrose and the acyclic polyol mannitol. Their metabolism, however, is different and their specific functions are uncertain. To compare their roles in carbon partitioning and sink-source transitions, developmental changes in (14)CO(2) labeling, pool sizes, and key enzyme activities in leaf tissues were examined. The proportion of label in mannitol increased dramatically with leaf maturation whereas that in sucrose remained fairly constant. Mannitol content, however, was high in all leaves and sucrose content increased as leaves developed. Activities of mannose-6-P reductase, cytoplasmic and chloroplastic fructose-1,6-bisphosphatases, sucrose phosphate synthase, and sucrose synthase increased with leaf maturation and decreased as leaves senesced. Ribulose bisphosphate carboxylase and nonreversible glyceraldehyde-3-P dehydrogenase activities rose as leaves developed but did not decrease. Thus, sucrose is produced in all photosynthetically active leaves whereas mannitol is synthesized primarily in mature leaves and stored in all leaves. Onset of sucrose export in celery may result from sucrose accumulation in expanding leaves, but mannitol export is clearly unrelated to mannitol concentration. Mannitol export, however, appears to coincide with increased mannitol biosynthesis. Although mannitol and sucrose arise from a common precursor in celery, subsequent metabolism and transport must be regulated separately.

  18. Biosynthesis of sucrose and mannitol as a function of leaf age in celery (Apium graveolens L. )

    SciTech Connect

    Davis, J.M.; Fellman, J.K.; Loescher, W.H.

    1988-01-01

    In celery (Apium graveolens L.), the two major translocated carbohydrates are sucrose and the acyclic polyol mannitol. Their metabolism, however, is different and their specific functions are uncertain. To compare their roles in carbon partitioning and sink-source transitions, developmental changes in /sup 14/CO/sub 2/ labeling, pool sizes, and key enzyme activities in leaf tissues were examined. The proportion of label in mannitol increased dramatically with leaf maturation whereas that in sucrose remained fairly constant. Mannitol content, however, was high in all leaves and sucrose content increased as leaves developed. Activities of mannose-6-P reductase, cytoplasmic and chloroplastic fructose-1,6-bis-phosphatases, sucrose phosphate synthase, and sucrose synthase increased with leaf maturation and decreased as leaves senesced. Ribulose bisphosphate carboxylase and nonreversible glyceraldehyde-3-P dehydrogenase activities rose as leaves developed but did not decrease. Thus, sucrose is produced in all photosynthetically active leaves whereas mannitol is synthesized primarily in mature leaves and stored in all leaves. Onset of sucrose export in celery may result from sucrose accumulation in expanding leaves, but mannitol export is clearly unrelated to mannitol concentration. Mannitol export, however, appears to coincide with increased mannitol biosynthesis. Although mannitol and sucrose arise from a common precursor in celery, subsequent metabolism and transport must be regulated separately.

  19. Effect of Greens and Soil Type, Sulfur Addition and Lithium Level on Leaf Constituents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A greenhouse experiment was conducted near Weslaco, Texas (Lat. 26o 8' N, Long. 97o 57' W) between Dec. 2006 and Feb 2007 to evaluate the effect of soil type, added sulfur and lithium level on the growth and leaf nutrients, particularly biofortified levels of Li and S, in spinach and mustard gree...

  20. Kinetic and structural characterization of spinach carbonic anhydrase.

    PubMed

    Rowlett, R S; Chance, M R; Wirt, M D; Sidelinger, D E; Royal, J R; Woodroffe, M; Wang, Y F; Saha, R P; Lam, M G

    1994-11-29

    We have carried out kinetics studies of spinach carbonic anhydrase (CA) using stopped-flow spectrophotometry at steady state and 13C-NMR exchange at chemical equilibrium. We found that the rate of CO2<-->HCO3- exchange catalyzed by spinach CA at pH 7.0 to be 3-5 times faster than the maximal kcat for either CO2 hydration or HCO3- dehydration at steady state, suggesting a rate-determining H+ transfer step in the catalytic mechanism. Correspondingly, we measured a pH-independent solvent deuterium isotope effect on kcat of approximately 2.0, and found that the rate of catalysis was significantly decreased at external buffer concentrations below 5 mM. Our results are consistent with a zinc-hydroxide mechanism of action with for spinach CA, similar to that of animal carbonic anhydrases. We have also collected X-ray absorption spectra of spinach CA. Analysis of the extended fine structure (EXAFS) suggests that the coordination sphere of Zn in spinach CA must have one or more sulfur ligands, in contrast to animal CAs which have only nitrogen and oxygen ligands. The models which best fit the data have average Zn-N(O) distances of 1.99-2.06 A, average Zn-S distances of 2.31--2.32 A, and a total coordination number of 4-6. We conclude that animal and spinach CAs are convergently evolved enzymes which are structurally quite different, but functionally equivalent. PMID:7947805

  1. Design, Synthesis, and Application of Spinach Molecular Beacons Triggered by Strand Displacement

    PubMed Central

    Bhadra, Sanchita; Ellington, Andrew D.

    2015-01-01

    We describe design parameters for the synthesis and analytical application of a label-free RNA molecular beacon, termed Spinach.ST. The RNA aptamer Spinach fluoresces upon binding the small-molecule fluorophore DFHBI ((Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-1,2-dimethyl-1H-imidazol-5(4H)-one). Spinach has been reengineered by extending its 5′- and 3′-ends to create Spinach.ST, which is predicted to fold into an inactive conformation that fails to bind DHFBI. Hybridization of a trigger oligonucleotide to a designed toehold on Spinach.ST initiates toehold-mediated strand displacement and restores the DFHBI-binding, fluorescence-enhancing conformation of Spinach. The versatile Spinach.ST sensor can detect DNA or RNA trigger sequences and can readily distinguish single-nucleotide mismatches in the trigger toehold. Primer design techniques are described that augment amplicons produced by enzymatic amplification with Spinach.ST triggers. Interaction between these triggers and Spinach.ST molecular beacons leads to the real-time, sequence-specific quantitation of these amplicons. The use of Spinach.ST with isothermal amplification reactions such as nucleic acid sequence-based amplification (NASBA) may enable point-of-care applications. The same design principles could also be used to adapt Spinach reporters to the assay of nonnucleic acid analytes in trans. PMID:25605388

  2. Design, synthesis, and application of Spinach molecular beacons triggered by strand displacement.

    PubMed

    Bhadra, Sanchita; Ellington, Andrew D

    2015-01-01

    We describe design parameters for the synthesis and analytical application of a label-free RNA molecular beacon, termed Spinach.ST. The RNA aptamer Spinach fluoresces upon binding the small-molecule fluorophore DFHBI ((Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-1,2-dimethyl-1H-imidazol-5(4H)-one). Spinach has been reengineered by extending its 5'- and 3'-ends to create Spinach.ST, which is predicted to fold into an inactive conformation that fails to bind DHFBI. Hybridization of a trigger oligonucleotide to a designed toehold on Spinach.ST initiates toehold-mediated strand displacement and restores the DFHBI-binding, fluorescence-enhancing conformation of Spinach. The versatile Spinach.ST sensor can detect DNA or RNA trigger sequences and can readily distinguish single-nucleotide mismatches in the trigger toehold. Primer design techniques are described that augment amplicons produced by enzymatic amplification with Spinach.ST triggers. Interaction between these triggers and Spinach.ST molecular beacons leads to the real-time, sequence-specific quantitation of these amplicons. The use of Spinach.ST with isothermal amplification reactions such as nucleic acid sequence-based amplification (NASBA) may enable point-of-care applications. The same design principles could also be used to adapt Spinach reporters to the assay of nonnucleic acid analytes in trans.

  3. Role of ascorbate in detoxifying ozone in the apoplast of spinach (Spinacia oleracea L. ) leaves

    SciTech Connect

    Luwe, M.W.F.; Takahama, Umeo; Heber, U. )

    1993-03-01

    Both reduced and oxidized ascorbate (AA and DHA) are present in the aqueous phase of the extracellular space, the apoplast, of spinach (Spinacia oleracea L.) leaves. Fumigation with 0.3 [mu]L L[sup [minus]1] of ozone resulted in ozone uptake by the leaves close to 0.9 pmol cm[sup [minus]2] of leaf surface area s[sup [minus]1]. Apoplastic AA was slowly oxidized by ozone. The initial decrease of apoplastic AA was <0.1 pmol cm[sup [minus]2] s[sup [minus]1]. The apoplastic ratio of AA to (AA + DHA) decreased within 6 h of fumigation from 0.9 to 0.1. Initially, the concentration of (AA + DHA) did not change in the apoplast, but when fumigation was continued, DHA increased and AA remained at a very low constant level. After fumigation was discontinued, DHA decreased very slowly in the apoplast, reaching control level after 70 h. Insufficient AA reached the apoplast from the cytosol to detoxify ozone in the apoplast when the ozone flux into the leaves was 0.9 pmol cm[sup [minus]2] s[sup [minus]1]. The transport of DHA back into the cytosol was slower than AA transport into the apoplast. No dehydroascorbate reductase activity could be detected in the apoplast of spinach leaves. In contrast to its extracellular redox state, the intracellular redox state of AA did not change appreciably during a 24-h fumigation period. However, intracellular glutathione became slowly oxidized. At the beginning of fumigation, 90% of the total glutathione was reduced. Only 10% was reduced after 24-h exposure of the leaves to 0.3 [mu]L L[sup [minus]1] of ozone. Necrotic leaf damage started to become visible when fumigation was extended beyond a 24-h period. A close correlation between the extent of damage, on the one hand, and the AA content and the ascorbate redox state of whole leaves, on the other, was observed after 48 h of fumigation. Only the youngest leaves that contained high ascorbate concentrations did not exhibit necrotic leaf damage after 48 h. 30 refs., 6 figs., 1 tab.

  4. Salt tolerance of spinach as related to seasonal climate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There is decreasing availability of fresh water for irrigated agriculture in semiarid regions throughout the world. Unfortunately most high value irrigated crops are relatively sensitive to salinity, mostly limiting use of saline waters for irrigation to use with low value crops. Spinach is an excep...

  5. Survival of pathogenic Escherichia coli on basil, lettuce, and spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The contamination of lettuce, spinach and basil with pathogenic E. coli has caused numerous illnesses over the past decade. E. coli O157:H7, E. coli O104:H4 and avian pathogenic E. coli (APECstx- and APECstx+) were inoculated on basil plants and in promix soiless substrate using drip and overhead ir...

  6. Quality indices of Jew's mallow and spinach during frozen storage.

    PubMed

    Labib, A A; Abd el-Latife, S A; Omran, H

    1997-01-01

    In this study, Jew's mallow and spinach were heat treated and kept frozen, at -18 degrees C, for 3 months. Analysis of some chemical components and enzymatic activities as well as a sensory evaluation were undertaken. At the end of the storage period, blanched spinach showed no PPO (polyphenyloxidase) activity at all, but still contained PO (peroxydase) and LO (lipoxygenase) activities. The untreated spinach stored under the same conditions showed high activities of PO, PPO and LO. The heat treated mallow still contained high enzyme activities which decreased slowly during frozen storage. Statistical analysis of the chemical composition and the sensory evaluation results indicated that heat treatments of mallow caused significant differences in total solids, pH, chlorophyll a and total chlorophyll, while frozen storage, of samples of the same treatment for 3 months caused no significant differences in moisture, ash, oxalic acid, pH, chlorophyll a and total chlorophyll. No significant differences could be noticed in the overall acceptability between mallow samples mixed with hot water and those mixed with a hot 0.1% MgCO3 solution. Significant differences were found between the heat treated and the untreated mallow. Changes of the chemical composition and the overall acceptability of blanched spinach during frozen storage showed almost the same trend noticed for the Jew's mallow.

  7. Isolation and Characterization of Phosphatidyl Choline from Spinach Leaves.

    ERIC Educational Resources Information Center

    Devor, Kenneth A.

    1979-01-01

    This inexpensive but informative experiment for undergraduate biochemistry students involves isolating phosphatidyl choline from spinach leaves. Emphasis is on introducing students to techniques of lipid extraction, separation of lipids, identification using thin layer chromatography, and identification of fatty acids. Three periods of three hours…

  8. Betaine Accumulation and Betaine-Aldehyde Dehydrogenase in Spinach Leaves 1

    PubMed Central

    Pan, Shu-Mei; Moreau, Robert A.; Yu, Charles; Huang, Anthony H. C.

    1981-01-01

    Spinach leaf discs accumulated betaine when exposed to a mannitol solution of −20 bars. The accumulation was 12 micromoles per gram original fresh weight in a 24-hour period. Betaine-aldehyde dehydrogenase (EC 1.2.1.8) was assayed in various subcellular fractions prepared from spinach leaves, and it was found only in the soluble fraction. This cytosolic enzyme was purified 175-fold, and its properties were studied. The enzyme was relatively specific for betaine aldehyde as the substrate with an apparent Km value of 2.08 × 10−4 molar. It also exerted activity on other aldehyde analogs tested, but with lower Vmax and higher Km values. The enzyme was relatively specific for nicotinamide adenine dinucleotide as the coenzyme, having an apparent Km value of 9.46 × 10−6 molar; lower activities were observed when nicotinamide adenine dinucleotide phosphate or 3-acetyl pyridine adenine dinucleotide were tested as electron acceptors. The activity was enhanced by dithiothreitol and inhibited by p-chloromercuribenzoate, and the inhibition by p-chloromercuribenzoate was partially reversed by the subsequent addition of dithiothreitol. The activity was inhibited by high concentrations of NaCl and, to a lesser extent, proline. The equilibrium of the enzymic reaction was strongly in favor of betaine formation. The in vitro activity of the enzyme under optimal assay conditions was high enough to account for the amount of betaine accumulated under water stress conditions. The enzyme activity was the same in unstressed leaves and in leaves that had been water stressed for 24 hours. PMID:16661818

  9. Biosynthesis of Sucrose and Mannitol as a Function of Leaf Age in Celery (Apium graveolens L.) 1

    PubMed Central

    Davis, Jeanine M.; Fellman, John K.; Loescher, Wayne H.

    1988-01-01

    In celery (Apium graveolens L.), the two major translocated carbohydrates are sucrose and the acyclic polyol mannitol. Their metabolism, however, is different and their specific functions are uncertain. To compare their roles in carbon partitioning and sink-source transitions, developmental changes in 14CO2 labeling, pool sizes, and key enzyme activities in leaf tissues were examined. The proportion of label in mannitol increased dramatically with leaf maturation whereas that in sucrose remained fairly constant. Mannitol content, however, was high in all leaves and sucrose content increased as leaves developed. Activities of mannose-6-P reductase, cytoplasmic and chloroplastic fructose-1,6-bisphosphatases, sucrose phosphate synthase, and sucrose synthase increased with leaf maturation and decreased as leaves senesced. Ribulose bisphosphate carboxylase and nonreversible glyceraldehyde-3-P dehydrogenase activities rose as leaves developed but did not decrease. Thus, sucrose is produced in all photosynthetically active leaves whereas mannitol is synthesized primarily in mature leaves and stored in all leaves. Onset of sucrose export in celery may result from sucrose accumulation in expanding leaves, but mannitol export is clearly unrelated to mannitol concentration. Mannitol export, however, appears to coincide with increased mannitol biosynthesis. Although mannitol and sucrose arise from a common precursor in celery, subsequent metabolism and transport must be regulated separately. PMID:16665852

  10. Expression and purification of spinach nitrite reductase in E. coli

    SciTech Connect

    Bellissimo, D.; Privalle, L. )

    1991-03-11

    The study of structure-function relationships in nitrite reductase (NiR) by site-directed mutagenesis requires an expression system from which suitable quantities of active enzyme can be purified. Spinach NiR cDNA was cloned into pUC18 and expressed in E.coli JM109 as a beta-galactosidase fusion protein. The IPTG-induced fusion protein contains five additional amino acids at the N-terminus. The expressed NiR in aerobic cultures was mostly insoluble and inactive indicating the presence of inclusion bodies. By altering growth conditions, active NiR could represent 0.5-1.0% of the total E.coli protein, Effects of the addition of delta-aminolevulinic acid, a heme precursor, and anaerobic growth were also examined. Spinach NiR was purified approximately 200 fold to homogeneity. When subjected to electrophoresis on SDS polyacrylamide gels, the NiR migrated as a single band with similar mobility to pure spinach enzyme. The expressed enzyme also reacted with rabbit anti-spinach NiR antibody as visualized by Western blot analysis. The absorption spectrum of the E.coli-expressed enzyme was identical to spinach enzyme with a Soret and alpha band a 386 and 573 nm, respectively, and an A{sub 278}/A{sub 386} = 1.9. The addition of nitrite produced the characteristic shifts in the spectrum. The E. coli-expressed NiR catalyzed the methylviologen-dependent reduction of nitrite. The specific activity was 100 U/mg. The K{sub m} determined for nitrite was 0.3 mM which is in agreement with values reported for the enzyme. These results indicate that the E.coli-expressed NiR is fully comparable to spinach NiR in purity, catalytic activity and physical state. Site-directed mutants have been made using PCR to examine structure-function relationships in this enzyme.

  11. Improving spinach, radish, and lettuce growth under red light-emitting diodes (LEDs) with blue light supplementation

    NASA Technical Reports Server (NTRS)

    Yorio, N. C.; Goins, G. D.; Kagie, H. R.; Wheeler, R. M.; Sager, J. C.

    2001-01-01

    Radish (Raphanus sativus L. cv. Cherriette), lettuce (Lactuca sativa L. cv. Waldmann's Green), and spinach (Spinacea oleracea L. cv. Nordic IV) plants were grown under 660-nm red light-emitting diodes (LEDs) and were compared at equal photosynthetic photon flux (PPF) with either plants grown under cool-white fluorescent lamps (CWF) or red LEDs supplemented with 10% (30 micromoles m-2 s-1) blue light (400-500 nm) from blue fluorescent (BF) lamps. At 21 days after planting (DAP), leaf photosynthetic rates and stomatal conductance were greater for plants grown under CWF light than for those grown under red LEDs, with or without supplemental blue light. At harvest (21 DAP), total dry-weight accumulation was significantly lower for all species tested when grown under red LEDs alone than when grown under CWF light or red LEDs + 10% BF light. Moreover, total dry weight for radish and spinach was significantly lower under red LEDs + 10% BF than under CWF light, suggesting that addition of blue light to the red LEDs was still insufficient for achieving maximal growth for these crops.

  12. Role of Cellulose and Colanic Acid in Attachment of Shiga Toxin-Producing Escherichia coli to Lettuce and Spinach in Different Water Hardness Environments.

    PubMed

    Lee, Chi-Ching; Chen, Jinru; Frank, Joseph F

    2015-08-01

    This study investigated the role of extracellular cellulose production by Shiga toxin-producing Escherichia coli (STEC) on attachment to lettuce and spinach in different water hardness environments. Two cellulose-producing wild-type STEC strains, 19 (O5:H-) and 49 (O103:H2), and their cellulose-deficient derivatives were used. Strain 49 also produced colanic acid as a constituent of its extracellular polymeric substances. Attached cells were determined by plate counts on the surface and cut edge of the leaves after an attachment period of 2 h at 4°C. Hydrophobicity and surface charge of the cells were determined. Strain 49 attached at levels 0.3 and 0.6 log greater to the surface and 0.9 and 0.4 log greater to the cut edges of spinach compared to strain 19 for both wild-type and cellulose-deficient cells (P > 0.05). Cellulose-producing cells attached more to the surface of lettuce but not of spinach than did cellulose-deficient cells. However, more cellulose-deficient cells attached (at levels 0.66 and 0.3 log greater) to the cut edge of lettuce (representing damaged tissue) than did cellulose-proficient cells (P > 0.05). Colanic acid production was associated with cell surfaces of low hydrophobicity. There was a decreasing level of attachment for the colanic acid-producing strain when water hardness increased from 200 to 1,000 pm on lettuce and spinach leaf surfaces, but no effects were seen for other cells. This decreased attachment was associated with a more negative surface charge. Cells that produced colanic acid were less hydrophobic and exhibited greater attachment to the surface and cut edge of spinach when compared to cells that did not produce colanic acid. Attachment of colanic acid-producing cells to leafy green surfaces was enhanced in higher water hardness environments. These data indicate that attachment of E. coli O157:H7 to leafy greens involves multiple mechanisms that are influenced by the type of leafy green, damage to the leaf, and the water

  13. Live-cell imaging of mammalian RNAs with Spinach2

    PubMed Central

    Strack, Rita L.; Jaffrey, Samie R.

    2015-01-01

    The ability to monitor RNAs of interest in living cells is crucial to understanding the function, dynamics, and regulation of this important class of molecules. In recent years, numerous strategies have been developed with the goal of imaging individual RNAs of interest in living cells, each with their own advantages and limitations. This chapter provides an overview of current methods of live-cell RNA imaging, including a detailed discussion of genetically encoded strategies for labeling RNAs in mammalian cells. This chapter then focuses on the development and use of “RNA mimics of GFP” or Spinach technology for tagging mammalian RNAs, and includes a detailed protocol for imaging 5S and CGG60 RNA with the recently described Spinach2 tag. PMID:25605384

  14. Photosynthate partitioning during flowering in relation to senescence of spinach

    SciTech Connect

    Sklensky, D.; Davies, P.J. )

    1990-05-01

    Male spinach plants are frequently cited as a counter-example to the nutrient drain hypothesis. Photosynthate partitioning in both male and female plants was examined. Leaves just below the inflorescences in plants at various stages of flowering were labelled with {sup 14}CO{sub 2} and the photosynthate allowed to partition for three hours. The leaves, flowers and stems of the inflorescence, and the other above ground vegetative tissue were harvested. These parts were combusted in a sample oxidizer for the collection of the {sup 14}CO{sub 2}. Allocation to the male and female flowers at very early stages are similar. As the flowers develop further, male flowers receive more photosynthate than do female flowers in early fruit production. Thus it is possible that nutrient drain to the flowers in male spinach plants is sufficient to account for senescence.

  15. Sizes of Mn-binding sites in spinach thylakoids

    SciTech Connect

    Takahashi, M.; Asada, K.

    1986-12-25

    The sizes of the Mn-binding sites in spinach thylakoids were estimated by target size analysis, assaying the membrane-bound Mn that was resistant to EDTA washing after radiation inactivation. The inactivation curve showed well the inactivation of two independent Mn-binding sites of different sizes: about two-thirds of the Mn coordinated to a binding site of 65 kDa, and the rest bound to a much smaller site of only about 3 kDa. In the large site, there was about 1 g atom of Mn/110 mol of chlorophyll in spinach thylakoids, which was constant in normally grown plants, although the Mn level in the small site depended on culture conditions. Thylakoids that had been incubated with hydroxylamine or in 0.8 M Tris lost Mn exclusively from the large binding site.

  16. Coupling Spore Traps and Quantitative PCR Assays for Detection of the Downy Mildew Pathogens of Spinach (Peronospora effusa) and Beet (P. schachtii)

    PubMed Central

    Klosterman, Steven J.; Anchieta, Amy; McRoberts, Neil; Koike, Steven T.; Subbarao, Krishna V.; Voglmayr, Hermann; Choi, Young-Joon; Thines, Marco; Martin, Frank N.

    2016-01-01

    Downy mildew of spinach (Spinacia oleracea), caused by Peronospora effusa, is a production constraint on production worldwide, including in California, where the majority of U.S. spinach is grown. The aim of this study was to develop a real-time quantitative polymerase chain reaction (qPCR) assay for detection of airborne inoculum of P. effusa in California. Among oomycete ribosomal DNA (rDNA) sequences examined for assay development, the highest nucleotide sequence identity was observed between rDNA sequences of P. effusa and P. schachtii, the cause of downy mildew on sugar beet and Swiss chard in the leaf beet group (Beta vulgaris subsp. vulgaris). Single-nucleotide polymorphisms were detected between P. effusa and P. schachtii in the 18S rDNA regions for design of P. effusa- and P. schachtii-specific TaqMan probes and reverse primers. An allele-specific probe and primer amplification method was applied to determine the frequency of both P. effusa and P. schachtii rDNA target sequences in pooled DNA samples, enabling quantification of rDNA of P. effusa from impaction spore trap samples collected from spinach production fields. The rDNA copy numbers of P. effusa were, on average, ≈3,300-fold higher from trap samples collected near an infected field compared with those levels recorded at a site without a nearby spinach field. In combination with disease-conducive weather forecasting, application of the assays may be helpful to time fungicide applications for disease management. PMID:24964150

  17. Coupling Spore Traps and Quantitative PCR Assays for Detection of the Downy Mildew Pathogens of Spinach (Peronospora effusa) and Beet (P. schachtii).

    PubMed

    Klosterman, Steven J; Anchieta, Amy; McRoberts, Neil; Koike, Steven T; Subbarao, Krishna V; Voglmayr, Hermann; Choi, Young-Joon; Thines, Marco; Martin, Frank N

    2014-12-01

    Downy mildew of spinach (Spinacia oleracea), caused by Peronospora effusa, is a production constraint on production worldwide, including in California, where the majority of U.S. spinach is grown. The aim of this study was to develop a real-time quantitative polymerase chain reaction (qPCR) assay for detection of airborne inoculum of P. effusa in California. Among oomycete ribosomal DNA (rDNA) sequences examined for assay development, the highest nucleotide sequence identity was observed between rDNA sequences of P. effusa and P. schachtii, the cause of downy mildew on sugar beet and Swiss chard in the leaf beet group (Beta vulgaris subsp. vulgaris). Single-nucleotide polymorphisms were detected between P. effusa and P. schachtii in the 18S rDNA regions for design of P. effusa- and P. schachtii-specific TaqMan probes and reverse primers. An allele-specific probe and primer amplification method was applied to determine the frequency of both P. effusa and P. schachtii rDNA target sequences in pooled DNA samples, enabling quantification of rDNA of P. effusa from impaction spore trap samples collected from spinach production fields. The rDNA copy numbers of P. effusa were, on average, ≈3,300-fold higher from trap samples collected near an infected field compared with those levels recorded at a site without a nearby spinach field. In combination with disease-conducive weather forecasting, application of the assays may be helpful to time fungicide applications for disease management.

  18. Spinach RNA aptamer detects lead (II) with high selectivity†

    PubMed Central

    DasGupta, Saurja; Shelke, Sandip A.; Li, Nan-sheng

    2015-01-01

    Spinach RNA aptamer contains a G-quadruplex motif that serves as a platform for binding and fluorescence activation of a GFP-like fluorophore. Here we show that Pb2+ induces formation of Spinach’s G-quadruplex and activates fluorescence with high selectivity and sensitivity. This device establishes the first example of an RNA-based sensor that provides a simple and inexpensive tool for Pb2+ detection. PMID:25940073

  19. Studies of GA sub 53 oxidase from spinach

    SciTech Connect

    Wilson, T.; Zeevaart, J.A.D. )

    1990-05-01

    GA{sub 53} oxidase was purified 1,750-fold with 1% recovery of activity from spinach after exposure to 8 long days. This preparation was injected into balb/c mice and hybridomas from spleen cells were produced. Upon preliminary screening by immunoprecipitation of enzyme activity, three positive cell lines were selected. These are being cloned to select a true monoclonal antibody cell line. This antibody will be used to study the light/dark regulation of this enzyme.

  20. Enantioselective degradation of metalaxyl in cucumber, cabbage, spinach and pakchoi.

    PubMed

    Wang, Meiyun; Zhang, Qing; Cong, Lujing; Yin, Wei; Wang, Minghua

    2014-01-01

    The enantioselective degradation behaviors of the acylamino acid fungicide metalaxyl in vegetables (cucumber, cabbage, spinach and pakchoi) under field and greenhouse conditions were elucidated by reversed-phase high-performance liquid chromatography (HPLC). A cellulose tris (3-chloro-4-methyl phenyl carbamate) chiral column (Lux Cellulose-2) was used for the chiral separation of the enantiomers using acetonitrile-0.1% formic acid solution (40:60, v/v) as mobile phase at a flow rate of 1.0 mL min(-1). The two enantiomers could be completely separated with a resolution (Rs) of 5.2. The linearity range, matrix effect, precision and recovery were evaluated. The method was successfully applied for the enantioselective analysis of metalaxyl in cucumber, cabbage, spinach and pakchoi under different conditions. The results showed that R-enantiomer dissipated faster than S-enantiomer in cucumber of greenhouse. Inversely, a preferential degradation of S-form was found in spinach and pakchoi under field and greenhouse conditions. No stereoselective degradation in cabbage was observed. These results can be applied for evaluating the environmental risk and food safety. PMID:24080002

  1. Enzyme-assisted extraction of stabilized chlorophyll from spinach.

    PubMed

    Özkan, Gülay; Ersus Bilek, Seda

    2015-06-01

    Zinc complex formation with chlorophyll derivatives in spinach pulp was studied by adding 300ppm Zn(2+) for production of stable food colorant, followed by the heating at 110°C for 15min. Zinc complex formation increased at pH values of 7.0 or greater. Pectinex Ultra SP-L was selected for enzyme-assisted release of zinc-chlorophyll derivatives from spinach pulp. Effect of enzyme concentration (1-9%), treatment temperature (30-60°C), and time (30-210min) on total chlorophyll content (TCC) were optimized using response surface methodology. A quadratic regression model (R(2)=0.9486) was obtained from the experimental design. Optimum treatment conditions were 8% enzyme concentration, 45°C, and 30min, which yielded a 50.747mgTCC/100g spinach pulp. Enzymatic treatment was followed by solvent extraction with ethanol at a solvent-to-sample ratio of 2.5:1 at 60°C for 45min for the highest TCC recovery. Pretreatment with enzyme and extraction in ethanol resulted in 39% increase in Zn-chlorophyll derivative yield.

  2. Comparative uptake of enteric viruses into spinach and green onions.

    PubMed

    Hirneisen, Kirsten A; Kniel, Kalmia E

    2013-03-01

    Root uptake of enteric pathogens and subsequent internalization has been a produce safety concern and is being investigated as a potential route of pre-harvest contamination. The objective of this study was to determine the ability of hepatitis A virus (HAV) and the human norovirus surrogate, murine norovirus (MNV), to internalize in spinach and green onions through root uptake in both soil and hydroponic systems. HAV or MNV was inoculated into soil matrices or into two hydroponic systems, floating and nutrient film technique systems. Viruses present within spinach and green onions were detected by RT-qPCR or infectivity assays after inactivating externally present viruses with Virkon(®). HAV and MNV were not detected in green onion plants grown up to 20 days and HAV was detected in only 1 of 64 spinach plants grown in contaminated soil substrate systems up to 20 days. Compared to soil systems, a drastic difference in virus internalization was observed in hydroponic systems; HAV or pressure-treated HAV and MNV were internalized up to 4 log RT-qPCR units and internalized MNV was shown to remain infectious. Understanding the interactions of human enteric viruses on produce can aid in the elucidation of the mechanisms of attachment and internalization, and aid in understanding risks associated with contamination events. PMID:23412715

  3. Leaf Activities.

    ERIC Educational Resources Information Center

    Mingie, Walter

    Leaf activities can provide a means of using basic concepts of outdoor education to learn in elementary level subject areas. Equipment needed includes leaves, a clipboard with paper, and a pencil. A bag of leaves may be brought into the classroom if weather conditions or time do not permit going outdoors. Each student should pick a leaf, examine…

  4. Effect of Photoperiod on the Metabolism of Deuterium-Labeled Gibberellin A53 in Spinach 1

    PubMed Central

    Gianfagna, Thomas; Zeevaart, Jan A. D.; Lusk, William J.

    1983-01-01

    Application of gibberellin A53 (GA53) to short-day (SD)-grown spinach (Spinacia oleracea L.) plants caused an increase in petiole length and leaf angle similar to that found in plants transferred to long days (LD). [2H] GA53 was fed to plants in SD, LD, and in a SD to LD transition experiment, and the metabolites were identified by gas chromatography with selected ion monitoring. After 2, 4, or 6 SD, [2H]GA53 was converted to [2H]GA19 and [2H]GA44. No other metabolites were detected. After 2 LD, only [2H] GA20 was identified. In the transition experiment in which plants were given 4 SD followed by 2 LD, all three metabolites were found. The results demonstrate unequivocally that GA19, GA20, and GA44 are metabolic products of GA53, and strongly suggest that photoperiod regulates GA metabolism, in part, by controlling the conversion of GA19 to GA20. PMID:16662988

  5. The plasma membrane-associated NADH oxidase of spinach leaves responds to blue light

    NASA Technical Reports Server (NTRS)

    Morre, D. James; Penel, Claude; Greppin, Hubert; Morre, Dorothy M.

    2002-01-01

    The plasma membrane-associated NADH oxidase (NOX) of spinach leaf disks is characterized by oscillations in activity with a regular period length of ca. 24 min. Within a single population of plants exposed to light at the same time, NOX activities of all plants function synchronously. Exposure of plants transferred from darkness to blue light (495 nm, 2 min, 50 micromoles m-2 s-1) resulted in a complex response pattern but with a new maximum in the rate of NOX activity 36 (24+12) min after illumination and then with maxima in the rate of NOX activity every 24 min thereafter. Transient maxima in NOX activity were observed as well after 9.3 + /- 1.4 and 20.7 +/- 2.1 min. The blue light response differed from the response to red (650 nm, 10 min, 50 micromoles m-2 s-1) or white light where activity maxima were initiated 12 min after the light exposure followed by maxima every 24 min thereafter. Green or yellow light was ineffective. The light response was independent of the time in the 24-min NOX cycle when the light was given. The net effects of blue and red light were ultimately the same with a new maximum in the rate of NOX activity at 12+24=36 min (and every 24 min thereafter), but the mechanisms appear to be distinct.

  6. Spinach downy mildew: overview of races and the development of molecular markers linked to major resistance genes.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spinach (Spinacia aleracea) has become an increasingly popular leafy vegetable crop, particularly in the United States. Recent trends have shown a substantial increase in per capita fresh market spinach consumption with a corresponding increase in production. As a result, spinach production practic...

  7. Biodegradable PLA (polylactic acid) hinged trays keep quality of fresh-cut and cooked spinach.

    PubMed

    Botondi, Rinaldo; Bartoloni, Serena; Baccelloni, Simone; Mencarelli, Fabio

    2015-09-01

    This work examines the effects of packaging using two different polymeric trays with hinged lids, polyethylene terephthalate (PET) and polylactic acid (PLA), on fresh-cut and cooked spinach (Spinacia oleracea). Samples were stored in a cold room for 16 days at 4 °C. Chemical (total pigments, total polyphenols, ascorbic acid, antioxidant activity), physical (water activity), technological (colour evaluation), sensorial (aroma, visual appearance and water accumulation) and microbial (total aerobic mesophilic and psychrotrophic counts) parameters were tested. Both polymeric trays maintained the overall quality of fresh spinach for 6 days but spinach stored in PLA trays maintained its flavour longer. A significant increase in total polyphenols, antiradical activity, total carotenoids as well as a decrease in ascorbic acid in fresh spinach was observed in the first 3 days of storage in both samples. Unfortunately, the PLA package accumulated condensed water. The total microbial load of fresh-cut spinach reached about 6.3-7.3 log CFU g(-1) within 8 days. Cooked spinach packed in PLA and PET polymeric hinged trays showed the same behaviour as fresh spinach in terms of quality and shelf life. In conclusion, PLA plastic hinged trays can be used for packaging fresh-cut and cooked cut spinach, but the problem of condensed water must be solved. PMID:26345011

  8. Plasmolysis and vital staining reveal viable oospores of Peronospora effusa in spinach seed lots

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Production of oospores by Peronospora effusa, the causal agent of downy mildew on spinach (Spinacia oleracea), was reported on spinach seed over three decades ago. In view of the rapid proliferation of new races of P. effusa worldwide, seed borne transmission has been suspected but methods to test ...

  9. Biodegradable PLA (polylactic acid) hinged trays keep quality of fresh-cut and cooked spinach.

    PubMed

    Botondi, Rinaldo; Bartoloni, Serena; Baccelloni, Simone; Mencarelli, Fabio

    2015-09-01

    This work examines the effects of packaging using two different polymeric trays with hinged lids, polyethylene terephthalate (PET) and polylactic acid (PLA), on fresh-cut and cooked spinach (Spinacia oleracea). Samples were stored in a cold room for 16 days at 4 °C. Chemical (total pigments, total polyphenols, ascorbic acid, antioxidant activity), physical (water activity), technological (colour evaluation), sensorial (aroma, visual appearance and water accumulation) and microbial (total aerobic mesophilic and psychrotrophic counts) parameters were tested. Both polymeric trays maintained the overall quality of fresh spinach for 6 days but spinach stored in PLA trays maintained its flavour longer. A significant increase in total polyphenols, antiradical activity, total carotenoids as well as a decrease in ascorbic acid in fresh spinach was observed in the first 3 days of storage in both samples. Unfortunately, the PLA package accumulated condensed water. The total microbial load of fresh-cut spinach reached about 6.3-7.3 log CFU g(-1) within 8 days. Cooked spinach packed in PLA and PET polymeric hinged trays showed the same behaviour as fresh spinach in terms of quality and shelf life. In conclusion, PLA plastic hinged trays can be used for packaging fresh-cut and cooked cut spinach, but the problem of condensed water must be solved.

  10. Effects of substrate type on plant growth and nitrogen and nitrate concentration in spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects of three commercial substrates (a mixture of forest residues, composted grape husks, and white peat; black peat; and coir) on plant growth and nitrogen (N) and nitrate (NO3) concentration and content were evaluated in spinach (Spinacia oleracea L. cv. Tapir). Spinach seedlings were trans...

  11. Characterization of spinach germplasm for resistance against two races of Verticillium dahliae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Historically, wilt disease caused by V. dahliae has not presented a problem in California spinach production because the crop is harvested well before the symptoms develop during the post-stem elongation (bolting) stage. However, infested spinach seeds introduce or increase inoculum in the soil for...

  12. Season-long dynamics of spinach downy mildew determined by spore trapping and disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peronospora effusa is an obligate oomycete pathogen, and the cause of downy mildew of spinach. Downy mildew threatens sustainable production of fresh market organic spinach in California, and routine fungicide sprays are often necessary for conventional production. In this study, airborne P. effus...

  13. Behavior of Escherichia coli O157:H7 on damaged leaves of spinach, lettuce, cilantro, and parsley stored at abusive temperatures.

    PubMed

    Khalil, Rowaida K; Frank, Joseph F

    2010-02-01

    Recent foodborne illness outbreaks associated with the consumption of leafy green produce indicates a need for additional information on the behavior of pathogenic bacteria on these products. Previous research indicates that pathogen growth and survival is enhanced by leaf damage. The objective of this study was to compare the behavior of Escherichia coli O157:H7 on damaged leaves of baby Romaine lettuce, spinach, cilantro, and parsley stored at three abusive temperatures (8, 12, and 15 degrees C). The damaged portions of leaves were inoculated with approximately 10(5) CFU E. coli O157:H7 per leaf. The pathogen grew on damaged spinach leaves held for 3 days at 8 and 12 degrees C (P < 0.05), with the population increasing by 1.18 and 2.08 log CFU per leaf, respectively. E. coli O157:H7 did not grow on damaged Romaine leaves at 8 or 12 degrees C, but growth was observed after 8 h of storage at 15 degrees C, with an increase of less than 1.0 log. Growth of E. coli O157:H7 on Romaine lettuce held at 8 or 12 degrees C was enhanced when inocula were suspended in 0.05% ascorbic acid, indicating the possibility of inhibition by oxidation reactions associated with tissue damage. Damaged cilantro and Italian parsley leaves held at 8 degrees C for 4 days did not support the growth of E. coli O157:H7. Behavior of the pathogen in leaf extracts differed from behavior on the damaged tissue. This study provides evidence that the damaged portion of a leafy green is a distinct growth niche that elicits different microbial responses in the various types of leafy greens.

  14. Survival of pathogenic Escherichia coli on basil, lettuce, and spinach.

    PubMed

    Markland, S M; Shortlidge, K L; Hoover, D G; Yaron, S; Patel, J; Singh, A; Sharma, M; Kniel, K E

    2013-12-01

    The contamination of lettuce, spinach and basil with pathogenic E. coli has caused numerous illnesses over the past decade. E. coli O157:H7, E. coli O104:H4 and avian pathogenic E. coli (APECstx- and APECstx+) were inoculated on basil plants and in promix substrate using drip and overhead irrigation. When overhead inoculated with 7 log CFU/ml of each strain, E. coli populations were significantly (P = 0.03) higher on overhead-irrigated plants than on drip-irrigated plants. APECstx-, E. coli O104:H4 and APECstx+ populations were recovered on plants at 3.6, 2.3 and 3.1 log CFU/g at 10 dpi (days post-inoculation), respectively. E. coli O157:H7 was not detected on basil after 4 dpi. The persistence of E. coli O157:H7 and APECstx- were similar when co-inoculated on lettuce and spinach plants. On spinach and lettuce, E. coli O157:H7 and APEC populations declined from 5.7 to 6.1 log CFU/g and 4.5 log CFU/g, to undetectable at 3 dpi and 0.6-1.6 log CFU/g at 7 dpi, respectively. The detection of low populations of APEC and E. coli O104:H4 strains 10 dpi indicates these strains may be more adapted to environmental conditions than E. coli O157:H7. This is the first reported study of E. coli O104:H4 on a produce commodity.

  15. Survival of pathogenic Escherichia coli on basil, lettuce, and spinach.

    PubMed

    Markland, S M; Shortlidge, K L; Hoover, D G; Yaron, S; Patel, J; Singh, A; Sharma, M; Kniel, K E

    2013-12-01

    The contamination of lettuce, spinach and basil with pathogenic E. coli has caused numerous illnesses over the past decade. E. coli O157:H7, E. coli O104:H4 and avian pathogenic E. coli (APECstx- and APECstx+) were inoculated on basil plants and in promix substrate using drip and overhead irrigation. When overhead inoculated with 7 log CFU/ml of each strain, E. coli populations were significantly (P = 0.03) higher on overhead-irrigated plants than on drip-irrigated plants. APECstx-, E. coli O104:H4 and APECstx+ populations were recovered on plants at 3.6, 2.3 and 3.1 log CFU/g at 10 dpi (days post-inoculation), respectively. E. coli O157:H7 was not detected on basil after 4 dpi. The persistence of E. coli O157:H7 and APECstx- were similar when co-inoculated on lettuce and spinach plants. On spinach and lettuce, E. coli O157:H7 and APEC populations declined from 5.7 to 6.1 log CFU/g and 4.5 log CFU/g, to undetectable at 3 dpi and 0.6-1.6 log CFU/g at 7 dpi, respectively. The detection of low populations of APEC and E. coli O104:H4 strains 10 dpi indicates these strains may be more adapted to environmental conditions than E. coli O157:H7. This is the first reported study of E. coli O104:H4 on a produce commodity. PMID:23280331

  16. Correlating Arsenic-Induced Morphological Change in Spinach Leaves With Leaf Spectral Characteristics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Arsenic (As) is a widely spread soil contaminant which can be accumulated into plant parts. The presence of As in edible portions of plants allows for potentially dangerous ingestion by humans and animals. The ability to detect As in plants is an important tool to minimize such risks. Remote sens...

  17. Pathway for the synthesis of triacylglycerols from monogalactosyldiacylglycerols in ozone-fumigated spinach leaves. [Spinacia oleracea L

    SciTech Connect

    Sakaki, Takeshi; Kondo, Noriaki; Yamada, Mitsuhiro Univ. of Tokyo )

    1990-10-01

    When the upper leaf surface of spinach (Spinacia oleracea L.) plants was treated with (1-{sup 14}C)acetate and grown for 2 days, {sup 14}C was effectively incorporated into acyl moieties of leaf lipids in ratios approximately their composition by mass. Fumigation of the plants with ozone (0.5 microliter per liter) caused a redistribution of {sup 14}C among lipid classes, i.e. a marked increase of {sup 14}C content in triacylglycerol (TG) and 1,2-diacylglycerol (1,2-DG) and a decrease of label in monogalactosyldiacylglycerol (MGDG) without affecting {sup 14}C distribution in leaf fatty acids. Label in both TG and 1,2-DG was found predominantly in their polyene molecular species. Since MGDG consists of similar polyene molecular species, the results indicate the synthesis of TG from MGDG via 1,2-DG. Label was also accumulated in tri- and tetragalactosyldiacylglycerol, products of galactolipid:galactolipid galactosyltransferase (GGGT). Moreover, there was a close relation between increases in the amounts of TG and the oligogalactolipids in ozone-treated leaves. These results indicate that MGDG was converted to 1,2-DG by GGGT and then to TG. In intact chloroplasts isolated from ozone-treated leaves, there was an enhanced production of free fatty acid (FFA), which was diminished by the addition of coenzyme A (CoA) and ATP, indicating that ozone stimulated the hydrolysis of MGDG to liberate FFA, which was in turn converted to acyl-CoA. The final step of TG synthesis, acylation of 1,2-DG with acyl-CoA, was confirmed by feeding with (1-{sup 14}C)linolenic acid in leaf discs excised from ozone-fumigated leaves; {sup 14}C was effectively incorporated into TG but not into 1,2-DG.

  18. The nucleotide sequence of spinach chloroplast tryptophan transfer RNA.

    PubMed Central

    Canaday, J; Guillemaut, P; Gloeckler, R; Weil, J H

    1981-01-01

    Spinach chloroplast tRNATrp, purified by column chromatography and two-dimensional gel electrophoresis, has been sequenced using in vitro labeling techniques. The sequence is : pG-C-G-C-U-C-U-U-A-G-U-U-C-A-G-U-U-C-Gm-G-D-A-G-A-A-C-m2G-psi-G-G-G-psi-C-U-C-A-A*-A-A-C-C-C-G-A-U-G-N-C-G-U-A-G-G-T-psi-C-A-A-G-U-C-C-U-A-C-A-G-A-G-C-G-U-G -C-C-AOH. Like the E. coli suppressor tRNA psu+UGA which translates both the opal terminator codon U-G-A and the tryptophan codon U-G-G, spinach chloroplast tRNATrp has C-C-A as an anticodon and contains an A-U pair in the D-stem. Images PMID:6907845

  19. Choline oxidation by intact spinach chloroplasts. [Spinacia oleracea L

    SciTech Connect

    Weigel, P.; Lerma, C.; Hanson, A.D.

    1988-01-01

    Plants synthesize betaine by a two-step oxidation of choline (choline ..-->.. betaine aldehyde ..-->.. betaine). Protoplast-derived chloroplasts of spinach (Spinacia oleracea L.) carry out both reactions, more rapidly in light than in darkness. We investigated the light-stimulated oxidation of choline, using spinach chloroplasts isolated directly from leaves. The rates of choline oxidation obtained (dark and light rates: 10-50 and 100-300 nanomoles per hour per milligram chlorophyll, respectively) were approximately 20-fold higher than for protoplast-derived chloroplasts. Betaine aldehyde was the main product. Choline oxidation in darkness and light was suppressed by hypoxia. Neither uncouplers not the Calvin cycle inhibitor glyceraldehyde greatly affected choline oxidation in the light, and maximal choline oxidation was attained far below light saturation of CO/sub 2/ fixation. The light stimulation of choline oxidation was abolished by the PSII inhibitors DCMU and dibromothymoquinone, and was partially restored by adding reduced diaminodurene, an electron donor to PSI. Both methyl viologen and phenazine methosulfate prevented choline oxidation. Adding dihydroxyacetone phosphate, which can generate NADPH in organello, doubled the dark rate of choline oxidation. These results indicate that choline oxidation in chloroplasts requires oxygen, and reducing power generated from PSI. Enzymic reactions consistent with these requirements are discussed.

  20. Assay, Purification, and Partial Characterization of Choline Monooxygenase from Spinach.

    PubMed

    Burnet, M.; Lafontaine, P. J.; Hanson, A. D.

    1995-06-01

    The osmoprotectant glycine betaine is synthesized via the path-way choline -> betaine aldehyde -> glycine betaine. In spinach (Spinacia oleracea), the first step is catalyzed by choline monooxygenase (CMO), and the second is catalyzed by betaine aldehyde dehydrogenase. Because betaine aldehyde is unstable and not easily detected, we developed a coupled radiometric assay for CMO. [14C]Choline is used as substrate; NAD+ and betaine aldehyde dehydrogenase prepared from Escherichia coli are added to oxidize [14C]betaine aldehyde to [14C]glycine betaine, which is isolated by ion exchange. The assay was used in the purification of CMO from leaves of salinized spinach. The 10-step procedure included polyethylene glycol precipitation, polyethyleneimine precipitation, hydrophobic interaction, anion exchange on choline-Sepharose, dimethyldiethanolamine-Sepharose, and Mono Q, hydroxyapatite, gel filtration, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Following gel filtration, overall purification was about 600-fold and recovery of activity was 0.5%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a polypeptide with a molecular mass of 45 kD. Taken with the value of 98 kD estimated for native CMO (R. Brouquisse, P. Weigel, D. Rhodes, C.F. Yocum, A.D. Hanson [1989] Plant Physiol 90: 322-329), this indicates that CMO is a homodimer. CMO preparations were red-brown, showed absorption maxima at 329 and 459 nm, and lost color upon dithionite addition, suggesting that CMO is an iron-sulfur protein.

  1. Effect of photoperiod on gibberellin biosynthetic enzymes in spinach

    SciTech Connect

    Gilmour, S.J.; Bleecker, A.B.; Zeevaart, J.A.D.

    1986-04-01

    The photoperiodic control of stem elongation in spinach, a long day (LD) rosette plant, is mediated by gibberellins (GAs). The early 13-hydroxylated GA biosynthetic pathway from GA/sub 12/ to GA/sub 20/ operates in spinach: GA/sub 12/ ..-->.. GA/sub 53/ ..-->.. GA/sub 44/ ..-->.. GA/sub 19/ ..-->.. GA/sub 20/. Two enzymes of this pathway, those converting GA/sub 53/ to GA/sub 44/ (GA/sub 53/ oxidase) and GA/sub 19/ to GA/sub 20/ (GA/sub 19/ oxidase), are regulated by light. The enzyme converting GA/sub 44/ to GA/sub 19/ (GA/sub 44/ oxidase) is not light-regulated. In the light GA/sub 53/ and GA/sub 18/ oxidase activities are increased, therefore causing the GA biosynthetic pathway to be turned on. This leads to the production of an active GA in LD, which causes an increase in stem elongation. Two the enzymes, GA/sub 44/ and GA/sub 53/ oxidases, can be separated from one another by anion exchange HPLC. Estimates of the molecular weights of these two enzymes based on gel filtration HPLC will be reported.

  2. Insect molting hormone and sterol biosynthesis in spinach

    SciTech Connect

    Grebenok, R.J.; Adler, J.H. )

    1990-05-01

    Insect molting hormones, which are produced by plants and are effective molecules in the control of insect crop pests, are biosynthesized in developing spinach leaves (Spinacia oleracea L.). The major sterols biosynthesized by spinach are avenasterol (24{alpha}-ethyl-5{alpha}-cholesta-7,24(28)-dien-3{beta}-ol), spinasterol (24{alpha}-ethyl-5{alpha}-cholesta-7,22-dien-3{beta}-ol), and 22-dihydrospinasterol (24{alpha}-ethyl-5{alpha}-cholest-7-en-3{beta}-ol). The major ecdysteroids biosynthesized are ecdysterone (2{beta},3{beta},14{alpha},20R,22R,25-hexahydroxy-5{beta}-cholest-7-en-6-one) and polypodine B (2{beta},3{beta},5{beta},14{alpha},20R,22R,25-heptahycroxycholest-7-en-6-one) and polypodine B (2{beta},3{beta},5{beta},14{alpha},20R,22R,25-heptahydroxycholest-7-en-6-one). When labeled 2-{sup 14}C-mevalonic acid was incorporated into young leaves isolated squalene, sterols and ecdysteroids contained the label. During a short (16 h) incorporation period in intact young leaves of 100 day old plants, the avenasterol has the highest specific activity in counts per minute per {mu}g of sterol followed by 22-dihydrospinasterol which is more highly labeled than spinasterol. The ecdysteroids synthesized, on an entire plant basis, account for 20% of the total steroid (sterol and ecdysteroid) isolated from the plant.

  3. Leaf Development

    PubMed Central

    2013-01-01

    Leaves are the most important organs for plants. Without leaves, plants cannot capture light energy or synthesize organic compounds via photosynthesis. Without leaves, plants would be unable perceive diverse environmental conditions, particularly those relating to light quality/quantity. Without leaves, plants would not be able to flower because all floral organs are modified leaves. Arabidopsis thaliana is a good model system for analyzing mechanisms of eudicotyledonous, simple-leaf development. The first section of this review provides a brief history of studies on development in Arabidopsis leaves. This history largely coincides with a general history of advancement in understanding of the genetic mechanisms operating during simple-leaf development in angiosperms. In the second section, I outline events in Arabidopsis leaf development, with emphasis on genetic controls. Current knowledge of six important components in these developmental events is summarized in detail, followed by concluding remarks and perspectives. PMID:23864837

  4. Effects of Growth Temperature on the Responses of Ribulose-1,5-Biphosphate Carboxylase, Electron Transport Components, and Sucrose Synthesis Enzymes to Leaf Nitrogen in Rice, and Their Relationships to Photosynthesis.

    PubMed

    Makino, A.; Nakano, H.; Mae, T.

    1994-08-01

    Effects of growth temperature on the photosynthetic gas-exchange rates and their underlying biochemical properties were examined in young, fully expanded leaves of rice (Oryza sativa L.). The plants were grown hydroponically under day/night temperature regimes of 18/15[deg]C, 23/18[deg]C, and 30/23[deg]C and all photosynthetic measurements were made at a leaf temperature of 25[deg]C and an irradiance of 1800 [mu]mol quanta m-2 s-1. Growth temperature affected the photosynthetic CO2 response curve. The relative ratio of the initial slope to the CO2-saturated photosynthesis increased with rising growth temperature. This was caused mainly by an increase in CO2-limited photosynthesis for a given leaf nitrogen content with rising growth temperature. However, there was no difference in ribulose-1,5-bisphosphate carboxylase (Rubisco) content at any given leaf nitrogen content among temperature treatments. In addition, the activation state and catalytic turnover rate of Rubisco were not affected by growth temperature. The increase in CO2-limited photosynthesis with rising growth temperature was the result of an increase in the CO2 transfer conductance between the intercellular airspaces and the carboxylation sites. The amounts of total chlorophyll and light-harvesting chlorophyll a/b protein II increased for the same leaf nitrogen content with rising growth temperature, but the amounts of cytochrome f and coupling factor 1 and the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were the same between plants grown at 23/18[deg]C and those grown at 30/23[deg]C. Similarly, CO2-saturated photosynthesis was not different for the same leaf nitrogen content between these treatments. For the 18/15[deg]C-grown plants, a slight decrease in the amounts of cytochrome f and coupling factor 1 and an increase in the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were found, but these were not reflected in CO2-saturated

  5. A real-time PCR assay for detection and quantification of Verticillium dahliae in spinach seed.

    PubMed

    Duressa, Dechassa; Rauscher, Gilda; Koike, Steven T; Mou, Beiquan; Hayes, Ryan J; Maruthachalam, Karunakaran; Subbarao, Krishna V; Klosterman, Steven J

    2012-04-01

    Verticillium dahliae is a soilborne fungus that causes Verticillium wilt on multiple crops in central coastal California. Although spinach crops grown in this region for fresh and processing commercial production do not display Verticillium wilt symptoms, spinach seeds produced in the United States or Europe are commonly infected with V. dahliae. Planting of the infected seed increases the soil inoculum density and may introduce exotic strains that contribute to Verticillium wilt epidemics on lettuce and other crops grown in rotation with spinach. A sensitive, rapid, and reliable method for quantification of V. dahliae in spinach seed may help identify highly infected lots, curtail their planting, and minimize the spread of exotic strains via spinach seed. In this study, a quantitative real-time polymerase chain reaction (qPCR) assay was optimized and employed for detection and quantification of V. dahliae in spinach germplasm and 15 commercial spinach seed lots. The assay used a previously reported V. dahliae-specific primer pair (VertBt-F and VertBt-R) and an analytical mill for grinding tough spinach seed for DNA extraction. The assay enabled reliable quantification of V. dahliae in spinach seed, with a sensitivity limit of ≈1 infected seed per 100 (1.3% infection in a seed lot). The quantification was highly reproducible between replicate samples of a seed lot and in different real-time PCR instruments. When tested on commercial seed lots, a pathogen DNA content corresponding to a quantification cycle value of ≥31 corresponded with a percent seed infection of ≤1.3%. The assay is useful in qualitatively assessing seed lots for V. dahliae infection levels, and the results of the assay can be helpful to guide decisions on whether to apply seed treatments.

  6. Rapid cloning and bioinformatic analysis of spinach Y chromosome-specific EST sequences.

    PubMed

    Deng, Chuan-Liang; Zhang, Wei-Li; Cao, Ying; Wang, Shao-Jing; Li, Shu-Fen; Gao, Wu-Jun; Lu, Long-Dou

    2015-12-01

    The genome of spinach single chromosome complement is about 1000 Mbp, which is the model material to study the molecular mechanisms of plant sex differentiation. The cytological study showed that the biggest spinach chromosome (chromosome 1) was taken as spinach sex chromosome. It had three alleles of sex-related X,X(m) and Y. Many researchers have been trying to clone the sex-determining genes and investigated the molecular mechanism of spinach sex differentiation. However,there are no successful cloned reports about these genes. A new technology combining chromosome microdissection with hybridization-specific amplification (HSA) was adopted. The spinach Y chromosome degenerate oligonucleotide primed-PCR (DOP-PCR) products were hybridized with cDNA of the male spinach flowers in florescence. The female spinach genome was taken as blocker and cDNA library specifically expressed in Y chromosome was constructed. Moreover, expressed sequence tag (EST) sequences in cDNA library were cloned, sequenced and bioinformatics was analysed. There were 63 valid EST sequences obtained in this study. The fragment size was between 53 and 486 bp. BLASTn homologous alignment indicated that 12 EST sequences had homologous sequences of nucleic acids, the rest were new sequences. BLASTx homologous alignment indicated that 16 EST sequences had homologous protein-encoding nucleic acid sequence. The spinach Y chromosome-specific EST sequences laid the foundation for cloning the functional genes, specifically expressed in spinach Y chromosome. Meanwhile, the establishment of the technology system in the research provided a reference for rapid cloning of other biological sex chromosome-specific EST sequences.

  7. Surface and internalized Escherichia coli O157:H7 on field-grown spinach and lettuce treated with spray-contaminated irrigation water.

    PubMed

    Erickson, Marilyn C; Webb, Cathy C; Diaz-Perez, Juan Carlos; Phatak, Sharad C; Silvoy, John J; Davey, Lindsey; Payton, Alison S; Liao, Jean; Ma, Li; Doyle, Michael P

    2010-06-01

    Numerous field studies have revealed that irrigation water can contaminate the surface of plants; however, the occurrence of pathogen internalization is unclear. This study was conducted to determine the sites of Escherichia coli O157:H7 contamination and its survival when the bacteria were applied through spray irrigation water to either field-grown spinach or lettuce. To differentiate internalized and surface populations, leaves were treated with a surface disinfectant wash before the tissue was ground for analysis of E. coli O157:H7 by direct plate count or enrichment culture. Irrigation water containing E. coli O157:H7 at 10(2), 10(4), or 10(6) CFU/ml was applied to spinach 48 and 69 days after transplantation of seedlings into fields. E. coli O157:H7 was initially detected after application on the surface of plants dosed at 10(4) CFU/ml (4 of 20 samples) and both on the surface (17 of 20 samples) and internally (5 of 20 samples) of plants dosed at 10(6) CFU/ml. Seven days postspraying, all spinach leaves tested negative for surface or internal contamination. In a subsequent study, irrigation water containing E. coli O157:H7 at 10(8) CFU/ml was sprayed onto either the abaxial (lower) or adaxial (upper) side of leaves of field-grown lettuce under sunny or shaded conditions. E. coli O157:H7 was detectable on the leaf surface 27 days postspraying, but survival was higher on leaves sprayed on the abaxial side than on leaves sprayed on the adaxial side. Internalization of E. coli O157:H7 into lettuce leaves also occurred with greater persistence in leaves sprayed on the abaxial side (up to 14 days) than in leaves sprayed on the adaxial side (2 days).

  8. Effect of antimycin a on photosynthesis of intact spinach chloroplasts.

    PubMed

    Schacter, B Z; Gibbs, M; Champigny, M L

    1971-10-01

    Low concentrations (0.5-10 mum) of antimycin A were shown to increase the rate of CO(2) fixation, O(2) evolution and inorganic phosphate esterification in intact spinach (Spinacia oleracea) chloroplasts. The increase was highest when the light intensity was saturating. Stimulation was independent of the bicarbonate concentration and was accompanied by an enhancement in the synthesis of glycerate 3-phosphate with a decrease in dihydroxyacetone phosphate. The antibiotic decreased the Michaelis constant of the chloroplast but not of ribulose 1,5-diphosphate carboxylase for bicarbonate. It was suggested that antimycin A is affecting that portion (outer envelope) of the intact chloroplast which contains the enzyme mechanism for controlling the pace of CO(2) fixation.

  9. Purification and characterization of ribulose-5-phosphate kinase from spinach

    SciTech Connect

    Porter, M.A.; Milanez, S.; Stringer, C.D.; Hartman, F.C.

    1986-02-15

    An efficient purification procedure utilizing affinity chromatography is described for spinach ribulose-5-phosphate kinase, a light-regulated chloroplastic enzyme. Gel filtration and polyacrylamide gel electrophoresis of the purified enzyme reveal a dimeric structure of 44,000 Mr subunits. Chemical crosslinking with dimethyl suberimidate confirms the presence of two subunits per molecule of native kinase, which are shown to be identical by partial NH2-terminal sequencing. Based on sulfhydryl titrations and on amino acid analyses, each subunit contains four to five cysteinyl residues. The observed slow loss of activity during spontaneous oxidation in air-saturated buffer correlates with the intramolecular oxidation of two sulfhydryl groups, presumably those involved in thioredoxin-mediated regulation.

  10. Influence of nutrient level on methylmercury content in water spinach.

    PubMed

    Greger, Maria; Dabrowska, Beata

    2010-08-01

    Widely consumed vegetables are often cultivated in sewage waters with high nutrient levels. They can contain high levels of methylmercury (MeHg), because they can form MeHg from inorganic Hg in their young shoots. We determined whether the MeHg uptake and the MeHg formation in the shoots of water spinach (Ipomoea aquatica) were affected by the presence of a high nutrient level in the growth medium. Water spinach shoots were rooted and pretreated in growth medium containing 7% (low) or 70% (high) Hoagland nutrient solution; thereafter, the plants were treated with either 0.02 microM MeHg or 0.2 microM HgCl2 for 3 d. Half the plants were then analyzed for total Hg and MeHg. The remaining plants were transferred to mercury-free medium with low or high nutrient levels and posttreated for 3 days before analysis of total Hg and MeHg in order to measure MeHg formation in the absence of external Hg. The results indicate that nutrient level did not influence MeHg uptake, but that a high nutrient level reduced the distribution of MeHg to the shoots 2.7-fold versus low nutrient level. After treatment with HgCl2, MeHg was found in roots and new shoots but not in old shoots. The MeHg:total-Hg ratio was higher in new shoots than in roots, being 13 times higher at high versus low nutrient levels. Thus, MeHg formation was the same in new shoots independent of inorganic Hg concentration, since the total Hg level decreased at a high nutrient level.

  11. Satiety effects of spinach in mixed meals: comparison with other vegetables.

    PubMed

    Gustafsson, K; Asp, N G; Hagander, B; Nyman, M

    1995-11-01

    The effects of vegetables in mixed meals on satiety were evaluated by varying the dose (portions 150 and 250 g containing 4.3 and 7.2 g of dietary fibre, respectively) and structure (cut and minced) of microwaved spinach. A control meal without spinach and the test meals with spinach were balanced regarding energy (2000 kJ) and digestible carbohydrates (59 g, 51 E%) and with protein and fat as far as possible similar. The meals were served in the morning, after an overnight fast, to ten healthy male volunteers. Satiety was registered and blood glucose as well as plasma insulin and C-peptide were analysed at regular intervals until 3.5 h postprandially. The largest spinach portions augmented satiety and reduced the postprandial glucose response. The total satiety scores seemed to be correlated positively to both the dietary fibre and the water content in the vegetable. Differences in structure had no influence on satiety scores.

  12. Reduction of cadmium uptake in spinach (Spinacia oleracea L.) by soil amendment with animal waste compost.

    PubMed

    Sato, Atsushi; Takeda, Hiroyuki; Oyanagi, Wataru; Nishihara, Eiji; Murakami, Masaharu

    2010-09-15

    A field experiment was conducted to evaluate the efficacy of animal waste compost (AWC) in reducing Cd uptake by spinach (Spinacia oleracea L.). Spinach was grown in a field that had been treated by having cattle, swine, or poultry waste compost incorporated into the soil before each crop throughout 4 years of rotational vegetable production. Cadmium concentration was 34-38% lower in spinach harvested from the AWC-treated soils than in the chemical fertilizer-treated soil. Although the repeated application of swine and poultry compost caused significant P accumulation in the cropped soils, that of cattle compost did not. These results indicate that cattle compost with high affinity for Cd and low P content should be the preferred soil amendment when used to reduce Cd uptake by spinach.

  13. Leaf Carbohydrate Status and Enzymes of Translocate Synthesis in Fruiting and Vegetative Plants of Cucumis sativus L. 1

    PubMed Central

    Pharr, David M.; Huber, Steven C.; Sox, Harriet N.

    1985-01-01

    Carbon partitioning in the leaves of Cucumis sativus L., a stachyose translocating plant, was influenced by the presence or absence of a single growing fruit on the plant. Fruit growth was very rapid with rates of fresh weight gain as high as 3.3 grams per hour. Fruit growth was highly competitive with vegetative growth as indicated by lower fresh weights of leaf blades, petioles, stem internodes and root systems on plants bearing a single growing fruit compared to plants not bearing a fruit. Carbon exchange rates, starch accumulation rates and carbon export rates were higher in leaves of plants bearing a fruit. Dry weight loss from leaves was higher at night from fruiting plants, and morning starch levels were consistently lower in leaves of fruiting than in leaves of vegetative plants indicating rapid starch mobilization at night from the leaves of fruiting plants. Galactinol, the galactosyl donor for stachyose biosynthesis, was present in the leaves of fruit-bearing plants at consistently lower concentration than in leaves of vegetative plants. Galactinol synthase, and sucrose phosphate synthase activities were not different on a per gram fresh weight basis in leaves from the two plant types; however, stachyose synthase activity was twice as high in leaves from fruiting plants. Thus, the lower galactinol pools may be associated with an activation of the terminal step in stachyose biosynthesis in leaves in response to the high sink demand of a growing cucumber fruit. PMID:16663989

  14. Bioavailability of iron from spinach using an in vitro/human Caco-2 cell bioassay model

    NASA Technical Reports Server (NTRS)

    Rutzke, Corinne J.; Glahn, Raymond P.; Rutzke, Michael A.; Welch, Ross M.; Langhans, Robert W.; Albright, Louis D.; Combs, Gerald F Jr; Wheeler, Raymond M.

    2004-01-01

    Spinach (Spinacia oleracea) cv Whitney was tested for iron bioavailabilty using an in vitro human intestinal cell culture ferritin bioassay technique previously developed. Spinach was cultured in a growth chamber for 33 days, harvested, and freeze-dried. Total iron in the samples was an average of 71 micrograms/g dry weight. Spinach was digested in vitro (pepsin and 0.1 M HCl followed by pancreatin and 0.1 M NaHCO3) with and without the addition of supplemental ascorbic acid. Caco-2 cell cultures were used to determine iron bioavailability from the spinach mixtures. Production of the iron-binding protein ferritin in the Caco-2 cells showed the supplemental ascorbic acid doubled bioavailability of iron from spinach. The data show fresh spinach is a poor source of iron, and emphasize the importance of evaluation of whole meals rather than single food items. The data support the usefulness of the in vitro/Caco-2 cell ferritin bioassay model for prescreening of space flight diets for bioavailable iron.

  15. Effect of spinach cultivar and bacterial adherence factors on survival of Escherichia coli O157:H7 on spinach leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Similarly to phytopathogens and epiphytic microorganisms, human bacterial pathogens have been shown to colonize on plant phylloplane. Along with environmental variables such as temperate, UV light, relative humidity, etc., plant cultivar and specifically the leaf blade morphological characteristics ...

  16. Inactivation of Escherichia coli O157:H7 attached to spinach harvester blade using bacteriophage.

    PubMed

    Patel, Jitendra; Sharma, Manan; Millner, Patricia; Calaway, Todd; Singh, Manpreet

    2011-04-01

    Outbreaks associated with leafy greens have focused attention on the transfer of human pathogens to these commodities during harvest with commercial equipment. Attachment of Escherichia coli O157:H7 on new or rusty spinach harvester blades immersed in spinach extract or 10% tryptic soy broth (TSB) was investigated. Bacteriophages specific for E. coli O157:H7 were evaluated to kill cells attached to blade. A cocktail of five nalidixic acid-resistant E. coli O157:H7 isolates was transferred to 25 mL of spinach extract or 10% TSB. A piece of sterilized spinach harvester blade (2×1") was placed in above spinach extract or 10% TSB and incubated at room (22 °C) or dynamic (30 °C day, 20 °C night) temperatures. E. coli O157:H7 populations attached to blade during incubation in spinach extract or 10% TSB were determined. When inoculated at 1 log CFU/mL, E. coli O157:H7 attachment to blades after 24 and 48 h incubation at dynamic temperature (6.09 and 6.37 log CFU/mL) was significantly higher than when incubated at 22 °C (4.84 and 5.68 log CFU/mL), respectively. After 48 h incubation, two blades were sprayed on each side with a cocktail of E. coli O157-specific bacteriophages before scraping the blade, and subsequent plating on Sorbitol MacConkey media-nalidixic acid. Application of bacteriophages reduced E. coli O157:H7 populations by 4.5 log CFU on blades after 2 h of phage treatment. Our study demonstrates that E. coli O157:H7 can attach to and proliferate on spinach harvester blades under static and dynamic temperature conditions, and bacteriophages are able to reduce E. coli O157:H7 populations adhered to blades. PMID:21453119

  17. Detection of genome DNA methylation change in spinach induced by 5-azaC.

    PubMed

    Gao, Wujun; Li, Shufen; Li, Zhongxia; Huang, Yingying; Deng, Chuanliang; Lu, Longdou

    2014-08-01

    DNA methylation has been implicated in the regulation of gene expression, genome imprinting, and chromatin remodeling in eukaryotes. In this study, we analyzed possible alterations in levels and patterns of cytosine methylation in male and female spinach plants after treatment with demethylation agent 5-azacytidine (5-azaC) using two methods: (1) direct determination of 5-methylcytidine (5 mC) amounts in genomic DNA by high-performance liquid chromatography (HPLC) separation and quantification of nucleosides and (2) methylation-sensitive inter-simple sequence repeat (MS-ISSR) technique. HPLC analysis revealed that the DNA methylation events in male and female spinach leaves markedly decreased upon 30 μM 5-azaC treatment, and the methylation level gradually decreased with the increase in 5-azaC concentration. To study the altered DNA methylation patterns in spinach after 5-azaC treatment, untreated and 500 μM 5-azaC-treated samples were analyzed by MS-ISSR assay. A total of 385 informative profiles were resolved using 35 ISSR primer sets. MS-ISSR analysis showed various altered methylation patterns between untreated and 5-azaC-treated spinach plants. These alterations were mainly demethylation events, which were largely consistent with the HPLC results. Both HPLC and MS-ISSR analyses showed that the changes in DNA methylation levels and patterns were similar in male and female spinach leaves, which implies that sex was not the main factor influencing DNA methylation levels and patterns in the vegetative organs of spinach. This study could provide a molecular basis of the altered DNA methylation induced by 5-azaC, and lay a foundation for further investigation of the relationship between methylation and sex determination and development in this dioecious plant spinach.

  18. Inactivation of Escherichia coli O157:H7 attached to spinach harvester blade using bacteriophage.

    PubMed

    Patel, Jitendra; Sharma, Manan; Millner, Patricia; Calaway, Todd; Singh, Manpreet

    2011-04-01

    Outbreaks associated with leafy greens have focused attention on the transfer of human pathogens to these commodities during harvest with commercial equipment. Attachment of Escherichia coli O157:H7 on new or rusty spinach harvester blades immersed in spinach extract or 10% tryptic soy broth (TSB) was investigated. Bacteriophages specific for E. coli O157:H7 were evaluated to kill cells attached to blade. A cocktail of five nalidixic acid-resistant E. coli O157:H7 isolates was transferred to 25 mL of spinach extract or 10% TSB. A piece of sterilized spinach harvester blade (2×1") was placed in above spinach extract or 10% TSB and incubated at room (22 °C) or dynamic (30 °C day, 20 °C night) temperatures. E. coli O157:H7 populations attached to blade during incubation in spinach extract or 10% TSB were determined. When inoculated at 1 log CFU/mL, E. coli O157:H7 attachment to blades after 24 and 48 h incubation at dynamic temperature (6.09 and 6.37 log CFU/mL) was significantly higher than when incubated at 22 °C (4.84 and 5.68 log CFU/mL), respectively. After 48 h incubation, two blades were sprayed on each side with a cocktail of E. coli O157-specific bacteriophages before scraping the blade, and subsequent plating on Sorbitol MacConkey media-nalidixic acid. Application of bacteriophages reduced E. coli O157:H7 populations by 4.5 log CFU on blades after 2 h of phage treatment. Our study demonstrates that E. coli O157:H7 can attach to and proliferate on spinach harvester blades under static and dynamic temperature conditions, and bacteriophages are able to reduce E. coli O157:H7 populations adhered to blades.

  19. Whole-Transcriptome Analysis of Verocytotoxigenic Escherichia coli O157:H7 (Sakai) Suggests Plant-Species-Specific Metabolic Responses on Exposure to Spinach and Lettuce Extracts

    PubMed Central

    Crozier, Louise; Hedley, Pete E.; Morris, Jenny; Wagstaff, Carol; Andrews, Simon C.; Toth, Ian; Jackson, Robert W.; Holden, Nicola J.

    2016-01-01

    Verocytotoxigenic Escherichia coli (VTEC) can contaminate crop plants, potentially using them as secondary hosts, which can lead to food-borne infection. Currently, little is known about the influence of the specific plant species on the success of bacterial colonization. As such, we compared the ability of the VTEC strain, E. coli O157:H7 ‘Sakai,’ to colonize the roots and leaves of four leafy vegetables: spinach (Spinacia oleracea), lettuce (Lactuca sativa), vining green pea (Pisum sativum), and prickly lettuce (Lactuca serriola), a wild relative of domesticated lettuce. Also, to determine the drivers of the initial response on interaction with plant tissue, the whole transcriptome of E. coli O157:H7 Sakai was analyzed following exposure to plant extracts of varying complexity (spinach leaf lysates or root exudates, and leaf cell wall polysaccharides from spinach or lettuce). Plant extracts were used to reduce heterogeneity inherent in plant–microbe interactions and remove the effect of plant immunity. This dual approach provided information on the initial adaptive response of E. coli O157:H7 Sakai to the plant environment together with the influence of the living plant during bacterial establishment and colonization. Results showed that both the plant tissue type and the plant species strongly influence the short-term (1 h) transcriptional response to extracts as well as longer-term (10 days) plant colonization or persistence. We show that propagation temperature (37 vs. 18°C) has a major impact on the expression profile and therefore pre-adaptation of bacteria to a plant-relevant temperature is necessary to avoid misleading temperature-dependent wholescale gene-expression changes in response to plant material. For each of the plant extracts tested, the largest group of (annotated) differentially regulated genes were associated with metabolism. However, large-scale differences in the metabolic and biosynthetic pathways between treatment types indicate

  20. Whole-Transcriptome Analysis of Verocytotoxigenic Escherichia coli O157:H7 (Sakai) Suggests Plant-Species-Specific Metabolic Responses on Exposure to Spinach and Lettuce Extracts.

    PubMed

    Crozier, Louise; Hedley, Pete E; Morris, Jenny; Wagstaff, Carol; Andrews, Simon C; Toth, Ian; Jackson, Robert W; Holden, Nicola J

    2016-01-01

    Verocytotoxigenic Escherichia coli (VTEC) can contaminate crop plants, potentially using them as secondary hosts, which can lead to food-borne infection. Currently, little is known about the influence of the specific plant species on the success of bacterial colonization. As such, we compared the ability of the VTEC strain, E. coli O157:H7 'Sakai,' to colonize the roots and leaves of four leafy vegetables: spinach (Spinacia oleracea), lettuce (Lactuca sativa), vining green pea (Pisum sativum), and prickly lettuce (Lactuca serriola), a wild relative of domesticated lettuce. Also, to determine the drivers of the initial response on interaction with plant tissue, the whole transcriptome of E. coli O157:H7 Sakai was analyzed following exposure to plant extracts of varying complexity (spinach leaf lysates or root exudates, and leaf cell wall polysaccharides from spinach or lettuce). Plant extracts were used to reduce heterogeneity inherent in plant-microbe interactions and remove the effect of plant immunity. This dual approach provided information on the initial adaptive response of E. coli O157:H7 Sakai to the plant environment together with the influence of the living plant during bacterial establishment and colonization. Results showed that both the plant tissue type and the plant species strongly influence the short-term (1 h) transcriptional response to extracts as well as longer-term (10 days) plant colonization or persistence. We show that propagation temperature (37 vs. 18°C) has a major impact on the expression profile and therefore pre-adaptation of bacteria to a plant-relevant temperature is necessary to avoid misleading temperature-dependent wholescale gene-expression changes in response to plant material. For each of the plant extracts tested, the largest group of (annotated) differentially regulated genes were associated with metabolism. However, large-scale differences in the metabolic and biosynthetic pathways between treatment types indicate

  1. Cedar leaf oil poisoning

    MedlinePlus

    Cedar leaf oil is made from some types of cedar trees. Cedar leaf oil poisoning occurs when someone swallows this substance. ... The substance in cedar leaf oil that can be harmful is thujone (a hydrocarbon).

  2. Influence of variations in soil copper on the yield and nutrition of spinach grown in microplots on two organic soils

    SciTech Connect

    Mathur, S.P.; Belanger, A.; Sanderson, R.B.; Valk, M.; Knibbe, E.N.

    1984-01-01

    Spinach (Spinacia oleracea L. cv. Symphony) was grown in spring 1982 in field microplots of an organic soil (site I-mucky peat) containing 81 to 1063 ..mu..g Cu.g/sup -1/ soil, and cv. America of the same crop taken in summer 1982 on a peaty organic soil (site II) varying in Cu content from 13 to 1659 ..mu..g.g/sup -1/. The variations in soil Cu were mainly due to three rates of Cu applications in 1978 at site II and in 1979 at site I. At site I, the diversity in soil-Cu had no effect on yield or foliar-Cu levels in the crop. At site II soil-Cu was positively correlated with yield and foliar Cu; and negatively with leaf Fe due to a dilution effect. Neither soil Cu nor foliar Cu had any significant effect on Mo in leaves at both sites, except that the increase in yield due to the highest level of Cu at site II was accompanied by an increased plant uptake of Mo. Also, foliar Cu was positively correlated with P, Mg and Mn levels in leaves at site I; and foliar Ca, Mg and Mn at site II. Residual soil Cu up to 1063 ..mu..g.g/sup -1/ in a mucky peat and 1659 ..mu..g./sup -1/ in a peat showed no signs of causing phytotoxocity or significant nutritional imbalance. 19 references, 4 tables.

  3. Purification of gibberellin sub 53 -oxidase from spinach

    SciTech Connect

    Wilson, T.M.; Zeevaart, J.A.D. )

    1989-04-01

    Spinach is a long-day rosette plants, in which stem growth is mediated by gibberellins. It has been shown that two enzymatic steps, GA{sub 53}-oxidase and GA{sub 19}-oxidase, are controlled by light. To develop an understanding into this light regulation, purification of GA{sub 53}-oxidase has been undertaken. The original assay relied on the HPLC separation of the product and substrate, but was considered too slow for the development of a purification scheme. A TLC system was developed which in conjunction with improvements to the assay conditions was sensitive and gave rapid results. The partial purification of the GA{sub 53}-oxidase is achieved by a high speed centrifugation, 40-55% ammonium sulfate precipitation, an hydroxyapatite column, Sephadex G-100 column and an anion exchange FPLC column, Mono Q HR10/10, yielding 1000-fold purification and 15% recovery. Monoclonal antibodies to the protein will be raised and used to further characterize the enzyme.

  4. Time-resolved fluorescence spectroscopy of spinach chloroplast.

    PubMed

    Yu, W; Pellegrino, F; Alfano, R R

    1977-04-11

    Picosecond fluorescent kinetics and time-resolved spectra of spinach chloroplast were measured at room temperature and low temperatures. The measurement is conducted with 530 nm excitation at an average intensity of 2-10(14) photons/cm2, pluse and at a pulse separation of 6 ns for the 100 pulses used. The 685 nm fluorescent kinetics was found to decay with two components, a fast component with a 56 ps lifetime, and a slow component with a 220 ps lifetime. The 730 nm fluorescent kinetics at room temperature is a single exponential decay with a 100 ps lifetime. The 730 nm fluorescence lifetime was found to increase by a factor of 6 when the temperature was lowered from room temperature to 90 K, while the 685 and 695 nm fluorescent kinetics were unchanged. The time-resolved spectra data obtained within 10 ps after excitation is consistent with the kinetic data reported here. A two-level fluorescence scheme is proposed to explain the kinetics. The effect of excitation with high light intensity and multiple pulses is discussed.

  5. Uptake of l-Ascorbate by Intact Spinach Chloroplasts

    PubMed Central

    Beck, Erwin; Burkert, Anette; Hofmann, Margit

    1983-01-01

    Uptake of l-[1-14C]ascorbate by intact ascorbate-free spinach (Spinacia oleracea L. cv Vitalr) chloroplasts has been investigated using the technique of silicone oil filtering. Rates greater than 100 micromoles per milligram chlorophyll per hour (external concentration, 10 millimolar) of ascorbate transport were observed. Ascorbate uptake into the sorbitol-impermeable space (stroma) followed the Michaelis-Menten-type characteristic for substrate saturation. A Km of 18 to 40 millimolar was determined. Transport of ascorbate across the chloroplast envelope resulted in an equilibrium of the ascorbate concentrations between stroma and medium. A pH optimum of 7.0 to 7.5 and the lack of alkalization of the medium upon ascorbate uptake suggest that only the monovalent ascorbate anion is able to cross the chloroplast envelope. The activation energy of ascorbate uptake was determined to be 65.8 kilojoules (16 kilocalories) per mole (8 to 20°C). Interference of ascorbate transport with substrates of the phosphate or dicarboxylate translocator could not be detected, but didehydroascorbate was a competitive inhibitor. Preloading of chloroplasts with didehydroascorbate resulted in an increase of Vmax but did not change the Km for ascorbate. Millimolar concentrations of the sulfhydryl reagent p-chloromercuriphenyl sulfonate inhibited ascorbate uptake. The data are interpreted in terms of ascorbate uptake into chloroplasts by the mechanism of facilitated diffusion mediated by a specific translocator. PMID:16663182

  6. Synthesis of Mono- and Digalactosyldiacylglycerol in Isolated Spinach Chloroplasts 1

    PubMed Central

    Heemskerk, Johan W. M.; Bögemann, Gerard; Helsper, Johannes P. F. G.; Wintermans, Jef F. G. M.

    1988-01-01

    Purified, intact chloroplasts of Spinacia oleracea L. synthesize galactose-labeled mono- and digalactosyldiacylglycerol (MGDG and DGDG) from UDP-[U-14C]galactose. In the presence of high concentrations of unchelated divalent cations they also synthesize tri- and tetra-galactosyldiacylglycerol. The acyl chains of galactose-labeled MGDG are strongly desaturated and such MGDG is a good precursor for DGDG and higher oligogalactolipids. The synthesis of MGDG is catalyzed by UDP-Gal:sn-1,2-diacylglycerol galactosyltransferase, and synthesis of DGDG and the oligogalactolipids is exclusively catalyzed by galactolipid:galactolipid galactosyltransferase. The content of diacylglycerol in chloroplasts remains low during UDP-Gal incorporation. This indicates that formation of diacylglycerol by galactolipid:galactolipid galactosyltransferase is balanced with diacylglycerol consumption by UDP-Gal:diacylglycerol galactosyltransferase for MGDG synthesis. Incubation of intact spinach chloroplasts with [2-14C]acetate or sn-[U-14C]glycerol-3-P in the presence of Mg2+ and unlabeled UDP-Gal resulted in high 14C incorporation into MGDG, while DGDG labeling was low. This de novo made MGDG is mainly oligoene. Its conversion into DGDG is also catalyzed, at least in part, by galactolipid:galactolipid galactosyltransferase. Images Fig. 1 PMID:16666019

  7. Spinach pyruvate kinase isoforms: partial purification and regulatory properties

    SciTech Connect

    Baysdorfer, C.; Bassham, J.A.

    1984-02-01

    Pyruvate kinase from spinach (Spinacea oleracea L.) leaves consists of two isoforms, separable by blue agarose chromatography. Both isoforms share similar pH profiles and substrate and alternate nucleotide K/sub m/ values. In addition, both isoforms are inhibited by oxalate and ATP and activated by AMP. The isoforms differ in their response to three key metabolites; citrate, aspartate, and glutamate. The first isoform is similar to previously reported plant pyruvate kinases in its sensitivity to citrate inhibition. The K/sub i/ for this inhibition is 1.2 millimolar citrate. The second isoform is not affected by citrate but is regulated by aspartate and glutamate. Aspartate is an activator with a K/sub a/ of 0.05 millimolar, and glutamate is an inhibitor with a K/sub i/ of 0.68 millimolar. A pyruvate kinase with these properties has not been previously reported. Based on these considerations, the authors suggest that the activity of the first isoform is regulated by respiratory metabolism. The second isoform, in contrast, may be regulated by the demand for carbon skeletons for use in ammonia assimilation.

  8. Phosphoglycolate phosphatase of spinach acts as a phosphoenzyme

    SciTech Connect

    Rose, Z.B.; Seal, S.N.

    1987-05-01

    When /sup 32/P-glycolate and phosphoglycolate phosphatase from spinach are mixed, /sup 32/P is incorporated into acid precipitated protein. Properties that relate this phosphorylation to the enzyme are: The K/sub m/ value for P-glycolate is similar for protein phosphorylation and substrate hydrolysis; the /sup 32/P appearing in the phosphoenzyme is diluted by unlabeled P-glycolate or the alternative substrate, ethyl-P; the activator Cl/sup -/ enhances the effectiveness of ethyl-P as a substrate and as an inhibitor of the formation of /sup 32/P-enzyme; and /sup 32/P is lost from the enzyme when /sup 32/P-glycolate is consumed. The acid denatured phosphorylated protein is a molecule of 34,000 Da, which is half of the molecular weight of the native protein and is similar in size to the labeled band that is seen on SDS-polyacrylamide gels. The enzyme-bound phosphoryl group appears to be an acyl-phosphate from its pH stability, being quite stable at pH 1, less stable at pH 5, and very unstable above pH 5. The bond is readily hydrolyzed in acid molybdate and it is sensitive to cleavage by hydroxylamine at pH 6.8. The demonstration of enzyme phosphorylation by /sup 32/P-glycolate resolves the dilemma presented by initial rate studies in which alternative substrates appeared to have different mechanisms.

  9. The activity of uridine diphosphate glucose–d-fructose 6-phosphate 2-glucosyltransferase in leaves

    PubMed Central

    Hawker, J. S.

    1967-01-01

    1. By using EDTA in reaction mixtures it was possible to determine the activity of sucrose phosphate synthetase in freshly prepared leaf extracts without the complications caused by sucrose phosphatase. 2. EDTA was found also to increase the activity of sucrose phosphate synthetase by as much as 100%. 3. High sucrose phosphate synthetase activities were found in leaf preparations in which sucrose phosphatase was inhibited by EDTA. By contrast with previous reports, the activities were sufficient to allow sucrose synthesis in leaves during photosynthesis to occur via sucrose phosphate. 4. Sugar-cane plants having different rates of photosynthesis also had different activities of sucrose phosphate synthetase in their leaves. 5. It is suggested that the activity of sucrose phosphate synthetase in leaves may play a role in the control of the rate of photosynthesis. PMID:16742569

  10. Pesticide residue analysis in parsley, lettuce and spinach by LC-MS/MS.

    PubMed

    Esturk, Okan; Yakar, Yasin; Ayhan, Zehra

    2014-03-01

    In this study, pesticide residues in parsley, lettuce and spinach (120 samples) were analyzed by the application of liquid chromatography-tandem mass spectrometry (LC-MS/MS). All samples of spinach, parsley or lettuce contained residues of three or more active substances. In parsley, carbendazim (100.0%), dichlorvos (100.0%), fenarimol (40.0%), pendimethalin (95.0%), in lettuce, diazinon (30.0%), dichlorvos (100.0%), pendimethalin (92.5%) phenthoate (12.5%), and in spinach, carbendazim (45.0%), cymoxanil (85.0%), dichlorvos (100.0%) and fenarimol (85.0%) were the significant active compounds. The maximum residue limits were exceeded in 28, 20 and 40 samples of parsley, lettuce and spinach, respectively. The results showed that there was a high occurrence of pesticide residues in parsley, lettuce and spinach samples from Hatay province, in which most of them were prohibited from use in Turkey for these vegetables. The contamination levels of these residues may be considered a serious public health problem according to the maximum residue limits (MRLs) of Turkey and the European Union (EU). PMID:24587520

  11. Shelf life and sensory characteristics of baby spinach subjected to electron beam irradiation.

    PubMed

    Neal, Jack A; Booren, Betsy; Cisneros-Zevallos, Luis; Miller, Rhonda K; Lucia, Lisa M; Maxim, Joseph E; Castillo, Alejandro

    2010-08-01

    The use of ionizing radiation for the control of foodborne pathogens and extending the shelf life of fresh iceberg lettuce and fresh spinach has recently been approved by the U.S. Food and Drug Administration. The efficacy of electron beam irradiation for controlling foodborne pathogens has been reported. For this experiment, the effectiveness of electron beam irradiation on the microbiological and sensory characteristics of fresh spinach was studied. Total aerobic plate counts were reduced by 2.6 and 3.2 log CFU/g at 0.7 and 1.4 kGy, respectively. Lactic acid bacteria were reduced at both doses of e-beam but grew slowly over the 35 d of the experiment. Yeasts and molds were not reduced in samples exposed to 0.7 kGy whereas 1.4 kGy significantly reduced microbial counts. Gas compositions (O(2) and CO(2)) were significantly different than controls. Oxygen levels inside the spinach sample bags decreased over time; however, O(2) levels did not drop below 1% that can induce anaerobic fermentation. CO(2) levels for all treatments increased through day 4; yet 7 d after irradiation, CO(2) level differences were not significant in both control and irradiated samples. Irradiation dose did not affect the basic tastes, aromatics, or mouth feels of fresh spinach, however; hardness attributes decreased as irradiated dose increased and slimy attributes of fresh spinach were higher in control samples compared to irradiated samples.

  12. An Improved Method for the Extraction and Thin-Layer Chromatography of Chlorophyll A and B from Spinach

    ERIC Educational Resources Information Center

    Quach, Hao T.; Steeper, Robert L.; Griffin, William G.

    2004-01-01

    A simple and fast method, which resolves chlorophyll a and b from spinach leaves on analytical plates while minimizing the appearance of chlorophyll degradation products is shown. An improved mobile phase for the Thin-layer chromatographic analysis of spinach extract that allows for the complete resolution of the common plant pigments found in…

  13. Quality of fresh-cut iceberg lettuce and spinach irradiated at doses up to 4kGy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to investigate radiation tolerance of fresh-cut Iceberg lettuce and spinach. Fresh-cut Iceberg lettuce packaged in modified atmosphere packages and spinach in perforated film bags were irradiated with gamma rays at doses of 0, 1, 2, 3, and 4 kGy. After irradiation, the sam...

  14. Evaluation of Postharvest Washing on Removal of Silver Nanoparticles (AgNPs) from Spinach Leaves.

    PubMed

    Zhang, Zhiyun; Guo, Huiyuan; Carlisle, Thomas; Mukherjee, Arnab; Kinchla, Amanda; White, Jason C; Xing, Baoshan; He, Lili

    2016-09-21

    There is increasing use of silver nanoparticles (AgNPs) as pesticides for fruits and vegetables due to the particles' unique antimicrobial and insecticidal properties. However, residual AgNPs in harvested produce may transfer through the food chain and pose a potential risk to public health. The objective of this study is to determine whether postharvest washing can effectively remove AgNPs that had accumulated on fresh produce. Ten microliters of commercially available 40 nm citrate coated AgNPs (0.4 mg/L) was dropped to a (1 × 1 cm(2)) spot on spinach leaves, followed by washing with deionized water (DI water), Tsunami 100 (80 mg/L), or Clorox bleach (200 mg/L). Then, the AgNP removal efficiency of the three treatments was evaluated by surface-enhanced Raman spectroscopy (SERS), scanning electron microscopy (SEM)-energy dispersive spectrometry (EDS), and inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS results showed that deionized water removed statistically insignificant amounts of total Ag (5%), whereas Tsunami 100 and Clorox bleach yielded 21 and 10% decreases in total Ag, respectively (P < 0.05). The increased removal efficiency resulted from AgNP dissolution and Ag(+) release upon contact with the oxidizing agents in Tsunami 100 (peroxyacetic acid, hydrogen peroxide) and Clorox bleach (sodium hypochlorite). According to the SERS results, the deionized water and Tsunami 100 treatments removed nonsignificant amounts of AgNPs. Clorox bleach decreased Ag NPs by >90% (P < 0.05); however, SEM-EDS images revealed the formation of large silver chloride (AgCl) crystals (162 ± 51 nm) on the leaf, which explained the low total Ag removal from ICP-MS. This study indicates current factory washing methods for fresh produce may not be effective in reducing AgNPs (by water and Tsunami 100) and total Ag (by all three means). This highlights the necessity to develop an efficient washing method for NP removal from food surfaces in the future. PMID:27548506

  15. Evaluation of Postharvest Washing on Removal of Silver Nanoparticles (AgNPs) from Spinach Leaves.

    PubMed

    Zhang, Zhiyun; Guo, Huiyuan; Carlisle, Thomas; Mukherjee, Arnab; Kinchla, Amanda; White, Jason C; Xing, Baoshan; He, Lili

    2016-09-21

    There is increasing use of silver nanoparticles (AgNPs) as pesticides for fruits and vegetables due to the particles' unique antimicrobial and insecticidal properties. However, residual AgNPs in harvested produce may transfer through the food chain and pose a potential risk to public health. The objective of this study is to determine whether postharvest washing can effectively remove AgNPs that had accumulated on fresh produce. Ten microliters of commercially available 40 nm citrate coated AgNPs (0.4 mg/L) was dropped to a (1 × 1 cm(2)) spot on spinach leaves, followed by washing with deionized water (DI water), Tsunami 100 (80 mg/L), or Clorox bleach (200 mg/L). Then, the AgNP removal efficiency of the three treatments was evaluated by surface-enhanced Raman spectroscopy (SERS), scanning electron microscopy (SEM)-energy dispersive spectrometry (EDS), and inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS results showed that deionized water removed statistically insignificant amounts of total Ag (5%), whereas Tsunami 100 and Clorox bleach yielded 21 and 10% decreases in total Ag, respectively (P < 0.05). The increased removal efficiency resulted from AgNP dissolution and Ag(+) release upon contact with the oxidizing agents in Tsunami 100 (peroxyacetic acid, hydrogen peroxide) and Clorox bleach (sodium hypochlorite). According to the SERS results, the deionized water and Tsunami 100 treatments removed nonsignificant amounts of AgNPs. Clorox bleach decreased Ag NPs by >90% (P < 0.05); however, SEM-EDS images revealed the formation of large silver chloride (AgCl) crystals (162 ± 51 nm) on the leaf, which explained the low total Ag removal from ICP-MS. This study indicates current factory washing methods for fresh produce may not be effective in reducing AgNPs (by water and Tsunami 100) and total Ag (by all three means). This highlights the necessity to develop an efficient washing method for NP removal from food surfaces in the future.

  16. iSpinach: a fluorogenic RNA aptamer optimized for in vitro applications

    PubMed Central

    Autour, Alexis; Westhof, Eric; Ryckelynck, Michael

    2016-01-01

    Using random mutagenesis and high throughput screening by microfluidic-assisted In Vitro Compartmentalization, we report the isolation of an order of magnitude times brighter mutants of the light-up RNA aptamers Spinach that are far less salt-sensitive and with a much higher thermal stability than the parent molecule. Further engineering gave iSpinach, a molecule with folding and fluorescence properties surpassing those of all currently known aptamer based on the fluorogenic co-factor 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI). We illustrate the potential of iSpinach in a new sensitive and high throughput-compatible fluorogenic assay that measures co-transcriptionally the catalytic constant (kcat) of a model ribozyme. PMID:26932363

  17. Systematic reconstruction of binding and stability landscapes of the fluorogenic aptamer spinach

    PubMed Central

    Ketterer, Simon; Fuchs, David; Weber, Wilfried; Meier, Matthias

    2015-01-01

    Fluorogenic RNAs that are based on the complex formed by 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) derivatives and the RNA aptamer named Spinach were used to engineer a new generation of in vitro and in vivo sensors for bioanalytics. With the resolved crystal structure of the RNA/small molecule complex, the engineering map becomes available, but comprehensive information regarding the thermodynamic profile of the molecule is missing. Here, we reconstructed the full thermodynamic binding and stability landscapes between DFHBI and a truncated sequence of first-generation Spinach. For this purpose, we established a systematic screening procedure for single- and double-point mutations on a microfluidic large-scale integrated chip platform for 87-nt long RNAs. The thermodynamic profile with single base resolution was used to engineer an improved fluorogenic spinach generation via a directed rather than evolutional approach. PMID:26400180

  18. Natural variation of folate content and composition in spinach (Spinacia oleracea) germplasm.

    PubMed

    Shohag, M J I; Wei, Yan-yan; Yu, Ning; Zhang, Jie; Wang, Kai; Patring, Johan; He, Zhen-li; Yang, Xiao-e

    2011-12-14

    Breeding to increase folate levels in edible parts of plants, termed folate biofortification, is an economical approach to fight against folate deficiency in humans, especially in the developing world. Germplasm with elevated folates are a useful genetic source for both breeding and direct use. Spinach is one of the well-know vegetables that contains a relatively high amount of folate. Currently, little is known about how much folate, and their composition varies in different spinach accessions. The aim of this study was to investigate natural variation in the folate content and composition of spinach genotypes grown under controlled environmental conditions. The folate content and composition in 67 spinach accessions were collected from the United States Department of Agriculture (USDA) and Asian Vegetable Research and Development Center (AVRDC) germplasm collections according to their origin, grown under control conditions to screen for natural diversity. Folates were extracted by a monoenzyme treatment and analyzed by a validated liquid chromatography (LC) method. The total folate content ranged from 54.1 to 173.2 μg/100 g of fresh weight, with 3.2-fold variation, and was accession-dependent. Four spinach accessions (PI 499372, NSL 6095, PI 261787, and TOT7337-B) have been identified as enriched folate content over 150 μg/100 g of fresh weight. The folate forms found were H(4)-folate, 5-CH(3)-H(4)-folate, and 5-HCO-H(4)-folate, and 10-CHO-folic acid also varied among different accessions and was responsible for variation in the total folate content. The major folate vitamer was represented by 5-CH(3)-H(4)-folate, which on average accounted for up to 52% of the total folate pool. The large variation in the total folate content and composition in diverse spinach accessions demonstrates the great genetic potential of diverse genotypes to be exploited by plant breeders.

  19. Nutritional Composition of Water Spinach (Ipomoea aquatica Forsk.) Leaves

    NASA Astrophysics Data System (ADS)

    Umar, K. J.; Hassan, L. G.; Dangoggo, S. M.; Ladan, M. J.

    Analyses of the nutritional composition of water spinach (Ipomoea aquatica) Forsk leaves were carried out using standard methods of food analysis. The proximate composition as well as mineral elements were determined. The leaves were found on dry weight basis to have high moisture (72.83±0.29%), ash (10.83±0.80%), crude lipid (11.00±0.50%), crude fibre (17.67±0.35%) and available carbohydrate (54.20±0.68%), but low in crude protein content (6.30±0.27%). The leaves also have energy value (300.94±5.31 kcal/100 g) that is within the range reported in some Nigerian leafy vegetables. The mineral element contents were high with remarkable concentration of K (5,458.33±954.70 mg/100 g) and Fe (210.30±2.47 mg/100 g). Also the leaves content moderate concentrations of Na (135.00±2.50 mg/100 g), calcium (416.70±5.77 mg/100 g), Magnesium (301.64±12.69 mg/100 g) and P (109.29±0.55 mg/100 g), with low Cu (0.36±0.01 mg/100 g), Mn (2.14±0.22 mg/100 g) and Zn (2.47±0.27 mg/100 g) contents. Comparing the mineral content with recommended dietary allowance, it was showed that the plant leaves is good sources of K, Mn and Fe for all categories of people, while Mg is adequate enough for adult female and children. From the result, Ipomoea aquatica Forsk leaves could be used for nutritional purposes, due to the amount and diversity of nutrients it contains.

  20. Availability to rats of iron from spinach: Effects of oxalic acid.

    PubMed

    Van Campen, D R; Welch, R M

    1980-08-01

    The availability to rats of iron from two varieties of spinach was determined. Also, the absorption of Fe was compared between FeCl3 and Fe-oxalate and the effects of adding 0.75% oxalate to the diet were determined. Absorption of iron from both varieties of spinach was comparable to that from FeCl3 and the iron was equally available from Fe-oxalate and FeCl3. The addition of 0.75% oxalic acid to the diet did not depress iron absorption and, if anything, appeared to enhance iron utilization by rats.

  1. Potential anticancer effect of red spinach (Amaranthus gangeticus) extract.

    PubMed

    Sani, Huzaimah Abdullah; Rahmat, Asmah; Ismail, Maznah; Rosli, Rozita; Endrini, Susi

    2004-01-01

    The objective of this study was to determine the anti cancer effects of red spinach (Amaranthus gangeticus Linn) in vitro and in vivo. For in vitro study, microtitration cytotoxic assay was done using 3-(4,5-dimethylthiazol-2-il)-2,5-diphenil tetrazolium bromide (MTT) kit assay. Results showed that aqueous extract of A gangeticus inhibited the proliferation of liver cancer cell line (HepG2) and breast cancer cell line (MCF-7). The IC(50) values were 93.8 mu g/ml and 98.8 mu g/ml for HepG2 and MCF-7, respectively. The inhibitory effect was also observed in colon cancer cell line (Caco-2), but a lower percentage compared to HepG2 and MCF-7. For normal cell line (Chang Liver), there was no inhibitory effect. In the in vivo study, hepatocarcinogenesis was monitored in rats according to Solt and Farber (1976) without partial hepatectomy. Assay of tumour marker enzymes such as glutathione S-transferase (GST), gamma-glutamyl transpeptidase (GGT), uridyl diphosphoglucuronyl transferase (UDPGT) and alkaline phosphatase (ALP) were carried out to determine the severity of hepatocarcinogenesis. The result found that supplementation of 5%, 7.5% and 10% of A. gangeticus aqueous extract to normal rats did not show any significant difference towards normal control (P <0.05). The exposure of the rats to chemical carcinogens diethylnitrosamine (DEN) and 2-acetylaminofluorene (AAF) showed a significant increase in specific enzyme activity of GGT, GST, UDPGT and ALP compared to normal control (P <0.05). However, it was found that the supplementation of A. gangeticus aqueous extract in 5%, 7.5% and 10% to cancer-induced rats could inhibit the activity of all tumour marker enzymes especially at 10% (P <0.05). Supplementation of anti cancer drug glycyrrhizin at suggested dose (0.005%) did not show any suppressive effect towards cancer control (P <0.05). In conclusion, A. gangeticus showed anticancer potential in in vitro and in vivo studies.

  2. Spectrophotometric studies on alkaline isomerization of spinach ferredoxin.

    PubMed

    Hasumi, H; Nagata, E; Nakamura, S

    1985-10-01

    The gross protein structure, the microenvironment of the iron-sulfur cluster, and the effect of neutral salts on the molecular structure of spinach ferredoxin were studied by CD and absorption spectroscopy in the alkaline pH range. In the pH range of 7-11, the existence of reversible isomerization which consisted of at least two proton dissociation processes was indicated by the statical CD and absorption spectra. The CD changes in the visible and far-UV regions were dramatic upon elevation of the pH from neutral to alkaline, indicating a significant alteration of the microenvironment of the cluster and a decrease in the ordered secondary structures. The absorption change in the visible region due to pH elevation was small but clearly observed with a high signal-to-noise ratio. The numbers of protons involved in the respective processes and the apparent pK values obtained from the pH-dependence of the CD changes were in good agreement with those obtained from the pH-dependence of the absorption changes in the visible region. In addition, the rate constants obtained from the time courses of the CD and absorption changes agreed with one another. By the addition of 1 M NaCl, the CD and absorption spectra at alkaline pH were reversed almost to those at neutral pH without significant pH change. On the other hand, above pH 11, ferredoxin was found to be irreversibly denatured. Based on analyses of the statical CD and absorption spectra and of the time courses of the CD changes, the probable mechanism of the isomerization was considered to be as follows: (Formula: see text) where H stands for a proton, N-form for native ferredoxin at neutral pH, N*-form for alkaline ferredoxin below pH 11 which still has the iron-sulfur cluster but with disordered secondary structures of the polypeptide chain, and D-form for completely denatured ferredoxin above pH 11. These results lead to the conclusions that (1) the interaction between the protein moiety and the iron-sulfur cluster is

  3. Differences in biofilm formation of produce and poultry Salmonella enterica isolates and their persistence on spinach plants.

    PubMed

    Patel, Jitendra; Singh, Manpreet; Macarisin, Dumitru; Sharma, Manan; Shelton, Daniel

    2013-12-01

    Spinach plants were irrigated biweekly with water containing 2.1 log CFU Salmonella/100 ml water (the maximum Escherichia coli MPN recommended by the Leafy Greens Marketing Agreement; LGMA), or 4.1 CFU Salmonella/100 ml water to determine Salmonella persistence on spinach leaves. Green Fluorescent protein expressing Salmonella were undetectable by most-probable number (MPN) at 24 h and 7 days following each irrigation event. This study indicates that Salmonella are unlikely to persist on spinach leaves when irrigation water is contaminated at a level below the LGMA standards. In a parallel study, persistence of Salmonella isolated from poultry or produce was compared following biweekly irrigation of spinach plants with water containing 6 log CFU Salmonella/100 ml. Produce Salmonella isolates formed greater biofilms on polystyrene, polycarbonate and stainless steel surfaces and persisted at significantly higher numbers on spinach leaves than those Salmonella from poultry origin during 35 days study. Poultry Salmonella isolates were undetectable (<1 log CFU/g) on spinach plants 7 days following each irrigation event when assayed by direct plating. This study indicates that Salmonella persistence on spinach leaves is affected by the source of contamination and the biofilm forming ability of the strain. PMID:24010621

  4. Isolation of a cDNA clone for spinach lipid transfer protein and evidence that the protein is synthesized by the secretory pathway

    SciTech Connect

    Bernhard, W.R.; Thoma, S.; Botella, J.; Somerville, C.R. )

    1991-01-01

    A cDNA clone encoding a nonspecific lipid transfer protein from spinach (Spinacia oleracea) was isolated by probing a library with synthetic oligonucleotides based on the amino acid sequence of the protein. Determination of the DNA sequence indicated a 354-nucleotide open reading frame which encodes a 118-amino acid residue polypeptide. The first 26 amino acids of the open reading frame, which are not present in the mature protein, have all the characteristics of a signal sequence which is normally associated with the synthesis of membrane proteins or secreted proteins. In vitro transcription of the cDNA and translation in the presence of canine pancreatic microsomes or microsomes from cultured maize endosperm cells indicated that proteolytic processing of the preprotein to the mature form was associated with cotranslational insertion into the microsomal membranes. Because there is no known mechanism by which the polypeptide could be transferred from the microsomal membranes to the cytoplasm, the proposed role of this protein in catalyzing lipid transfer between intracellular membranes is in doubt. Although the lipid transfer protein is one of the most abundant proteins in leaf cells, the results of genomic Southern analysis were consistent with the presence of only one gene. Analysis of the level of mRNA by Northern blotting indicated that the transcript was several-fold more abundant than an actin transcript in leaf and petiole tissue, but was present in roots at less than 1% of the level in petioles.

  5. Multispectral fluorescence imaging for detection of bovine feces on Romaine lettuce and baby spinach leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hyperspectral fluorescence imaging with ultraviolet-A excitation was used to evaluate the feasibility of two-waveband fluorescence algorithms for the detection of bovine fecal contaminants on the abaxial and adaxial surfaces of Romaine lettuce and baby spinach leaves. Correlation analysis was used t...

  6. Perchlorate uptake in spinach as related to perchlorate, nitrate and chloride concentrations in irrigation water

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several studies have reported on the detection of perchlorate in edible leafy vegetables irrigated with Colorado River water. However, there is no information on spinach as related to perchlorate in irrigation water nor on the effect of other anions on perchlorate uptake. A greenhouse perchlorate up...

  7. Acute Effects of a Spinach Extract Rich in Thylakoids on Satiety: A Randomized Controlled Crossover Trial

    PubMed Central

    Rebello, Candida J.; Chu, Jessica; Beyl, Robbie; Edwall, Dan; Erlanson-Albertsson, Charlotte; Greenway, Frank L.

    2015-01-01

    Objective: By retarding fat digestion, thylakoids, the internal photosynthetic membrane system of green plants, promote the release of satiety hormones. This study examined the effect of consuming a single dose of concentrated extract of thylakoids from spinach on satiety, food intake, lipids, and glucose compared to a placebo. Design: Sixty overweight and obese individuals enrolled in a double-blind randomized crossover study consumed the spinach extract or placebo in random order at least a week apart. Blood was drawn for assessments of lipids and glucose before a standard breakfast meal, followed 4 hours later by a 5 g dose of the extract and a standard lunch. Visual analog scales were administered before lunch and at intervals until an ad libitum pizza dinner served 4 hours later. Two hours after lunch a second blood draw was conducted. Mixed models were used to analyze response changes. Results: Compared to placebo, consuming the spinach extract reduced hunger (p < 0.01) and longing for food over 2 hours (p < 0.01) and increased postprandial plasma glucose concentrations (p < 0.01). There were no differences in plasma lipids and energy intake at dinner, but males showed a trend toward decreased energy intake (p = 0.08). Conclusions: At this dose, the spinach extract containing thylakoids increases satiety over a 2-hour period compared to a placebo. Thylakoid consumption may influence gender-specific food cravings. PMID:26029978

  8. Microdissection and painting of the Y chromosome in spinach (Spinacia oleracea).

    PubMed

    Deng, Chuan-Liang; Qin, Rui-Yun; Cao, Ying; Gao, Jun; Li, Shu-Fen; Gao, Wu-Jun; Lu, Long-Dou

    2013-07-01

    Spinach has long been used as a model for genetic and physiological studies of sex determination and expression. Although trisomic analysis from a cross between diploid and triploid plants identified the XY chromosome as the largest chromosome, no direct evidence has been provided to support this at the molecular level. In this study, the largest chromosomes of spinach from mitotic metaphase spreads were microdissected using glass needles. Degenerate oligonucleotide primed polymerase chain reaction was used to amplify the dissected chromosomes. The amplified products from the Y chromosome were identified using the male-specific marker T11A. For the first time, the largest spinach chromosome was confirmed to be a sex chromosome at the molecular level. PCR products from the isolated chromosomes were used in an in situ probe mixture for painting the Y chromosome. The fluorescence signals were mainly distributed on all chromosomes and four pair of weaker punctate fluorescence signal sites were observed on the terminal region of two pair of autosomes. These findings provide a foundation for the study of sex chromosome evolution in spinach.

  9. Polycations Globally Enhance Binding of 14-3-3 omega to Target Proteins in Spinach Leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The binding of 14-3-3' to phosphorylated NR (pNR) is stimulated by cations such as Mg2+ or spermine, and decreased by 5'-AMP. In order to determine whether binding to other cellular proteins is affected similarly, Far-Western overlays of extracts prepared from light- or dark-treated spinach (Spinac...

  10. Optimization of an installation angle of a root-cutting blade for an automatic spinach harvester

    NASA Astrophysics Data System (ADS)

    Fujisawa, A.; Chida, Y.

    2016-09-01

    This paper presents an optimization of the installation angle of a root-cutting blade relative to the arm of an automatic spinach harvester. In the harvesting operation, the blade, which is a rigid body, moves under the planted rows in soil of powder consistency to cut the roots of the spinach and to harvest the spinach on a conveyor. Therefore, the interaction between a rigid body and powder is an important consideration. Experiments were conducted on the design of the harvester. The experiments revealed that a certain path of the blade is more favorable for both harvesting spinach easily and minimizing the amount of soil removed by the blade. In this paper, without revising the favorable path, the optimum installation angle of the blade is derived. To derive the installation angle, a nonlinear optimization problem is solved as an evaluation function consisting of the volume of soil pushed by the blade and the installation angle, which is a design parameter. The utility of the installation angle is confirmed by the Discrete Element Method (DEM), which analyzes the interaction between a rigid body and powder.

  11. Responses of spinach to salinity and nutrient deficiency in growth, physiology and nutritional value

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salinity and nutrient depleted soil are major constraints to crop production, especially for vegetable crops. The effects of salinity and nutrient deficiency on spinach were evaluated in sand cultures under greenhouse conditions. Plants were watered every day with Hoagland nutrition solution, depriv...

  12. SNP association analysis of resistance to Verticillium wilt (Verticillium dahliae Kleb.) in spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Verticillium wilt, caused by Verticillium dahliae Kleb. is one of important fungus diseases in spinach (Spinacia oleracea L.) and the most economical method of control this disease is through the use of genetic resistance, especially for organic growers. The objective of this research is to evaluate...

  13. Adherence of curli producing Shiga-toxigenic Escherichia coli to baby spinach leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cellular appendages, such as curli fibers have been suggested to be involved in STEC persistence in fresh produce as these curli are critical in biofilm formation and adherence to animal cells. We determined the role of curli in attachment of STEC on spinach leaves. The curli expression by wild-ty...

  14. Internalization of E. coli O157:H7 in spinach cultivated in soil and hydroponic media

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Internalization of E. coli O157:H7 into spinach plants through root uptake is a potential route of contamination. Previous studies that have investigated uptake of E. coli O157:H7 into leafy greens have expressed green fluorescent protein (gfp) from a plasmid, possibly limiting detecti...

  15. Iron bioavailability of rats fed liver, lentil, spinach and their mixtures.

    PubMed

    Rewashdeh, Abdullah Y; El-Qudah, Jafar M; Al-Dmoor, Hanee; Al-Qudah, Maisa M; Mamkagh, Amer M; Tarawneh, Khaled A; Hawari, Azmi D; Dababneh, Basem F; Al-Bakheit, Alaa A; Haddad, Moawya A

    2009-02-15

    To study the effects of dietary iron source (basal diet-FeSO4 x 7H2O, liver, lentil, spinach, liver + lentil, liver+spinach and lentil+spinach) on iron bioavailability, fifty-six Albino Sprague Dawley derived male 21 days old rats were fed on iron-deficient diet (7.8 mg Fe kg(-1) diet) and the mentioned seven iron containing diets (40 mg Fe kg(-1) diet) for 10 days. Rats fed liver diet showed higher iron apparent absorption (52.1%), hemoglobin (Hb) gain (0.94 g/100 mL), Hb-iron gain (1.2 mg), Hb-regeneration efficiency (HRE%) (50.8%), relative efficiency of HRE% (106.5%), packed cell volume gain (2.22%) and mean corpuscular hemoglobin concentration (0.64 g dL(-1)). Liver resulted in an increase in these parameters when mixed with lentil and spinach diets. However, rats fed iron free diet showed the higher dry matter absorption.

  16. Survival of Salmonella on spinach leaves treated with contaminated irrigation water

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Salmonella outbreaks have been associated with the consumption of fresh produce. The produce may be contaminated with Salmonella during on-farm contact with contaminated water. Transmission of Salmonella from contaminated irrigation water to spinach plants in growth chamber settings ...

  17. 24-epibrassinolide and 20-hydroxyecdysone affect photosynthesis differently in maize and spinach.

    PubMed

    Rothová, Olga; Holá, Dana; Kočová, Marie; Tůmová, Lenka; Hnilička, František; Hniličková, Helena; Kamlar, Marek; Macek, Tomáš

    2014-07-01

    The aim of the work was to examine the effect of brassinosteroid (24-epibrassinolide; 24E) and ecdysteroid (20-hydroxyecdysone; 20E) on various parts of primary photosynthetic processes in maize and spinach. Additionally, the effect of steroids on gaseous exchange, pigment content and biomass accumulation was studied. The efficiency of the photosynthetic whole electron-transport chain responded negatively to the 24E or 20E treatment in both species, but there were interspecific differences regarding Photosystem (PS) II response. A positive effect on its oxygen-evolving complex and a slightly better energetical connectivity between PSII units were observed in maize whereas the opposite was true for spinach. The size of the pool of the PSI end electron acceptors was usually diminished due to 24E or 20E treatment. The treatment of plants with 24E or 20E applied individually positively influenced the content of photosynthetic pigments in maize (not in spinach). On the other hand, it did not affect gaseous exchange in maize but resulted in its reduction in spinach. Plants treated with combination of both steroids mostly did not significantly differ from the control plants. We have demonstrated for the first time that 20E applied in low (10nM) concentration can affect various parts of photosynthetic processes similarly to 24E and that brassinosteroids regulate not only PSII but also other parts of the photosynthetic electron transport chain - but not necessarily in the same way.

  18. Genetic diversity and association analysis of leafminer (Liriomyza langei) resistance in spinach (Spinacia oleracea)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafminer (Liriomyza spp.) is a major insect pest of many important agricultural crops, including spinach (Spinacia oleracea). Use of genetic resistance is an efficient, economic and environment-friendly method to control this pest. The objective of this research was to conduct association analysis ...

  19. Using Spinach aptamer to correlate mRNA and protein levels in Escherichia coli.

    PubMed

    Pothoulakis, Georgios; Ellis, Tom

    2015-01-01

    In vivo gene expression measurements have traditionally relied on fluorescent proteins such as green fluorescent protein (GFP) with the help of high-sensitivity equipment such as flow cytometers. However, fluorescent proteins report only on the protein level inside the cell without giving direct information about messenger RNA (mRNA) production. In 2011, an aptamer termed Spinach was presented that acts as an RNA mimic of GFP when produced in Escherichia coli and mammalian cells. It was later shown that coexpression of a red fluorescent protein (mRFP1) and the Spinach aptamer, when included into the same gene expression cassette, could be utilized for parallel in vivo measurements of mRNA and protein production. As accurate characterization of component biological parts is becoming increasingly important for fields such as synthetic biology, Spinach in combination with mRFP1 provide a great tool for the characterization of promoters and ribosome binding sites. In this chapter, we discuss how live-cell imaging and flow cytometry can be used to detect and measure fluorescence produced in E. coli cells by different constructs that contain the Spinach aptamer and the mRFP1 gene.

  20. De novo and comparative transcriptome analysis of cultivated and wild spinach.

    PubMed

    Xu, Chenxi; Jiao, Chen; Zheng, Yi; Sun, Honghe; Liu, Wenli; Cai, Xiaofeng; Wang, Xiaoli; Liu, Shuang; Xu, Yimin; Mou, Beiquan; Dai, Shaojun; Fei, Zhangjun; Wang, Quanhua

    2015-12-04

    Spinach (Spinacia oleracea L.) is an economically important green leafy vegetable crop. In this study, we performed deep transcriptome sequencing for nine spinach accessions: three from cultivated S. oleracea, three from wild S. turkestanica and three from wild S. tetrandra. A total of approximately 100 million high-quality reads were generated, which were de novo assembled into 72,151 unigenes with a total length of 46.5 Mb. By comparing sequences of these unigenes against different protein databases, nearly 60% of them were annotated and 50% could be assigned with Gene Ontology terms. A total of 387 metabolic pathways were predicted from the assembled spinach unigenes. From the transcriptome sequencing data, we were able to identify a total of ~320,000 high-quality single nucleotide polymorphisms (SNPs). Phylogenetic analyses using SNPs as well as gene expression profiles indicated that S. turkestanica was more closely related to the cultivated S. oleracea than S. tetrandra. A large number of genes involved in responses to biotic and abiotic stresses were found to be differentially expressed between the cultivated and wild spinach. Finally, an interactive online database (http://www.spinachbase.org) was developed to allow the research community to efficiently retrieve, query, mine and analyze our transcriptome dataset.

  1. Tandem Spinach Array for mRNA Imaging in Living Bacterial Cells.

    PubMed

    Zhang, Jichuan; Fei, Jingyi; Leslie, Benjamin J; Han, Kyu Young; Kuhlman, Thomas E; Ha, Taekjip

    2015-11-27

    Live cell RNA imaging using genetically encoded fluorescent labels is an important tool for monitoring RNA activities. A recently reported RNA aptamer-fluorogen system, the Spinach, in which an RNA aptamer binds and induces the fluorescence of a GFP-like 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) ligand, can be readily tagged to the RNA of interest. Although the aptamer-fluorogen system is sufficient for imaging highly abundant non-coding RNAs (tRNAs, rRNAs, etc.), it performs poorly for mRNA imaging due to low brightness. In addition, whether the aptamer-fluorogen system may perturb the native RNA characteristics has not been systematically characterized at the levels of RNA transcription, translation and degradation. To increase the brightness of these aptamer-fluorogen systems, we constructed and tested tandem arrays containing multiple Spinach aptamers (8-64 aptamer repeats). Such arrays enhanced the brightness of the tagged mRNA molecules by up to ~17 fold in living cells. Strong laser excitation with pulsed illumination further increased the imaging sensitivity of Spinach array-tagged RNAs. Moreover, transcriptional fusion to the Spinach array did not affect mRNA transcription, translation or degradation, indicating that aptamer arrays might be a generalizable labeling method for high-performance and low-perturbation live cell RNA imaging.

  2. De novo and comparative transcriptome analysis of cultivated and wild spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study, we performed deep transcriptome sequencing for nine spinach (Spinacia oleracea L., 2n = 2× = 12) accessions, three from cultivated S. oleracea, three from wild S. turkestanica and three from wild S. tetrandra, using the Illumina sequencing technology. A total of approximately 100 mill...

  3. Evaluation of Oxalate Concentration in the U.S. Spinach Germplasm Collection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In addition to its high nutrient content, spinach (Spinacia oleracea L.) is also known to have greater amount of oxalic acid than most crops. Oxalic acid may form crystals with minerals to reduce the bioavailability and absorption of calcium and iron in diets, and calcium oxalate may deposit in the...

  4. Generic Escherichia coli Contamination of Spinach at the Preharvest Stage: Effects of Farm Management and Environmental Factors

    PubMed Central

    Navratil, Sarah; Gregory, Ashley; Bauer, Arin; Srinath, Indumathi; Jun, Mikyoung; Szonyi, Barbara; Nightingale, Kendra; Anciso, Juan; Ivanek, Renata

    2013-01-01

    The objective of this study was to determine the effects of farm management and environmental factors on preharvest spinach contamination with generic Escherichia coli as an indicator of fecal contamination. A repeated cross-sectional study was conducted by visiting spinach farms up to four times per growing season over a period of 2 years (2010 to 2011). Spinach samples (n = 955) were collected from 12 spinach farms in Colorado and Texas as representative states of the Western and Southwestern United States, respectively. During each farm visit, farmers were surveyed about farm-related management and environmental factors using a questionnaire. Associations between the prevalence of generic E. coli in spinach and farm-related factors were assessed by using a multivariable logistic regression model including random effects for farm and farm visit. Overall, 6.6% of spinach samples were positive for generic E. coli. Significant risk factors for spinach contamination with generic E. coli were the proximity (within 10 miles) of a poultry farm, the use of pond water for irrigation, a >66-day period since the planting of spinach, farming on fields previously used for grazing, the production of hay before spinach planting, and the farm location in the Southwestern United States. Contamination with generic E. coli was significantly reduced with an irrigation lapse time of >5 days as well as by several factors related to field workers, including the use of portable toilets, training to use portable toilets, and the use of hand-washing stations. To our knowledge, this is the first report of an association between field workers' personal hygiene and produce contamination with generic E. coli at the preharvest level. Collectively, our findings support that practice of good personal hygiene and other good farm management practices may reduce produce contamination with generic E. coli at the preharvest level. PMID:23666336

  5. Antioxidant Effects of Spinach (Spinacia oleracea L.) Supplementation in Hyperlipidemic Rats.

    PubMed

    Ko, Sang-Heui; Park, Jae-Hee; Kim, So-Yun; Lee, Seon Woo; Chun, Soon-Sil; Park, Eunju

    2014-01-01

    Increased consumption of fresh vegetables that are high in polyphenols has been associated with a reduced risk of oxidative stress-induced disease. The present study aimed to evaluate the antioxidant effects of spinach in vitro and in vivo in hyperlipidemic rats. For measurement of in vitro antioxidant activity, spinach was subjected to hot water extraction (WE) or ethanol extraction (EE) and examined for total polyphenol content (TPC), oxygen radical absorbance capacity (ORAC), cellular antioxidant activity (CAA), and antigenotoxic activity. The in vivo antioxidant activity of spinach was assessed using blood and liver lipid profiles and antioxidant status in rats fed a high fat-cholesterol diet (HFCD) for 6 weeks. The TPC of WE and EE were shown as 1.5±0.0 and 0.5±0.0 mg GAE/g, respectively. Increasing the concentration of the extracts resulted in increased ORAC value, CAA, and antigenotoxic activity for all extracts tested. HFCD-fed rats displayed hyperlipidemia and increased oxidative stress, as indicated by a significant rise in blood and liver lipid profiles, an increase in plasma conjugated diene concentration, an increase in liver thiobarbituric acid reactive substances (TBARS) level, and a significant decrease in manganese superoxide dismutase (Mn-SOD) activity compared with rats fed normal diet. However, administration of 5% spinach showed a beneficial effect in HFCD rats, as indicated by decreased liver TBARS level and DNA damage in leukocyte and increased plasma conjugated dienes and Mn-SOD activity. Thus, the antioxidant activity of spinach may be an effective way to ameliorate high fat and cholesterol diet-induced oxidative stress.

  6. Growing and processing conditions lead to changes in the carotenoid profile of spinach.

    PubMed

    Heymann, Thomas; Westphal, Lore; Wessjohann, Ludger; Glomb, Marcus A

    2014-05-28

    This study aimed to evaluate the influence of different light regimens during spinach cultivation on the isomeric composition of β-carotene. Irradiation with a halogen lamp, which has a wavelength spectrum close to that of daylight, was used to mimic field-grown conditions. The additional use of optical filters was established as a model system for greenhouse cultivation. Field-grown model systems led to a preferential increase of 9-cis-β-carotene, whereas 13-cis-β-carotene was just formed at the beginning of irradiation. Additionally 9,13-di-cis-β-carotene decreased significantly in the presence of energy-rich light. Isomerization of β-carotene was strongly suppressed during irradiation in greenhouse-grown model systems and led to significant differences. These results were verified in biological samples. Authentic field-grown spinach (Spinacia oleracea L.) showed among changes of other isomers a significantly higher level of 9-cis-isomers (7.52 ± 0.14%) and a significantly lower level of 9,13-di-cis-isomers (0.25 ± 0.03%) compared to authentic greenhouse-grown spinach (6.49 ± 0.11 and 0.76 ± 0.05%). Almost all analyzed commercial spinach samples (fresh and frozen) were identified as common field-grown cultivation. Further investigations resulted in a clear differentiation of frozen commercial samples from fresh spinach, caused by significantly higher levels of 13-cis- and 15-cis-β-carotene as a result of industrial blanching processes.

  7. Antioxidant Effects of Spinach (Spinacia oleracea L.) Supplementation in Hyperlipidemic Rats

    PubMed Central

    Ko, Sang-Heui; Park, Jae-Hee; Kim, So-Yun; Lee, Seon Woo; Chun, Soon-Sil; Park, Eunju

    2014-01-01

    Increased consumption of fresh vegetables that are high in polyphenols has been associated with a reduced risk of oxidative stress-induced disease. The present study aimed to evaluate the antioxidant effects of spinach in vitro and in vivo in hyperlipidemic rats. For measurement of in vitro antioxidant activity, spinach was subjected to hot water extraction (WE) or ethanol extraction (EE) and examined for total polyphenol content (TPC), oxygen radical absorbance capacity (ORAC), cellular antioxidant activity (CAA), and antigenotoxic activity. The in vivo antioxidant activity of spinach was assessed using blood and liver lipid profiles and antioxidant status in rats fed a high fat-cholesterol diet (HFCD) for 6 weeks. The TPC of WE and EE were shown as 1.5±0.0 and 0.5±0.0 mg GAE/g, respectively. Increasing the concentration of the extracts resulted in increased ORAC value, CAA, and antigenotoxic activity for all extracts tested. HFCD-fed rats displayed hyperlipidemia and increased oxidative stress, as indicated by a significant rise in blood and liver lipid profiles, an increase in plasma conjugated diene concentration, an increase in liver thiobarbituric acid reactive substances (TBARS) level, and a significant decrease in manganese superoxide dismutase (Mn-SOD) activity compared with rats fed normal diet. However, administration of 5% spinach showed a beneficial effect in HFCD rats, as indicated by decreased liver TBARS level and DNA damage in leukocyte and increased plasma conjugated dienes and Mn-SOD activity. Thus, the antioxidant activity of spinach may be an effective way to ameliorate high fat and cholesterol diet-induced oxidative stress. PMID:24772405

  8. Effect of γ-irradiation on the thermomechanical and morphological properties of chitosan obtained from prawn shell: Evaluation of potential for irradiated chitosan as plant growth stimulator for Malabar spinach

    NASA Astrophysics Data System (ADS)

    Rahman, Mohammed Mizanur; Kabir, Shahriar; Rashid, Taslim Ur; Nesa, Bodrun; Nasrin, Romana; Haque, Papia; Khan, Mubarak A.

    2013-01-01

    In the present study we have synthesized chitosan from waste prawn shell via γ-irradiation of chitin and subsequent alkaline treatment. The detailed experimental studies demonstrated that nonirradiated chitin deacetylated by 40% NaOH solution showed 72% degree of deacetylation (DD), however 50 kGy irradiated chitin, deacetylated by 20% NaOH demonstrated 81.5% DD. Chitosan in solid state as obtained from γ-irradiation of chitin was further irradiated by different doses (2-100 kGy) of gamma irradiation and the effects of irradiation on the molecular weight, thermo-mechanical by differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA), and antimicrobial properties were evaluated with respect to nonirradiated chitosan sample. Gamma irradiation of chitosan with a dose of 100 kGy caused a decrease in average molecular weight from 1.9×105 to 6.5×104 Da and thus increased its solubility in water. Nonirradiated and γ-irradiated chitosan at concentration 1% (w/w) in water were prepared and used to evaluate of its potentiality for growth stimulation of Malabar spinach. The chitosan solution was sprayed on the specimen plants and neighboring soil where germinations were taken place and various plant growth parameters such as plant height, number of leaves, leaf areas, dry and wet weight of the plants and roots were investigated. The details study revealed that application of 30 kGy irradiated chitosan yielded 60% higher growth of the Malabar spinach than that obtained from nonirradiated chitosan. The data are consistent with preliminary results from field experiments and unambiguously confirms that a minor amount of chitosan has a profound effect on the growth and development of Malabar spinach.

  9. Structure and Stability of the Spinach Aquaporin SoPIP2;1 in Detergent Micelles and Lipid Membranes

    PubMed Central

    Plasencia, Inés; Survery, Sabeen; Ibragimova, Sania; Hansen, Jesper S.; Kjellbom, Per; Helix-Nielsen, Claus; Johanson, Urban; Mouritsen, Ole G.

    2011-01-01

    Background SoPIP2;1 constitutes one of the major integral proteins in spinach leaf plasma membranes and belongs to the aquaporin family. SoPIP2;1 is a highly permeable and selective water channel that has been successfully overexpressed and purified with high yields. In order to optimize reconstitution of the purified protein into biomimetic systems, we have here for the first time characterized the structural stability of SoPIP2;1. Methodology/Principal Finding We have characterized the protein structural stability after purification and after reconstitution into detergent micelles and proteoliposomes using circular dichroism and fluorescence spectroscopy techniques. The structure of SoPIP2;1 was analyzed either with the protein solubilized with octyl-β-D-glucopyranoside (OG) or reconstituted into lipid membranes formed by E. coli lipids, diphytanoylphosphatidylcholine (DPhPC), or reconstituted into lipid membranes formed from mixtures of 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPE), 1-palmitoyl-2oleoyl-phosphatidylethanolamine (POPE), 1-palmitoyl-2-oleoyl-phosphatidylserine (POPS), and ergosterol. Generally, SoPIP2;1 secondary structure was found to be predominantly α-helical in accordance with crystallographic data. The protein has a high thermal structural stability in detergent solutions, with an irreversible thermal unfolding occurring at a melting temperature of 58°C. Incorporation of the protein into lipid membranes increases the structural stability as evidenced by an increased melting temperature of up to 70°C. Conclusion/Significance The results of this study provide insights into SoPIP2;1 stability in various host membranes and suggest suitable choices of detergent and lipid composition for reconstitution of SoPIP2;1 into biomimetic membranes for biotechnological applications. PMID:21339815

  10. Spatial and Temporal Factors Associated with an Increased Prevalence of Listeria monocytogenes in Spinach Fields in New York State

    PubMed Central

    Weller, Daniel; Wiedmann, Martin

    2015-01-01

    While rain and irrigation events have been associated with an increased prevalence of foodborne pathogens in produce production environments, quantitative data are needed to determine the effects of various spatial and temporal factors on the risk of produce contamination following these events. This study was performed to quantify these effects and to determine the impact of rain and irrigation events on the detection frequency and diversity of Listeria species (including L. monocytogenes) and L. monocytogenes in produce fields. Two spinach fields, with high and low predicted risks of L. monocytogenes isolation, were sampled 24, 48, 72, and 144 to 192 h following irrigation and rain events. Predicted risk was a function of the field's proximity to water and roads. Factors were evaluated for their association with Listeria species and L. monocytogenes isolation by using generalized linear mixed models (GLMMs). In total, 1,492 (1,092 soil, 334 leaf, 14 fecal, and 52 water) samples were collected. According to the GLMM, the likelihood of Listeria species and L. monocytogenes isolation from soil samples was highest during the 24 h immediately following an event (odds ratios [ORs] of 7.7 and 25, respectively). Additionally, Listeria species and L. monocytogenes isolates associated with irrigation events showed significantly lower sigB allele type diversity than did isolates associated with precipitation events (P = <0.001), suggesting that irrigation water may be a point source of L. monocytogenes contamination. Small changes in management practices (e.g., not irrigating fields before harvest) may therefore reduce the risk of L. monocytogenes contamination of fresh produce. PMID:26116668

  11. Spatial and Temporal Factors Associated with an Increased Prevalence of Listeria monocytogenes in Spinach Fields in New York State.

    PubMed

    Weller, Daniel; Wiedmann, Martin; Strawn, Laura K

    2015-09-01

    While rain and irrigation events have been associated with an increased prevalence of foodborne pathogens in produce production environments, quantitative data are needed to determine the effects of various spatial and temporal factors on the risk of produce contamination following these events. This study was performed to quantify these effects and to determine the impact of rain and irrigation events on the detection frequency and diversity of Listeria species (including L. monocytogenes) and L. monocytogenes in produce fields. Two spinach fields, with high and low predicted risks of L. monocytogenes isolation, were sampled 24, 48, 72, and 144 to 192 h following irrigation and rain events. Predicted risk was a function of the field's proximity to water and roads. Factors were evaluated for their association with Listeria species and L. monocytogenes isolation by using generalized linear mixed models (GLMMs). In total, 1,492 (1,092 soil, 334 leaf, 14 fecal, and 52 water) samples were collected. According to the GLMM, the likelihood of Listeria species and L. monocytogenes isolation from soil samples was highest during the 24 h immediately following an event (odds ratios [ORs] of 7.7 and 25, respectively). Additionally, Listeria species and L. monocytogenes isolates associated with irrigation events showed significantly lower sigB allele type diversity than did isolates associated with precipitation events (P = <0.001), suggesting that irrigation water may be a point source of L. monocytogenes contamination. Small changes in management practices (e.g., not irrigating fields before harvest) may therefore reduce the risk of L. monocytogenes contamination of fresh produce.

  12. De novo transcriptome sequencing and gene expression profiling of spinach (Spinacia oleracea L.) leaves under heat stress

    PubMed Central

    Yan, Jun; Yu, Li; Xuan, Jiping; Lu, Ying; Lu, Shijun; Zhu, Weimin

    2016-01-01

    Spinach (Spinacia oleracea) has cold tolerant but heat sensitive characteristics. The spinach variety ‘Island,’ is suitable for summer periods. There is lack molecular information available for spinach in response to heat stress. In this study, high throughput de novo transcriptome sequencing and gene expression analyses were carried out at different spinach variety ‘Island’ leaves (grown at 24 °C (control), exposed to 35 °C for 30 min (S1), and 5 h (S2)). A total of 133,200,898 clean reads were assembled into 59,413 unigenes (average size 1259.55 bp). 33,573 unigenes could match to public databases. The DEG of controls vs S1 was 986, the DEG of control vs S2 was 1741 and the DEG of S1 vs S2 was 1587. Gene Ontology (GO) and pathway enrichment analysis indicated that a great deal of heat-responsive genes and other stress-responsive genes were identified in these DEGs, suggesting that the heat stress may have induced an extensive abiotic stress effect. Comparative transcriptome analysis found 896 unique genes in spinach heat response transcript. The expression patterns of 13 selected genes were verified by RT-qPCR (quantitative real-time PCR). Our study found a series of candidate genes and pathways that may be related to heat resistance in spinach. PMID:26857466

  13. Heavy metals phyto-assessment in commonly grown vegetables: water spinach (I. aquatica) and okra (A. esculentus).

    PubMed

    Ng, Chuck Chuan; Rahman, Md Motior; Boyce, Amru Nasrulhaq; Abas, Mhd Radzi

    2016-01-01

    The growth response, metal tolerance and phytoaccumulation properties of water spinach (Ipomoea aquatica) and okra (Abelmoschus esculentus) were assessed under different contaminated spiked metals: control, 50 mg Pb/kg soil, 50 mg Zn/kg soil and 50 mg Cu/kg soil. The availability of Pb, Zn and Cu metals in both soil and plants were detected using flame atomic absorption spectrometry. The concentration and accumulation of heavy metals from soil to roots and shoots (edible parts) were evaluated in terms of translocation factor, accumulation factor and tolerance index. Okra recorded the highest accumulation of Pb (80.20 mg/kg) in its root followed by Zn in roots (35.70 mg/kg) and shoots (34.80 mg/kg) of water spinach, respectively. Different accumulation trends were observed with, Pb > Zn > Cu in okra and Zn > Pb > Cu in water spinach. Significant differences (p < 0.01) of Pb, Zn and Cu accumulation were found in both water spinach and okra cultivated among tested treatments. However, only the accumulation of Pb metal in the shoots of water spinach and okra exceeded the maximum permissible levels of the national Malaysian Food Act 1983 and Food Regulations 1985 (2006) as well as the international Codex Alimentarius Commission limits. This study has shown that both water spinach and okra have good potential as Pb and Zn phytoremediators.

  14. Heavy metals phyto-assessment in commonly grown vegetables: water spinach (I. aquatica) and okra (A. esculentus).

    PubMed

    Ng, Chuck Chuan; Rahman, Md Motior; Boyce, Amru Nasrulhaq; Abas, Mhd Radzi

    2016-01-01

    The growth response, metal tolerance and phytoaccumulation properties of water spinach (Ipomoea aquatica) and okra (Abelmoschus esculentus) were assessed under different contaminated spiked metals: control, 50 mg Pb/kg soil, 50 mg Zn/kg soil and 50 mg Cu/kg soil. The availability of Pb, Zn and Cu metals in both soil and plants were detected using flame atomic absorption spectrometry. The concentration and accumulation of heavy metals from soil to roots and shoots (edible parts) were evaluated in terms of translocation factor, accumulation factor and tolerance index. Okra recorded the highest accumulation of Pb (80.20 mg/kg) in its root followed by Zn in roots (35.70 mg/kg) and shoots (34.80 mg/kg) of water spinach, respectively. Different accumulation trends were observed with, Pb > Zn > Cu in okra and Zn > Pb > Cu in water spinach. Significant differences (p < 0.01) of Pb, Zn and Cu accumulation were found in both water spinach and okra cultivated among tested treatments. However, only the accumulation of Pb metal in the shoots of water spinach and okra exceeded the maximum permissible levels of the national Malaysian Food Act 1983 and Food Regulations 1985 (2006) as well as the international Codex Alimentarius Commission limits. This study has shown that both water spinach and okra have good potential as Pb and Zn phytoremediators. PMID:27119073

  15. De novo transcriptome sequencing and gene expression profiling of spinach (Spinacia oleracea L.) leaves under heat stress.

    PubMed

    Yan, Jun; Yu, Li; Xuan, Jiping; Lu, Ying; Lu, Shijun; Zhu, Weimin

    2016-01-01

    Spinach (Spinacia oleracea) has cold tolerant but heat sensitive characteristics. The spinach variety 'Island,' is suitable for summer periods. There is lack molecular information available for spinach in response to heat stress. In this study, high throughput de novo transcriptome sequencing and gene expression analyses were carried out at different spinach variety 'Island' leaves (grown at 24 °C (control), exposed to 35 °C for 30 min (S1), and 5 h (S2)). A total of 133,200,898 clean reads were assembled into 59,413 unigenes (average size 1259.55 bp). 33,573 unigenes could match to public databases. The DEG of controls vs S1 was 986, the DEG of control vs S2 was 1741 and the DEG of S1 vs S2 was 1587. Gene Ontology (GO) and pathway enrichment analysis indicated that a great deal of heat-responsive genes and other stress-responsive genes were identified in these DEGs, suggesting that the heat stress may have induced an extensive abiotic stress effect. Comparative transcriptome analysis found 896 unique genes in spinach heat response transcript. The expression patterns of 13 selected genes were verified by RT-qPCR (quantitative real-time PCR). Our study found a series of candidate genes and pathways that may be related to heat resistance in spinach.

  16. Bioavailability of total iron from meat, spinach (Spinacea oleracea L.) and meat-spinach mixtures by anaemic and non-anaemic rats.

    PubMed

    Zhang, D; Hendricks, D G; Mahoney, A W

    1989-03-01

    1. Bioavailability of Fe from beef, spinach (Spinacea oleracea L.) and their mixtures was studied using anaemic and non-anaemic rats by haemoglobin regeneration efficiency (HRE) and apparent Fe absorption in two trials. 2. The initial haemoglobin levels of severely anaemic, mildly anaemic and non-anaemic rats were 63, 88 and 113 g/l, respectively. The Fe level in diets was about 30 mg/kg. All other nutrients equalled or exceeded the requirement of the growing rat. 3. The spinach Fe was well utilized by the rats with average HRE of 0.41, 0.53 and 0.36, and apparent Fe absorptions averaging 0.48, 0.59 and 0.37 for the severely anaemic, mildly anaemic and non-anaemic animals respectively. 4. Beef Fe was efficiently used by rats as reported by others. Average HRE were 0.42, 0.51 and 0.44, and average apparent Fe absorptions were 0.44, 0.47 and 0.46 for the severely anaemic, mildly anaemic and non-anaemic rats respectively. 5. When the percentage of meat Fe was increased from 0 to 25, 50, 75 or 100 of the dietary Fe, HRE and apparent Fe absorption were not increased significantly. A meat enhancement effect on total Fe absorption, reported by others for non-haem-Fe, did not occur in the present experiment. 6. Negative correlation coefficients between initial haemoglobin and HRE (r -0.79), and initial haemoglobin and apparent Fe absorption (r -0.73) were seen with the rats fed on dietary Fe from FeSO4.7H2O. This was not seen with the rats fed on dietary Fe from beef or spinach. 7. The Fe absorption pattern for the different Fe sources is evidence for a third Fe pool, a pool made up of highly soluble inorganic Fe salt, in addition to haem-Fe and non-haem-Fe complex pools. FeSO4.7H2O is not in the same gastrointestinal pool as non-haem-Fe complex such as spinach Fe. A suggested mechanism of absorption is discussed.

  17. Increasing total and biologically active chromium in wheat grain and spinach by spraying with chromium salts

    SciTech Connect

    Vicini, F.A.; Ellis, B.G.

    1981-06-01

    Recently, chromium has been shown to be necessary for glucose metabolism in man. But most plant species greatly restrict the uptake of Cr. This study was conducted to determine if both total and biologically active Cr could be increased in wheat grain or spinach by spraying the plants with either Cr/sub 2/(SO/sub 4/)/sub 3/ or Cr-EDTA. Concentrations of Cr in wheat grain were about doubled in a greenhouse experiment by spraying with either Cr source. Biologically active Cr (estimated by extraction with ethanol or NH/sub 4/OH) was increased from about 40 to greater than 50% of total Cr when wheat was sprayed with Cr salts. Total Cr in spinach leaves was increased by as much as 10-fold by spraying, with the sulfate source being more effective than the EDTA.

  18. Enhancement of iron content in spinach plants stimulated by magnetic nano particles

    NASA Astrophysics Data System (ADS)

    Yulianto, Agus; Astuti, Budi; Amalia, Saptaria Rosa

    2016-04-01

    In our previous study, the iron content in spinach plants could be detected by magnetic susceptibility values. In the present work, magnetic nano particles were found from the iron sand. The magnetic nano particles are synthesis by using co-precipitation process and sol-gel technique. The stimulation of magnetic nano particles in the plant has been done by the provision of magnetic nano particles in growing media. After certain time, plant samples was characterized using susceptibility-meter MS2B and atomic absorption spectroscopy to measure the magnetic susceptibility and the amount of iron content that absorbed of the plant, respectively. The iron content in the spinach plants was increased when the magnetic nano particles was injected in the growing media.

  19. Fat Metabolism in Higher Plants: LXII. Stearl-acyl Carrier Protein Desaturase from Spinach Chloroplasts.

    PubMed

    Jacobson, B S; Jaworski, J G; Stumpf, P K

    1974-10-01

    Stearyl-acyl carrier protein desaturase (EC 1.14.99.6), present in the stroma fraction of spinach (Spinacia oleracea) chloroplasts, rapidly desaturated enzymatically prepared stearyl-acyl carrier protein to oleic acid. No other substrates were desaturated. In addition to stearyl-acyl carrier protein, reduced ferredoxin was an essential component of the system. The electron donor systems were either ascorbate, dichlorophenolindophenol, photosystem I and light, or NADPH and ferredoxin-NADP reductase. The desaturase was more active in extracts prepared from chloroplasts obtained from immature spinach leaves than from mature leaves. Stearyl-acyl carrier protein desaturase also occurs in soluble extracts of avocado (Persea americana Mill.) mesocarp and of developing safflower (Carthamus tinctorius) seeds.

  20. Impact of coal mine dump contaminated soils on elemental uptake by Spinacia oleracea (spinach)

    SciTech Connect

    Chunilall, V.; Kindness, A.; Jonnalagadda, S.B.

    2006-07-01

    The elemental uptake and the growth response of Spinacia oleracea (spinach) to the soil contaminated with the South African bituminous coal mine dump soil, viz. 0%, 5%, 15%, and 25% w/w, was investigated. The contaminated soils were analyzed for pH, cation exchange capacity (CEC), soil organic matter (SOM), and concentrations of selected heavy metals. The pH, SOM, and CEC decreased with an increase in contamination indicating the acidic nature of coal mine soil and the raise in the soil binding sites. The distribution of Fe, Mn, Ni, Cd, and Pb in the roots and leaves of the plants was determined in two stages of plant growth. Spinach showed high accumulation of Fe and increased levels of Ni and Cd with an increase in contamination. No plant growth was recorded with 25% contamination.

  1. The role of leaves in sex expression in hemp and spinach.

    PubMed

    Chailakhyan, M K; Khryanin, V N

    1979-01-01

    When shoots of young plants of hemp (Cannabis sativa L.) and spinach (Spinacea oleracea L.) were cultured as cuttings and allowed to regenerate advenitious roots, ca. 80-85% became female (formed pistillate flowers) regardless of whether the leaves were left on the plants or were cut off (except for the 2-3 uppermost ones) after the beginning of adventitious-root formation. But when the leaves were cut off and the cuttings treated with gibberellic acid (GA3, 25 mg/l) ca. 77-80% of the plants became male (formed staminate flowers). The result was quite similar when roots and leaves of young hemp plants were removed at the same time and the cuttings treated with GA3. It is suggested that the leaves play an essential role in sex expression in hemp and spinach and that this role is related to gibberellin synthesis in the leaves.

  2. Interactions between food-borne pathogens and protozoa isolated from lettuce and spinach.

    PubMed

    Gourabathini, Poornima; Brandl, Maria T; Redding, Katherine S; Gunderson, John H; Berk, Sharon G

    2008-04-01

    The survival of Salmonella enterica was recently shown to increase when the bacteria were sequestered in expelled food vacuoles (vesicles) of Tetrahymena. Because fresh produce is increasingly linked to outbreaks of enteric illness, the present investigation aimed to determine the prevalence of protozoa on spinach and lettuce and to examine their interactions with S. enterica, Escherichia coli O157:H7, and Listeria monocytogenes. Glaucoma sp., Colpoda steinii, and Acanthamoeba palestinensis were cultured from store-bought spinach and lettuce and used in our study. A strain of Tetrahymena pyriformis previously isolated from spinach and a soil-borne Tetrahymena sp. were also used. Washed protozoa were allowed to graze on green fluorescent protein- or red fluorescent protein-labeled enteric pathogens. Significant differences in interactions among the various protist-enteric pathogen combinations were observed. Vesicles were produced by Glaucoma with all of the bacterial strains, although L. monocytogenes resulted in the smallest number per ciliate. Vesicle production was observed also during grazing of Tetrahymena on E. coli O157:H7 and S. enterica but not during grazing on L. monocytogenes, in vitro and on leaves. All vesicles contained intact fluorescing bacteria. In contrast, C. steinii and the amoeba did not produce vesicles from any of the enteric pathogens, nor were pathogens trapped within their cysts. Studies of the fate of E. coli O157:H7 in expelled vesicles revealed that by 4 h after addition of spinach extract, the bacteria multiplied and escaped the vesicles. The presence of protozoa on leafy vegetables and their sequestration of enteric bacteria in vesicles indicate that they may play an important role in the ecology of human pathogens on produce. PMID:18310421

  3. Spinach - A software library for simulation of spin dynamics in large spin systems

    NASA Astrophysics Data System (ADS)

    Hogben, H. J.; Krzystyniak, M.; Charnock, G. T. P.; Hore, P. J.; Kuprov, Ilya

    2011-02-01

    We introduce a software library incorporating our recent research into efficient simulation algorithms for large spin systems. Liouville space simulations (including symmetry, relaxation and chemical kinetics) of most liquid-state NMR experiments on 40+ spin systems can now be performed without effort on a desktop workstation. Much progress has also been made with improving the efficiency of ESR, solid state NMR and Spin Chemistry simulations. Spinach is available for download at http://spindynamics.org.

  4. Chemical accessibility of the 4.5S RNA in spinach chloroplast ribosomes.

    PubMed Central

    Kumagai, I; Bartsch, M; Subramanian, A R; Erdmann, V A

    1983-01-01

    We have examined the accessibility to diethylpyrocarbonate of spinach chloroplast 4.5S ribosomal RNA when free and when it is part of the ribosomal structure. The modifications in free 4.5S RNA were found mostly in single-stranded regions of the secondary structure model proposed in our previous paper (Kumagai, I. et al. (1982) J.B.C. 257, 12924-28): adenines at positions 17, 19, 33, 36, 54, 55, 60, 64, 68, 72, 77, 86 and 87 were identified as the reactive residues. On the other hand, in 4.5S RNA in 70S ribosomes or 50S subunits, adenine 33 was exclusively modified, and its reactivity was much higher than in free 4.5S RNA. This highly accessible A33 of spinach 4.5S RNA is located within a characteristic seven nucleotide sequence, which is found in the 4.5S rRNAs from spinach, tobacco and a fern but deleted in 4.5S RNAs from maize and wheat. Images PMID:6828382

  5. Relationship of components in wheat bran and spinach to iron bioavailability in the anemic rat.

    PubMed

    Gordon, D T; Chao, L S

    1984-03-01

    Components found in wheat bran and spinach were evaluated as to their affect on the bioavailability of ferrous iron (i.e., FeSO4) by using the criteria of hemoglobin regeneration in anemic rats. The relative biological value (RBV) of iron in wheat bran and spinach (FeSO4 = 100%) were determined to be 124 and 53%, respectively. Control diets with graded levels of FeSO4 did not contain dietary fiber (i.e., cellulose). Adding cellulose (1.74%) or phytic acid (0.66%) at levels contained in the wheat bran diet, significantly increased (P less than 0.05) the RBV f the ferrous iron to 126 and 124%, respectively. The addition of 2.10% oxalic acid, the amount in the spinach diet, caused the highest increase in RBV to 164%. Combining these dietary components, plus lignin (0.67%) and pectin (0.63%), in various combinations, resulted in RBVs equivalent or significantly higher than 100%. The bioavailability of iron in plant foods appears to be dependent on how this nutrient is presented to the mucosa. Cellulose, phytate or oxalate added to a purified diet containing ferrous iron significantly enhanced the bioavailability of this element. Relative biological values for iron were also calculated based on food intake and growth rate. The latter parameters are believed to have greater utility in determining RBV when food intake and/or growth rate may vary among animals consuming different sources of test iron.

  6. Uptake, translocation, and transformation of pentachlorophenol in soybean and spinach plants

    SciTech Connect

    Casterline, J.L. Jr.; Barnett, N.M.; Ku, Y.

    1985-06-01

    Soybean plants were grown for 90 days and spinach plants for 64 days in a mixture of sterilized greenhouse soil and sand containing 10 ppm pentachlorophenol. All plant parts and soil samples were extracted and separated into nonpolar and polar fractions. Major nonpolar and polar metabolites were identified by gas-liquid chromatography and mass spectrometry. Nonpolar fractions from both soybean and spinach plants were found to contain pentachlorophenol and its metabolites, 2,3,4,6-tetrachlorophenol, methoxytetrachlorophenol, 2,3,4,6-tetrachloroanisole, and pentachloroanisole. Cleavage of polar metabolites from the soybean plants by acid hydrolysis yielded organic solvent-extractable products. These products were identified as pentachlorophenol, 2,3,4,6-tetrachlorophenol, and methoxytetrachlorophenol. Cleavage of polar materials from spinach plants yielded only pentachlorophenol. The polar metabolites from the soybean plants were also subjected to enzymatic cleavage by beta-glucosidase. The conjugates consisted mostly of O-glucosides of the same metabolites released by acid hydrolysis. Failure of hydrolysis by aryl sulfatase indicated that very little or no sulfates were present. The metabolites found in the plants were not detected in soil samples obtained from pots immediately after the plants were harvested.

  7. Conversion of monogalactosyldiacylglycerols to triacylglycerols in ozone-fumigated spinach leaves. [Spinacia oleracea L

    SciTech Connect

    Sakaki, Takeshi; Saito, Kazuki; Kawaguchi, Akihiko; Kondo, Noriaki; Yamada, Mitsuhiro Keio Univ., Tokyo Univ. of Tokyo )

    1990-10-01

    Molecular species and fatty acid distribution of triacylglycerol (TG) accumulated in spinach (Spinacia oleracea L.) leaves fumigated with ozone (0.5 microliter per liter) were compared with those of monogalactosyldiacylglyerol (MGDG). Analysis of positional distribution of the fatty acids in MGDG and the accumulated TG by the enzymatic digestion method showed that hexadecatrienoate (16:3) was restricted to sn-2 position of the glycerol backbone in both MGDG and TG, whereas {alpha}-linolenate (18:3) was preferentially located at sn-1 position in MGDG, and sn-1 and/or sn-3 positions in TG, suggesting that 1,2-diacylglycerol moieties of MGDG are the direct precursor of TG in ozone-fumigated leaves. Further analysis of TG molecular species by argentation chromatography and mass spectrometry showed that TG increased with ozone fumigation consisted of approximately an equal molar ratio of sn-1,3-18:3-2-16:3 and sn-1,2,3-18:3. Because the molecular species of MGDG in spinach leaves is composed of a similar molar ratio of sn-1-18:3-2-16:3 and sn-1,2-18:3, we conducted that MGDG was converted to 1,2-diacylglycerol and acylated with 18:3 to TG in ozone-fumigated spinach leaves.

  8. Gamma irradiation dose: Effects on spinach baby-leaf ascorbic acid, carotenoids, folate, alpha-tocopherol, and phylloquinone concentrations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ionizing radiation of fruits and vegetables, in the form of gamma rays or electron beams, is effective in overcoming quarantine barriers in trade, decontamination, disinfestation and prolonging shelf life, but a void of information persists on ionizing radiation effects of vitamin profiles in indivi...

  9. Employing response surface methodology for the optimization of ultrasound assisted extraction of lutein and β-carotene from spinach.

    PubMed

    Altemimi, Ammar; Lightfoot, David A; Kinsel, Mary; Watson, Dennis G

    2015-04-14

    The extraction of lutein and β-carotene from spinach (Spinacia oleracea L.) leaves is important to the dietary supplement industry. A Box-Behnken design and response surface methodology (RSM) were used to investigate the effect of process variables on the ultrasound-assisted extraction (UAE) of lutein and β-carotene from spinach. Three independent variables, extraction temperature (°C), extraction power (%) and extraction time (min) were studied. Thin-layer chromatography (TLC) followed by UV visualization and densitometry was used as a simple and rapid method for both identification and quantification of lutein and β-carotene during UAE. Methanol extracts of leaves from spinach and authentic standards of lutein and β-carotene were separated by normal-phase TLC with ethyl acetate-acetone (5:4 (v/v)) as the mobile phase. In this study, the combination of TLC, densitometry, and Box-Behnken with RSM methods were effective for the quantitative analysis of lutein and β-carotene from spinach extracts. The resulting quadratic polynomial models for optimizing lutein and β-carotene from spinach had high coefficients of determination of 0.96 and 0.94, respectively. The optimal UAE settings for output of lutein and β-carotene simultaneously from spinach extracts were an extraction temperature of 40 °C, extraction power of 40% (28 W/cm3) and extraction time of 16 min. The identity and purity of each TLC spot was measured using time-of-flight mass spectrometry. Therefore, UAE assisted extraction of carotenes from spinach can provide a source of lutein and β-carotene for the dietary supplement industry.

  10. Simple quantitative analysis of Escherichia coli K-12 internalized in baby spinach using Fourier Transform Infrared spectroscopy.

    PubMed

    Wang, Jun; Kim, Kyung Ho; Kim, Sungkyun; Kim, Yong Soo; Li, Qing X; Jun, Soojin

    2010-11-15

    Bacterial contamination continues to be a serious concern for food safety. Although washing fresh produce helps in reducing pathogen levels, pathogen internalization often limits the effectiveness of washing. When pathogens internalize in leafy vegetables, the method of identification and quantitative measurement would be called into question. This study was aimed to use Fourier Transform Infrared (FTIR) spectroscopy integrated with an attenuated total reflectance kit for quantification of Escherichia coli K-12 internalized in baby spinach. The bacteria were inoculated into vascular and intracellar tissues of spinach leaves by syringe injection and the distribution of internalized E. coli K-12 cells was confirmed under scanning electron microscopy (SEM). FTIR measurement following the preparation of bacterial suspension from spinach leaves with high speed pulverizing enabled to detect the absorbance peaks in the amide II region between 1590 and 1490 cm⁻¹ as a fingerprint for the microbes. It was found that the estimated concentrations of E. coli K-12 agreed well with the concentrations determined by plate counting with R² values of 0.98 and 0.97 in peptone water and spinach extracts, respectively. The results demonstrated that FTIR can identify and quantify E. coli K-12 in baby spinach extracts at a limit of detection of approximately 100 CFU/mL in 5 min. The developed method is expected to be suitable for the analysis of pathogenic E. coli strains and other bacterial species in fresh vegetables.

  11. Assessment of microbial risk factors and impact of meteorological conditions during production of baby spinach in the Southeast of Spain.

    PubMed

    Castro-Ibáñez, I; Gil, M I; Tudela, J A; Ivanek, R; Allende, A

    2015-08-01

    There is a timely need to evaluate the effect agricultural factors and meteorological conditions on fresh produce contamination. This study evaluated those risk factors and described, for the first time, the distribution of indicator microorganisms (Escherichia coli, Enterococcus, coliforms, and Enterobacteriaceae) and the prevalence of foodborne pathogens (Enterohaemorrhagic E. coli, Listeria monocytogenes and Salmonella spp.) in baby spinach grown in the Southeast of Spain. A longitudinal study was conducted on three farms (2011-2013). Results obtained for E. coli highlighted soil and irrigation water as important factors affecting the microbial safety of baby spinach. Significant differences in the proportion of E. coli positive samples were found between treated (46.1%) and untreated (100%) irrigation water. However, the microbial quality of irrigation water didn't affect E. coli prevalence in produce. All E. coli positive spinach samples were detected at the highest observed temperature range suggesting that ambient temperature affects the probability and extent of spinach contamination. Salmonella spp. was detected by RT-PCR in manure, soil, irrigation water and baby spinach but only two of them (manure and irrigation water) were confirmed by isolation in culture media. Salmonella RT-PCR positive samples showed higher levels of E. coli than Salmonella negative samples. This preliminary finding supports recent identification of E. coli as a suitable parameter for the hygiene criterion at the primary production of leafy greens. PMID:25846928

  12. Effect of electrolyzed oxidizing water treatment on the reduction of nitrite levels in fresh spinach during storage.

    PubMed

    Hao, Jianxiong; Li, Huiying; Wan, Yangfang; Liu, Haijie

    2015-03-01

    Leafy vegetables are the major source of nitrite intake in the human diet, and technological processing to control nitrite levels in harvested vegetables is necessary. In the current work, the effect of electrolyzed oxidizing water (EOW) on the nitrite and nitrate levels in fresh spinach during storage was studied. EOW treatment, including slightly acidic electrolyzed water and acidic electrolyzed water, was found to effectively reduce nitrite levels in fresh spinach during storage; levels in the late period were 30 to 40% lower than that of the control. However, the nitrate levels in fresh spinach during storage were not influenced by EOW treatment. The reduction of nitrite levels in EOW-treated fresh spinach during storage can be attributed to the inactivation of nitrate reductase directly and to the reduction of bacterial populations. Our results suggest that treatment with slightly acidic electrolyzed water may be a better choice to control nitrite levels in fresh vegetables during storage. This study provided a useful method to reduce nitrite levels in fresh spinach.

  13. Assessment of microbial risk factors and impact of meteorological conditions during production of baby spinach in the Southeast of Spain.

    PubMed

    Castro-Ibáñez, I; Gil, M I; Tudela, J A; Ivanek, R; Allende, A

    2015-08-01

    There is a timely need to evaluate the effect agricultural factors and meteorological conditions on fresh produce contamination. This study evaluated those risk factors and described, for the first time, the distribution of indicator microorganisms (Escherichia coli, Enterococcus, coliforms, and Enterobacteriaceae) and the prevalence of foodborne pathogens (Enterohaemorrhagic E. coli, Listeria monocytogenes and Salmonella spp.) in baby spinach grown in the Southeast of Spain. A longitudinal study was conducted on three farms (2011-2013). Results obtained for E. coli highlighted soil and irrigation water as important factors affecting the microbial safety of baby spinach. Significant differences in the proportion of E. coli positive samples were found between treated (46.1%) and untreated (100%) irrigation water. However, the microbial quality of irrigation water didn't affect E. coli prevalence in produce. All E. coli positive spinach samples were detected at the highest observed temperature range suggesting that ambient temperature affects the probability and extent of spinach contamination. Salmonella spp. was detected by RT-PCR in manure, soil, irrigation water and baby spinach but only two of them (manure and irrigation water) were confirmed by isolation in culture media. Salmonella RT-PCR positive samples showed higher levels of E. coli than Salmonella negative samples. This preliminary finding supports recent identification of E. coli as a suitable parameter for the hygiene criterion at the primary production of leafy greens.

  14. Ascorbic acid, carotenoids, and visual quality of baby spinach as affected by shade netting and postharvest storage.

    PubMed

    Bergquist, Sara A M; Gertsson, Ulla E; Nordmark, Lotta Y G; Olsson, Marie E

    2007-10-17

    Baby spinach ( Spinacia oleracea L.) was grown under three types of shade netting (high transmittance, spectrum-altering, and low transmittance) to study the effect on the concentrations of vitamin C (ascorbic acid and dehydroascorbic acid), carotenoids, and chlorophyll and on the visual quality of the leaves. The spinach was sown in April and August and harvested at two growth stages. After harvest, leaves were stored in polypropylene bags at 2 and 10 degrees C. Shading significantly decreased the ascorbic acid concentration of April-sown spinach by 12-33%, but in the August-sown spinach, the response was inconsistent. Concentrations of total carotenoids and total chlorophylls were significantly higher under the nettings in many cases, especially under the spectrum-altering and low-transmittance nettings. Postharvest visual quality and postharvest persistence of the compounds analyzed were not greatly affected by shading. We conclude that these shade nettings are acceptable to use in baby spinach production when it comes to the studied aspects of internal and external quality of the produce.

  15. 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.

  16. Effect of organic and conventional cropping systems on ascorbic acid, vitamin C, flavonoids, nitrate, and oxalate in 27 varieties of spinach (Spinacia oleracea L.).

    PubMed

    Koh, Eunmi; Charoenprasert, Suthawan; Mitchell, Alyson E

    2012-03-28

    This study was undertaken to compare the levels of ascorbic acid, vitamin C, flavonoids, nitrate, and oxalate in 27 spinach varieties grown in certified organic and conventional cropping systems. Liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-(ESI)MS/MS) of methanolic extracts of spinach demonstrated 17 flavonoids, including glucuronides and acylated di- and triglycosides of methylated and methylenedioxyderivatives of 6-oxygenated flavonoids. The mean levels of ascorbic acid and flavonoids were significantly (p < 0.001) higher in the organically grown [40.48 ± 6.16 and 2.83 ± 0.03 mg/kg of fresh weight (FW)] spinach compared to the conventionally grown spinach (25.75 ± 6.12 and 2.27 ± 0.02 mg/kg of FW). Conversely, the mean levels of nitrate were significantly (p < 0.001) higher in the conventionally grown spinach compared to the organically grown spinach. No significant effects were observed in the oxalate content of spinach from either production system. The levels of nitrate correlated negatively with those of ascorbic acid, vitamin C, and total flavonoids and showed a positive correlation with the oxalate content. These results suggest that organic cropping systems result in spinach with lower levels of nitrates and higher levels of flavonoids and ascorbic acid.

  17. In vitro bioavailability of iron from spinach (Spinacea oleracea) cultivated in soil fortified with graded levels of iron and zinc.

    PubMed

    Reddy, N S; Sondge, C V; Khan, T N

    1993-11-01

    A pot-culture experiment was conducted to assess the bioavailability of iron from spinach cultivated in soil fortified with graded levels of iron and zinc (FeSO4 x 7H2(0) and ZnSO4 x 7H2(0), respectively). Applications of varying levels of iron to soil increased the total iron and phosphorus contents and decreased the zinc content (P < 0.05). The effect of applying varying levels of zinc was the opposite of on the minerals in spinach. The ascorbic acid content was remarkably reduced with varying levels of iron and zinc. Higher levels of zinc and lower levels of iron in the soil increased the bioavailability of iron from spinach (P < 0.05). In conclusion, the interactions of 15 ppm zinc with 30 ppm iron significantly enhanced the bioavailability of iron, total iron and zinc contents.

  18. Biosynthesis and Desaturation of Prokaryotic Galactolipids in Leaves and Isolated Chloroplasts from Spinach 1

    PubMed Central

    Heemskerk, Johan W. M.; Schmidt, Hermann; Hammer, Ute; Heinz, Ernst

    1991-01-01

    Mono- and digalactosyldiacylglycerol (MGDG and DGDG) were isolated from the leaves of sixteen 16:3 plants. In all of these plant species, the sn-2 position of MGDG was more enriched in C16 fatty acids than sn-2 of DGDG. The molar ratios of prokaryotic MGDG to prokaryotic DGDG ranged from 4 to 10. This suggests that 16:3 plants synthesize more prokaryotic MGDG than prokaryotic DGDG. In the 16:3 plant Spinacia oleracea L. (spinach), the formation of prokaryotic galactolipids was studied both in vivo and in vitro. In intact spinach leaves as well as in chloroplasts isolated from these leaves, radioactivity from [1-14C]acetate accumulated 10 times faster in MGDG than in DGDG. After 2 hours of incorporation, most labeled galactolipids from leaves and all labeled galactolipids from isolated chloroplasts were in the prokaryotic configuration. Both in vivo and in vitro, the desaturation of labeled palmitate and oleate to trienoic fatty acids was higher in MGDG than in DGDG. In leaves, palmitate at the sn-2 position was desaturated in MGDG but not in DGDG. In isolated chloroplasts, palmitate at sn-2 similarly was desaturated only in MGDG, but palmitate and oleate at the sn-1 position were desaturated in MGDG as well as in DGDG. Apparently, palmitate desaturase reacts with sn-1 palmitate in either galactolipid, but does not react with the sn-2 fatty acid of DGDG. These results demonstrate that isolated spinach chloroplasts can synthesize and desaturate prokaryotic MGDG and DGDG. The finally accumulating molecular species, MGDG(18:3/16:3) and DGDG(18:3/16:0), are made by the chloroplasts in proportions similar to those found in leaves. PMID:16668143

  19. Serum antioxidant capacity is increased by consumption of strawberries, spinach, red wine or vitamin C in elderly women.

    PubMed

    Cao, G; Russell, R M; Lischner, N; Prior, R L

    1998-12-01

    It is often assumed that antioxidant nutrients contribute to the protection afforded by fruits, vegetables, and red wine against diseases of aging. However, the effect of fruit, vegetable and red wine consumption on the overall antioxidant status in human is unclear. In this study we investigated the responses in serum total antioxidant capacity following comsumption of strawberries (240 g), spinach (294 g), red wine (300 ml) or vitamin C (1250 mg) in eight elderly women. Total antioxidant capacity was determined using different methods: oxygen radical absorbance capacity (ORAC) assay, Trolox equivalent antioxidant capacity (TEAC) assay and ferric reducing ability (FRAP) assay. The results showed that the total antioxidant capacity of serum determined as ORAC, TEAC and FRAP, using the area under the curve, increased significantly by 7-25% during the 4-h period following consumption of red wine, strawberries, vitamin C or spinach. The total antioxidant capacity of urine determined as ORAC increased (P < 0.05) by 9.6, 27.5, and 44.9% for strawberries, spinach, and vitamin C, respectively, during the 24-h period following these treatments. The plasma vitamin C level after the strawberry drink, and the serum urate level after the strawberry and spinach treatments, also increased significantly. However, the increased vitamin C and urate levels could not fully account for the increased total antioxidant capacity in serum following the consumption of strawberries, spinach or red wine. We conclude that the consumption of strawberries, spinach or red wine, which are rich in antioxidant phenolic compounds, can increase the serum antioxidant capacity in humans. J. Nutr. 2383-2390, 1998 PMID:9868185

  20. Leaf growth is conformal

    NASA Astrophysics Data System (ADS)

    Alim, Karen; Armon, Shahaf; Shraiman, Boris I.; Boudaoud, Arezki

    2016-10-01

    Growth pattern dynamics lie at the heart of morphogenesis. Here, we investigate the growth of plant leaves. We compute the conformal transformation that maps the contour of a leaf at a given stage onto the contour of the same leaf at a later stage. Based on the mapping we predict the local displacement field in the leaf blade and find it to agree with the experimentally measured displacement field to 92%. This approach is applicable to any two-dimensional system with locally isotropic growth, enabling the deduction of the whole growth field just from observation of the tissue contour.

  1. Leaf growth is conformal.

    PubMed

    Alim, Karen; Armon, Shahaf; Shraiman, Boris I; Boudaoud, Arezki

    2016-01-01

    Growth pattern dynamics lie at the heart of morphogenesis. Here, we investigate the growth of plant leaves. We compute the conformal transformation that maps the contour of a leaf at a given stage onto the contour of the same leaf at a later stage. Based on the mapping we predict the local displacement field in the leaf blade and find it to agree with the experimentally measured displacement field to 92%. This approach is applicable to any two-dimensional system with locally isotropic growth, enabling the deduction of the whole growth field just from observation of the tissue contour. PMID:27597439

  2. Radiosensitization of Salmonella spp. and Listeria spp. in ready-to-eat baby spinach leaves.

    PubMed

    Gomes, Carmen; Moreira, Rosana G; Castell-Perez, Elena

    2011-01-01

    The FDA recently approved irradiation treatment of leafy greens such as spinach up to 1 kGy; however, it is important to reduce the dose required to decontaminate the produce while maintaining its quality. Thus, the objectives of this study were: (1) to assess the radiation sensitivities of Salmonella spp. and Listeria spp. inoculated in ready-to-eat baby spinach leaves under modified atmosphere packaging (MAP) and irradiated using a 1.35-MeV Van de Graff accelerator (the leaves were irradiated both at room temperature and at -5 °C); and (2) to understand and optimize the synergistic effect of MAP and irradiation by studying the radiolysis of ozone formation under different temperatures, the effect of dose rate on its formation, and its decomposition. Results showed that increased concentrations of oxygen in the packaging significantly increased the radiation sensitivity of the test organisms, ranging from 7% up to 25% reduction in D(10)-values. In particular, radiosensitization could be effected (P < 0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Radiosensitization was demonstrated for both microorganisms with irradiation of either fresh or frozen (-5 °C) baby spinach. These results suggest that low-dose (below 1 kGy) e-beam radiation under modified atmosphere packaging (100% O(2) and N(2):O(2)[1:1]) may be a viable tool for reducing microbial populations or eliminating Salmonella spp. and Listeria spp. from baby spinach. A suggested treatment to achieve a 5-log reduction of the test organisms would be irradiation at room temperature under 100% O(2) atmosphere at a dose level of 0.7 kGy. Practical Application: Decontamination of minimally processed fruits and vegetables from food-borne pathogens presents technical and economical challenges to the produce industry. Internalized microorganisms cannot be eliminated by the current procedure (water-washed or treated with 200-ppm chlorine

  3. Thermal inactivation of human norovirus surrogates in spinach and measurement of its uncertainty.

    PubMed

    Bozkurt, Hayriye; D'souza, Doris H; Davidson, P Michael

    2014-02-01

    Leafy greens, including spinach, have potential for human norovirus transmission through improper handling and/or contact with contaminated water. Inactivation of norovirus prior to consumption is essential to protect public health. Because of the inability to propagate human noroviruses in vitro, murine norovirus (MNV-1) and feline calicivirus (FCV-F9) have been used as surrogates to model human norovirus behavior under laboratory conditions. The objectives of this study were to determine thermal inactivation kinetics of MNV-1 and FCV-F9 in spinach, compare first-order and Weibull models, and measure the uncertainty associated with the process. D-values were determined for viruses at 50, 56, 60, 65, and 72 °C in 2-ml vials. The D-values calculated from the first-order model (50 to 72 °C) ranged from 0.16 to 14.57 min for MNV-1 and 0.15 to 17.39 min for FCV-9. Using the Weibull model, the tD for MNV-1 and FCV-F9 to destroy 1 log (D ≈ 1) at the same temperatures ranged from 0.22 to 15.26 and 0.27 to 20.71 min, respectively. The z-values determined for MNV-1 were 11.66 ± 0.42 °C using the Weibull model and 10.98 ± 0.58 °C for the first-order model and for FCV-F9 were 10.85 ± 0.67 °C and 9.89 ± 0.79 °C, respectively. There was no difference in D- or z-value using the two models (P > 0.05). Relative uncertainty for dilution factor, personal counting, and test volume were 0.005, 0.0004, and ca. 0.84%, respectively. The major contribution to total uncertainty was from the model selected. Total uncertainties for FCV-F9 for the Weibull and first-order models were 3.53 to 7.56% and 11.99 to 21.01%, respectively, and for MNV-1, 3.10 to 7.01% and 13.14 to 16.94%, respectively. Novel and precise information on thermal inactivation of human norovirus surrogates in spinach was generated, enabling more reliable thermal process calculations to control noroviruses. The results of this study may be useful to the frozen food industry in designing blanching processes for

  4. Effects of spinach nitrate on insulin resistance, endothelial dysfunction markers and inflammation in mice with high-fat and high-fructose consumption

    PubMed Central

    Li, Ting; Lu, Xinshan; Sun, Yanfei; Yang, Xingbin

    2016-01-01

    Background Insulin resistance, which is associated with an increased risk of cardiovascular morbidity and mortality, has become a leading nutrition problem. Inorganic nitrate enriched in spinach has been demonstrated to reverse the pathological features of insulin resistance and endothelial dysfunction. However, the effects of a direct intake of nitrate-enriched spinach on insulin resistance and endothelial dysfunction have not been studied. Objective To investigate the effects of spinach nitrate on insulin resistance, lipid metabolism, endothelial function, and inflammation in mice fed with a high-fat and high-fructose diet. Design A diet intervention of spinach with or without nitrate was performed in mice. A high-fat and high-fructose diet was used to cause insulin resistance, endothelial dysfunction, and inflammation in mice. The impacts of spinach nitrate on lipid profile, insulin resistance, markers of endothelial function, and inflammation were determined in mice. Results Spinach nitrate improved the vascular endothelial function of the mice with high-fat and high-fructose consumption, as evidenced by the elevated plasma nitrite level, increased serum nitric oxide (NO) level and decreased serum ET-1 level after spinach nitrate intervention. Spinach nitrate also reduced serum triglycerides, total cholesterol, and low-density lipoprotein-cholesterol levels and elevated serum high-density lipoprotein-cholesterol levels in the mice fed with a high-fat and high-fructose diet. Mice receiving spinach with 60 mg/kg of nitrate (1.02±0.34) showed a significantly low homeostasis model assessment-insulin resistance index as compared with the model mice (2.05±0.58), which is indicating that spinach nitrate could effectively improve the insulin resistance. In addition, spinach nitrate remarkably decreased the elevated serum C-reactive protein, tumor necrosis factor α, and interleukin-6 levels induced by a high-fat and high-fructose diet. Conclusions The intake of

  5. Thermoluminescence study of photosystem II activity in Haberlea rhodopensis and spinach leaves during desiccation.

    PubMed

    Peeva, V; Maslenkova, L

    2004-05-01

    Thermoluminescence glow curve parameters were used to access the functional features of PS II in the Balkan endemic Haberlea rhodopensis. This representative of the higher desiccation-tolerant plants is unique for the European flora. An unusual high temperature of TL emission from Haberlea leaves after excitation by one flash at 5 degrees C was observed. The position of the main TL B band (S (2)Q (B)(-)) was at 45 - 47 degrees C, while this temperature was 30 - 32 degrees C in drought-sensitive mesophytic spinach. Consistent with the up-shift in TL emission, the lifetime of the S (2) state was also increased, showing a stabilization of charge storage in PS II complex in this resurrection plant. In addition, a part of PS II centres was less susceptible to DCMU. We consider the observed unusual TL characteristics of Haberlea rhodopensis reflect some structural modifications in PS II (especially in D1 protein), which could be related to the desiccation tolerance of this plant. This suggestion was supported by the different manner in which dehydration affected the TL properties in desiccation-tolerant Haberlea and desiccation-sensitive spinach plants.

  6. Possible association of actin filaments with chloroplasts of spinach mesophyll cells in vivo and in vitro.

    PubMed

    Kumatani, T; Sakurai-Ozato, N; Miyawaki, N; Yokota, E; Shimmen, T; Terashima, I; Takagi, S

    2006-11-01

    In palisade mesophyll cells of spinach (Spinacia oleracea L.) kept under low-intensity white light, chloroplasts were apparently immobile and seemed to be surrounded by fine bundles of actin filaments. High-intensity blue light induced actin-dependent chloroplast movement concomitant with the appearance of a couple of long, straight bundles of actin filaments in each cell, whereas high-intensity red light was essentially ineffective in inducing these responses. The actin organization observed under low-intensity white light has been postulated to function in anchoring chloroplasts at proper intracellular positions through direct interaction with the chloroplasts. Intact chloroplasts, which retained their outer envelopes, were isolated after homogenization of leaves and Percoll centrifugation. No endogenous actin was detected by immunoblotting in the final intact-chloroplast fraction prepared from the leaves kept under low-intensity white light or in darkness. In cosedimentation assays with exogenously added skeletal muscle filamentous actin, however, actin was detected in the intact-chloroplast fraction precipitated after low-speed centrifugation. The association of actin with chloroplasts was apparently dependent on incubation time and chloroplast density. After partial disruption of the outer envelope of isolated chloroplasts by treatment with trypsin, actin was no longer coprecipitated. The results suggest that chloroplasts in spinach leaves can directly interact with actin, and that this interaction may be involved in the regulation of intracellular positioning of chloroplasts.

  7. Influences of nano-TiO2 on the chloroplast aging of spinach under light.

    PubMed

    Hong, Fashui; Yang, Fan; Liu, Chao; Gao, Qing; Wan, Zhigang; Gu, Fugen; Wu, Cheng; Ma, Zhenni; Zhou, Juan; Yang, Ping

    2005-06-01

    The effects of nano-TiO2 (rutile) on the chloroplast aging of spinach under light were studied. The results showed that when the chloroplasts were illuminated for 1, 5, and 10 min with 500 micromol/cm2/min light intensity, respectively, the evolution oxygen rate was rapidly increased; when the chloroplasts were treated for 20, 30, and 40 min with 500 micromol/cm2/min light intensity, respectively, the evolution oxygen rate was gradually decreased. While spinach was treated with 0.25% nano-TiO2, the rate of evolution oxygen of chloroplasts in different illumination times (1, 5, 10, 20, 30, and 40 min) was higher than that of control, and when the illumination time was over 10 min, the reduction of the evolution oxygen rate was lower than that of control. It suggested that nano-TiO2 treatment could protect chloroplasts from aging for long-time illumination. The mechanism researches indicated that nano-TiO2 treatment could significantly increase the activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), decrease accumulation of reactive oxygen free radicals and the level of malondialdehyde (MDA), and maintain stability of membrane structure of chloroplast under light.

  8. Effects of Pb 2+ on energy distribution and photochemical activity of spinach chloroplast

    NASA Astrophysics Data System (ADS)

    Wu, Xiao; Hong, Fashui; Liu, Chao; Su, Mingyu; Zheng, Lei; Gao, Fengqing; Yang, Fan

    2008-03-01

    Lead (Pb 2+) is a well-known highly toxic element. The mechanisms of the Pb 2+ toxicity are not well understood for photosynthesis. In this paper, we reported the effect of Pb 2+ on light absorption, distribution and conversion of spinach chloroplast by spectroscopy, and photochemical reaction activities. Several effects of Pb 2+ were observed: (1) the absorption peak intensity of chloroplast obviously decreased in red and blue region and produced optical flattering; (2) fluorescence quantum yield nearby 680 nm of chloroplast greatly declined; (3) the excitation band nearby 440 nm of chloroplast significantly descended; (4) Pb 2+ treatments reduced of the rate of whole chain electron transport, photochemical activities of PSII DCPIP photoreduction and oxygen evolution, but the photoreduction activities of PSI were little changed. Together, the studies of the experiments showed that Pb 2+ decreased absorption of light on spinach chloroplast and inhibited excitation energy to be absorbed by LHCII and transferred to PSII, then reduced the conversion from light energy to electron energy, and decelerated electron transport, water photolysis and oxygen evolution.

  9. Effects of Pb2+ on energy distribution and photochemical activity of spinach chloroplast.

    PubMed

    Wu, Xiao; Hong, Fashui; Liu, Chao; Su, Mingyu; Zheng, Lei; Gao, Fengqing; Yang, Fan

    2008-03-01

    Lead (Pb(2+)) is a well-known highly toxic element. The mechanisms of the Pb(2+) toxicity are not well understood for photosynthesis. In this paper, we reported the effect of Pb(2+) on light absorption, distribution and conversion of spinach chloroplast by spectroscopy, and photochemical reaction activities. Several effects of Pb(2+) were observed: (1) the absorption peak intensity of chloroplast obviously decreased in red and blue region and produced optical flattering; (2) fluorescence quantum yield nearby 680 nm of chloroplast greatly declined; (3) the excitation band nearby 440 nm of chloroplast significantly descended; (4) Pb(2+) treatments reduced of the rate of whole chain electron transport, photochemical activities of PSII DCPIP photoreduction and oxygen evolution, but the photoreduction activities of PSI were little changed. Together, the studies of the experiments showed that Pb(2+) decreased absorption of light on spinach chloroplast and inhibited excitation energy to be absorbed by LHCII and transferred to PSII, then reduced the conversion from light energy to electron energy, and decelerated electron transport, water photolysis and oxygen evolution.

  10. Effects of Mg(2+)on spectral characteristics and photosynthetic functions of spinach photosystem II.

    PubMed

    Liang, Chen; Xiao, Wu; Hao, Huang; Xiaoqing, Liu; Chao, Liu; Lei, Zheng; Fashui, Hong

    2009-03-01

    In the present paper we report the results obtained with the photosystem II (PSII) isolated from spinach treated by MgCl(2), and studied the effect of Mg(2+) on spectral characteristics and photosynthetic functions of PSII. The results showed that Mg(2+) treatment at a suitable concentration could significantly increase the absorption intensity of PSII and the intensity ratio of Soret band to Q band of chlorophyll-a. The treatment also elevated the excited peak intensity at 230, 278 and 343 nm, and the emitted peak intensity at 304 and 682 nm, and the ratio of F(278)/F(230), respectively. The results implied that Mg(2+) increased absorbance for visible light, improving energy transfer among amino acids within PSII protein complex and accelerating energy transport from tyrosine residue to chlorophyll-a. The photochemical activity and oxygen evolving rate of PSII were also enhanced by Mg(2+). This is viewed as evidence that Mg(2+) can promote energy transfer and oxygen evolution in PSII of spinach.

  11. Using EDDS and NTA for enhanced phytoextraction of Cd by water spinach.

    PubMed

    Hseu, Zeng-Yei; Jien, Shih-Hao; Wang, Shih-Hui; Deng, Hong-Wei

    2013-03-15

    A greenhouse experiment was used to test the applicability of [S,S]-Ethylenediaminedisuccinic acid (EDDS) and nitrilotriacetic acid (NTA) at rates of 2.5 mmol kg(-1) and 5.0 mmol kg(-1), respectively, to increase the uptake of Cd by water spinach (Ipomoea aquatic Forsk) in soils with 2.5-30 mg Cd kg(-1). The addition of EDDS and NTA significantly increased water soluble Cd in soils. However, the Cd concentration in the root and shoot was higher in the NTA treatment than in the EDDS treatment. No instance of Cd hyperaccumulation was observed; however, the 5.0 mmol kg(-1) NTA treatment for soil with 30 mg Cd kg(-1) caused the Cd concentration to increase to 86 mg kg(-1), which is close to the critical concentration (100 mg kg(-1)) of a hyperaccumulator. The total Cd uptake in the treatments of EDDS and NTA for soils with 2.5, 5.0, and 10 mg Cd kg(-1) was acceptable, and was higher than the control. The level of 5.0 mmol kg(-1) EDDS was excessively high for enhanced phytoextraction in soils with 20 and 30 mg Cd kg(-1). Water spinach for Cd phytoextraction is a viable alternative to using herbaceous hyperaccumulators.

  12. The polar auxin transport inhibitor TIBA inhibits endoreduplication in dark grown spinach hypocotyls.

    PubMed

    Amijima, Makoto; Iwata, Yuji; Koizumi, Nozomu; Mishiba, Kei-Ichiro

    2014-08-01

    We addressed the question of whether an additional round of endoreduplication in dark-grown hypocotyls is a common feature in dicotyledonous plants having endopolyploid tissues. Ploidy distributions of hypocotyl tissues derived from in vitro-grown spinach (Spinacia oleracea L. cv. Atlas) seedlings grown under different light conditions were analyzed by flow cytometry. An additional round of endoreduplication (represented by 32C cells) was found in the dark-grown hypocotyl tissues. This response was inhibited by light, the intensity of which is a crucial factor for the inhibition of endoreduplication. The higher ploidy cells in cortical tissues of the dark-grown hypocotyls had larger cell sizes, suggesting that the additional round of endoreduplication contributes to hypocotyl elongation. More importantly, a polar auxin transport inhibitor, 2,3,5-triiodobenzoic acid (TIBA), strongly inhibits endoreduplication, not only in spinach but also in Arabidopsis. Because other polar auxin transport inhibitors or an auxin antagonist show no or mild effects, TIBA may have a specific feature that inhibits endoreduplication.

  13. Effects of Mg 2+on spectral characteristics and photosynthetic functions of spinach photosystem II

    NASA Astrophysics Data System (ADS)

    Liang, Chen; Xiao, Wu; Hao, Huang; Xiaoqing, Liu; Chao, Liu; Lei, Zheng; Fashui, Hong

    2009-03-01

    In the present paper we report the results obtained with the photosystem II (PSII) isolated from spinach treated by MgCl 2, and studied the effect of Mg 2+ on spectral characteristics and photosynthetic functions of PSII. The results showed that Mg 2+ treatment at a suitable concentration could significantly increase the absorption intensity of PSII and the intensity ratio of Soret band to Q band of chlorophyll-a. The treatment also elevated the excited peak intensity at 230, 278 and 343 nm, and the emitted peak intensity at 304 and 682 nm, and the ratio of F278/ F230, respectively. The results implied that Mg 2+ increased absorbance for visible light, improving energy transfer among amino acids within PSII protein complex and accelerating energy transport from tyrosine residue to chlorophyll-a. The photochemical activity and oxygen evolving rate of PSII were also enhanced by Mg 2+. This is viewed as evidence that Mg 2+ can promote energy transfer and oxygen evolution in PSII of spinach.

  14. Active site histidine in spinach ribulosebisphosphate carboxylase/oxygenase modified by diethyl pyrocarbonate

    SciTech Connect

    Igarashi, Y.; McFadden, B.A.; el-Gul, T.

    1985-07-16

    (TH) Diethyl pyrocarbonate was synthesized from (TH) ethanol prepared by the reduction of acetaldehyde by NaB3H4. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) from spinach was inactivated with this reagent at pH 7.0 the presence of 20 mM MgS , and tryptic peptides that contained modified histidine residues were isolated by reverse-phase high-performance liquid chromatography. Labeling of the enzyme was conducted in the presence and absence of the competitive inhibitor sedoheptulose 1,7-bisphosphate. The amount of one peptide that was heavily labeled in the absence of this compound was reduced 10-fold in its presence. The labeled residue was histidine-298. This result, in combination with earlier experiments, suggests that His-298 in spinach RuBisCO is located in the active site domain and is essential to enzyme activity. This region of the primary structure is strongly conserved in seven other ribulosebisphosphate carboxylases from divergent sources.

  15. Deer predation on leaf miners via leaf abscission

    NASA Astrophysics Data System (ADS)

    Yamazaki, Kazuo; Sugiura, Shinji

    2008-03-01

    The evergreen oak Quercus gilva Blume sheds leaves containing mines of the leaf miner Stigmella sp. (Lepidoptera: Nepticulidae) earlier than leaves with no mines in early spring in Nara, central Japan. The eclosion rates of the leaf miner in abscised and retained leaves were compared in the laboratory to clarify the effects of leaf abscission on leaf miner survival in the absence of deer. The leaf miner eclosed successfully from both fallen leaves and leaves retained on trees. However, sika deer ( Cervus nippon centralis Kishida) feed on the fallen mined leaves. Field observations showed that deer consume many fallen leaves under Q. gilva trees, suggesting considerable mortality of leaf miners due to deer predation via leaf abscission. This is a previously unreported relationship between a leaf miner and a mammalian herbivore via leaf abscission.

  16. Aluminium resistance requires resistance to acid stress: a case study with spinach that exudes oxalate rapidly when exposed to Al stress.

    PubMed

    Yang, Jian Li; Zheng, Shao Jian; He, Yun Feng; Matsumoto, Hideaki

    2005-04-01

    Spinach is a vegetable with a high oxalate concentration in its tissues. Oxalate efflux from spinach (Spinacia oleracea L. cv. Quanneng) roots was rapidly stimulated (within 30 min) by aluminium (Al) treatment. The efflux was constant within 6 h, but increased with increasing Al concentration. The efflux was confined to the root tip (0-5 mm), which showed a 5-fold greater efflux than the root zone distal to the tip (5-10 mm). Oxalate efflux could not be triggered by treatment with the trivalent cation lanthanum or by phosphorus deficiency, indicating that the efflux was specific to the Al treatment. All this evidence suggested that spinach possesses Al-resistance mechanisms. However, spinach was found to be as sensitive to Al toxicity as the Al-sensitive wheat line ES8, which had no Al-dependent organic acids efflux. The Al accumulated in the apical 5 mm of the roots of spinach which was also similar to that in the Al-sensitive wheat after 24 h treatment with 50 microM AlCl(3), indicating a non-exclusion mechanism. In addition, root elongation in spinach was significantly inhibited at pH 4.5, compared with that at pH 6.5. Based on this evidence, it is concluded that the sensitivity to acid stress in spinach could mask the potential role for oxalate to protect the plant roots from Al toxicity.

  17. Bioactive compounds during storage of fresh-cut spinach: the role of endogenous ascorbic acid in the improvement of product quality.

    PubMed

    Bottino, Antonella; Degl'Innocenti, Elena; Guidi, Lucia; Graziani, Giulia; Fogliano, Vincenzo

    2009-04-01

    Spinach is rich in bioactive constituents such as vitamin C, flavonoids and phenolic acids. In this work, the biochemical modifications occurring during one week of storage at 4 degrees C were evaluated both in intact and in fresh-cut spinach. Results showed that vitamin C concentration is less affected by storage in fresh-cut spinach with respect to intact spinach. MS/MS analysis showed that the main flavonoids are not modified during storage in intact leaves, while some of them increased significantly during storage in the fresh-cut samples. Fresh-cut spinach did not show color alteration even if PPO activity increased significantly during storage. This finding was related to the high ascorbic acid content, which delays the subsequent polymerization events. This finding was confirmed by the unaltered concentration of phenolic compounds in fresh-cut spinach during storage. In conclusion, data about nutritional content and visual performance concurrently suggest that spinach is a suitable species for marketing as a fresh-cut product. PMID:19253961

  18. Inactivation of Escherichia coli O157:H7 in vitro and on the surface of spinach leaves by biobased surfactants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to evaluate the effect of biosurfactants on the populations of Escherichia coli O157:H7 in suspension and on spinach leaves. Eight surfactants including four soybean oil-based biosurfactants, sodium dodecyl sulfate (SDS), polyoxyethylene sorbitan monooleate (Tween 80), sopho...

  19. Effect of lead stress on mineral content and growth of wheat (Triticum aestivum) and spinach (Spinacia oleracea) seedlings.

    PubMed

    Lamhamdi, Mostafa; El Galiou, Ouiam; Bakrim, Ahmed; Nóvoa-Muñoz, Juan Carlos; Arias-Estévez, Manuel; Aarab, Ahmed; Lafont, René

    2013-01-01

    Lead (Pb) is the most common heavy metal contaminant in the environment. Pb is not an essential element for plants, but they absorb it when it is present in their environment, especially in rural areas when the soil is polluted by automotive exhaust and in fields contaminated with fertilizers containing heavy metal impurities. To investigate lead effects on nutrient uptake and metabolism, two plant species, spinach (Spinacia oleracea) and wheat (Triticum aestivum), were grown under hydroponic conditions and stressed with lead nitrate, Pb(NO3)2, at three concentrations (1.5, 3, and 15 mM). Lead is accumulated in a dose-dependent manner in both plant species, which results in reduced growth and lower uptake of all mineral ions tested. Total amounts and concentrations of most mineral ions (Na, K, Ca, P, Mg, Fe, Cu and Zn) are reduced, although Mn concentrations are increased, as its uptake is reduced less relative to the whole plant's growth. The deficiency of mineral nutrients correlates in a strong decrease in the contents of chlorophylls a and b and proline in both species, but these effects are less pronounced in spinach than in wheat. By contrast, the effects of lead on soluble proteins differ between species; they are reduced in wheat at all lead concentrations, whereas they are increased in spinach, where their value peaks at 3 mM Pb. The relative lead uptake by spinach and wheat, and the different susceptibility of these two species to lead treatment are discussed.

  20. Quality of fresh-cut Iceberg lettuce and spinach irradiated at doses up to 4 kGy

    NASA Astrophysics Data System (ADS)

    Fan, Xuetong; Guan, Wenqiang; Sokorai, Kimberly J. B.

    2012-08-01

    Fresh-cut Iceberg lettuce packaged in modified atmosphere packages and spinach in perforated film bags were irradiated with gamma rays at doses of 0, 1, 2, 3, and 4 kGy. After irradiation, the samples were stored for 14 days at 4 °C. O2 levels in the packages of fresh-cut Iceberg lettuce decreased and CO2 levels increased with increasing radiation dose, suggesting that irradiation increased respiration rates of lettuce. Tissue browning of irradiated cut lettuce was less severe than that of non-irradiated, probably due to the lower O2 levels in the packages. However, samples irradiated at 3 and 4 kGy had lower maximum force and more severe sogginess than the non-irradiated control. In addition, ascorbic acid content of irradiated lettuce was 22-40% lower than the non-irradiated samples after 14 days of storage. The visual appearance of spinach was not affected by irradiation even at a dose of 4 kGy. Consumer acceptance suggested that more people would dislike and would not buy spinach that was treated at 3 and 4 kGy as compared to the non-irradiated sample. Overall, irradiation at doses of 1 and 2 kGy may be employed to enhance microbial safety of fresh-cut Iceberg lettuce and spinach while maintaining quality.

  1. Survival and transfer of murine norovirus 1, a surrogate for human noroviruses, during the production process of deep-frozen onions and spinach.

    PubMed

    Baert, Leen; Uyttendaele, Mieke; Vermeersch, Mattias; Van Coillie, Els; Debevere, Johan

    2008-08-01

    The reduction of murine norovirus 1 (MNV-1) on onions and spinach by washing was investigated as was the risk of contamination during the washing procedure. To decontaminate wash water, the industrial sanitizer peracetic acid (PAA) was added to the water, and the survival of MNV-1 was determined. In contrast to onions, spinach undergoes a heat treatment before freezing. Therefore, the resistance of MNV-1 to blanching of spinach was examined. MNV-1 genomic copies were detected with a real-time reverse transcription PCR assay in PAA-treated water and blanched spinach, and PFUs (representing infectious MNV-1 units) were determined with a plaque assay. A < or = 1-log reduction in MNV-1 PFUs was achieved by washing onion bulbs and spinach leaves. More than 3 log PFU of MNV-1 was transmitted to onion bulbs and spinach leaves when these vegetables were washed in water containing approximately 5 log PFU/ml. No decline of MNV-1 occurred in used industrial spinach wash water after 6 days at room temperature. A concentration of 20 ppm of PAA in demineralized water (pH 4.13) and in potable water (pH 7.70) resulted in reductions of 2.88 +/- 0.25 and 2.41 +/- 0.18 log PFU, respectively, after 5 min of exposure, but no decrease in number of genomic copies was observed. No reduction of MNV-1 PFUs was observed on frozen onions or spinach during storage for 6 months. Blanching spinach (80 degrees C for 1 min) resulted in at least 2.44-log reductions of infectious MNV-1, but many genomic copies were still present.

  2. Survival and transfer of murine norovirus 1, a surrogate for human noroviruses, during the production process of deep-frozen onions and spinach.

    PubMed

    Baert, Leen; Uyttendaele, Mieke; Vermeersch, Mattias; Van Coillie, Els; Debevere, Johan

    2008-08-01

    The reduction of murine norovirus 1 (MNV-1) on onions and spinach by washing was investigated as was the risk of contamination during the washing procedure. To decontaminate wash water, the industrial sanitizer peracetic acid (PAA) was added to the water, and the survival of MNV-1 was determined. In contrast to onions, spinach undergoes a heat treatment before freezing. Therefore, the resistance of MNV-1 to blanching of spinach was examined. MNV-1 genomic copies were detected with a real-time reverse transcription PCR assay in PAA-treated water and blanched spinach, and PFUs (representing infectious MNV-1 units) were determined with a plaque assay. A < or = 1-log reduction in MNV-1 PFUs was achieved by washing onion bulbs and spinach leaves. More than 3 log PFU of MNV-1 was transmitted to onion bulbs and spinach leaves when these vegetables were washed in water containing approximately 5 log PFU/ml. No decline of MNV-1 occurred in used industrial spinach wash water after 6 days at room temperature. A concentration of 20 ppm of PAA in demineralized water (pH 4.13) and in potable water (pH 7.70) resulted in reductions of 2.88 +/- 0.25 and 2.41 +/- 0.18 log PFU, respectively, after 5 min of exposure, but no decrease in number of genomic copies was observed. No reduction of MNV-1 PFUs was observed on frozen onions or spinach during storage for 6 months. Blanching spinach (80 degrees C for 1 min) resulted in at least 2.44-log reductions of infectious MNV-1, but many genomic copies were still present. PMID:18724752

  3. Damped leaf flexure hinge

    NASA Astrophysics Data System (ADS)

    Chen, Zhong; Chen, Guisheng; Zhang, Xianmin

    2015-05-01

    Flexure-based mechanism like compliant actuation system embeds complex dynamics that will reduce the control bandwidth and limits their dynamic positioning precision. This paper presents a theoretical model of a leaf flexure hinge with damping layers using strain energy method and Kelvin damping model. The modified loss factor of the damped leaf flexure hinge is derived, and the equivalent viscous damping coefficient of the damped leaf hinge is obtained, which could be used to improve the pseudo-rigid-model. The free vibration signals of the hinge in three different damping configurations are measured. The experimental modal analysis also is performed on the three kinds of damped leaf flexure hinges in order to evaluate their 1st order bending natural frequency and vibration-suppressing effects. The evaluation of modified loss factor model also is performed. The experimental results indicate that the constrained layer damping can enhance the structure damping of the hinge even if only single damping layer each side, the modified loss factor model can get good predicts of a damped leaf flexure hinge in the frequency range below 1st order natural frequency, and it is necessary that the dimensional parameters of the damping layers and basic layer of the hinge should be optimized for simplification at the mechanism's design stage.

  4. Damped leaf flexure hinge.

    PubMed

    Chen, Zhong; Chen, Guisheng; Zhang, Xianmin

    2015-05-01

    Flexure-based mechanism like compliant actuation system embeds complex dynamics that will reduce the control bandwidth and limits their dynamic positioning precision. This paper presents a theoretical model of a leaf flexure hinge with damping layers using strain energy method and Kelvin damping model. The modified loss factor of the damped leaf flexure hinge is derived, and the equivalent viscous damping coefficient of the damped leaf hinge is obtained, which could be used to improve the pseudo-rigid-model. The free vibration signals of the hinge in three different damping configurations are measured. The experimental modal analysis also is performed on the three kinds of damped leaf flexure hinges in order to evaluate their 1st order bending natural frequency and vibration-suppressing effects. The evaluation of modified loss factor model also is performed. The experimental results indicate that the constrained layer damping can enhance the structure damping of the hinge even if only single damping layer each side, the modified loss factor model can get good predicts of a damped leaf flexure hinge in the frequency range below 1st order natural frequency, and it is necessary that the dimensional parameters of the damping layers and basic layer of the hinge should be optimized for simplification at the mechanism's design stage. PMID:26026549

  5. Population dynamics of Escherichia coli inoculated by irrigation into the phyllosphere of spinach grown under commercial production conditions.

    PubMed

    Wood, J D; Bezanson, G S; Gordon, R J; Jamieson, R

    2010-10-15

    Recent outbreaks of food-borne illnesses associated with the consumption of fresh produce have increased attention on irrigation water as a potential source of pathogen contamination. A better understanding of the behaviour of enteric pathogens introduced into agricultural systems during irrigation will aid in risk assessments and support the development of appropriate farm-level water management practices. For this reason, the survival dynamics of two nalidixic acid resistant strains of Escherichia coli after their spray inoculation into the phyllosphere and soil of field spinach were examined over two growing seasons. E. coli strains NAR, an environmental isolate, and DM3n, a non-pathogenic serotype O157:H7, were applied at rates of 10⁴ to 10⁷ cfu/100ml to the fully developed spinach plants that arose subsequent to the harvesting of their upper leafy portions for commercial purposes (secondary-growth plants). After 72 h, E. coli on spinach were reduced by 3-5 logs. Culturable E. coli were recovered from plants up to 6 days post-inoculation. Survival in soil was greater than in the phyllosphere. Under ambient conditions, the mean 72 h first order decay constant computed by Chick's Law was 0.1 h⁻¹. Although light reduction studies indicated UV irradiation negatively influenced the persistence of E. coli, a simple relationship between UV exposure and phyllosphere E. coli densities could not be established. E. coli introduced to the leafy portions of spinach via spray irrigation displayed rapid declines in their culturability under the open environmental conditions experienced during this study. A 6 day period between the last irrigation and harvest would minimize the risks of E. coli survival in the spinach phyllosphere. E. coli NAR was identified as a possible surrogate for the O157:H7 strain, DM3n.

  6. Combination treatment of chlorine dioxide gas and aerosolized sanitizer for inactivating foodborne pathogens on spinach leaves and tomatoes.

    PubMed

    Park, Sang-Hyun; Kang, Dong-Hyun

    2015-08-17

    The objective of this study was to evaluate the antimicrobial effect of chlorine dioxide (ClO2) gas and aerosolized sanitizer, when applied alone or in combination, on the survival of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes inoculated onto spinach leaves and tomato surfaces. Spinach leaves and tomatoes were inoculated with a cocktail of three strains each of the three foodborne pathogens. ClO2 gas (5 or 10 ppmv) and aerosolized peracetic acid (PAA) (80 ppm) were applied alone or in combination for 20 min. Exposure to 10 ppmv of ClO2 gas for 20 min resulted in 3.4, 3.3, and 3.4 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes on spinach leaves, respectively. Treatment with 80 ppm of aerosolized PAA for 20 min caused 2.3, 1.9, and 0.8 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. Combined treatment of ClO2 gas (10 ppmv) and aerosolized PAA (80 ppm) for 20 min caused 5.4, 5.1, and 4.1 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. E. coli O157:H7, S. Typhimurium, and L. monocytogenes on tomatoes experienced similar reduction patterns to those on spinach leaves. As treatment time increased, most combinations of ClO2 gas and aerosolized PAA showed additive effects in the inactivation of the three pathogens. Combined treatment of ClO2 gas and aerosolized PAA produced injured cells of three pathogens on spinach leaves while generally did not produce injured cells of these pathogens on tomatoes. Combined treatment of ClO2 gas (10 ppmv) and aerosolized PAA (80 ppm) did not significantly (p>0.05) affect the color and texture of samples during 7 days of storage.

  7. Colonization of spinach by Verticillium dahliae and effects of pathogen localization on the efficacy of seed treatments.

    PubMed

    Maruthachalam, Karunakaran; Klosterman, Steven J; Anchieta, Amy; Mou, Beiquan; Subbarao, Krishna V

    2013-03-01

    Verticillium wilt on spinach (Spinacia oleracea) is caused by the soilborne fungus Verticillium dahliae. The pathogen is seedborne and transmission through seed is a major concern because of the dispersal of the pathogen to areas where fresh and processing spinach crops are grown in rotation with susceptible crops. Reduction in seedborne inoculum minimizes pathogen spread; therefore, knowledge of pathogen localization in seed is critical to develop methods to reduce seedborne inoculum. Spinach seedlings were inoculated with conidial suspensions of a green fluorescent protein-tagged strain of V. dahliae and colonization events were followed through seed production by confocal laser-scanning microscopy. Between 24 to 96 h postinoculation (PI), conidia germinated and formed hyphal colonies on root tips and in root elongation zones. Hyphae colonized root cortical tissues both intra and intercellularly by 2 weeks, and colonized the taproot xylem with abundant mycelia and conidia that led to vascular discoloration coincident with foliar symptom expression by 8 weeks PI. At 10 weeks PI, the xylem of the upper stem, inflorescence, and spinach seed parts, including the pericarp, seed coat, cotyledons, and radicle, had been colonized by the pathogen but not the perisperm (the diploid maternal tissue). Maximum concentration of the fungus was in the seed coat, the outermost layer of the vasculature. Infection of V. dahliae in spinach seed was systemic and transmissible to developing seedlings. Additional analyses indicated that fungicide and steam seed treatments reduced detectable levels of the pathogen but did not eliminate the pathogen from the seed. This information will assist in the development of seed treatments that will reduce the seedborne inoculum transmission to crop production fields.

  8. Combination treatment of chlorine dioxide gas and aerosolized sanitizer for inactivating foodborne pathogens on spinach leaves and tomatoes.

    PubMed

    Park, Sang-Hyun; Kang, Dong-Hyun

    2015-08-17

    The objective of this study was to evaluate the antimicrobial effect of chlorine dioxide (ClO2) gas and aerosolized sanitizer, when applied alone or in combination, on the survival of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes inoculated onto spinach leaves and tomato surfaces. Spinach leaves and tomatoes were inoculated with a cocktail of three strains each of the three foodborne pathogens. ClO2 gas (5 or 10 ppmv) and aerosolized peracetic acid (PAA) (80 ppm) were applied alone or in combination for 20 min. Exposure to 10 ppmv of ClO2 gas for 20 min resulted in 3.4, 3.3, and 3.4 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes on spinach leaves, respectively. Treatment with 80 ppm of aerosolized PAA for 20 min caused 2.3, 1.9, and 0.8 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. Combined treatment of ClO2 gas (10 ppmv) and aerosolized PAA (80 ppm) for 20 min caused 5.4, 5.1, and 4.1 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. E. coli O157:H7, S. Typhimurium, and L. monocytogenes on tomatoes experienced similar reduction patterns to those on spinach leaves. As treatment time increased, most combinations of ClO2 gas and aerosolized PAA showed additive effects in the inactivation of the three pathogens. Combined treatment of ClO2 gas and aerosolized PAA produced injured cells of three pathogens on spinach leaves while generally did not produce injured cells of these pathogens on tomatoes. Combined treatment of ClO2 gas (10 ppmv) and aerosolized PAA (80 ppm) did not significantly (p>0.05) affect the color and texture of samples during 7 days of storage. PMID:26001524

  9. Gibberellin A[sub 1] is required for stem elongation in spinach

    SciTech Connect

    Zeevaart, J.A.D.; Gage, D.A.; Talon, M. )

    1993-08-01

    The effects of the growth retardants 2'-isopropyl-4'-(trimethylammonium chloride)-5'-methylphenyl piperidine-1-carboxylate (AMO-1618) and calcium 3,5-dioxo-4-propionylcyclohexanecarboxylate (BX-112) on stem elongation were investigated in the rosette plant spinach (Spinacia oleracea L.) under long-day (LD) conditions. Stem growth induced by a LD treatment was prevented by both retardants. The inhibition caused by AMO-1618 was reversed by gibberellin A[sub 1] (GA[sub 1]) and GA[sub 20], whereas the effects of BX-112 were reversed by GA[sub 1] only. Six GAs (GA[sub 53], GA[sub 44], GA[sub 19], GA[sub 20], GA[sub 1], and GA[sub 8]) were quantified by gas chromatography-selected ion monitoring using internal standards. Plants treated with BX-112 had reduced levels of GA[sub 1], and GA[sub 8] and accumulated GA[sub 53], GA[sub 44], GA[sub 19], and GA[sub 20]. The relative levels of four additional GAs (3-epi-GA[sub 1], GA[sub 29], GA[sub 60], and GA[sub 81]) were compared by ion intensities only. Relative to GA[sub 81], the level of GA[sub 29] was decreased by BX-112, whereas the levels of GA[sub 6] and 3-epi-GA[sub 1] were increased. Transfer of spinach from short-day conditions to LD conditions caused an increase in all identified GAs of the early 13-hydroxylation pathway with GA[sub 20], GA[sub 1], and GA[sub 8] showing the largest increases. These findings support the position that, of the GA[sub s] belonging to the early 13-hydroxylation pathway, GA[sub 1] is the primary GA active per se for stem elongation in spinach. The increase in endogenous GA[sub 1] in plants in LD conditions is most likely the primary factor for stem elongation. 23 refs., 7 figs., 3 tabs.

  10. Investigation of Detergent Effects on the Solution Structure of Spinach Light Harvesting Complex II

    SciTech Connect

    Cardoso, Mateus B; Smolensky, Dmitriy; Heller, William T; O'Neill, Hugh Michael

    2010-01-01

    The properties of spinach light harvesting complex II (LHC II), stabilized in the detergents Triton X-100 (TX100) and n-Octyl-{beta}-D-Glucoside (BOG), were investigated by small-angle neutron scattering (SANS). The LHC II-BOG scattering curve overlaid well with the theoretical scattering curve generated from the crystal structure of LHC II indicating that the protein preparation was in its native functional state. On the other hand, the simulated LHC II curve deviated significantly from the LHC II-TX100 experimental data. Analysis by circular dichroism spectroscopy supported the SANS analysis and showed that LHC II-TX100 is inactivated. This investigation has implications for extracting and stabilizing photosynthetic membrane proteins for the development of biohybrid photoconversion devices.

  11. Visible light absorption and photo-sensitizing properties of spinach leaves and beetroot extracted natural dyes.

    PubMed

    Sengupta, D; Mondal, B; Mukherjee, K

    2015-09-01

    Herein, chlorophyll and betalain dyes are extracted from fresh spinach leaves and beetroots. Fourier transform infrared spectra are used to identify the characteristic peaks of the extracted dyes. UV-vis light absorption characteristics of the dyes and their mixed counterpart are investigated by varying their pH and temperature. These dyes are used as photo sensitizer for fabrication of zinc oxide photo-anode based dye sensitized solar cells (DSSCs). The photo-voltaic characteristics of the developed DSSCs are measured under simulated solar light (power of incident light 100 mW cm(-2) from Air Mass 1.5G). The solar to electric conversion efficiencies for the chlorophyll, betalain and mixed dye based solar cells are estimated as 0.148%, 0.197% and 0.294% respectively. The highest conversion efficiency for mixed dye based solar cell is attributed due to the absorption of wider range of solar spectrum.

  12. Isolation of lumenal proteins from spinach thylakoid membranes by triton X-114 phase partitioning.

    PubMed

    Bricker, T M; Prevost, M; Vu, V; Laborde, S; Womack, J; Frankel, L K

    2001-01-19

    The proteins present in the thylakoid lumen of higher plant chloroplasts have not been rigorously examined. In this communication we present a simple and rapid procedure for the isolation of the soluble proteins and extrinsic membrane proteins present in the thylakoid lumen from spinach. Our procedure involves extensive washing of the thylakoid membranes followed by Triton X-114 phase partitioning. When analyzed by one-dimensional polyacrylamide gel electrophoresis (PAGE), we obtain results which are very similar to those obtained by Kieselbach et al. using more classical methods [T. Kieselbach, A. Hagman, B. Andersson, W.P. Schroder, J. Biol. Chem. 273 (1998) 6710-6716]. About 25 major proteins are observed upon Coomassie blue staining. Upon two-dimensional isoelectric focusing-sodium dodecyl sulfate-PAGE and either Coomassie blue or silver staining, however, numerous other protein components are resolved. Our findings indicate that the total number of proteins (soluble and extrinsic membrane) present in the lumen may exceed 150.

  13. The role of glycinebetaine in the protection of spinach thylakoids against freezing stress.

    PubMed

    Coughlan, S J; Heber, U

    1982-11-01

    The quaternary ammonium compound glycinebetaine has been tested for cryoprotective properties, using isolated spinach thylakoids as a model membrane system. The effect of a 3-h,-20°C freezing regime on different photosynthetic parameters was measured. These parameters were the light-stimulated ΔpH formation and dark ΔpH decay, light-stimulated proton uptake, electron flow through photosystem II, photosystem I and total linear electron flow, and pyocyanine-mediated cyclic photophosphorylation. It was shown that below 100 mM glycinebetaine was superior as a cryoprotectant to sucrose on a molar, a molal and an activity basis. At higher concentrations, glycinebetaine was less efficient in preventing inactivation of thylakoids during freezing than sucrose. These observations are discussed in relation to the permeability of biomembranes to glycinebetaine and the colligative theory of cryoprotection. It is concluded that colligative protection is modified by direct interaction between cryoprotectant and membranes.

  14. Fast isolation of highly active photosystem II core complexes from spinach.

    PubMed

    Wang, Zhao-Gai; Xu, Tian-Hua; Liu, Cheng; Yang, Chun-Hong

    2010-09-01

    Purification of photosystem II (PSII) core complexes is a time-consuming and low-efficiency process. In order to isolate pure and active PSII core complexes in large amounts, we have developed a fast method to isolate highly active monomeric and dimeric PSII core complexes from spinach leaves by using sucrose gradient ultracentrifugation. By using a vertical rotor the process was completed significantly faster compared with a swing-out rotor. In order to keep the core complexes in high activity, the whole isolation procedure was performed in the presence of glycine betain and pH at 6.3. The isolated pigment-protein complexes were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, absorption spectroscopy, 77 K fluorescence spectroscopy and high performance liquid chromatography. Our results show that this method is a better choice for quick and efficient isolation of functionally active PSII core complexes. PMID:20738723

  15. Split Spinach Aptamer for Highly Selective Recognition of DNA and RNA at Ambient Temperatures.

    PubMed

    Kikuchi, Nanami; Kolpashchikov, Dmitry M

    2016-09-01

    Split spinach aptamer (SSA) probes for fluorescent analysis of nucleic acids were designed and tested. In SSA design, two RNA or RNA/DNA strands hybridized to a specific nucleic acid analyte and formed a binding site for low-fluorescent 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) dye, which resulted in up to a 270-fold increase in fluorescence. The major advantage of the SSA over state-of-the art fluorescent probes is high selectivity: it produces only background fluorescence in the presence of a single-base-mismatched analyte, even at room temperature. SSA is therefore a promising tool for label-free analysis of nucleic acids at ambient temperatures. PMID:27305425

  16. Visible light absorption and photo-sensitizing properties of spinach leaves and beetroot extracted natural dyes

    NASA Astrophysics Data System (ADS)

    Sengupta, D.; Mondal, B.; Mukherjee, K.

    2015-09-01

    Herein, chlorophyll and betalain dyes are extracted from fresh spinach leaves and beetroots. Fourier transform infrared spectra are used to identify the characteristic peaks of the extracted dyes. UV-vis light absorption characteristics of the dyes and their mixed counterpart are investigated by varying their pH and temperature. These dyes are used as photo sensitizer for fabrication of zinc oxide photo-anode based dye sensitized solar cells (DSSCs). The photo-voltaic characteristics of the developed DSSCs are measured under simulated solar light (power of incident light 100 mW cm-2 from Air Mass 1.5G). The solar to electric conversion efficiencies for the chlorophyll, betalain and mixed dye based solar cells are estimated as 0.148%, 0.197% and 0.294% respectively. The highest conversion efficiency for mixed dye based solar cell is attributed due to the absorption of wider range of solar spectrum.

  17. Combining Spinach-tagged RNA and gene localization to image gene expression in live yeast

    PubMed Central

    Guet, David; Burns, Laura T.; Maji, Suman; Boulanger, Jérôme; Hersen, Pascal; Wente, Susan R.; Salamero, Jean; Dargemont, Catherine

    2015-01-01

    Although many factors required for the formation of export-competent mRNPs have been described, an integrative view of the spatiotemporal coordinated cascade leading mRNPs from their site of transcription to their site of nuclear exit, at a single cell level, is still partially missing due to technological limitations. Here we report that the RNA Spinach aptamer is a powerful tool for mRNA imaging in live S. cerevisiae with high spatial-temporal resolution and no perturbation of the mRNA biogenesis properties. Dedicated image processing workflows are developed to allow detection of very low abundance of transcripts, accurate quantitative dynamic studies, as well as to provide a localization precision close to 100 nm at consistent time scales. Combining these approaches has provided a state-of-the-art analysis of the osmotic shock response in live yeast by localizing induced transcription factors, target gene loci and corresponding transcripts. PMID:26582123

  18. Fabrication of Organic Light-Emitting Diodes Using Photosynthetic Pigments Extracted from Spinach

    NASA Astrophysics Data System (ADS)

    Ohtani, Naoki; Kitagawa, Natsuko; Matsuda, Takashi

    2011-01-01

    We fabricated organic light-emitting diodes (OLEDs) containing chlorophylls in the active region, which were extracted from spinach using a chemical method. Photoluminescence (PL) cannot be observed in the thin film of the extracted chlorophylls owing to concentration quenching. To overcome the concentration quenching, a host material, poly[(m-phenylenevinylene)-alt-(2,5-dihexyloxy-p-phenylenevinylene)] (PPV) was added in the active region. This leads to the observaton of electroluminescence (EL) signals originating from chlorophyll a. We also evaluated the lifetime of the PL and EL. Consequently, the OLEDs containing carotenoids in the active region exhibit the light-emission much longer time than that without carotenoidos. This is assigned to the antioxidant activities of carotenoids. OLEDs containing a large amount of carotenoids are resistant to the oxidation damage.

  19. Characterization of elemental sulfur in isolated intact spinach chloroplasts. [Spinacia oleracea L

    SciTech Connect

    Joyard, J.; Douce, R. ); Forest, E. ); Blee, E. )

    1988-12-01

    Incubation of intact spinach (Spinacia oleracea L.) chloroplasts in the presence of {sup 35}SO{sub 4}{sup 2{minus}} resulted in the light-dependent formation of a chloroform-soluble sulfur-containing compound distinct from sulfolipid. The authors have identified this compound as the most stable form (S{sub 8}) of elemental sulfur (S{sup 0}, valence state for S = O) by mass spectrometry. It is possible that elemental sulfur (S{sup 0}) was formed by oxidation of bound sulfide, i.e. after the photoreduction of sulfate to sulfide by intact chloroplasts, and released as S{sub 8} under the experimental conditions used for analysis.

  20. Investigation of detergent effects on the solution structure of spinach Light Harvesting Complex II

    NASA Astrophysics Data System (ADS)

    Cardoso, Mateus B.; Smolensky, Dmitriy; Heller, William T.; O'Neill, Hugh

    2010-11-01

    The properties of spinach light harvesting complex II (LHC II), stabilized in the detergents Triton X-100 (TX100) and n-Octyl-β-D-Glucoside (BOG), were investigated by small-angle neutron scattering (SANS). The LHC II-BOG scattering curve overlaid well with the theoretical scattering curve generated from the crystal structure of LHC II indicating that the protein preparation was in its native functional state. On the other hand, the simulated LHC II curve deviated significantly from the LHC II-TX100 experimental data. Analysis by circular dichroism spectroscopy supported the SANS analysis and showed that LHC II-TX100 is inactivated. This investigation has implications for extracting and stabilizing photosynthetic membrane proteins for the development of biohybrid photoconversion devices.

  1. Temperature dependent steady state and picosecond kinetic fluorescence measurements of a photosystem I preparation from spinach

    SciTech Connect

    Mukerji, I.; Sauer, K.

    1988-08-01

    The fluorescence properties of a photosystem I (PSI) preparation from spinach containing approximately 200 chlorophyll (Chl) per reaction center were investigated. The preparation, characterized both spectroscopically and biochemically, contained the peripheral light harvesting antenna associated with PSI. In this study steady state fluorescence measurements were performed as a function of temperature. An emission maximum at 690 nm and a long wavelength shoulder from 710 to 740 nm were observed. The fluorescence yield at 690 nm is temperature independent, while the yield of the long wavelength shoulder increases dramatically with decreasing temperature. Additionally, kinetic measurements using the technique of single photon counting were done at room temperature and 77K. At 295K a four component fit was needed to describe the fluorescence decay; whereas at 77K, an additional 40-50 ps rise component indicative of fluorescence induction was necessary. 28 refs., 13 figs., 1 tab.

  2. Contribution of vitamin K1 to the electron spin polarization in spinach photosystem I

    SciTech Connect

    Rustandi, R.R.; Snyder, S.W.; Feezel, L.L.; Michalski, T.J.; Norris, J.R.; Thurnauer, M.C.; Biggins, J. )

    1990-09-04

    The electron spin polarized (ESP) electron paramagnetic resonance (EPR) signal observed in spinach photosystem I (PSI) particles was examined in preparations depleted of vitamin K1 by solvent extraction and following biological reconstitution by the quinone. The ESP EPR signal was not detected in the solvent-extracted PSI sample but was restored upon reconstitution with either protonated or deuterated vitamin K1 under conditions that also restored electron transfer to the terminal PSI acceptors. Reconstitution using deuterated vitamin K1 resulted in a line narrowing of the ESP EPR signal, supporting the conclusion that the ESP EPR signals in the reconstituted samples arise from a radical pair consisting of the oxidized PSI primary donor, P700+, and reduced vitamin K1.

  3. Electron spin resonance studies of urea-ferricyanide inactivated spinach photosystem I particles

    SciTech Connect

    Golbeck, J.H.; Warden, J.T.

    1981-09-01

    The photosystem I acceptor system of a subchloroplast particle from spinach was investigated by optical and electron spin resonance (ESR) spectroscopy following graduated inactivation of the bound iron-sulfur proteins by urea-ferricyanide. The chemical analysis of iron and sulfur and the ESR properties of centers A, B, and X are consistent with the participation of three iron-sulfur centers in photosystem I. A differential decrease in centers A, B, and X is observed under conditions which induce S= ..-->.. S/sup 0/ conversion in the bound iron-sulfur proteins. Center B is shown to be the most susceptible, while center X is the least susceptible component to oxidative denaturation. Stepwise inactivation experiments suggest that electron transport in photosystem I does not occur sequentially from X ..-->.. B ..-->.. A since there is quantitative photoreduction of center A in the absence of center B. We propose that center A is directly reduced by X.

  4. Photosynthetic activity of spinach chloroplasts after isopycnic centrifugation in gradients of silica.

    PubMed

    Morgenthaler, J J; Price, C A

    1974-10-01

    Chloroplast suspensions from spinach (Spinacia oleracea L.) were clearly resolved into intact and stripped chloroplasts by isopycnic centrifugation in density gradients of silica sol ("Ludox") and polyethlene glycol. The intact chloroplasts fixed CO(2) and evolved O(2) more rapidly than the crude suspensions; the stripped chloroplasts were inactive. During the photosynthetic fixation of (14)CO(2) in the intact chloroplasts recovered from the gradient, the (14)C label was observed to spread through the photosynthetic intermediate pools, as well as into starch, which indicates that the purified chloroplasts are metabolically competent. This appears to be the first report of the retention of photosynthetic activity following the purification of chloroplasts in density gradients. PMID:16658922

  5. Flavin and iron-sulfur containing ferredoxin-linked glutamate synthase from spinach leaves.

    PubMed

    Hirasawa, M; Tamura, G

    1984-04-01

    Ferredoxin-dependent glutamate synthase (native enzyme) [EC 1.4.7.1] of spinach has been purified to homogeneity in the presence of 2-oxoglutarate and sodium chloride and the properties of the enzyme have been studied. The molecular weight of the enzyme was estimated to be 140,000 by gel filtration. Subunit analysis by SDS-gel electrophoresis yielded a single protein band whose molecular weight was about 170,000. This purified enzyme showed a flavo-protein-like absorption spectrum having maxima at 279 and 438 nm with shoulders at 415 and 460 nm and a broad band around 360 nm. Fluorometric data indicated the presence of 2 mol of flavin per mol of the enzyme. Preliminary paper chromatography results indicated the presence of FAD and FMN in the purified enzyme. The enzyme also contained 4 mol of acid-labile sulfide and 4 g-atoms iron per mol of enzyme. In the absence of 2-oxoglutarate and/or sodium chloride, the purified enzyme was separated by either DE-52 cellulose chromatography or gel filtration with Ultrogel AcA 34 into two molecular forms (modified enzymes) with considerable inactivation. When reduced methyl viologen plus ferredoxin was used as the electron donor, the purified (native) enzyme showed high ferredoxin-dependent activity with a specific activity of 100 units/mg protein. Methyl viologen-dependent activity was negligible in the absence of ferredoxin. Kinetic properties and results of ESR studies were described. The results indicate that ferredoxin-linked glutamate synthase of spinach leaves is an iron-sulfur flavoprotein.

  6. Free Fatty Acids Regulate Two Galactosyltransferases in Chloroplast Envelope Membranes Isolated from Spinach Leaves

    PubMed Central

    Sakaki, Takeshi; Kondo, Noriaki; Yamada, Mitsuhiro

    1990-01-01

    Effects of MgCl2 and free fatty acids (FFA) on galactolipid:galactolipid galactosyltransferase (GGGT) and UDP-galactose: 1,2-diacylglycerol galactosyltransferase (UDGT) in chloroplast envelope membranes isolated from spinach (Spinacia oleracea L.) leaves were examined. GGGT activity was sigmoidally stimulated by MgCl2 with a saturated concentration of more than 5 millimolar. Free α-linolenic acid (18:3) caused a drastic increase in GGGT activity under limiting concentrations of MgCl2, without affecting its maximum activity at higher MgCl2 concentrations. Free 18:3 alone did not affect the GGGT activity. The effective species of FFA for the stimulation of GGGT activity in the presence of MgCl2 were unsaturated 16- and 18-carbon fatty acids. GGGT activity was also stimulated by 18:3 in the presence of MnCl2, CaCl2 and a high concentration of KCl in place of MgCl2. UDGT activity was hyperbolically enhanced by MgCl2 with a saturated concentration of 1 to 2 millimolar. In contrast to GGGT, UDGT was severely inhibited by 18:3, and MgCl2-induced stimulation was completely abolished by 18:3. Unsaturated 16- and 18-carbon fatty acids were more inhibitory to UDGT than the saturated acids. The dependence of GGGT activity on monogalactosyldiacylglycerol (MGDG) and MgCl2 concentrations was identical in the envelope membranes isolated from non- and ozone (0.5 microliter/liter)-fumigated spinach leaves, indicating that GGGT remained active in the leaves during ozone fumigation. The results are discussed in relation to the regulation of galactolipid biosynthesis by the endogenous FFA in the envelopes and to the involvement of GGGT in the triacylglycerol synthesis from MGDG in ozone-fumigated leaves. PMID:16667779

  7. Probing the donor side of photosystem II in spinach chloroplasts and algae using electron paramagnetic resonance

    SciTech Connect

    Boska, M.D.

    1985-11-01

    this work concerns electron transfer reactions in photosystem II (PS II). Investigations carried out in this work examine the redox reaction rates in PS II using EPR. In Tris-washed PS II preparations from spinach, it is observed that the oxidation kinetics of S II/sub f/, the EPR signal formed by Z/sup +/ after deactivation of oxygen evolution, mirror the reduction kinetics of P680/sup +/ seen by EPR in samples poised at a variety of pH's. These data agree with previous data on the optically measured reduction kinetics of P680/sup +/. The oxidation kinetics of S II/sub vf/, the EPR transient seen from Z/sup +/ in samples active in O/sub 2/ evolving samples, were instrument limited (t/sub 1/2/ less than 4 ..mu..s) and thus could not be directly measured. These results taken together support a model where Z donates electrons directly to P680/sup +/. The examination of the oxidation and reduction kinetics of S II in monovalent and divalent salt-washed PS II preparations from spinach correlated most of the change of Z oxidation and re-reduction kinetics seen upon Tris-treatment with the loss of a 33 kDa polypeptide associated with the donor side of PS II. These data coupled with observations of stead-state light-induced amplitude changes in S II give evidence for the existance of an electron carrier between the water-splitting enzyme and Z. Observation of S II amplitude and kinetics in highly resolved PS II protein complexes from Synechoccus sp., consisting of either a 5 polypeptide PS II core complex (E-1) or a 4 polypeptide PS II core complex (CP2b), localize Z and P680 within the 4 polypeptide complex. 187 refs., 17 figs., 7 tabs.

  8. Effect of growth regulators and role of roots in sex expression in spinach.

    PubMed

    Chailakhyan, M K; Khryanin, V N

    1978-01-01

    When 7-d-old plantlets of spinach (Spinacia oleracea L.) were immersed with their roots for 24 h in 25 mg/l gibberellic acid (GA3), or 15 mg/l 6-benzylaminopurine (6-BAP), or 15 mg/l indole-3-acetic acid (IAA), or 10 mg/l abscisic acid (ABA) and subsequently grown on long (18-h) days, the ratio of plants with male and female flowers, which in the controls was almost 1:1 (48 and 52%, respectively), was greatly altered. The treatments with 6-BAP, IAA and ABA raised the percentage of female plants to 88, 76 and 71%, respectively; the GA3 treatment increased the percent of male plants to 79%. When young, vegetative spinach plants (3 visible leaves) grown in 18-h days were cut a the root neck, and the shoots grown with their bases in nutrient solution, with adventitious roots either being allowed to develop or being systematically removed, 85% of the plants without roots became males, 85% of those with roots became females. But if the cut shoots were first, for 28 h, placed in a 15-mg/l 6-BAP solution and then grown in the absence of roots, the percent of female plants was restored to 84. These results fully agree with those obtained previously with hemp, namely, that plant growth regulators exert a regulating effect on the sex expression of dioecious plants when applied through the roots in early stages of development; that the root system plays an important role in determining the sex of these plants, that this role of the roots is associated with the synthesis of cytokinins in them. Dioecious short- and long-day plants do not differ in these respects.

  9. Maple Leaf Outdoor Centre.

    ERIC Educational Resources Information Center

    Maguire, Molly; Gunton, Ric

    2000-01-01

    Maple Leaf Outdoor Centre (Ontario) has added year-round outdoor education facilities and programs to help support its summer camp for disadvantaged children. Schools, youth centers, religious groups, and athletic teams conduct their own programs, collaborate with staff, or use staff-developed programs emphasizing adventure education and personal…

  10. Bacterial leaf spot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial leaf spot has been reported in Australia (Queensland), Egypt, El Salvador, India, Japan, Nicaragua, Sudan, and the United States (Florida, Iowa, Kansas, Maryland, and Wisconsin). It occasionally causes locally severe defoliation and post-emergence damping-off and stunting. The disease is...

  11. Comparative leaf development in angiosperms.

    PubMed

    Tsukaya, Hirokazu

    2014-02-01

    Recent accumulation of our knowledge on basic leaf development mechanisms in model angiosperm species has allowed us to pursue evolutionary development (evo/devo) studies of various kinds of leaf development. As a result, unexpected findings and clues have been unearthed aiding our understanding of the mechanisms involved in the diversity of leaf morphology, although the covered remain limited. In this review, we highlight recent findings of diversified leaf development in angiosperms.

  12. The acute effect of flavonoid-rich apples and nitrate-rich spinach on cognitive performance and mood in healthy men and women.

    PubMed

    Bondonno, Catherine P; Downey, Luke A; Croft, Kevin D; Scholey, Andrew; Stough, Con; Yang, Xingbin; Considine, Michael J; Ward, Natalie C; Puddey, Ian B; Swinny, Ewald; Mubarak, Aidilla; Hodgson, Jonathan M

    2014-05-01

    Flavonoids and nitrate in a fruit and vegetable diet may be protective against cardiovascular disease and cognitive decline through effects on nitric oxide (NO) status. The circulating NO pool is increased via distinct pathways by dietary flavonoids and nitrate. Our aim was to investigate the acute effects of apples, rich in flavonoids, and spinach, rich in nitrate, independently and in combination on NO status, cognitive function and mood in a randomised, controlled, cross-over trial with healthy men and women (n = 30). The acute effects of four energy-matched treatments (control, apple, spinach and apple + spinach) were compared. Endpoints included plasma nitric oxide status (determined by measuring S-nitrosothiols + other nitroso species (RXNO)), plasma nitrate and nitrite, salivary nitrate and nitrite, urinary nitrate and nitrite as well as cognitive function (determined using the Cognitive Drug Research (CDR) computerized cognitive assessment battery) and mood. Relative to control, all treatments resulted in higher plasma RXNO. A significant increase in plasma nitrate and nitrite, salivary nitrate and nitrite as well as urinary nitrate and nitrite was observed with spinach and apple + spinach compared to control. No significant effect was observed on cognitive function or mood. In conclusion, flavonoid-rich apples and nitrate-rich spinach augmented NO status acutely with no concomitant improvements or deterioration in cognitive function and mood.

  13. Effects of mercury on visible/near-infrared reflectance spectra of mustard spinach plants (Brassica rapa P.).

    PubMed

    Dunagan, Sarah C; Gilmore, Martha S; Varekamp, Johan C

    2007-07-01

    Mustard spinach plants were grown in mercury-spiked and contaminated soils collected in the field under controlled laboratory conditions over a full growth cycle to test if vegetation grown in these soils has discernible characteristics in visible/near-infrared (VNIR) spectra. Foliar Hg concentrations (0.174-3.993ppm) of the Mustard spinach plants were positively correlated with Hg concentration of soils and varied throughout the growing season. Equations relating foliar Hg concentration to spectral reflectance, its first derivative, and selected vegetation indices were generated using stepwise multiple linear regression. Significant correlations are found for limited wavelengths for specific treatments and dates. Ratio Vegetation Index (RVI) and Red Edge Position (REP) values of plants in Hg-spiked and field-contaminated soils are significantly lower relative to control plants during the early and middle portions of the growth cycle which may be related to lower chlorophyll abundance or functioning in Hg-contaminated plants.

  14. The broad spectrum 2-oxoglutarate oxygenase inhibitor N-oxalylglycine is present in rhubarb and spinach leaves.

    PubMed

    Al-Qahtani, Khalid; Jabeen, Bushra; Sekirnik, Rok; Riaz, Naheed; Claridge, Timothy D W; Schofield, Christopher J; McCullagh, James S O

    2015-09-01

    2-Oxoglutarate (2OG) and ferrous iron dependent oxygenases are involved in many biological processes in organisms ranging from humans (where some are therapeutic targets) to plants. These enzymes are of significant biomedicinal interest because of their roles in hypoxic signaling and epigenetic regulation. Synthetic N-oxalylglycine (NOG) has been identified as a broad-spectrum 2OG oxygenase inhibitor and is currently widely used in studies on the hypoxic response and chromatin modifications in animals. We report the identification of NOG as a natural product present in Rheum rhabarbarum (rhubarb) and Spinach oleracea (spinach) leaves; NOG was not observed in Escherchia coli or human embryonic kidney cells (HEK 293T). The finding presents the possibility that NOG plays a natural role in regulating gene expression by inhibiting 2OG dependent oxygenases. This has significance because tricarboxylic acid cycle (TCA) intermediate inhibition of 2OG dependent oxygenases has attracted major interest in cancer research.

  15. Leaf absorbance and photosynthesis

    NASA Technical Reports Server (NTRS)

    Schurer, Kees

    1994-01-01

    The absorption spectrum of a leaf is often thought to contain some clues to the photosynthetic action spectrum of chlorophyll. Of course, absorption of photons is needed for photosynthesis, but the reverse, photosynthesis when there is absorption, is not necessarily true. As a check on the existence of absorption limits we measured spectra for a few different leaves. Two techniques for measuring absorption have been used, viz. the separate determination of the diffuse reflectance and the diffuse transmittance with the leaf at a port of an integrating sphere and the direct determination of the non-absorbed fraction with the leaf in the sphere. In a cross-check both methods yielded the same results for the absorption spectrum. The spectrum of a Fuchsia leaf, covering the short-wave region from 350 to 2500 nm, shows a high absorption in UV, blue and red, the well known dip in the green and a steep fall-off at 700 nm. Absorption drops to virtually zero in the near infrared, with subsequent absorptions, corresponding to the water absorption bands. In more detailed spectra, taken at 5 nm intervals with a 5 nm bandwidth, differences in chlorophyll content show in the different depths of the dip around 550 nm and in a small shift of the absorption edge at 700 nm. Spectra for Geranium (Pelargonium zonale) and Hibiscus (with a higher chlorophyll content) show that the upper limit for photosynthesis can not be much above 700 nm. No evidence, however, is to be seen of a lower limit for photosynthesis and, in fact, some experiments down to 300 nm still did not show a decrease of the absorption although it is well recognized that no photosynthesis results with 300 nm wavelengths.

  16. Effects of stresses on the growth and Cytotoxicity of Shiga-Toxin producing Escherichia coli in ground beef and spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were to examine the effect of stresses on the growth and cytotoxicity of pathogenic Escherichia coli in beef and spinach. A mixture of three strains of Shiga toxin-producing E. coli (STEC) O157:H7 or four strains of non-O157 STEC O26:H11, O103:H1, O104:H4, and O145:NM wa...

  17. Characterization of spinach ribulose-1,5-bisphosphate carboxylase/oxygenase activase isoforms reveals hexameric assemblies with increased thermal stability.

    PubMed

    Keown, Jeremy R; Pearce, Frederick Grant

    2014-12-15

    Most plants contain two isoforms of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (Rca), a chloroplast protein that maintains the activity of Rubisco during photosynthesis. The longer (α-) Rca isoform has previously been shown to regulate the activity of Rubisco in response to both the ADP:ATP ratio and redox potential via thioredoxin-f. We have characterized the arrangement of the different spinach (Spinacia oleracea) isoforms in solution, and show how the presence of nucleotides changes the oligomeric state. Although the shorter (β-) isoform from both tobacco (Nicotiana tabacum) and spinach tend to form a range of oligomers in solution, the size of which are relatively unaffected by the addition of nucleotide, the spinach α-isoform assembles as a hexamer in the presence of adenosine 5'-[γ-thio]triphosphate (ATPγS). These hexamers have significantly higher heat stability, and may play a role in optimizing photosynthesis at higher temperatures. Hexamers were also observed for mixtures of the two isoforms, suggesting that the α-isoform can act as a structural scaffold for hexamer formation by the β-isoform. Additionally, it is shown that a variant of the tobacco β-isoform acts in a similar fashion to the α-isoform of spinach, forming thermally stable hexamers in the presence of ATPγS. Both isoforms had similar rates of ATP hydrolysis, suggesting that a propensity for hexamer formation may not necessarily be correlated with activity. Modelling of the hexameric structures suggests that although the N-terminus of Rca forms a highly dynamic, extended structure, the C-terminus is located adjacent to the intersubunit interface.

  18. Interactions of plant zinc and plant species on the bioavailability of plant cadmium to Japanese quail fed lettuce and spinach

    SciTech Connect

    McKenna, I.M.; Keach, R.M. Jr; Williams, F.M. ); Chaney, R.L. Dept. of Agriculture, Beltsville, MD ); Tao, Shyy-Hwa )

    1992-02-01

    Many cadmium-contaminated environments contain high levels of zinc. The effects of plant Zn and plant species on plant Cd bioavailability were tested in Japanese quail fed lettuce and spinach. Four groups of birds received 10% of their diets as lettuce or spinach leaves intrinsically labeled with {sup 109}Cd and containing low or high intrinsic Zn. Two other groups were fed control diets containing {sup 109}Cd as CdSO{sub 4} and low or high Zn as ZnCO{sub 3}. Cadmium concentrations in diets ranged from 0.857 to 1.05 {mu}g/g dry wt. Zinc concentrations in low-Zn diets ranged from 21.2 to 22.8, and in high-Zn diets from 56.0 to 63.3 {mu}g/g dry wt. Increased lettuce and spinach Zn decreased plant Cd retention in kidney, liver, and jejunum-ileum of Japanese quail. Spinach Cd was less absorbed than lettuce Cd at both Zn levels. Inorganic Zn produced a lesser decrease in Cd retention in kidney, liver, and jejunum-ileum than did plant Zn. The authors conclude that (1) crops that transport Zn and Cd readily into edible tissues show lower Cd bioavailability when grown in Zn-Cd contaminated environments than in Cd-only polluted sites, (2) plant species differ in Cd bioavailability for identical concentrations of Zn and Cd in edible tissues, and (3) toxicological studies with animals exposed to Cd salts and Zn supplements do not assess Cd bioavailability of Zn-Cd contaminated crops.

  19. Live Cell Imaging Using Riboswitch-Spinach tRNA Fusions as Metabolite-Sensing Fluorescent Biosensors.

    PubMed

    Kellenberger, Colleen A; Hallberg, Zachary F; Hammond, Ming C

    2015-01-01

    The development of fluorescent biosensors is motivated by the desire to monitor cellular metabolite levels in real time. Most genetically encodable fluorescent biosensors are based on receptor proteins fused to fluorescent protein domains. More recently, small molecule-binding riboswitches have been adapted for use as fluorescent biosensors through fusion to the in vitro selected Spinach aptamer, which binds a pro-fluorescent, cell-permeable small molecule mimic of the GFP chromophore, DFHBI. Here we describe methods to prepare and analyze riboswitch-Spinach tRNA fusions for ligand-dependent activation of fluorescence in vivo. Example procedures describe the use of the Vc2-Spinach tRNA biosensor to monitor perturbations in cellular levels of cyclic di-GMP using either fluorescence microscopy or flow cytometry. The relative ease of cloning and imaging of these biosensors, as well as their modular nature, should make this method appealing to other researchers interested in utilizing riboswitch-based biosensors for metabolite sensing.

  20. Level 2 validation of a flow cytometric method for detection of Escherichia coli O157:H7 in raw spinach.

    PubMed

    Williams, Anna J; Cooper, Willie M; Summage-West, Christine V; Sims, Lillie M; Woodruff, Robert; Christman, Jessica; Moskal, Ted J; Ramsaroop, Shawn; Sutherland, John B; Alusta, Pierre; Wilkes, Jon G; Buzatu, Dan A

    2015-12-23

    The Bacteriological Analytical Manual (BAM) method currently used by the United States Food and Drug Administration (FDA) to detect Escherichia coli O157:H7 in spinach was systematically compared to a new flow cytometry based method. This Food and Drug Administration (FDA) level 2 external laboratory validation study was designed to determine the latter method's sensitivity and speed for analysis of this pathogen in raw spinach. Detection of target cell inoculations with a low cell count is critical, since enterohemorrhagic strains of E. coli require an infective dose of as few as 10 cells (Schmid-Hempel and Frank, 2007). Although, according to the FDA, the infectious dose is unknown (Food and Drug Administration, 1993). Therefore, the inoculation level into the spinach, a total of 2.0±2.6 viable E. coli O157 cells, was specified to yield between 25% and 75% detection by the new method, out of 20 samples (10 positives and 10 negatives). This criterion was met in that the new method detected 60% of the nominally positive samples; the corresponding sensitivity of the reference method was 50%. For both methods the most likely explanation for false negatives was that no viable cells were actually introduced into the sample. In this validation study, the flow cytometry method was equal to the BAM in sensitivity and far superior in speed.

  1. Effect of sulfur and nitrogen fertilization on the content of nutritionally relevant carotenoids in spinach ( Spinacia oleracea ).

    PubMed

    Reif, Constance; Arrigoni, Eva; Neuweiler, Reto; Baumgartner, Daniel; Nyström, Laura; Hurrell, Richard F

    2012-06-13

    Spinach is an important dietary source of lutein and β-carotene. Their synthesis is closely linked to chlorophyll synthesis and dependent upon an adequate supply of sulfur and nitrogen. Soils may become sulfur-deficient during winter because microorganisms convert atmospheric SO2 less efficiently to sulfate. The influence of sulfur and nitrogen fertilization on the carotenoid and chlorophyll contents of spinach grown in summer or winter was investigated. Carotenoid and chlorophyll levels were positively correlated. Lutein and β-carotene were 25% higher in summer than in winter. Winter levels were increased by 35-40% by sulfur fertilization in one location but not in the other, with the impact depending upon soil type, growing location, and atmospheric conditions. Carotenoids were little or not affected by nitrogen addition in winter or sulfur addition in summer. It is concluded that sulfur fertilization of spinach in winter may modestly increase carotenoids but high carotenoid levels are best assured with carotenoid-rich cultivars grown in summer.

  2. Differences in responses of summer and winter spinach to elevated UV-B at varying soil NPK levels.

    PubMed

    Singh, Suruchi; Agrawal, Madhoolika; Agrawal, S B

    2014-05-01

    Seasonal variations in response of spinach to elevated ultraviolet-B (UV-B) during summer and winter were assessed with respect to growth, biomass, yield, NPK uptake and NPK use efficiencies at varying NPK levels. The nutrient amendments were recommended NPK (RNPK) and 1.5 times recommended NPK (1.5 RNPK). Season significantly affected the measured parameters except the number of leaves. Under ambient UV-B, the growth performance of summer spinach was better in both the NPK levels, higher being at 1.5 RNPK leading to higher nutrient uptake. However, more reduction in biomass under elevated UV-B in 1.5 RNPK was recorded during summer, while during winter in RNPK. Reduction in biomass under elevated UV-B was accompanied by the modification in its partitioning with more biomass allocation to root during summer compared to winter at both the NPK levels. NPK uptake was higher in summer, while NPK use efficiencies were higher during winter. At higher than recommended NPK level, better NPK use efficiencies were displayed during both the seasons. Increased NPK supply during winter enabled spinach to capitalize light more efficiently and hence increased biomass accumulation. Strategies for surviving elevated UV-B in winter differ from those that provided protection from the same stress when it occurs in summer.

  3. The metabolic significance of octulose phosphates in the photosynthetic carbon reduction cycle in spinach

    PubMed Central

    MacLeod, John K.

    2006-01-01

    14C-Labelled octulose phosphates were formed during photosynthetic 14CO2 fixation and were measured in spinach leaves and chloroplasts. Because mono- and bisphosphates of d-glycero-d-ido-octulose are the active 8-carbon ketosugar intermediates of the L-type pentose pathway, it was proposed that they may also be reactants in a modified Calvin–Benson–Bassham pathway reaction scheme. This investigation therefore initially focussed only on the ido-epimer of the octulose phosphates even though 14C-labelled d-glycero-d-altro-octulose mono- and bisphosphates were also identified in chloroplasts and leaves. 14CO2 predominantly labelled positions 5 and 6 of d-glycero-d-ido-octulose 1,8-P2 consistent with labelling predictions of the modified scheme. The kinetics of 14CO2 incorporation into ido-octulose was similar to its incorporation into some traditional intermediates of the path of carbon, while subsequent exposure to 12CO2 rapidly displaced the 14C isotope label from octulose with the same kinetics of label loss as some of the confirmed Calvin pathway intermediates. This is consistent with octulose phosphates having the role of cyclic intermediates rather than synthesized storage products. (Storage products don’t rapidly exchange isotopically labelled carbons with unlabelled CO2.) A spinach chloroplast extract, designated stromal enzyme preparation (SEP), catalysed and was used to measure rates of CO2 assimilation with Calvin cycle intermediates and octulose and arabinose phosphates. Only pentose (but not arabinose) phosphates and sedoheptulose 7-phosphate supported CO2 fixation at rates in excess of 120 μmol h−1 mg−1 Chl. Rates for octulose, sedoheptulose and fructose bisphosphates, octulose, hexose and triose monophosphates were all notably less than the above rate and arabinose 5-phosphate was inactive. Altro-octulose phosphates were more active than phosphate esters of the ido-epimer. The modified scheme proposed a specific phosphotransferase and SEP

  4. Functional analysis of B and C class floral organ genes in spinach demonstrates their role in sexual dimorphism

    PubMed Central

    2010-01-01

    Background Evolution of unisexual flowers entails one of the most extreme changes in plant development. Cultivated spinach, Spinacia oleracea L., is uniquely suited for the study of unisexual flower development as it is dioecious and it achieves unisexually by the absence of organ development, rather than by organ abortion or suppression. Male staminate flowers lack fourth whorl primordia and female pistillate flowers lack third whorl primordia. Based on theoretical considerations, early inflorescence or floral organ identity genes would likely be directly involved in sex-determination in those species in which organ initiation rather than organ maturation is regulated. In this study, we tested the hypothesis that sexual dimorphism occurs through the regulation of B class floral organ gene expression by experimentally knocking down gene expression by viral induced gene silencing. Results Suppression of B class genes in spinach resulted in the expected homeotic transformation of stamens into carpels but also affected the number of perianth parts and the presence of fourth whorl. Phenotypically normal female flowers developed on SpPI-silenced male plants. Suppression of the spinach C class floral organ identity gene, SpAG, resulted in loss of reproductive organ identity, and indeterminate flowers, but did not result in additional sex-specific characteristics or structures. Analysis of the genomic sequences of both SpAP3 and SpPI did not reveal any allelic differences between males and females. Conclusion Sexual dimorphism in spinach is not the result of homeotic transformation of established organs, but rather is the result of differential initiation and development of the third and fourth whorl primordia. SpAG is inferred to have organ identity and meristem termination functions similar to other angiosperm C class genes. In contrast, while SpPI and SpAP3 resemble other angiosperms in their essential functions in establishing stamen identity, they also appear to have

  5. Efficient Photoelectrochemical Energy Conversion using Spinach Photosystem II (PSII) in Lipid Multilayer Films.

    PubMed

    Zhang, Yun; Magdaong, Nikki M; Shen, Min; Frank, Harry A; Rusling, James F

    2015-04-01

    The need for clean, renewable energy has fostered research into photovoltaic alternatives to silicon solar cells. Pigment-protein complexes in green plants convert light energy into chemical potential using redox processes that produce molecular oxygen. Here, we report the first use of spinach protein photosystem II (PSII) core complex in lipid films in photoelectrochemical devices. Photocurrents were generated from PSII in a ∼2 μm biomimetic dimyristoylphosphatidylcholine (DMPC) film on a pyrolytic graphite (PG) anode with PSII embedded in multiple lipid bilayers. The photocurrent was ∼20 μA cm(-2) under light intensity 40 mW cm(-2). The PSII-DMPC anode was used in a photobiofuel cell with a platinum black mesh cathode in perchloric acid solution to give an output voltage of 0.6 V and a maximum output power of 14 μW cm(-2). Part of this large output is related to a five-unit anode-cathode pH gradient. With catholytes at higher pH or no perchlorate, or using an MnO2 oxygen-reduction cathode, the power output was smaller. The results described raise the possibility of using PSII-DMPC films in small portable power conversion devices.

  6. X-ray absorption spectroscopy and EPR studies of oriented spinach thylakoid preparations

    SciTech Connect

    Andrews, J.C. |

    1995-08-01

    In this study, oriented Photosystem II (PS II) particles from spinach chloroplasts are studied with electron paramagnetic resonance (EPR) and x-ray absorption spectroscopy (XAS) to determine more details of the structure of the oxygen evolving complex (OEC). The nature of halide binding to Mn is also studied with Cl K-edge and Mn EXAFS (extended x-ray absorption fine structure) of Mn-Cl model compounds, and with Mn EXAFS of oriented PS II in which Br has replaced Cl. Attention is focused on the following: photosynthesis and the oxygen evolving complex; determination of mosaic spread in oriented photosystem II particles from signal II EPR measurement; oriented EXAFS--studies of PS II in the S{sub 2} state; structural changes in PS II as a result of treatment with ammonia: EPR and XAS studies; studies of halide binding to Mn: Cl K-edge and Mn EXAFS of Mn-Cl model compounds and Mn EXAFS of oriented Br-treated photosystem II.

  7. Survival of Salmonella enterica in Dried Turkey Manure and Persistence on Spinach Leaves.

    PubMed

    Oni, Ruth A; Sharma, Manan; Buchanan, Robert L

    2015-10-01

    Concerns about the microbiological safety of fresh produce have attracted attention in the past three decades due to multiple foodborne outbreaks. Animal manure contaminated with enteric pathogens has been identified as an important preharvest pathogen source. This study investigated the survival of Salmonella enterica in dust particles of dehydrated turkey manure and how association with manure dust may enhance the survival of salmonellae on leafy greens in the field. The survival of a cocktail of multiple Salmonella serotypes in the dried fecal material of various particle sizes (125 to 500 μm) was examined at varying moisture contents (5, 10, and 15%). Survival times of the pathogen were inversely related to moisture content and particle size of manure dust, with viable Salmonella still detectable for up to 291 days in the smallest particle size (125 μm) with 5% moisture. Association with manure dust particles increased the survival of Salmonella when subjected to UV light both under laboratory conditions and on the surface of spinach leaves in a greenhouse setting. The results of this study suggest that aerosolized manure particles could be a potential vehicle for Salmonella dispersal to leafy greens if the microorganism is present in the dry manure.

  8. Spinach thylakoid polyphenol oxidase isolation, activation, and properties of the native chloroplast enzyme

    SciTech Connect

    Golbeck, J.H.; Cammarata, K.V.

    1981-05-01

    Polyphenol oxidase activity (E.C. 1.14,18.1) has been found in two enzyme species isolated from thylakoid membranes of spinach chloroplasts. The proteins were released from the membrane by sonication and purified >900-fold by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatography. The enzymes appear to be the tetramer and monomer of a subunit with a molecular weight of 42,500 as determined by lithium dodecyl sulfate gel electrophoresis. Sonication releases polyphenol oxidase from the membrane largely in the latent state. In the absence of added fatty acids, the isolated enzyme spontaneously, but slowly, activates with time. Purified polyphenol oxidase utilizes o-diphenols as substrates and shows no detectable levels of monophenol or p-diphenol oxidase activities. Suitable substrates include chlorogenic acid, catechol, caffeic acid, pyrogallol, and dopamine; however, the enzyme is substrate-inhibited by the last four at concentrations near their K/sub m/. A large seasonal variation in polyphenol oxidase activity may result from a decrease in enzyme content rather than inhibition of the enzyme present.

  9. Cryo-EM structure of the large subunit of the spinach chloroplast ribosome

    PubMed Central

    Ahmed, Tofayel; Yin, Zhan; Bhushan, Shashi

    2016-01-01

    Protein synthesis in the chloroplast is mediated by the chloroplast ribosome (chloro-ribosome). Overall architecture of the chloro-ribosome is considerably similar to the Escherichia coli (E. coli) ribosome but certain differences are evident. The chloro-ribosome proteins are generally larger because of the presence of chloroplast-specific extensions in their N- and C-termini. The chloro-ribosome harbours six plastid-specific ribosomal proteins (PSRPs); four in the small subunit and two in the large subunit. Deletions and insertions occur throughout the rRNA sequence of the chloro-ribosome (except for the conserved peptidyl transferase center region) but the overall length of the rRNAs do not change significantly, compared to the E. coli. Although, recent advancements in cryo-electron microscopy (cryo-EM) have provided detailed high-resolution structures of ribosomes from many different sources, a high-resolution structure of the chloro-ribosome is still lacking. Here, we present a cryo-EM structure of the large subunit of the chloro-ribosome from spinach (Spinacia oleracea) at an average resolution of 3.5 Å. High-resolution map enabled us to localize and model chloro-ribosome proteins, chloroplast-specific protein extensions, two PSRPs (PSRP5 and 6) and three rRNA molecules present in the chloro-ribosome. Although comparable to E. coli, the polypeptide tunnel and the tunnel exit site show chloroplast-specific features. PMID:27762343

  10. Toxicity and Bioaccumulation of Heavy Metals in Spinach (Spinacia oleracea) Grown in a Controlled Environment.

    PubMed

    Alia, Naz; Sardar, Khan; Said, Muhammad; Salma, Khalid; Sadia, Alam; Sadaf, Siddique; Toqeer, Ahmed; Miklas, Scholz

    2015-07-01

    The impact of heavy metal toxicity on the shoot and root lengths, total protein, fiber characteristics, moisture content and nutrient composition of spinach (Spinacia oleracea) was evaluated. Plants were grown in pots containing soil and treated with different concentrations (mg/kg) of lead (Pb; 300, 400 and 500), cadmium (Cd; 0.5, 1 and 1.5) and zinc (Zn; 250, 500, and 700) as well as mixtures of Cd and Pb (0.5/300, 1/400, 1.5/500), Cd and Zn (0.5/250, 1/500, 1.5/700), and Pb and Zn (300/250, 400/500, 500/700). Soil contaminated by long-term irrigation with wastewater containing heavy metals was simulated. An increase in concentrations of heavy metals both individually and as mixtures significantly (p < 0.05) reduced the growth parameters and nutrient contents of S. oleracea. The uptake patterns of heavy metals in mixtures showed antagonistic impacts on each other. The toxicities of the mixtures Cd and Pb, Cd and Zn as well as Pb and Zn were higher than those observed in separate heavy metal applications but less than their additive sums. The toxicity caused by individual heavy metals was the highest for Cd followed by Pb and Zn. The highest toxicity was observed in plants grown in soil contaminated by Cd and Pb. PMID:26133131

  11. Sub-plastidial localization of two different phage-type RNA polymerases in spinach chloroplasts

    PubMed Central

    Azevedo, Jacinthe; Courtois, Florence; Lerbs-Mache, Silva

    2006-01-01

    Plant plastids contain a circular genome of ∼150 kb organized into ∼35 transcription units. The plastid genome is organized into nucleoids and attached to plastid membranes. This relatively small genome is transcribed by at least two different RNA polymerases, one being of the prokaryotic type and plastid-encoded (PEP), the other one being of the phage-type and nucleus-encoded (NEP). The presumed localization of a second phage-type RNA polymerase in plastids is still questionable. There is strong evidence for a sequential action of NEP and PEP enzymes during plant development attributing a prevailing role of NEP during early plant and plastid development, although NEP is present in mature chloroplasts. In the present paper, we have analysed two different NEP enzymes from spinach with respect to subcellular and intra-plastidial localization in mature chloroplasts with the help of specific antibodies. Results show the presence of the two different NEP enzymes in mature chloroplasts. Both enzymes are entirely membrane bound but, unlike previously thought, this membrane binding is not mediated via DNA. This finding indicates that NEP enzymes are not found as elongating transcription complexes on the template DNA in mature chloroplasts and raises the question of their function in mature chloroplasts. PMID:16421271

  12. Toxicity and Bioaccumulation of Heavy Metals in Spinach (Spinacia oleracea) Grown in a Controlled Environment.

    PubMed

    Alia, Naz; Sardar, Khan; Said, Muhammad; Salma, Khalid; Sadia, Alam; Sadaf, Siddique; Toqeer, Ahmed; Miklas, Scholz

    2015-06-30

    The impact of heavy metal toxicity on the shoot and root lengths, total protein, fiber characteristics, moisture content and nutrient composition of spinach (Spinacia oleracea) was evaluated. Plants were grown in pots containing soil and treated with different concentrations (mg/kg) of lead (Pb; 300, 400 and 500), cadmium (Cd; 0.5, 1 and 1.5) and zinc (Zn; 250, 500, and 700) as well as mixtures of Cd and Pb (0.5/300, 1/400, 1.5/500), Cd and Zn (0.5/250, 1/500, 1.5/700), and Pb and Zn (300/250, 400/500, 500/700). Soil contaminated by long-term irrigation with wastewater containing heavy metals was simulated. An increase in concentrations of heavy metals both individually and as mixtures significantly (p < 0.05) reduced the growth parameters and nutrient contents of S. oleracea. The uptake patterns of heavy metals in mixtures showed antagonistic impacts on each other. The toxicities of the mixtures Cd and Pb, Cd and Zn as well as Pb and Zn were higher than those observed in separate heavy metal applications but less than their additive sums. The toxicity caused by individual heavy metals was the highest for Cd followed by Pb and Zn. The highest toxicity was observed in plants grown in soil contaminated by Cd and Pb.

  13. Efficient Photoelectrochemical Energy Conversion using Spinach Photosystem II (PSII) in Lipid Multilayer Films

    PubMed Central

    Zhang, Yun; Magdaong, Nikki M; Shen, Min; Frank, Harry A; Rusling, James F

    2015-01-01

    The need for clean, renewable energy has fostered research into photovoltaic alternatives to silicon solar cells. Pigment–protein complexes in green plants convert light energy into chemical potential using redox processes that produce molecular oxygen. Here, we report the first use of spinach protein photosystem II (PSII) core complex in lipid films in photoelectrochemical devices. Photocurrents were generated from PSII in a ∼2 μm biomimetic dimyristoylphosphatidylcholine (DMPC) film on a pyrolytic graphite (PG) anode with PSII embedded in multiple lipid bilayers. The photocurrent was ∼20 μA cm−2 under light intensity 40 mW cm−2. The PSII–DMPC anode was used in a photobiofuel cell with a platinum black mesh cathode in perchloric acid solution to give an output voltage of 0.6 V and a maximum output power of 14 μW cm−2. Part of this large output is related to a five-unit anode–cathode pH gradient. With catholytes at higher pH or no perchlorate, or using an MnO2 oxygen-reduction cathode, the power output was smaller. The results described raise the possibility of using PSII–DMPC films in small portable power conversion devices. PMID:25969807

  14. Crystallographic structure of the turbine C-ring from spinach chloroplast F-ATP synthase

    PubMed Central

    Balakrishna, Asha Manikkoth; Seelert, Holger; Marx, Sven-Hendric; Dencher, Norbert A.; Grüber, Gerhard

    2014-01-01

    In eukaryotic and prokaryotic cells, F-ATP synthases provide energy through the synthesis of ATP. The chloroplast F-ATP synthase (CF1FO-ATP synthase) of plants is integrated into the thylakoid membrane via its FO-domain subunits a, b, b’ and c. Subunit c with a stoichiometry of 14 and subunit a form the gate for H+-pumping, enabling the coupling of electrochemical energy with ATP synthesis in the F1 sector. Here we report the crystallization and structure determination of the c14-ring of subunit c of the CF1FO-ATP synthase from spinach chloroplasts. The crystals belonged to space group C2, with unit-cell parameters a=144.420, b=99.295, c=123.51 Å, and β=104.34° and diffracted to 4.5 Å resolution. Each c-ring contains 14 monomers in the asymmetric unit. The length of the c-ring is 60.32 Å, with an outer ring diameter 52.30 Å and an inner ring width of 40 Å. PMID:24521269

  15. Toxicity and Bioaccumulation of Heavy Metals in Spinach (Spinacia oleracea) Grown in a Controlled Environment

    PubMed Central

    Alia, Naz; Sardar, Khan; Said, Muhammad; Salma, Khalid; Sadia, Alam; Sadaf, Siddique; Toqeer, Ahmed; Miklas, Scholz

    2015-01-01

    The impact of heavy metal toxicity on the shoot and root lengths, total protein, fiber characteristics, moisture content and nutrient composition of spinach (Spinacia oleracea) was evaluated. Plants were grown in pots containing soil and treated with different concentrations (mg/kg) of lead (Pb; 300, 400 and 500), cadmium (Cd; 0.5, 1 and 1.5) and zinc (Zn; 250, 500, and 700) as well as mixtures of Cd and Pb (0.5/300, 1/400, 1.5/500), Cd and Zn (0.5/250, 1/500, 1.5/700), and Pb and Zn (300/250, 400/500, 500/700). Soil contaminated by long-term irrigation with wastewater containing heavy metals was simulated. An increase in concentrations of heavy metals both individually and as mixtures significantly (p < 0.05) reduced the growth parameters and nutrient contents of S. oleracea. The uptake patterns of heavy metals in mixtures showed antagonistic impacts on each other. The toxicities of the mixtures Cd and Pb, Cd and Zn as well as Pb and Zn were higher than those observed in separate heavy metal applications but less than their additive sums. The toxicity caused by individual heavy metals was the highest for Cd followed by Pb and Zn. The highest toxicity was observed in plants grown in soil contaminated by Cd and Pb. PMID:26133131

  16. Structural insights into energy regulation of light-harvesting complex CP29 from spinach.

    PubMed

    Pan, Xiaowei; Li, Mei; Wan, Tao; Wang, Longfei; Jia, Chenjun; Hou, Zhiqiang; Zhao, Xuelin; Zhang, Jiping; Chang, Wenrui

    2011-03-01

    CP29, one of the minor light-harvesting complexes of higher-plant photosystem II, absorbs and transfers solar energy for photosynthesis and also has important roles in photoprotection. We have solved the crystal structure of spinach CP29 at 2.80-Å resolution. Each CP29 monomer contains 13 chlorophyll and 3 carotenoid molecules, which differs considerably from the major light-harvesting complex LHCII and the previously proposed CP29 model. The 13 chlorophyll-binding sites are assigned as eight chlorophyll a sites, four chlorophyll b and one putative mixed site occupied by both chlorophylls a and b. Based on the present X-ray structure, an integrated pigment network in CP29 is constructed. Two special clusters of pigment molecules, namely a615-a611-a612-Lut and Vio(Zea)-a603-a609, have been identified and might function as potential energy-quenching centers and as the exit or entrance in energy-transfer pathways.

  17. CARBONIC ANHYDRASE ACTIVITY OF INTEGRAL-FUNCTIONAL COMPLEXES OF THYLAKOID MEMBRANES OF SPINACH CHLOROPLASTS.

    PubMed

    Semenihin, A V; Zolotareva, O K

    2015-01-01

    Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1) within gel was determined using the color reaction for ATPase activity. Due to the presence of cytochromes, the red band in unstained gels corresponded to the cytochrome b6f complex. Localization of the cytochrome b6f complex, ATP synthase and coupling CF1 in the native gel was confirmed by their subunit composition determined after SDS-electrophoretic analysis. Carbonic anhydrase (CA) activity in polypeptide zones of PS II, cytochrome b6f complex, and ATP-synthase CF1 was identified in native gels using indicator bromothymol blue. CA activity of isolated CF1 in solution was determined by infrared gas analysis as the rate of bicarbonate dehydration. The water-soluble acetazolamide, an inhibitor of CA, unlike lipophilic ethoxyzolamide inhibited CA activity of CF1 Thus, it was shown for the first time that ATP-synthase has a component which is capable of catalyzing the interconversion of forms of carbonic acid associated with proton exchange. The data obtained suggest the presence of multiple forms of carbonic anhydrase in the thylakoid membranes of spinach chloroplasts and confirm their involvement in the proton transfer to the ATP synthase. PMID:26502699

  18. Interactions between iron and titanium metabolism in spinach: a chlorophyll fluorescence study in hydropony.

    PubMed

    Cigler, Petr; Olejnickova, Julie; Hruby, Martin; Csefalvay, Ladislav; Peterka, Jiri; Kuzel, Stanislav

    2010-12-15

    One of the elements showing strong beneficial effect on plants at low concentrations and toxic effects at higher concentrations is titanium (Ti). We investigated the interconnection between the Fe uptake and the Ti intoxication in model experiment on Fe-deficient spinach (Spinacia oleracea) plants to help to elucidate the mechanism of the biological activity of titanium in plants. The two different Ti (0 and 20 mg L⁻¹) and two different Fe (0 and 1.35 mg L⁻¹) concentrations in hydroponic medium were used in all four possible combinations. We compared chemical analysis of Ti and Fe in roots and shoots with the changes of the in vivo chlorophyll fluorescence. Although Fe and Ti concentration found in shoots of Ti-non-treated Fe-deficient plants was comparable with that in Ti-treated Fe-deficient plants, the soluble form of Ti present in the growth media had a negative effect on photosynthetic activity monitored by chlorophyll fluorescence measurements. The presence of Fe in growth medium significantly decreased the Ti concentration in shoots and increased the photosynthetic activity. Here, we propose that Ti affect components of electron transport chain containing Fe in their structure (particularly photosystem I) and decrease the photosystem II efficiency. PMID:20708819

  19. Partial purification of gibberellin oxidases from spinach leaves. [Spinacia oleracea L

    SciTech Connect

    Gilmour, S.J.; Bleecker, A.B.; Zeevaart, J.A.D.

    1987-09-01

    Four enzyme activities catalyzing the following oxidative steps in the gibberellin (GA) biosynthetic pathway have been extracted from spinach (Spinacia oleracea L.) leaves after exposure to 8 long days: GA/sub 12/ ..-->.. GA/sub 53/ ..-->.. GA/sub 44/ ..-->.. GA/sub 19/ ..-->.. GA/sub 20/. Two of these, GA/sub 53/ oxidase and GA/sup 19/ oxidase, were separable from the other two, GA/sub 44/ oxidase and GA/sub 12/ 13-hydroxylase, by anion exchange high performance liquid chromatography (HPLC). Apparent molecular weights of the four enzymes as determined by gel filtration HLPL are: GA/sub 12/ 13-hydroxylase, 28,400; GA/sub 43/ oxidase, 42,500; GA/sub 44/ oxidase, 38,100; GA/sub 19/ oxidase, 39,500. GA/sub 44/ oxidase was purified approximately 100-fold in 0.3% yield by a combination of ammonium sulfate fractionation, anion exchange HPLC, phenyl-Sepharose chromatography and gel filtration HLPC.

  20. Spectral characterization in a supersonic beam of neutral chlorophyll a evaporated from spinach leaves

    NASA Astrophysics Data System (ADS)

    Shafizadeh, N.; Ha-Thi, M. H.; Soep, B.; Gaveau, M. A.; Piuzzi, F.; Pothier, C.

    2011-09-01

    The observation of the light absorption of neutral biomolecules has been made possible by a method implemented for their preparation in the gas phase, in supersonically cooled molecular beams, based upon the work of Focsa et al. [C. Mihesan, M. Ziskind, B. Chazallon, E. Therssen, P. Desgroux, S. Gurlui, and C. Focsa, Appl. Surf. Sci. 253, 1090 (2006)], 10.1016/j.apsusc.2006.01.082. The biomolecules diluted in frozen water solutions are entrained in the gas plume of evaporated ice generated by an infrared optical parametric oscillators (OPO) laser tuned close to its maximum of absorption, at ˜3 μm. The biomolecules are then picked up in the flux of a supersonic expansion of argon. The method was tested with indole dissolved in water. The excitation spectrum of indole was found cold and large clusters of indole with water were observed up to n = 75. Frozen spinach leaves were examined with the same method to observe the chlorophyll pigments. The Qy band of chlorophyll a has been observed in a pump probe experiment. The Qy bands of chlorophyll a is centred at 647 nm, shifted by 18 nm from its position in toluene solutions. The ionization threshold could also be determined as 6.1 ± 0.05 eV.

  1. Structure of spinach photosystem II-LHCII supercomplex at 3.2 Å resolution.

    PubMed

    Wei, Xuepeng; Su, Xiaodong; Cao, Peng; Liu, Xiuying; Chang, Wenrui; Li, Mei; Zhang, Xinzheng; Liu, Zhenfeng

    2016-06-01

    During photosynthesis, the plant photosystem II core complex receives excitation energy from the peripheral light-harvesting complex II (LHCII). The pathways along which excitation energy is transferred between them, and their assembly mechanisms, remain to be deciphered through high-resolution structural studies. Here we report the structure of a 1.1-megadalton spinach photosystem II-LHCII supercomplex solved at 3.2 Å resolution through single-particle cryo-electron microscopy. The structure reveals a homodimeric supramolecular system in which each monomer contains 25 protein subunits, 105 chlorophylls, 28 carotenoids and other cofactors. Three extrinsic subunits (PsbO, PsbP and PsbQ), which are essential for optimal oxygen-evolving activity of photosystem II, form a triangular crown that shields the Mn4CaO5-binding domains of CP43 and D1. One major trimeric and two minor monomeric LHCIIs associate with each core-complex monomer, and the antenna-core interactions are reinforced by three small intrinsic subunits (PsbW, PsbH and PsbZ). By analysing the closely connected interfacial chlorophylls, we have obtained detailed insights into the energy-transfer pathways between the antenna and core complexes.

  2. Nanodomains of cytochrome b6f and photosystem II complexes in spinach grana thylakoid membranes.

    PubMed

    Johnson, Matthew P; Vasilev, Cvetelin; Olsen, John D; Hunter, C Neil

    2014-07-01

    The cytochrome b6f (cytb6f) complex plays a central role in photosynthesis, coupling electron transport between photosystem II (PSII) and photosystem I to the generation of a transmembrane proton gradient used for the biosynthesis of ATP. Photosynthesis relies on rapid shuttling of electrons by plastoquinone (PQ) molecules between PSII and cytb6f complexes in the lipid phase of the thylakoid membrane. Thus, the relative membrane location of these complexes is crucial, yet remains unknown. Here, we exploit the selective binding of the electron transfer protein plastocyanin (Pc) to the lumenal membrane surface of the cytb6f complex using a Pc-functionalized atomic force microscope (AFM) probe to identify the position of cytb6f complexes in grana thylakoid membranes from spinach (Spinacia oleracea). This affinity-mapping AFM method directly correlates membrane surface topography with Pc-cytb6f interactions, allowing us to construct a map of the grana thylakoid membrane that reveals nanodomains of colocalized PSII and cytb6f complexes. We suggest that the close proximity between PSII and cytb6f complexes integrates solar energy conversion and electron transfer by fostering short-range diffusion of PQ in the protein-crowded thylakoid membrane, thereby optimizing photosynthetic efficiency.

  3. beta. -carotene synthesis in spinach chloroplasts is tightly linked to photosynthetic carbon metabolism

    SciTech Connect

    Schulze-Siebert, D.; Schultz, G.

    1987-04-01

    When purified, highly intact spinach chloroplasts were illuminated in the presence of NaH/sup 14/CO/sub 3/, the largest portion of acetate derived compounds formed was ..beta..-carotene and not fatty acids (20 and 2.5 natoms C incorporated/mg chlorophyll x h when 5 mM NaH /sup 14/CO/sub 3/ was used). From isotopic dilution experiments applying glyceraldehyde 3-P, dihydroxyacetone-P, 3-phosphoglycerate (3-PGA), 2-PGA, phosphoenolpyruvate (PEP) and pyruvate, respectively, evidence was obtained that acetyl-CoA to form isopentenyl-PP (IPP) originates from a low capacity but highly effective flow from photosynthetic CO/sub 2/-fixation via 3-PGA - 2-PGA - PEP - pyruvate. Phosphoglycerate mutase in chloroplasts, of which the presence hitherto was not unequivocally proven, was detected by (i) latency technique and (ii) feeding/1-/sup 14/C/glycerate and following 3-PGA, 2-PGA, PEP and pyruvate level in chloroplasts and suspension medium. From highly effective synthesis of ..beta..-carotene from CO/sub 2/ conclusion may be drawn that pathways for both, acetyl compounds and IPP, to form isoprenoids must exist in chloroplasts.

  4. Evidence for a reactive cysteine at the nucleotide binding site of spinach ribulose-5-phosphate kinase

    SciTech Connect

    Omnaas, J.; Porter, M.A.; Hartman, F.C.

    1985-02-01

    Ribulose-5-phosphate kinase from spinach was rapidly inactivated by N-bromoacetylethanolamine phosphate in a bimolecular fashion with a k2 of 2.0 m s at 2C and pH 8.0. Ribulose 5-phosphate had little effect on the rate of inactivation, whereas complete protection was afforded by ADP or ATP. The extent of incorporation as determined with UC-labeled reagent was about 1 molar equivalent per subunit in the presence of ATP with full retention of enzymatic activity, and about 2 molar equivalents per subunit in the completely inactivated enzyme. Amino acid analyses of enzyme derivatized with UC-labeled reagent reveal that all of the covalently incorporated reagent was associated with cysteinyl residues. Hence, two sulfhydryls are reactive, but the inactivation correlates with alkylation of one cysteinyl residue at or near the enzyme's nucleotide binding site. The kinase was also extremely sensitive to the sulfhydryl reagents 5,5'-dithiobis(2-nitrobenzoic acid) and N-ethylmaleimide. The reactive sulfhydryl groups are likely to be those generated by reduction of a disulfide during activation. 20 references, 3 figures, 2 tables.

  5. FTIR spectroscopy shows structural similarities between photosystems II from cyanobacteria and spinach.

    PubMed

    Remy, André; Niklas, Jens; Kuhl, Helena; Kellers, Petra; Schott, Thomas; Rögner, Matthias; Gerwert, Klaus

    2004-02-01

    Photosystem II (PSII), an essential component of oxygenic photosynthesis, is a membrane-bound pigment protein complex found in green plants and cyanobacteria. Whereas the molecular structure of cyanobacterial PSII has been resolved with at least medium resolution [Zouni, A., Witt, H.-T., Kern, J., Fromme, P., Krauss, N., Saenger, W. & Orth, P. (2001) Nature (London) 409, 739-743; Kamiya, N. & Shen, J.R. (2003) Proc. Natl Acad. Sci. USA 100, 98-103], the structure of higher plant PSII is only known at low resolution. Therefore Fourier transform infrared (FTIR) difference spectroscopy was used to compare PSII from both Thermosynechococcus elongatus and Synechocystis PCC6803 core complexes with PSII-enriched membranes from spinach (BBY). FTIR difference spectra of T. elongatus core complexes are presented for several different intermediates. As the FTIR difference spectra show close similarities among the three species, the structural arrangement of cofactors in PSII and their interactions with the protein microenvironment during photosynthetic charge separation must be very similar in higher plant PSII and cyanobacterial PSII. A structural model of higher plant PSII can therefore be predicted from the structure of cyanobacterial PSII.

  6. Histamine, cadaverine, and putrescine produced in vitro by enterobacteriaceae and pseudomonadaceae isolated from spinach.

    PubMed

    Lavizzari, T; Breccia, M; Bover-Cid, S; Vidal-Carou, M C; Veciana-Nogués, M T

    2010-02-01

    A total of 364 bacterial isolates, obtained from spinach leaves, were assayed in a decarboxylase broth containing histidine, lysine, and ornithine to check their ability to produce biogenic amines, and then quantified by high-performance liquid chromatography. Among these isolates, 240 formed cadaverine, 208 formed putrescine, and 196 formed histamine, in widely varying amounts. They frequently produced more than one biogenic amine. Klebsiella pneumoniae subsp. pneumoniae and Morganella morganii were the main histamine producers, with mean values of 1,600 and 2,440 mg/liter, respectively, followed by Pantoea spp. 3 (1,710 mg/liter) and Hafnia alvei (2,500 mg/liter). Enterobacter amnigenus and Enterobacter cloacae produced particularly high amounts of putrescine, with mean values of 2,340 and 2,890 mg/liter, respectively. The strongest cadaverine formation was shown by Serratia liquefaciens (3,300 mg/liter), Serratia marcescens (3,280 mg/liter), and Stenotrophomonas maltophilia (1,000 mg/liter). PMID:20132689

  7. Effects of inorganic phosphate on the light dependent thylakoid energization of intact spinach chloroplasts

    SciTech Connect

    Heineke, D.; Heldt, H.W. ); Stitt, M. )

    1989-09-01

    The light dependent energization of the thylakoid membrane was analyzed in isolated intact spinach (Spinacia oleracea L.) chloroplasts incubated with different concentrations of inorganic phosphate (Pi). Two independent methods were used: (a) the accumulation of ({sup 14}C)5,5-dimethyl-2,4-oxazolidinedione and ({sup 14}C)methylamine; (b) the energy dependent chlorophyll fluorescence quenching. The inhibition of CO{sub 2} fixation by superoptimal medium Pi or by adding glyceraldehyde - an inhibitor of the Calvin cycle - leads to an increased energization of the thylakoid membrane; however, the membrane energization decreases when chloroplasts are inhibited by suboptimal Pi. This specific low phosphate effect could be partially reversed by adding oxaloacetate, which regenerates the electron acceptor NADP{sup +} and stimulates linear electron transport. The energization seen in low Pi is, however, always lower than in superoptimal Pi, even in the presence of oxaloacetate. Energization recovers in the presence of low amounts of N,N{prime}-dicyclohexylcarbodiimide, which reacts with proton channels including the coupling factor 1 ATP synthase. N,N{prime}-Dicyclohexylcarbodiimide has no effect on energization of chloroplasts in superoptimal Pi. These results suggest there is a specific low phosphate proton leak in the thylakoids, and its origin is discussed.

  8. Synthesis of poly(A)-containing RNA by isolated spinach chloroplasts.

    PubMed

    Bartolf, M; Price, C A

    1979-05-01

    Chloroplasts were isolated from spinach leaves and the intact chloroplasts separated by centrifugation on gradients of silica sol. Chloroplasts prepared in this way were almost completely free of cytoplasmic rRNA. The purified chloroplasts were incubated with 32PO4 in the light. The nucleic acids were then extracted and the RNA was fractionated into poly(A)-lacking RNA and poly(A)-containing RNA (poly(A)-RNA) via oligo(dT)-cellulose chromatography. The poly(A)-RNA had a mean size of approximately 18--20 S as determined by polyacrylamide gel electrophoresis. The poly(A)-RNA was digested with RNase A and RNase T1, and the resulting poly(A) segments were subjected to electrophoresis on a 10% w/v polyacrylamide gel 98% v/v formamide). Radioactivity was incorporated into both poly(A)-RNA and poly(A)-lacking RNA and into the poly(A) segments themselves. The poly(A) segments were between 10 and 45 residues long and alkaline hydrolysis of poly(A) segments followed by descending paper chromatography showed that they were composed primarily of adenine residues. There was no 32PO4 incorporation into acid-insoluble material in the dark. We conclude that isolated chloroplasts are capable of synthesizing poly(A)-RNA. PMID:435477

  9. 7 CFR 29.2528 - Leaf.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf. 29.2528 Section 29.2528 Agriculture Regulations...-Cured Tobacco (u.s. Types 22, 23, and Foreign Type 96) § 29.2528 Leaf. Whole, unstemmed leaf. Leaf, when applied to tobacco in strip form, shall describe the divided unit of a whole leaf....

  10. 7 CFR 29.3525 - Leaf.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf. 29.3525 Section 29.3525 Agriculture Regulations... Type 95) § 29.3525 Leaf. Whole, unstemmed leaf. Leaf, when applied to tobacco in strip form, shall describe the divided unit of a whole leaf....

  11. 7 CFR 29.1028 - Leaf.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf. 29.1028 Section 29.1028 Agriculture Regulations... Type 92) § 29.1028 Leaf. Whole, unstemmed leaf. Leaf, when applied to tobacco in strip form, shall describe the divided unit of a whole leaf....

  12. 7 CFR 29.3033 - Leaf.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf. 29.3033 Section 29.3033 Agriculture Regulations... Leaf. Whole, unstemmed leaf. Leaf, when applied to tobacco in strip form, shall describe the divided unit of a whole leaf....

  13. Analytical method for 44 pesticide residues in spinach using multi-plug-filtration cleanup based on multiwalled carbon nanotubes with liquid chromatography and tandem mass spectrometry detection.

    PubMed

    Qin, Yuhong; Huang, Baoyong; Zhang, Jingru; Han, Yongtao; Li, Yanjie; Zou, Nan; Yang, Jianguo; Pan, Canping

    2016-05-01

    Spinach is one of the most commonly planted vegetables worldwide. A high chlorophyll content makes spinach a complicated matrix in pesticide residue analysis. In this study, a rapid clean-up method was developed for the analysis of pesticide multi-residues in spinach followed by liquid chromatography with tandem mass spectrometry. A modified QuEChERS method with multiwalled carbon nanotubes and carbon material was adopted in the multi-Plug Filtration Cleanup procedure. This method was validated for 44 representative pesticides spiked at two concentration levels of 10 and 100 μg/kg. The pesticides of different physicochemical properties were registered on spinach in China. The recoveries were between 76 and 114% for major pesticides with relative standard deviations of less than 15%, except for quizalofop-P-ethyl, pyrimethanil, and carbendazim. Matrix-matched calibration curves were performed with the coefficients of determination higher than 0.995 for the studied pesticides for concentration levels of 10-500 μg/kg. The limits of quantitation ranged from 2 to 10 μg/kg. The developed method was successfully applied to determine pesticide residues in Chinese market spinach samples. PMID:26968118

  14. Influence of cold stress on contents of soluble sugars, vitamin C and free amino acids including gamma-aminobutyric acid (GABA) in spinach (Spinacia oleracea).

    PubMed

    Yoon, Young-Eun; Kuppusamy, Saranya; Cho, Kye Man; Kim, Pil Joo; Kwack, Yong-Bum; Lee, Yong Bok

    2017-01-15

    The contents of soluble sugars (sucrose, fructose, glucose, maltose and raffinose), vitamin C and free amino acids (34 compounds, essential and non-essential) were quantified in open-field and greenhouse-grown spinaches in response to cold stress using liquid chromatography. In general, greenhouse cultivation produced nutritionally high value spinach in a shorter growing period, where the soluble sugars, vitamin C and total amino acids concentrations, including essential were in larger amounts compared to those grown in open-field scenarios. Further, low temperature exposure of spinach during a shorter growth period resulted in the production of spinach with high sucrose, ascorbate, proline, gamma-aminobutyric acid, valine and leucine content, and these constitute the most important energy/nutrient sources. In conclusion, cultivation of spinach in greenhouse at a low temperature (4-7°C) and exposure for a shorter period (7-21days) before harvest is recommended. This strategy will produce a high quality product that people can eat. PMID:27542466

  15. Influence of cold stress on contents of soluble sugars, vitamin C and free amino acids including gamma-aminobutyric acid (GABA) in spinach (Spinacia oleracea).

    PubMed

    Yoon, Young-Eun; Kuppusamy, Saranya; Cho, Kye Man; Kim, Pil Joo; Kwack, Yong-Bum; Lee, Yong Bok

    2017-01-15

    The contents of soluble sugars (sucrose, fructose, glucose, maltose and raffinose), vitamin C and free amino acids (34 compounds, essential and non-essential) were quantified in open-field and greenhouse-grown spinaches in response to cold stress using liquid chromatography. In general, greenhouse cultivation produced nutritionally high value spinach in a shorter growing period, where the soluble sugars, vitamin C and total amino acids concentrations, including essential were in larger amounts compared to those grown in open-field scenarios. Further, low temperature exposure of spinach during a shorter growth period resulted in the production of spinach with high sucrose, ascorbate, proline, gamma-aminobutyric acid, valine and leucine content, and these constitute the most important energy/nutrient sources. In conclusion, cultivation of spinach in greenhouse at a low temperature (4-7°C) and exposure for a shorter period (7-21days) before harvest is recommended. This strategy will produce a high quality product that people can eat.

  16. Retrieval of spinach crop parameters by microwave remote sensing with back propagation artificial neural networks: A comparison of different transfer functions

    NASA Astrophysics Data System (ADS)

    Prasad, Rajendra; Pandey, A.; Singh, K. P.; Singh, V. P.; Mishra, R. K.; Singh, D.

    2012-08-01

    Back propagation artificial natural network (BPANN) is a well known and widely used machine learning methodology in the field of remote sensing. In this paper an attempt is made to retrieve the spinach crop parameters like biomass, leaf area index, average plant height and soil moisture content by using the X-band scattering coefficients with BPANN at different growth stages of this crop. The maturity age of this crop was found to be 45 days from the date of sowing. After 45 days from the date of sowing, this crop was cut at a certain height for production. Then, it is a point of interest to investigate the microwave response of variation in production. Significant variations in all the crop parameters were observed after cutting the crop and consequently made the problem more critical. Our work confirms the utility of BPANN in handling such a non-linear data set. The BPANN is essentially a network of simple processing nodes arranged into different layers as input, hidden and the output. The input layer propagates components of a particular input vector after weighting these with synaptic weights to each node in the hidden layer. At each node, these weighted input vector components are added. Each hidden layer computes output corresponding to these weighted sum through a non-linear/linear function (e.g. LOGSIG, TANSIG and PURLIN). These functions are known as transfer functions. Thus, each of the hidden layer nodes compute output values, which become inputs to the nodes of the output layer. At nodes of output layer also a weighted sum of outputs of previous layer (hidden layer) are obtained and processed through a transfer function. Thus, the output layer nodes compute the network output for the particular input vector. In this paper, output nodes use linear transfer function. Different transfer functions e.g. TANSIG, LOGSIG and PURELIN were used and the performance of the ANN was optimized by changing the number of neurons in the hidden layers. The present

  17. Calcium oxalate nephrolithiasis and tubular necrosis in recent metamorphs of Rana sylvatica (Lithobates sylvaticus) fed spinach during the premetamorphic (tadpole) stage.

    PubMed

    Forzán, M J; Ferguson, L V; Smith, T G

    2015-03-01

    Amphibians in the family Ranidae (true frogs) seem highly susceptible to oxalosis, particularly when fed a diet high in oxalic acid during the premetamorphic (tadpole) stage. The authors describe the mortality of 150 captive-raised wood frogs (Rana sylvatica or Lithobates sylvaticus) from oxalate nephrolithiasis and renal tubular necrosis caused by consumption of boiled spinach during tadpole development. Renal lesions were due to intraluminal transparent crystals which were birefringent under polarized light and were identified morphologically and histochemically as composed of calcium oxalate. Evidence of early fibrosis or squamous metaplasia, and a presentation at least 2 weeks after spinach consumption had ended, suggested a subacute course. Tadpole-feeding protocols should avoid plants with high oxalate content (eg, spinach and rhubarb leaves), and any episode of high mortality in captive amphibians along with nephrolithiasis should prompt an evaluation of the feed sources for material with high oxalate content.

  18. Multifactorial Effects of Ambient Temperature, Precipitation, Farm Management, and Environmental Factors Determine the Level of Generic Escherichia coli Contamination on Preharvested Spinach

    PubMed Central

    Navratil, Sarah; Gregory, Ashley; Bauer, Arin; Srinath, Indumathi; Szonyi, Barbara; Nightingale, Kendra; Anciso, Juan; Jun, Mikyoung; Han, Daikwon; Ivanek, Renata

    2015-01-01

    A repeated cross-sectional study was conducted to identify farm management, environment, weather, and landscape factors that predict the count of generic Escherichia coli on spinach at the preharvest level. E. coli was enumerated for 955 spinach samples collected on 12 farms in Texas and Colorado between 2010 and 2012. Farm management and environmental characteristics were surveyed using a questionnaire. Weather and landscape data were obtained from National Resources Information databases. A two-part mixed-effect negative binomial hurdle model, consisting of a logistic and zero-truncated negative binomial part with farm and date as random effects, was used to identify factors affecting E. coli counts on spinach. Results indicated that the odds of a contamination event (non-zero versus zero counts) vary by state (odds ratio [OR] = 108.1). Odds of contamination decreased with implementation of hygiene practices (OR = 0.06) and increased with an increasing average precipitation amount (mm) in the past 29 days (OR = 3.5) and the application of manure (OR = 52.2). On contaminated spinach, E. coli counts increased with the average precipitation amount over the past 29 days. The relationship between E. coli count and the average maximum daily temperature over the 9 days prior to sampling followed a quadratic function with the highest bacterial count at around 24°C. These findings indicate that the odds of a contamination event in spinach are determined by farm management, environment, and weather factors. However, once the contamination event has occurred, the count of E. coli on spinach is determined by weather only. PMID:25636850

  19. Multifactorial effects of ambient temperature, precipitation, farm management, and environmental factors determine the level of generic Escherichia coli contamination on preharvested spinach.

    PubMed

    Park, Sangshin; Navratil, Sarah; Gregory, Ashley; Bauer, Arin; Srinath, Indumathi; Szonyi, Barbara; Nightingale, Kendra; Anciso, Juan; Jun, Mikyoung; Han, Daikwon; Lawhon, Sara; Ivanek, Renata

    2015-04-01

    A repeated cross-sectional study was conducted to identify farm management, environment, weather, and landscape factors that predict the count of generic Escherichia coli on spinach at the preharvest level. E. coli was enumerated for 955 spinach samples collected on 12 farms in Texas and Colorado between 2010 and 2012. Farm management and environmental characteristics were surveyed using a questionnaire. Weather and landscape data were obtained from National Resources Information databases. A two-part mixed-effect negative binomial hurdle model, consisting of a logistic and zero-truncated negative binomial part with farm and date as random effects, was used to identify factors affecting E. coli counts on spinach. Results indicated that the odds of a contamination event (non-zero versus zero counts) vary by state (odds ratio [OR] = 108.1). Odds of contamination decreased with implementation of hygiene practices (OR = 0.06) and increased with an increasing average precipitation amount (mm) in the past 29 days (OR = 3.5) and the application of manure (OR = 52.2). On contaminated spinach, E. coli counts increased with the average precipitation amount over the past 29 days. The relationship between E. coli count and the average maximum daily temperature over the 9 days prior to sampling followed a quadratic function with the highest bacterial count at around 24°C. These findings indicate that the odds of a contamination event in spinach are determined by farm management, environment, and weather factors. However, once the contamination event has occurred, the count of E. coli on spinach is determined by weather only.

  20. Electron nuclear double resonance evidence supporting a monomeric nature for P700 in spinach chloroplasts.

    PubMed

    O'malley, P J; Babcock, G T

    1984-02-01

    Proton electron nuclear double resonance (ENDOR) spectra of P700(+) in spinach chloroplasts and in photosystem I particles have been obtained and compared with the corresponding ENDOR spectrum of monomeric chlorophyl a(+) (Chla(+)) cation radical. The hyperfine couplings for P700(+) can be interpreted in terms of those expected for a monomer Chla(+) radical. The reduction in alpha-carbon spin densities observed for the in vivo species when compared to the in vitro radical is attributed to differences in the composition of the ground-state orbital for the two systems. For P700(+), a mixture of 75% D(0)/25% D(1), in which D(0) and D(1) represent the ground-and first excited-state orbitals calculated by Petke et al. for Chla(+) [Petke, J. D., Maggiora, G. M., Shipman, L. L. & Christoffersen, R. E. (1980) Photochem. Photobiol. 31, 243-257], gives good agreement between calculated and experimental spin-density reduction factors. Interaction of the pigment ion with its protein environment such as through ligation of the central Mg atom, hydrogen bonding to the 9-keto-carbonyl group, and electrostatic interactions with charged amino acid residues are proposed as factors responsible for the lowering in energy of the D(1) level in vivo. Combined with similar previous proposals for P680(+) of photosystem II, the data suggest that both primary donor cation radicals of green plant photosynthesis can be viewed as monomeric Chla(+) species in which the D(1) orbital makes a significant contribution to the spin-density distribution. PMID:16593417

  1. [Chemical characteristics of the rhizosphere soil of water spinach cultivars differing in Cd accumulation].

    PubMed

    Gong, Yu-Lian; Yang, Zhong-Yi

    2014-08-01

    A rhizobox experiment was conducted to investigate the chemical characteristics of the rhizosphere soils of two water spinach cultivars differing in Cd accumulation, QLQ (a low-Cd cultivar) and T308 (a high-Cd cultivar). The results showed that the diethylenetriamine pentacetate acid extractable Cd (DTPA-Cd) concentration in the rhizos-phere soil of QLQ was significantly higher than that of T308 (P < 0.05). pH and Eh in the rhizosphere soil of QLQ were significantly higher than those of T308 (P < 0.05), while EC was opposite. Contents of organic matter and dissolved organic matter (DOM) in the rhizosphere soil of QLQ were both higher than those of T308. In contaminated soil, the composition and concentration of low molecular weight organic acids in the rhizosphere between the two cultivars were both different. Acetic, propionic, citric and fumaric acids were detected in the rhizosphere soil of T308, and only citric and fumaric acids were detected in that of QLQ. The total concentration of low molecular weight organic acids in the rhizosphere soil of QLQ (1.93 nmol x g(-1) DM) was lower than that of T308 (15.11 nmol x g(-1) DM) (P < 0.01). Compared with the high-Cd cultivar (T308), the chemical characteristics of the rhizosphere soil of the low-Cd cultivar (QLQ) were obviously distinct, i. e., the relatively higher content of organic matter, the lower content of low molecular weight organic acids with a specific composition, less acidification of soil, and a lower ability in reduction, correspondingly lowering the mobility of Cd in soil and reducing Cd accumulation by plant.

  2. Molecular dissection of the epsilon subunit of the chloroplast ATP synthase of spinach.

    PubMed Central

    Cruz, J A; Harfe, B; Radkowski, C A; Dann, M S; McCarty, R E

    1995-01-01

    The gene encoding the epsilon subunit (atpE) of the chloroplast ATP synthase of Spinacia oleracea has been overexpressed in Escherichia coli. The recombinant protein can be solubilized in 8 M urea and directly diluted into buffer containing ethanol and glycerol to obtain epsilon that is as biologically active as epsilon purified from chloroplast-coupling factor 1 (CF1). Recombinant epsilon folded in this manner inhibits the ATPase activity of soluble and membrane-bound CF1 deficient in epsilon and restores proton impermeability to thylakoid membranes reconstituted with CF1 deficient in epsilon. Site-directed mutagenesis was used to generate truncations and single amino acid substitutions in the primary structure of epsilon. In the five mutants tested, alterations that weaken ATPase inhibition by recombinant epsilon affect its ability to restore proton impermeability to a similar extent, with one exception. Substitution of histidine-37 with arginine appears to uncouple ATPase inhibition and the restoration of proton impermeability. As in the case of E. coli, it appears that N-terminal truncations of the epsilon subunit have more profound effects than C-terminal deletions on the function of epsilon. Recombinant epsilon with six amino acids deleted from the C terminus, which is the only region of significant mismatch between the epsilon of spinach and the epsilon of Pisum sativum, inhibits ATPase activity with a reduced potency similar to that of purified pea epsilon. Four of the six amino acids are serine or threonine. These hydroxylated amino acids may be important in epsilon-CF1 interactions. PMID:8539297

  3. Dihydroxy acid dehydratase from spinach contains a [2Fe-2S] cluster.

    PubMed

    Flint, D H; Emptage, M H

    1988-03-15

    Dihydroxy acid dehydratase, the third enzyme in the branched-chain amino acid biosynthetic pathway, has been purified to homogeneity (5000-fold) from spinach leaves. The molecular weights of dihydroxy acid dehydratase as determined by sodium dodecyl sulfate and native gel electrophoresis are 63,000 and 110,000, respectively, suggesting the native enzyme is a dimer. 2 moles of iron were found per mol of protein monomer. Chemical analyses of iron and labile sulfide gave an Fe/S2- ratio of 0.95. The EPR spectrum of dithionite-reduced enzyme (gavg = 1.91) is similar to spectra characteristic of Rieske Fe-S proteins and has a spin concentration of 1 spin/1.9 irons. These results strongly suggest that dihydroxy acid dehydratase contains a [2Fe-2S] cluster, a novel finding for enzymes of the hydrolyase class. In contrast to the Rieske Fe-S proteins, the redox potential of the Fe-S cluster is quite low (-470 mV). Upon addition of substrate, the EPR signal of the reduced enzyme changes to one typical of 2Fe ferredoxins (gavg = 1.95), and the visible absorption spectrum of the native enzyme shows substantial changes between 400 and 600 nm. Reduction of the Fe-S cluster decreases the enzyme activity by 6-fold under Vmax conditions. These results suggest the direct involvement of the [2Fe-2S] cluster of dihydroxy acid dehydratase in catalysis. Similar conclusions have been reached for the catalytic involvement of the [4Fe-4S] cluster of the hydrolyase aconitase (Emptage, M. H., Kent, T. A., Kennedy, M. C., Beinert, H., and Münck, E. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 4674-4678).

  4. Synthesis and bioaccessibility of Fe-pheophytin derivatives from crude spinach extract.

    PubMed

    Nelson, R E; Ferruzzi, M G

    2008-06-01

    Heme iron is recognized as a highly bioavailable source of iron suitable for treatment of iron deficiency anemia. However, the animal origin of purified heme limits its broad applicability due to religious, personal, and food safety issues. Development of chlorophyll-derived heme mimetics offers opportunities to expand current iron fortification strategies. The objective of this study was the synthesis of Fe-pheophytin (FePhe) derivatives from natural chlorophyll and subsequent evaluation of their digestive behavior and bioaccessibility in vitro. FePhe a and a' were synthesized from crude spinach extracts by treatment with 1.3 M iron(II)chloride and 0.25 M Na-acetate dissolved in glacial acetic acid at 80 degrees C for 30 min. FePhe-rich extracts (approximately 1 mM) were formulated into corn starch based test meals (7.5% lipid) and subjected to a 2-step in vitro digestion designed to simulate in vivo gastric and small intestinal conditions. Recovery of FePhe following digestion and transfer of FePhe and pheophytins (Phe) from test meal matrix to mixed micelles was assessed by RP C18-HPLC to determine the digestive stability and micellarization efficiency (bioaccessibility). FePhe a and a' derivatives were moderately stable to digestive conditions with recoveries of 52.3% and 58.7%, respectively. Residual Phe a was stable to digestion. Micellarization efficiency of FePhe a (4%) and a' (3.4%) was significantly (P < 0.05) lower than Phe a (25.8%) from test meals. While digestive stability and micellarization efficiency are limiting, the presence of lipophilic FePhe derivatives in mixed micelles suggests that these compounds would be available for subsequent absorption in the intestinal tract.

  5. Analyses of optical absorption and circular dichroism spectra of spinach ferredoxin at alkaline pH.

    PubMed

    Hasumi, H

    1982-10-01

    The whole protein structure and the microenvironments of the iron-sulfur cluster and of the side chains of amino acid residues of spinach ferredoxin were studied by optical absorption and circular dichroism (CD) spectroscopy in the alkaline pH range. From the pH-dependence of the optical absorption changes at 245 nm, the four tyrosyl residues of ferredoxin were classified into three groups: one exposed residue with a normal apparent pK value of 10.1, two exposed residues with abnormal apparent pK values of 12.0, and one buried residue showing time-dependent ionization. The absorption in the visible region disappeared gradually with the ionization of the buried residue rather than that of the three exposed residues. The apparent pK value of 10.0 was obtained from the rapid CD changes at 258 nm caused by pH elevation from neutral to alkaline pH. The structural alteration associated with the CD change had no effect on the secondary structure of the protein moiety other than the iron-sulfur cluster and the microenvironment of the cluster. The rate constants obtained from the time courses of the CD changes in the near-ultraviolet and visible regions were in good agreement with those obtained from the time courses of the optical absorption changes. These results lead to the conclusions that (1) the native ferredoxin structure is maintained through the interaction with the iron-sulfur cluster and (2) the protein structure in the neighborhood of the cluster, important for the physiological activity, is not perturbed even though the exposed tyrosyl residues are ionized.

  6. Temperature and pH effects on chloroplastic respiration of glucose and fructose in spinach

    SciTech Connect

    Singh, K.K.; Gibbs, M. )

    1993-05-01

    Respiration was monitored principally as CO[sub 2] release in the darkened intact spinach chloroplast supplied with [sup 14]C-glucose and [sup 14]C-fructose. The rate of flucose respiration, optimum pH 7.5, increased from 15[degrees]C up to 40[degrees]C and then decreased in the presence of added ATP. In the absence of ATP, the optimum temperature for CO[sub 2] release was 25 [degrees]C and then decreased. At optimum pH 8.5, both in the absence and presence of ATP, the rate increased up to 25[degrees]C and then decreased. The negative effect of high temperature was not reversed when the chloroplast was returned to 25[degrees]C. Higher temperature (40[degrees]C vs 15[degrees]C) and higher pH (8.5 vs 7.5) increased radioactivity into starch and decreased radioactivity in CO[sub 2]. The rate of fructose respiration, optimum pH 7.5 but also at pH 8.5, increased CO[sub 2] release from 15[degrees]C to 40[degrees]C and then decreased both in the absence and presence of externally supplied ATP. Temperature and pH has no effect on radioactivity in starch and CO[sub 2] when fructose was substrate. The difference in results between glucose and fructose may reflect the localization of fructokinase in the stroma and glucokinase both in the stroma and cytosolic side of the outer chloroplastic membrane. It may be also reflect the equilibrium of phosphohexose isomerase favoring fructose-6-P.

  7. Formation of the Fe-S cluster of ferredoxin in lysed spinach chloroplasts. [Spinacia oleracea

    SciTech Connect

    Takahashi, Yasuhiro; Mitsui, Akira; Matsubara, Hiroshi )

    1991-01-01

    In vitro formation of the {sup 35}S-labeled Fe-S cluster of ferredoxin (Fd) has been achieved by incubating apo-Fd and ({sup 35}S)cysteine with osmotically lysed chloroplasts of spinach (Spinacia oleracea). Correct integration of the {sup 35}S-labeled Fe-S cluster into Fd was verified on the basis of the following: (a) Under nondenaturing conditions, {sup 35}S-labeled holo-Fd showed the same electrophoretic mobility as authentic holo-Fd; (b) {sup 35}S-labeled holo-Fd showed an ability to bind Fd-NADP{sup +} reductase; (c) the {sup 35}S-labeled moiety was removed from the Fd polypeptide by TCA treatment but not by 2-mercaptoethanol treatment; (d) externally added pea II apo-Fd was converted to {sup 35}S-labeled holo-Fd. This reconstitution was dependent on both ATP and light, and formation of the {sup 35}S-labeled Fe-S cluster was observed upon addition of ATP or when an ATP generation-system was constructed in the light. In contrast, ATP-consuming systems abolished the Fe-S cluster formation. A non-hydrolyzable ATP analog was unable to serve as an ATP substitute, indicating the requirement of ATP hydrolysis for cluster formation. GTP was able to substitute for ATP, but CTP and UTP were less effective. Fe-S cluster formation in lysed chloroplasts was stimulated by light even in the presence of added ATP. Light stimulation was inhibited by DCMU or methyl viologen but not by NH{sub 4}{sup +}. NADPH was able to substitute for light, indicating that light energy is required for the production of reducing compounds such as NADPH in addition to the generation of ATP.

  8. Effects of ethanol volume percent on fluorescein-labeled spinach apo- and holocalmodulin.

    PubMed

    Doody, M A; Baker, G A; Pandey, S; Bright, F V

    2000-01-01

    We report the effects of EtOH volume percent (0-70%) on spinach apo- and holocalmodulin that have been site-selectively labeled with fluorescein (F). In these experiments, calmodulin (CaM) has one F reporter group attached to Cys-26, and this site is located immediately adjacent to one of the four primary Ca(2+)-binding sites (EF hands). The optimum analytical CaM-F sensitivity to Ca2+ occurs between approximately 10 and 30% EtOH. Our results also show that added EtOH causes changes in CaM and these changes are surprisingly different for apo- and holo-CaM. Apo-CaM-F appears to lose one of its two waters of hydration at approximately 20% EtOH and retains one water of hydration between approximately 20 and 70% EtOH. In apo-CaM-F, the semiangle that describes the range over which the fluorescein reporter group can precess remains essentially constant (42 +/- 2 degrees) between 0 and 70% EtOH. This shows that the fluorescein reporter group precessional freedom in apo-CaM-F is not affected significantly by EtOH. Holo-CaM-F also appears to lose one water of hydration at approximately 20-30% EtOH but then appears to denature as the EtOH volume percent increases. The fluorescein reporter group semiangle within holo-CaM-F decreases from 43 +/- 1 degrees in neat aqueous buffer to 36 +/- 1 degrees at 70% EtOH. This shows that holo-CaM-F is less nativelike and the EF hand "closes down" about the fluorescein reporter group in holo-CaM-F as the EtOH volume percent increases.

  9. Differential sensitivity of spinach and amaranthus to enhanced UV-B at varying soil nutrient levels: association with gas exchange, UV-B-absorbing compounds and membrane damage.

    PubMed

    Singh, Suruchi; Agrawal, Madhoolika; Agrawal, S B

    2013-07-01

    The metabolic reasons associated with differential sensitivity of C3 and C4 plant species to enhanced UV-B under varying soil nutrient levels are not well understood. In the present study, spinach (Spinacia oleracea L. var All Green), a C3 and amaranthus (Amaranthus tricolor L. var Pusa Badi Chaulai), a C4 plant were subjected to enhanced UV-B (280-315 nm; 7.2 kJ m(-2) day(-1)) over ambient under varying soil nutrient levels. The nutrient amendments were recommended Nitrogen (N), Phosphorus (P), Potassium (K), 1.5× recommended NPK, 1.5× recommended N and 1.5× recommended K. Enhanced UV-B negatively affected both the species at all nutrient levels, but the reductions varied with nutrient concentration and combinations. Reductions in photosynthetic rate, stomatal conductance and chlorophyll content were significantly more in spinach compared with amaranthus. The reduction in photosynthetic rate was maximum at 1.5× recommended K and minimum in 1.5× NPK amended plants. The oxidative damage to membranes measured in terms of malondialdehyde content was significantly higher in spinach compared with amaranthus. Enhanced UV-B reduced SOD activity in both the plants except in amaranthus at 1.5× recommended K. POX activity increased under enhanced UV-B at all nutrient levels in amaranthus, but only at 1.5× K in spinach. Amaranthus had significantly higher UV-B-absorbing compounds than spinach even under UV-B stress. Lowest reductions in yield and total biomass under enhanced UV-B compared with ambient were observed in amaranthus grown at 1.5× recommended NPK. Enhanced UV-B did not significantly change the nitrogen use efficiency in amaranthus at all NPK levels, but reduced in spinach except at 1.5× K. These findings suggest that the differential sensitivity of the test species under enhanced UV-B at varying nutrient levels is due to varying antioxidative and UV-B screening capacity, and their ability to utilize nutrients. Amaranthus tolerated enhanced UV-B stress

  10. Leaf hydraulics II: vascularized tissues.

    PubMed

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

    2014-01-01

    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.

  11. Leaf Relative Water Content Estimated from Leaf Reflectance and Transmittance

    NASA Technical Reports Server (NTRS)

    Vanderbilt, Vern; Daughtry, Craig; Dahlgren, Robert

    2016-01-01

    Remotely sensing the water status of plants and the water content of canopies remain long term goals of remote sensing research. In the research we report here, we used optical polarization techniques to monitor the light reflected from the leaf interior, R, as well as the leaf transmittance, T, as the relative water content (RWC) of corn (Zea mays) leaves decreased. Our results show that R and T both change nonlinearly. The result show that the nonlinearities cancel in the ratio R/T, which appears linearly related to RWC for RWC less than 90%. The results suggest that potentially leaf water status and perhaps even canopy water status could be monitored starting from leaf and canopy optical measurements.

  12. Regulation of Compound Leaf Development

    PubMed Central

    Wang, Yuan; Chen, Rujin

    2013-01-01

    Leaf morphology is one of the most variable, yet inheritable, traits in the plant kingdom. How plants develop a variety of forms and shapes is a major biological question. Here, we discuss some recent progress in understanding the development of compound or dissected leaves in model species, such as tomato (Solanum lycopersicum), Cardamine hirsuta and Medicago truncatula, with an emphasis on recent discoveries in legumes. We also discuss progress in gene regulations and hormonal actions in compound leaf development. These studies facilitate our understanding of the underlying regulatory mechanisms and put forward a prospective in compound leaf studies. PMID:27135488

  13. Deriving leaf chlorophyll content of green-leafy vegetables from hyperspectral reflectance

    NASA Astrophysics Data System (ADS)

    Xue, Lihong; Yang, Linzhang

    Different nitrogen (N) treatments of four common green-leafy vegetable varieties with different leaf color: lettuce ( Lactuca sativa L. var. crispa L.) with yellow green leaves, pakchoi ( Brassica chinensis L.) var. aijiaohuang in Chinese (AJH) with middle green leaves, spinach ( Spinacia oleracea L.) with green leaves and pakchoi ( B. chinensis L.) var. shanghaiqing in Chinese (SHQ) with dark green leaves, were carried out to achieve a wide range of chlorophyll content. The relationship of vegetable leaf hyperspectral response to its chlorophyll content was examined in this study. Almost all reported successful leaf chlorophyll indices in the literature were evaluated for their ability to predict the chlorophyll content in vegetable leaves. Some new indices based on the first derivative curve were also developed, and compared with the chlorophyll indices published. The results showed that most of the indices showed a strong relation with leaf chlorophyll content. In general, modified indices with the blue or near red edge wavelength performed better than their simple counterpart without modification, ratio indices performed a little better than normalized indices when chlorophyll expressed on area basis and reversed when chlorophyll expressed on fresh weight basis. A normalized derivative difference ratio (BND: (D722-D700)/(D722+D700) calibrated by Maire et al. [Maire, G., Francois, C., Dufrene, E., 2004. Towards universal broad leaf chlorophyll indices using PROSPECT simulated database and hyperspectral reflectance measurements. Remote Sensing of Environment 89 (1), 1-28]) gave the best results among all published indices in this study (RMSE=22.1 mg m -2), then the mSR-like indices with the RMSE between 22.6 and 23.0 mg m -2. The new indices EBAR (ratio of the area of red and blue, ∑ dRE/∑ dB), EBFN (normalized difference of the amplitude of red and blue, (dRE-dB)/(dRE+dB)) and EBAN (normalized difference of the area of red and blue, (∑ dRE-∑ dB)/(∑ d

  14. Sucrose phosphate synthase activity rises in correlation with high-rate cellulose synthesis in three heterotrophic systems.

    PubMed

    Babb, V M; Haigler, C H

    2001-11-01

    Based on work with cotton fibers, a particulate form of sucrose (Suc) synthase was proposed to support secondary wall cellulose synthesis by degrading Suc to fructose and UDP-glucose. The model proposed that UDP-glucose was then channeled to cellulose synthase in the plasma membrane, and it implies that Suc availability in cellulose sink cells would affect the rate of cellulose synthesis. Therefore, if cellulose sink cells could synthesize Suc and/or had the capacity to recycle the fructose released by Suc synthase back to Suc, cellulose synthesis might be supported. The capacity of cellulose sink cells to synthesize Suc was tested by analyzing the Suc phosphate synthase (SPS) activity of three heterotrophic systems with cellulose-rich secondary walls. SPS is a primary regulator of the Suc synthesis rate in leaves and some Suc-storing, heterotrophic organs, but its activity has not been previously correlated with cellulose synthesis. Two systems analyzed, cultured mesophyll cells of Zinnia elegans L. var. Envy and etiolated hypocotyls of kidney beans (Phaseolus vulgaris), contained differentiating tracheary elements. Cotton (Gossypium hirsutum L. cv Acala SJ-1) fibers were also analyzed during primary and secondary wall synthesis. SPS activity rose in all three systems during periods of maximum cellulose deposition within secondary walls. The Z. elegans culture system was manipulated to establish a tight linkage between the timing of tracheary element differentiation and rising SPS activity and to show that SPS activity did not depend on the availability of starch for degradation. The significance of these findings in regard to directing metabolic flux toward cellulose will be discussed. PMID:11706202

  15. Hormonal Regulation of Leaf Abscission

    PubMed Central

    Jacobs, William P.

    1968-01-01

    A review is given of the progress made during the last 6 years in elucidating the nature, locus of action, and transport properties of the endogenous hormones that control leaf abscission. PMID:16657014

  16. Experiments in Whole Leaf Photosynthesis

    ERIC Educational Resources Information Center

    Stewart, J. C.; And Others

    1974-01-01

    Described is a simple experimental system, which uses radioactive carbon dioxide to study whole leaf photosynthesis under a variety of conditions. Other experiments and simple apparatus for the experiments are also described. (Author/RH)

  17. 7 CFR 29.3036 - Leaf surface.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Leaf surface. The smoothness or roughness of the web or lamina of a tobacco leaf. Leaf surface is... 7 Agriculture 2 2013-01-01 2013-01-01 false Leaf surface. 29.3036 Section 29.3036 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  18. 7 CFR 29.3036 - Leaf surface.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Leaf surface. The smoothness or roughness of the web or lamina of a tobacco leaf. Leaf surface is... 7 Agriculture 2 2014-01-01 2014-01-01 false Leaf surface. 29.3036 Section 29.3036 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  19. 7 CFR 29.3036 - Leaf surface.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Leaf surface. The smoothness or roughness of the web or lamina of a tobacco leaf. Leaf surface is... 7 Agriculture 2 2012-01-01 2012-01-01 false Leaf surface. 29.3036 Section 29.3036 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  20. 7 CFR 29.3036 - Leaf surface.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Leaf surface. The smoothness or roughness of the web or lamina of a tobacco leaf. Leaf surface is... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf surface. 29.3036 Section 29.3036 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  1. 7 CFR 29.3036 - Leaf surface.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Leaf surface. The smoothness or roughness of the web or lamina of a tobacco leaf. Leaf surface is... 7 Agriculture 2 2011-01-01 2011-01-01 false Leaf surface. 29.3036 Section 29.3036 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  2. Molecular characterization of PsbW, a nuclear-encoded component of the photosystem II reaction center complex in spinach.

    PubMed Central

    Lorković, Z J; Schröder, W P; Pakrasi, H B; Irrgang, K D; Herrmann, R G; Oelmüller, R

    1995-01-01

    We describe the isolation and characterization of cDNAs encoding the precursor polypeptide of the 6.1-kDa polypeptide associated with the reaction center core of the photosystem II complex from spinach. PsbW, the gene encoding this polypeptide, is present in a single copy per haploid genome. The mature polypeptide with 54 amino acid residues is characterized by a hydrophobic transmembrane segment, and, although an intrinsic membrane protein, it carries a bipartite transit peptide of 83 amino acid residues which directs the N terminus of the mature protein into the chloroplast lumen. Thylakoid integration of this polypeptide does not require a delta pH across the membrane, nor is it azide-sensitive, suggesting that the polypeptide chain inserts spontaneously in an as yet unknown way. The PsbW mRNA levels are light regulated. Similar to cytochrome b559 and PsbS, but different from the chlorophyll-complexing polypeptides D1, D2, CP43, and CP47 of photosystem II, PsbW is present in etiolated spinach seedlings. Images Fig. 1 Fig. 4 Fig. 5 Fig. 6 PMID:7568046

  3. In vivo tracing of organochloride and organophosphorus pesticides in different organs of hydroponically grown malabar spinach (Basella alba L.).

    PubMed

    Qiu, Junlang; Chen, Guosheng; Xu, Jianqiao; Luo, Erlun; Liu, Yan; Wang, Fuxin; Zhou, Hong; Liu, Yuan; Zhu, Fang; Ouyang, Gangfeng

    2016-10-01

    An in vivo uptake and elimination tracing study based on solid phase microextraction (SPME) was conducted to investigate the accumulation, persistence and distribution of organochloride pesticides (OCPs) and organophosphorus pesticides (OPPs) in malabar spinach (Basella alba L.) plants. Uptake and elimination of the pesticides were traced in leaves, stems and roots of living malabar spinach plants. Root concentration factor (RCF), distribution concentration factor (DCF) and transpiration stream concentration factor (TSCF) were calculated based on the in vivo tracing data. The tracing data showed that the OCPs were much more accumulative and persistent than the OPPs in roots, while they were similarly accumulative and persistent in leaves and stems. RCF values of the OPPs or OCPs were likely to increase with the increase in LogKow values except fenthion. Obtained DCF values indicated that OPPs and OCPs were more accumulative in the organs containing more lipids. TSCF values showed that the translocation of OPPs and OCPs from roots to foliage was firstly dependent on the hydrophobicity of the compounds, but also significantly affected by the water solubility. This is the first study of generating RCF, DCF and TSCF data in living plants by in vivo sampling method, which provides a foundation to promote the application of in vivo SPME and improve understanding of contaminant behaviors in living plants.

  4. [Fast separation and analysis of water-soluble vitamins in spinach by capillary electrophoresis with high voltage].

    PubMed

    Hu, Xiaoqin; You, Huiyan

    2009-11-01

    In capillary electrophoresis, 0-40 kV (even higher) voltage can be reached by a connecting double-model high voltage power supply. In the article, water-soluble vitamins, VB1, VB2, VB6, VC, calcium D-pantothenate, D-biotin, nicotinic acid and folic acid in vegetable, were separated by using the high voltage power supply under the condition of electrolyte water solution as running buffer. The separation conditions, such as voltage, the concentration of buffer and pH value etc. , were optimized during the experiments. The results showed that eight water-soluble vitamins could be baseline separated in 2.2 min at 40 kV applied voltage, 25 mmol/L sodium tetraborate buffer solution (pH 8.8). The water-soluble vitamins in spinach were quantified and the results were satisfied. The linear correlation coefficients of the water-soluble vitamins ranged from 0.9981 to 0.9999. The detection limits ranged from 0.2 to 0.3 mg/L. The average recoveries ranged from 88.0% to 100.6% with the relative standard deviations (RSD) range of 1.15%-4.13% for the spinach samples. PMID:20352941

  5. Automatic detection of regions in spinach canopies responding to soil moisture deficit using combined visible and thermal imagery.

    PubMed

    Raza, Shan-e-Ahmed; Smith, Hazel K; Clarkson, Graham J J; Taylor, Gail; Thompson, Andrew J; Clarkson, John; Rajpoot, Nasir M

    2014-01-01

    Thermal imaging has been used in the past for remote detection of regions of canopy showing symptoms of stress, including water deficit stress. Stress indices derived from thermal images have been used as an indicator of canopy water status, but these depend on the choice of reference surfaces and environmental conditions and can be confounded by variations in complex canopy structure. Therefore, in this work, instead of using stress indices, information from thermal and visible light imagery was combined along with machine learning techniques to identify regions of canopy showing a response to soil water deficit. Thermal and visible light images of a spinach canopy with different levels of soil moisture were captured. Statistical measurements from these images were extracted and used to classify between canopies growing in well-watered soil or under soil moisture deficit using Support Vector Machines (SVM) and Gaussian Processes Classifier (GPC) and a combination of both the classifiers. The classification results show a high correlation with soil moisture. We demonstrate that regions of a spinach crop responding to soil water deficit can be identified by using machine learning techniques with a high accuracy of 97%. This method could, in principle, be applied to any crop at a range of scales.

  6. Mild Fe-deficiency improves biomass production and quality of hydroponic-cultivated spinach plants (Spinacia oleracea L.).

    PubMed

    Jin, Chong-Wei; Liu, Yue; Mao, Qian-Qian; Wang, Qian; Du, Shao-Ting

    2013-06-15

    It is of great practical importance to improve yield and quality of vegetables in soilless cultures. This study investigated the effects of iron-nutrition management on yield and quality of hydroponic-cultivated spinach (Spinacia oleracea L.). The results showed that mild Fe-deficient treatment (1 μM FeEDTA) yielded a greater biomass of edible parts than Fe-omitted treatment (0 μM FeEDTA) or Fe-sufficient treatments (10 and 50 μM FeEDTA). Conversely, mild Fe-deficient treatment had the lowest nitrate concentration in the edible parts out of all the Fe treatments. Interestingly, all the concentrations of soluble sugar, soluble protein and ascorbate in mild Fe-deficient treatments were higher than Fe-sufficient treatments. In addition, both phenolic concentration and DPPH scavenging activity in mild Fe-deficient treatments were comparable with those in Fe-sufficient treatments, but were higher than those in Fe-omitted treatments. Therefore, we concluded that using a mild Fe-deficient nutrition solution to cultivate spinach not only would increase yield, but also would improve quality.

  7. Automatic Detection of Regions in Spinach Canopies Responding to Soil Moisture Deficit Using Combined Visible and Thermal Imagery

    PubMed Central

    Raza, Shan-e-Ahmed; Smith, Hazel K.; Clarkson, Graham J. J.; Taylor, Gail; Thompson, Andrew J.; Clarkson, John; Rajpoot, Nasir M.

    2014-01-01

    Thermal imaging has been used in the past for remote detection of regions of canopy showing symptoms of stress, including water deficit stress. Stress indices derived from thermal images have been used as an indicator of canopy water status, but these depend on the choice of reference surfaces and environmental conditions and can be confounded by variations in complex canopy structure. Therefore, in this work, instead of using stress indices, information from thermal and visible light imagery was combined along with machine learning techniques to identify regions of canopy showing a response to soil water deficit. Thermal and visible light images of a spinach canopy with different levels of soil moisture were captured. Statistical measurements from these images were extracted and used to classify between canopies growing in well-watered soil or under soil moisture deficit using Support Vector Machines (SVM) and Gaussian Processes Classifier (GPC) and a combination of both the classifiers. The classification results show a high correlation with soil moisture. We demonstrate that regions of a spinach crop responding to soil water deficit can be identified by using machine learning techniques with a high accuracy of 97%. This method could, in principle, be applied to any crop at a range of scales. PMID:24892284

  8. In vivo tracing of organochloride and organophosphorus pesticides in different organs of hydroponically grown malabar spinach (Basella alba L.).

    PubMed

    Qiu, Junlang; Chen, Guosheng; Xu, Jianqiao; Luo, Erlun; Liu, Yan; Wang, Fuxin; Zhou, Hong; Liu, Yuan; Zhu, Fang; Ouyang, Gangfeng

    2016-10-01

    An in vivo uptake and elimination tracing study based on solid phase microextraction (SPME) was conducted to investigate the accumulation, persistence and distribution of organochloride pesticides (OCPs) and organophosphorus pesticides (OPPs) in malabar spinach (Basella alba L.) plants. Uptake and elimination of the pesticides were traced in leaves, stems and roots of living malabar spinach plants. Root concentration factor (RCF), distribution concentration factor (DCF) and transpiration stream concentration factor (TSCF) were calculated based on the in vivo tracing data. The tracing data showed that the OCPs were much more accumulative and persistent than the OPPs in roots, while they were similarly accumulative and persistent in leaves and stems. RCF values of the OPPs or OCPs were likely to increase with the increase in LogKow values except fenthion. Obtained DCF values indicated that OPPs and OCPs were more accumulative in the organs containing more lipids. TSCF values showed that the translocation of OPPs and OCPs from roots to foliage was firstly dependent on the hydrophobicity of the compounds, but also significantly affected by the water solubility. This is the first study of generating RCF, DCF and TSCF data in living plants by in vivo sampling method, which provides a foundation to promote the application of in vivo SPME and improve understanding of contaminant behaviors in living plants. PMID:27209519

  9. Photoregulation of fructose and glucose respiration in the intact chloroplasts of Chlamydomonas reinhardtii F-60 and spinach

    SciTech Connect

    Singh, K.K.; Changguo Chen; Gibbs, M. )

    1993-04-01

    The photoregulation of chloroplastic respiration was studied by monitoring in darkness and in light the release of [sup 14]CO[sub 2] from whole chloroplasts of Chlamydomonas reinhardtii F-60 and spinach (Spinacia oleracea L.) supplied externally with [[sup 14]C]glucose and [[sup 14]C]fructose, respectively. CO[sub 2] release was inhibited more than 90% in both chloroplasts by a light intensity of 4 W m[sup [minus]2]. Oxidants, oxaloacetate in Chlamydomonas, nitrite in spinach, and phenazine methosulfate in both chloroplasts, reversed the inhibition. The onset of the photoinhibitory effect on CO[sub 2] release was relatively rapid compared to the restoration of CO[sub 2] release following illumination. In both darkened chloroplasts, dithiothreitol inhibited release. Of the four enzymes (fructokinase, phosphoglucose isomerase, glucose-6-P dehydrogenase, and gluconate-6-P dehydrogenase) in the pathway catalyzing the release of CO[sub 2] from fructose, only glucose-6-P dehydrogenase was deactivated by light and by dithiothreitol. 33 refs., 3 figs., 4 tabs.

  10. Automatic detection of regions in spinach canopies responding to soil moisture deficit using combined visible and thermal imagery.

    PubMed

    Raza, Shan-e-Ahmed; Smith, Hazel K; Clarkson, Graham J J; Taylor, Gail; Thompson, Andrew J; Clarkson, John; Rajpoot, Nasir M

    2014-01-01

    Thermal imaging has been used in the past for remote detection of regions of canopy showing symptoms of stress, including water deficit stress. Stress indices derived from thermal images have been used as an indicator of canopy water status, but these depend on the choice of reference surfaces and environmental conditions and can be confounded by variations in complex canopy structure. Therefore, in this work, instead of using stress indices, information from thermal and visible light imagery was combined along with machine learning techniques to identify regions of canopy showing a response to soil water deficit. Thermal and visible light images of a spinach canopy with different levels of soil moisture were captured. Statistical measurements from these images were extracted and used to classify between canopies growing in well-watered soil or under soil moisture deficit using Support Vector Machines (SVM) and Gaussian Processes Classifier (GPC) and a combination of both the classifiers. The classification results show a high correlation with soil moisture. We demonstrate that regions of a spinach crop responding to soil water deficit can be identified by using machine learning techniques with a high accuracy of 97%. This method could, in principle, be applied to any crop at a range of scales. PMID:24892284

  11. Why do leaf-tying caterpillars abandon their leaf ties?

    PubMed

    Sliwinski, Michelle; Sigmon, Elisha

    2013-01-01

    Leaf-tying caterpillars act as ecosystem engineers by building shelters between overlapping leaves, which are inhabited by other arthropods. Leaf-tiers have been observed to leave their ties and create new shelters (and thus additional microhabitats), but the ecological factors affecting shelter fidelity are poorly known. For this study, we explored the effects of resource limitation and occupant density on shelter fidelity and assessed the consequences of shelter abandonment. We first quantified the area of leaf material required for a caterpillar to fully develop for two of the most common leaf-tiers that feed on white oak, Quercus alba. On average, Psilocorsis spp. caterpillars consumed 21.65 ± 0.67 cm(2) leaf material to complete development. We also measured the area of natural leaf ties found in a Maryland forest, to determine the distribution of resources available to caterpillars in situ. Of 158 natural leaf ties examined, 47% were too small to sustain an average Psilocorsis spp. caterpillar for the entirety of its development. We also manipulated caterpillar densities within experimental ties on potted trees to determine the effects of cohabitants on the likelihood of a caterpillar to leave its tie. We placed 1, 2, or 4 caterpillars in ties of a standard size and monitored the caterpillars twice daily to track their movement. In ties with more than one occupant, caterpillars showed a significantly greater propensity to leave their tie, and left sooner and at a faster rate than those in ties as single occupants. To understand the consequences of leaf tie abandonment, we observed caterpillars searching a tree for a site to build a shelter in the field. This is a risky behavior, as 17% of the caterpillars observed died while searching for a shelter site. Caterpillars that successfully built a shelter traveled 110 ± 20 cm and took 28 ± 7 min to find a suitable site to build a shelter. In conclusion, leaf-tying caterpillars must frequently abandon their leaf

  12. What determines a leaf's shape?

    PubMed

    Dkhar, Jeremy; Pareek, Ashwani

    2014-01-01

    The independent origin and evolution of leaves as small, simple microphylls or larger, more complex megaphylls in plants has shaped and influenced the natural composition of the environment. Significant contributions have come from megaphyllous leaves, characterized usually as flat, thin lamina entrenched with photosynthetic organelles and stomata, which serve as the basis of primary productivity. During the course of evolution, the megaphylls have attained complexity not only in size or venation patterns but also in shape. This has fascinated scientists worldwide, and research has progressed tremendously in understanding the concept of leaf shape determination. Here, we review these studies and discuss the various factors that contributed towards shaping the leaf; initiated as a small bulge on the periphery of the shoot apical meristem (SAM) followed by asymmetric outgrowth, expansion and maturation until final shape is achieved. We found that the underlying factors governing these processes are inherently genetic: PIN1 and KNOX1 are indicators of leaf initiation, HD-ZIPIII, KANADI, and YABBY specify leaf outgrowth while ANGUSTIFOLIA3 and GROWTH-REGULATING FACTOR5 control leaf expansion and maturation; besides, recent research has identified new players such as APUM23, known to specify leaf polarity. In addition to genetic control, environmental factors also play an important role during the final adjustment of leaf shape. This immense amount of information available will serve as the basis for studying and understanding innovative leaf morphologies viz. the pitchers of the carnivorous plant Nepenthes which have evolved to provide additional support to the plant survival in its nutrient-deficient habitat. In hindsight, formation of the pitcher tube in Nepenthes might involve the recruitment of similar genetic mechanisms that occur during sympetaly in Petunia. PMID:25584185

  13. What determines a leaf's shape?

    PubMed

    Dkhar, Jeremy; Pareek, Ashwani

    2014-01-01

    The independent origin and evolution of leaves as small, simple microphylls or larger, more complex megaphylls in plants has shaped and influenced the natural composition of the environment. Significant contributions have come from megaphyllous leaves, characterized usually as flat, thin lamina entrenched with photosynthetic organelles and stomata, which serve as the basis of primary productivity. During the course of evolution, the megaphylls have attained complexity not only in size or venation patterns but also in shape. This has fascinated scientists worldwide, and research has progressed tremendously in understanding the concept of leaf shape determination. Here, we review these studies and discuss the various factors that contributed towards shaping the leaf; initiated as a small bulge on the periphery of the shoot apical meristem (SAM) followed by asymmetric outgrowth, expansion and maturation until final shape is achieved. We found that the underlying factors governing these processes are inherently genetic: PIN1 and KNOX1 are indicators of leaf initiation, HD-ZIPIII, KANADI, and YABBY specify leaf outgrowth while ANGUSTIFOLIA3 and GROWTH-REGULATING FACTOR5 control leaf expansion and maturation; besides, recent research has identified new players such as APUM23, known to specify leaf polarity. In addition to genetic control, environmental factors also play an important role during the final adjustment of leaf shape. This immense amount of information available will serve as the basis for studying and understanding innovative leaf morphologies viz. the pitchers of the carnivorous plant Nepenthes which have evolved to provide additional support to the plant survival in its nutrient-deficient habitat. In hindsight, formation of the pitcher tube in Nepenthes might involve the recruitment of similar genetic mechanisms that occur during sympetaly in Petunia.

  14. Increasing leaf hydraulic conductance with transpiration rate minimizes the water potential drawdown from stem to leaf.

    PubMed

    Simonin, Kevin A; Burns, Emily; Choat, Brendan; Barbour, Margaret M; Dawson, Todd E; Franks, Peter J

    2015-03-01

    Leaf hydraulic conductance (k leaf) is a central element in the regulation of leaf water balance but the properties of k leaf remain uncertain. Here, the evidence for the following two models for k leaf in well-hydrated plants is evaluated: (i) k leaf is constant or (ii) k leaf increases as transpiration rate (E) increases. The difference between stem and leaf water potential (ΔΨstem-leaf), stomatal conductance (g s), k leaf, and E over a diurnal cycle for three angiosperm and gymnosperm tree species growing in a common garden, and for Helianthus annuus plants grown under sub-ambient, ambient, and elevated atmospheric CO₂ concentration were evaluated. Results show that for well-watered plants k leaf is positively dependent on E. Here, this property is termed the dynamic conductance, k leaf(E), which incorporates the inherent k leaf at zero E, which is distinguished as the static conductance, k leaf(0). Growth under different CO₂ concentrations maintained the same relationship between k leaf and E, resulting in similar k leaf(0), while operating along different regions of the curve owing to the influence of CO₂ on g s. The positive relationship between k leaf and E minimized variation in ΔΨstem-leaf. This enables leaves to minimize variation in Ψleaf and maximize g s and CO₂ assimilation rate over the diurnal course of evaporative demand.

  15. Increasing leaf hydraulic conductance with transpiration rate minimizes the water potential drawdown from stem to leaf

    PubMed Central

    Simonin, Kevin A.; Burns, Emily; Choat, Brendan; Barbour, Margaret M.; Dawson, Todd E.; Franks, Peter J.

    2015-01-01

    Leaf hydraulic conductance (k leaf) is a central element in the regulation of leaf water balance but the properties of k leaf remain uncertain. Here, the evidence for the following two models for k leaf in well-hydrated plants is evaluated: (i) k leaf is constant or (ii) k leaf increases as transpiration rate (E) increases. The difference between stem and leaf water potential (ΔΨstem–leaf), stomatal conductance (g s), k leaf, and E over a diurnal cycle for three angiosperm and gymnosperm tree species growing in a common garden, and for Helianthus annuus plants grown under sub-ambient, ambient, and elevated atmospheric CO2 concentration were evaluated. Results show that for well-watered plants k leaf is positively dependent on E. Here, this property is termed the dynamic conductance, k leaf(E), which incorporates the inherent k leaf at zero E, which is distinguished as the static conductance, k leaf(0). Growth under different CO2 concentrations maintained the same relationship between k leaf and E, resulting in similar k leaf(0), while operating along different regions of the curve owing to the influence of CO2 on g s. The positive relationship between k leaf and E minimized variation in ΔΨstem–leaf. This enables leaves to minimize variation in Ψleaf and maximize g s and CO2 assimilation rate over the diurnal course of evaporative demand. PMID:25547915

  16. Natural Occurrence of Tomato leaf curl New Delhi virus in Iranian Cucurbit Crops

    PubMed Central

    Yazdani-Khameneh, Sara; Aboutorabi, Samaneh; Shoori, Majid; Aghazadeh, Azin; Jahanshahi, Parastoo; Golnaraghi, Alireza; Maleki, Mojdeh

    2016-01-01

    The main areas for field-grown vegetable production in Iran were surveyed during the years of 2012–2014 to determine the occurrence of begomoviruses infecting these crops. A total of 787 leaf samples were collected from vegetables and some other host plants showing virus-like symptoms and tested by an enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies produced against Tomato yellow leaf curl virus (TYLCV). According to the ELISA results, 81 samples (10.3%) positively reacted with the virus antibodies. Begomovirus infections were confirmed by polymerase chain reaction (PCR) using previously described TYLCV-specific primer pair TYLCV-Sar/TYLCV-Isr or universal primer pair Begomo-F/Begomo-R. The PCR tests using the primer pair TYLCV-Sar/TYLCV-Isr resulted in the amplification of the expected fragments of ca. 0.67-kb in size for ELISA-positive samples tested from alfalfa, pepper, spinach and tomato plants, confirming the presence of TYLCV. For one melon sample, having a week reaction in ELISA and no reaction in PCR using TYLCV-specific primers, the PCR reaction using the primer pair Begomo-F/Begomo-R resulted in the amplification fragments of the expected size of ca. 2.8 kb. The nucleotide sequences of the DNA amplicons derived from the isolate, Kz-Me198, were determined and compared with other sequences available in GenBank. BLASTN analysis confirmed the begomovirus infection of the sample and showed 99% identities with Tomato leaf curl New Delhi virus (ToLCNDV); phylogenetic analysis supported the results of the database searches. This study reports the natural occurrence of TYLCV in different hosts in Iran. Our results also reveal the emergence of ToLCNDV in Iranian cucurbit crops. PMID:27298595

  17. Natural Occurrence of Tomato leaf curl New Delhi virus in Iranian Cucurbit Crops.

    PubMed

    Yazdani-Khameneh, Sara; Aboutorabi, Samaneh; Shoori, Majid; Aghazadeh, Azin; Jahanshahi, Parastoo; Golnaraghi, Alireza; Maleki, Mojdeh

    2016-06-01

    The main areas for field-grown vegetable production in Iran were surveyed during the years of 2012-2014 to determine the occurrence of begomoviruses infecting these crops. A total of 787 leaf samples were collected from vegetables and some other host plants showing virus-like symptoms and tested by an enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies produced against Tomato yellow leaf curl virus (TYLCV). According to the ELISA results, 81 samples (10.3%) positively reacted with the virus antibodies. Begomovirus infections were confirmed by polymerase chain reaction (PCR) using previously described TYLCV-specific primer pair TYLCV-Sar/TYLCV-Isr or universal primer pair Begomo-F/Begomo-R. The PCR tests using the primer pair TYLCV-Sar/TYLCV-Isr resulted in the amplification of the expected fragments of ca. 0.67-kb in size for ELISA-positive samples tested from alfalfa, pepper, spinach and tomato plants, confirming the presence of TYLCV. For one melon sample, having a week reaction in ELISA and no reaction in PCR using TYLCV-specific primers, the PCR reaction using the primer pair Begomo-F/Begomo-R resulted in the amplification fragments of the expected size of ca. 2.8 kb. The nucleotide sequences of the DNA amplicons derived from the isolate, Kz-Me198, were determined and compared with other sequences available in GenBank. BLASTN analysis confirmed the begomovirus infection of the sample and showed 99% identities with Tomato leaf curl New Delhi virus (ToLCNDV); phylogenetic analysis supported the results of the database searches. This study reports the natural occurrence of TYLCV in different hosts in Iran. Our results also reveal the emergence of ToLCNDV in Iranian cucurbit crops. PMID:27298595

  18. The orientation of membrane bound radicals: an EPR investigation of magnetically ordered spinach chloroplasts.

    PubMed

    Dismukes, G C; Sauer, K

    1978-12-01

    The orientation of membrane-bound radicals in spinach chloroplasts is examined by electron paramagnetic resonance (EPR) spectroscopy of chloroplasts oriented by magnetic fields. Several of the membrane-bound radicals which possess g-tensor anisotropy display EPR signals with a marked dependence on the orientation of the membranes relative to the applied EPR field. The fraction of oxidized and reduced plastocyanin, P-700, iron-sulfur proteins A and B, and the X center, an early acceptor of Photosystem I, can be controlled by the light intensity during steady-state illumination and can be trapped by cooling. The X center can be photoreduced and trapped in the absence of strong reductants and high pH, conditions previously found necessary for its detection. These results confirm its role as an early electron acceptor in P-700 photo-oxidation. X is oriented with its smallest principal g-tensor axis (gx) predominantly parallel to the normal to the thylakoid membrane, the same orientation as was found for an early electron acceptor based on time-resolved electron spin polarization studies. We propose that the X center is the first example of a high potential iron-sulfur protein which functions in electron transfer in its 'superreduced' state. We present evidence which suggests that iron-sulfur proteins A and B are 4Fe-4S clusters in an 8Fe-8S protein. Center B is oriented with gy predominantly normal to the membrane plane. The spectra of center A and plastocyanin do not show significant changes with sample orientation. In the case of plastocyanin, this may indicate a lack of molecular orientation. The absence of an orientation effect for reduced center A is reconcilable with a 4Fe-4S geometry, provided that the electron obtained upon reduction can be shared between any pair of Fe atoms in the center. Orientation of the 'Rieske' iron-sulfur protein is also observed. It has axial symmetry with g parallel close to the plane of the membrane. A model is proposed for the

  19. Spinach nitrite reductase. Purification and properties of a siroheme-containing iron-sulfur enzyme.

    PubMed

    Vega, J M; Kamin, H

    1977-02-10

    Ferredoxin-nitrite reductase (EC 1.7.7.1.) from spinach has been purified to homogeneity with a specific activity of 110 units/mg of protein. The enzyme, Mr = 61,000 has 3 iron atoms (of which one is in siroheme) and 2 labile sulfides, i.e. 1 (Fe2-S2) per molecule, with absorption maxima at 276, 386 (Soret), 573 (alpha), and 690 nm, with an E386 of 3.97 X 10(4) M-1-cm-1, and A276/A386 absorptivity ratio of 1.8. Anaerobic addition of dithionite results in the loss of the 690 nm peak and the splitting of the 573 nm absorption band into two broad peaks at 545 and 585 nm. Reduction by dithionite is enhanced by cyanide (Fig. 7) and requires about 3 electron eq per mol of enzyme. With nitrite or hydroxylamine (substrates of the enzyme), cyanide (a competitive inhibitor with respect to nitrite), or sulfite, the 690 nm absorption band of substrate-free enzyme disappears and the absorbance in the Soret and alpha region are altered. The high spin EPR signals disappear (J. M. Vega, H. Kamin, N. R. Orme-Johnson, and W. H. Orme-Johnson, unpublished observations). Titration permits calculation of 1 mol of nitrite bound/mol of enzyme with a Kdiss of 3.2 X 10(-6) M. Dithionite-reduced enzyme also forms complexes with added nitrite, hydroxylamine, or cyanide, characterized by marked alterations in the 573 (alpha) absorption band. THus, substrates or competitive inhibitors can be bound to the oxidized or reduced enzyme forms. CO inhibits nitrite reductase and forms a complex with reduced enzyme (epsilonmax at 395, 543, and 585 nm). Formation or dissociation of the spectrophotometrically detectable CO complex correlates with inhibition or inhibition-reversal of nitrite reduction catalysis. During steady state turnover with dithionite and nitrite, the enzyme forms a complex with added nitrite with absorption difference maxima at 445, 538, and 580 nm with respect to reduced enzyme. When nearly all substrate is depleted the spectrum of a new species appears, indicating that nitrite

  20. Leaf exsertion, leaf elongation, and leaf senescence in Eriophorum vaginatum and Carex Bigelowii

    SciTech Connect

    Shaver, G.R.; Yandow, T.; Laundre, J.

    1990-01-01

    Most of the common sedges of arctic vegetation show a pattern of leaf production in which the exsertion and elongation of new leaves is more or less simultaneous with the senescence of old leaves. The present study was designed to increase our understanding of the variability sequential leaf production by arctic sedges, and to determine some of the controls on that variability. We did this in two ways: first, we compared the sequential patterns of leaf growth and senescence in E. vaginatum with those of Carex Bigelowii Torr. at two tussock tundra sites near Toolik Lake on the North Slope of Alaska. Second, we compared the responses of leaf growth in these species in control and fertilized plots and in two microenvironments thought to differ sharply in nutrient availability and total productivity. 29 refs., 28 figs., 2 tabs.

  1. Effect of gamma radiation on the reduction of Salmonella strains, Listeria monocytogenes, and Shiga toxin-producing Escherichia coli and sensory evaluation of minimally processed spinach (Tetragonia expansa).

    PubMed

    Rezende, Ana Carolina B; Igarashi, Maria Crystina; Destro, Maria Teresa; Franco, Bernadette D G M; Landgraf, Mariza

    2014-10-01

    This study evaluated the effects of irradiation on the reduction of Shiga toxin-producing Escherichia coli (STEC), Salmonella strains, and Listeria monocytogenes, as well as on the sensory characteristics of minimally processed spinach. Spinach samples were inoculated with a cocktail of three strains each of STEC, Salmonella strains, and L. monocytogenes, separately, and were exposed to gamma radiation doses of 0, 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. Samples that were exposed to 0.0, 1.0, and 1.5 kGy and kept under refrigeration (4°C) for 12 days were submitted to sensory analysis. D10 -values ranged from 0.19 to 0.20 kGy for Salmonella and from 0.20 to 0.21 for L. monocytogenes; for STEC, the value was 0.17 kGy. Spinach showed good acceptability, even after exposure to 1.5 kGy. Because gamma radiation reduced the selected pathogens without causing significant changes in the quality of spinach leaves, it may be a useful method to improve safety in the fresh produce industry. PMID:25285495

  2. Mathematical modeling and numerical analysis of the growth of Non-O157 shiga toxin-producing Escherichia coli in spinach leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to investigate the growth of non-O157 Shiga toxin-producing Escherichia coli (STEC) in spinach leaves and to develop kinetic models to describe the bacterial growth. Six serogroups of non-O157 STEC, including O26, O45, O103, O111, O121, and O145, were used in the growth stu...

  3. Use of zero-valent iron biosand filters to reduce E. coli O157:H12 in irrigation water applied to spinach plants in a field setting

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Zero-valent iron (ZVI) filters may provide an efficient method to mitigate the contamination of produce crops through irrigation water. Purpose: To evaluate the use of ZVI-filtration in decontaminating E. coli O157:H12 in irrigation water and on spinach plants in a small, field-scale...

  4. Isolation of Campylobacter from feral swine (Sus scrofa) on the ranch associated with the 2006 Escherichia coli O157:H7 spinach outbreak investigation in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report the isolation of Campylobacter species from the same population of feral swine that was investigated in San Benito County, California during the 2006 spinach-related Escherichia coli O157:H7 outbreak. This is the first survey of Campylobacter in a free-ranging feral swine population in the...

  5. The effect of total organic carbon content and repeated irrigation on the persistence of E. coli O157:H7 on baby spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Contaminated fresh-cut spinach and other leafy greens have caused foodborne illness in the United States. In response, growers are adopting recommendations stated in the California Leafy Greens Marketing Agreement (LGMA). The LGMA permits a maximum population of 126 Most Probable Nu...

  6. The Effect of Repeated Irrigation with Water Containing Varying Levels of Total Organic Carbon on the Persistence of Escherichia coli O157:H7 on Baby Spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The California lettuce and leafy greens industry has adopted the Leafy Greens Marketing Agreement (LGMA), which allows for 126 Most Probable Number (MPN) generic E. coli/100ml in irrigation water. Repeat irrigation of baby spinach plants with water containing E. coli O157:H7 and different levels of...

  7. The effect of repeated irrigation with varying total organic carbon content on the persistence of E. coli O157:H7 on baby spinach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In response to U.S. foodborne illnesses caused by contaminated spinach, growers have adopted regulations stated in the California Leafy Greens Marketing Agreement (LGMA). The LGMA permits a maximum population mean of 126 Most Probable Number (MPN) generic E. coli per 100 ml irrigation water. These...

  8. Organic chelants-mediated enhanced lead (Pb) uptake and accumulation is associated with higher activity of enzymatic antioxidants in spinach (Spinacea oleracea L.).

    PubMed

    Khan, Imran; Iqbal, Muhammad; Ashraf, Muhammad Yasin; Ashraf, Muhammad Arslan; Ali, Shafaqat

    2016-11-01

    The spinach was tested in the present studies for its phytoextraction potential. Furthermore, the study assessed whether organic chelants could reduce oxidative stress, and thus enhance growth of spinach plants under 2.42 and 4.83mM Pb regimes. Different organic chelates viz. ethylenediamine tetra acetic acid, (EDTA), citric acid (CA), oxalic acid (OA), tartaric acid (TA) and malic acid (MA) were applied separately in addition to control (without chelating agents) under different Pb regimes. The low (2.42mM) Pb regime increased biological yield (kgha(-1)). All the chelates except OA increased biological yield under low Pb regime. In contrast, TA caused less decrease in biomass under high (4.83mM) Pb regime. The chelate-assisted rise in the antioxidant activities substantially contributed to reactive oxygen species (ROS) neutralization. Of the chelates, TA was the most effective in improving Pb uptake and its root to shoot translocation. Overall, the chelate-assisted buildup of Pb in the spinach did not exhibit inhibitory effects on the plant growth possibly due to their potential to decrease Pb-induced oxidative damage. The results elaborated the potential of TA in increasing root to shoot translocation of Pb, biomass, and thus suggested its use for phytoextraction of Pb using spinach in Pb contaminated environments. PMID:27318732

  9. A novel approach to investigate the uptake and internalization of Escherichia coli O157:H7 in spinach cultivated in soil and hydroponic media

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Internalization of E. coli O157:H7 into spinach plants through root uptake is a potential route of contamination. A Tn7-based plasmid vector was used to insert the green fluorescent protein (gfp) gene into the attTn7 site in the E. coli chromosome. Three gfp-labeled E. coli inocula, O157:H7 strains ...

  10. D-ribulose-5-phosphate 3-epimerase: Cloning and heterologous expression of the spinach gene, and purification and characterization of the recombinant enzyme

    SciTech Connect

    Chen, Y.R.; Hartman, F.C.; Lu, T.Y.S.; Larimer, F.W.

    1998-09-01

    The authors have achieved, to their knowledge, the first high-level heterologous expression of the gene encoding D-ribulose-5-phosphate 3-epimerase from any source, thereby permitting isolation and characterization of the epimerase as found in photosynthetic organisms. The extremely labile recombinant spinach (Spinacia oleracea L.) enzyme was stabilized by DL-{alpha}-glycerophosphate or ethanol and destabilized by D-ribulose-5-phosphate or 2-mercaptoethanol. Despite this lability, the unprecedentedly high specific activity of the purified material indicates that the structural integrity of the enzyme is maintained throughout isolation. Ethylenediaminetetraacetate and divalent metal cations did not affect epimerase activity, thereby excluding a requirement for the latter in catalysis. As deduced from the sequence of the cloned spinach gene and the electrophoretic mobility under denaturing conditions of the purified recombinant enzyme, its 25-kD subunit size was about the same as that of the corresponding epimerases of yeast and mammals. However, in contrast to these other species, the recombinant spinach enzyme was octameric rather than dimeric, as assessed by gel filtration and polyacrylamide gel electrophoresis under nondenaturing conditions. Western-blot analyses with antibodies to the purified recombinant enzyme confirmed that the epimerase extracted from spinach leaves is also octameric.

  11. Effect of Nd{sup 3+} ion on carboxylation activity of ribulose-1,5-bisphosphate carboxylase/oxygenase of spinach

    SciTech Connect

    Liu Chao; Hong Fashui . E-mail: Hongfsh_cn@sina.com; Wu Kang; Ma, Hong-bing; Zhang Xueguang; Hong Chengjiao; Wu Cheng; Gao Fengqing; Yang Fan; Zheng Lei; Wang Xuefeng; Liu Tao; Xie Yaning; Xu Jianhua; Li Zhongrui

    2006-03-31

    Neodymium (Nd), as a member of rare earth elements, proved to enhance the photosynthesis rate and organic substance accumulation of spinach through the increase in carboxylation activity of Rubisco. Although the oxygenase activity of spinach Rubisco was slightly changed with the Nd{sup 3+} treatment, the specific factor of Rubisco was greatly increased. It was partially due to the promotion of Rubisco activase (R-A) activity but mainly to the formation of Rubisco-Rubisco activase super-complex, a heavier molecular mass protein (about 1200 kD) comprising both Rubisco and Rubisco activase. This super-complex was found during the extraction procedure of Rubisco by the gel electrophoresis and Western-blot studies. The formation of Rubisco-R-A super-complex suggested that the secondary structure of the protein purified from the Nd{sup 3+}-treated spinach was different from that of the control. Extended X-ray absorption fine structure study of the 'Rubisco' purified from the Nd{sup 3+}-treated spinach revealed that Nd was bound with four oxygen atoms and two sulfur atoms of amino acid residues at the Nd-O and Nd-S bond lengths of 2.46 and 2.89 A, respectively.

  12. Influence of the interaction between light intensity and CO2 concentration on productivity and quality of spinach (Spinacia oleracea L.) grown in fully controlled environment

    NASA Astrophysics Data System (ADS)

    Proietti, Simona; Moscatello, Stefano; Giacomelli, Gene A.; Battistelli, Alberto

    2013-09-01

    The effects of the factorial combination of two light intensities (200 and 800 μmol m-2 s-1) and two CO2 concentrations (360 and 800 ppm) were studied on the productivity and nutritional quality of spinach (Spinacia oleracea L.) grown under controlled environment. After 6 weeks within a growth chamber, spinach plants were sampled and analyzed for productivity and quality. There were no statistically significant interactions between the effects of light and CO2 for all of the variables studied, except for the nitrate and oxalic acid content of the leaves. High light and high CO2 independently one from the other, promoted spinach productivity, and the accumulation of ascorbic acid, while their interactive effect limited the accumulation of nitrate and oxalic acid in the spinach leaves. The results highlight the importance of considering the effects of the interaction among environmental variables on maximizing production and the nutritional quality of the food when cultivating and modeling the plant response in controlled environment systems such as for bioregenerative life support.

  13. Respiration of sugars in spinach (Spinacia oleraces), maize (Zea mays), and Chlamydomonas reinhardtii F-60 chloroplasts with emphasis on the hexose kinases

    SciTech Connect

    Singh, K.K.; Chen, C.; Epstein, D.K.; Gibbs, M. )

    1993-06-01

    The role of hexokinase in carbohydrate degradation in isolated, intact chloroplasts was evaluated. This was accomplished by monitoring the evolution of [sup 14]CO[sub 2] from darkened spinach (Spinacia oleracea), maize (Zea mays) mesophyll, and Chlamydomonas reinhardtii chloroplasts externally supplied with [sup 14]C-labeled fructose, glucose, mannose, galactose, maltose, and ribose. Glucose and ribose were the preferred substrates with the Chlamydomonas and maize chloroplasts, respectively. The rate of CO[sub 2] release from fructose was about twice that from glucose in the spinach chloroplast. externally supplied ATP stimulated the rate of CO[sub 2] release. The pH optimum for CO[sub 2] release was 7.5 with ribose and fructose and 8.5 with glucose as substrates. Probing the outer membrane polypeptides of the intact spinach chloroplast with two proteases, trypsin and thermolysin, decreased [sup 14]CO[sub 2] release from glucose about 50% but had little effect when fructose was the substrate. Tryptic digestion decreased CO[sub 2] release from glucose in the Chlamydomonas chloroplast about 70%. [sup 14]CO[sub 2] evolution from [1-[sup 14]C]-glucose-6-phosphate in both chloroplasts was unaffected by treatment with trypsin. Enzymic analysis of the supernatant (stroma) of the lysed spinach chloroplast indicated a hexokinase active primarily with fructose but with some affinity for glucose. The pellet (membranal fraction) contained a hexokinase utilizing both glucose and fructose but with considerably less total activity than the stormal enzyme. Treatment with trypsin and thermolysin eliminated more than 50% of the glucokinase activity but had little effect on fructokinase activity in the spinach chloroplast. Tryptic digestion of the Chlamydomonas chloroplast resulted in a loss of about 90% of glucokinase activity. 34 refs., 2 figs., 6 tabs.

  14. Spectroscopic and kinetic properties of a recombinant form of the flavin domain of spinach NADH: nitrate reductase.

    PubMed

    Quinn, G B; Trimboli, A J; Prosser, I M; Barber, M J

    1996-03-01

    The C-terminal 268 residues of the spinach assimilatory NADH:nitrate reductase amino acid sequence that correspond to the flavin-containing domain of the enzyme have been selectively amplified and expressed as a recombinant protein in Escherichia coli. The recombinant protein, which was produced in both soluble and insoluble forms, was purified to homogeneity using a combination of ammonium sulfate precipitation, affinity chromatography on 5'-ADP-agarose and FPLC gel filtration. The purified domain exhibited a molecular weight of approximately 30 kDa, estimated by polyacrylamide gel electrophoresis, and a molecular mass of 30,169 for the apoprotein determined by mass spectrometry, which also confirmed the presence of FAD. The UV/visible spectrum was typical of a flavoprotein, with maxima at 272, 386, and 461 nm in the oxidized form while CD spectroscopy yielded both positive and negative maxima at 313 and 382 nm and 461 and 484 nm, respectively. The purified domain showed immunological cross-reactivity with anti-spinach nitrate reductase polyclonal antibodies while both N-terminal and internal amino acid sequencing of isolated peptides confirmed the fidelity of the domain's primary sequence. The protein retained NADH-ferricyanide reductase activity (Vmax=84 micromol NADH consumer/min/nmol FAD) with Km's of 17 and 34 microM for NADH and ferricyanide, respectively, with a pH optimum of approximately 6.5 A variety of NADH-analogs could also function as electron donors, though with decreased efficiency, the most effective being reduced nicotinamide hypoxanthine dinucleotide (V(max) = 35 micromol NHDH consumer/min/nmol FAD) and Km = 22 microM). NAD+ was demonstrated to be a competitive inhibitor (Ki = 1.9 mM) while analysis of inhibition by a variety of NAD+-analogs indicated the most efficient inhibitor to be ADP (Ki = 0.2 mM), with analogs devoid of either the phosphate, ribose, or adenine moieties proving to be markedly less-efficient inhibitors. The isolated domain

  15. How to pattern a leaf.

    PubMed

    Bolduc, N; O'Connor, D; Moon, J; Lewis, M; Hake, S

    2012-01-01

    Leaf development presents a tremendous resource for tackling the question of patterning in biology. Leaves can be simple or highly dissected. They may have elaborated parts such as the tendrils of a pea leaf or the rolled blade of a carnivorous pitcher plant. Despite the variation in size, shape, and function, all leaves initiate in the same manner: from the flanks of a meristem. The maize leaf is useful for analysis of patterning due to the wealth of mutants and the distinct tissues along the proximal distal axis. The blade is distal, the sheath is proximal, and the ligule forms at the blade/sheath boundary. Establishment of this boundary involves the transcription factors LIGULELESS1 and LIGULELESS2 and the kinase LIGULELESS NARROW. The meristem-specific protein KNOTTED1 (KN1) binds and modulates the lg2 gene. Given the localization of KN1 at the proximal end of the leaf from the time of inception, we hypothesize that KN1 has a role in establishing the very proximal end of the leaf, whereas an auxin maximum guides the growing distal tip. PMID:23174765

  16. [Studies on the light-induced reversible xanthophyll-conversions in Chlorella and spinach leaves].

    PubMed

    Hager, A

    1967-06-01

    1. Using new methods in thin-layer chromatography, experiments were carried out to prove the light-induced changes in the quantity of various xanthophylls in Chlorella and spinach leaves. The probable connection of these interconversions to electron transport in photosynthesis was demonstrated. 2. The kinetics of these xanthophyll conversions were investigated during strong illumination and in the succeeding dark period (Chlorella). Already after illumination of 1 min one can detect a decrease of the di-epoxide xanthophyll violaxanthin and a corresponding increase of the epoxide-free zeaxanthin. The intermediate of this interconversion is the mono-epoxide antheraxanthin. Neoxanthin exhibits no change in concentration under the given light intensity and an illumination time of 60 min and more; the same result can be observed with the other carotenoids (α-carotene, β-carotene, lutein, lutein-5,6-epoxyd) and the chlorophylls a and b. 3. The light-induced formation of zeaxanthin is not correlated with those pigment interconversions which are photooxidative in their nature and which may be detected only after long illuminations. However, by using damaged, e.g., briefly heated Chlorella cells, a photooxidative-induced decrease of carotenes and chlorophyll a and a smaller decrease of xanthophylls and chlorophyll b could already be demonstrated after illumination of 15 min. In this case the ratio xanthophylls/ carotenes increases. 4. The transformation violaxanthin → antheraxanthin → zeaxanthin ("forward-reaction") is induced not only by an illumination with white light (point 2) but also with red light (>600 nm); that means the reaction proceeds at a wavelength which cannot be absorbed by the xanthophylls themselves. Chlorophyll acts as light-acceptor. 5. The "forward-reaction" does not proceed after the cells have been heated for a short time. The presence of inhibitors of light-reaction II in photosynthesis such as o-phenanthroline, hydroxylamine and DCMU entirely

  17. Complementary nutrient effects of separately collected human faeces and urine on the yield and nutrient uptake of spinach (Spinacia oleracea).

    PubMed

    Kutu, Funso R; Muchaonyerwa, Pardon; Mnkeni, Pearson N S

    2011-05-01

    A glasshouse experiment was conducted to evaluate the combined use of separately collected human faeces and urine as fertilizer for spinach (Spinacia oleracea) production. Seven human faeces N : urine N combinations (1 : 7 to 7 : 1) each supplying 200 kg N ha(-1) were evaluated along with sole human faeces, sole urine, inorganic fertilizer and an unamended control. Complementary application of the two resources, human faeces and urine, increased fresh and dry matter yields only in treatments having high proportions of urine. Nitrogen uptake followed the same trend but the opposite trend occurred for P uptake indicating that urine was a better source of N whereas human faeces were the better source of P. Potassium uptake was not influenced by the two resources. The minimal improvement observed in the fertilizer value of human faeces when co-applied with urine suggested that co-application of the two resources may not give an added yield advantage when compared with sole human faeces.

  18. Biochemical and proton NMR characterization of the isolated functional beta-subunit of coupling factor one from spinach chloroplasts

    SciTech Connect

    Roux-Fromy, M.; Neumann, J.M.; Andre, F.; Berger, G.; Girault, G.; Galmiche, J.M.; Remy, R.

    1987-04-29

    Beta subunits have been dissociated from CF1 of spinach chloroplasts, purified by HPLC and characterized by two-dimensional electrophoresis and fluorescence emission. The solutions of isolated beta subunits are able to hydrolyze MgATP; this ATPase activity is an intrinsic property of the beta molecule. From proton NMR at 300 and 500 MHz, it is shown that the preparations are fully reproducible and that beta subunits remain monomeric with 75% aliphatic protons associated with rigid parts of the molecule. The other 25% give rise to separate resonances and belong to mobile side-chains and/or to flexible regions. The measurement of the transverse relaxation times T2 has permitted a detailed characterization of the molecular dynamics of the isolated beta subunits.

  19. Biophysical control of leaf temperature

    NASA Astrophysics Data System (ADS)

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

    2014-12-01

    In principle sunlit leaves can maintain their temperatures within a narrower range than ambient temperatures. This is an important and long-known (but now overlooked) prediction of energy balance theory. Net radiation at leaf surface in steady state (which is reached rapidly) must be equal to the combination of sensible and latent heat exchanges with surrounding air, the former being proportional to leaf-to-air temperature difference (ΔT), the latter to the transpiration rate. We present field measurements of ΔT which confirm the existence of a 'crossover temperature' in the 25-30˚C range for species in a tropical savanna and a tropical rainforest environment. This finding is consistent with a simple representation of transpiration as a function of net radiation and temperature (Priestley-Taylor relationship) assuming an entrainment factor (ω) somewhat greater than the canonical value of 0.26. The fact that leaves in tropical forests are typically cooler than surrounding air, often already by solar noon, is consistent with a recently published comparison of MODIS day-time land-surface temperatures with air temperatures. Theory further predicts a strong dependence of leaf size (which is inversely related to leaf boundary-layer conductance, and therefore to absolute magnitude of ΔT) on moisture availability. Theoretically, leaf size should be determined by either night-time constraints (risk of frost damage to active leaves) or day-time constraints (risk of heat stress damage),with the former likely to predominate - thereby restricting the occurrence of large leaves - at high latitudes. In low latitudes, daytime maximum leaf size is predicted to increase with temperature, provided that water is plentiful. If water is restricted, however, transpiration cannot proceed at the Priestley-Taylor rate, and it quickly becomes advantageous for plants to have small leaves, which do not heat up much above the temperature of their surroundings. The difference between leaf

  20. Application of Metagenomic Sequencing to Food Safety: Detection of Shiga Toxin-Producing Escherichia coli on Fresh Bagged Spinach

    PubMed Central

    Leonard, Susan R.; Mammel, Mark K.; Lacher, David W.

    2015-01-01

    Culture-independent diagnostics reduce the reliance on traditional (and slower) culture-based methodologies. Here we capitalize on advances in next-generation sequencing (NGS) to apply this approach to food pathogen detection utilizing NGS as an analytical tool. In this study, spiking spinach with Shiga toxin-producing Escherichia coli (STEC) following an established FDA culture-based protocol was used in conjunction with shotgun metagenomic sequencing to determine the limits of detection, sensitivity, and specificity levels and to obtain information on the microbiology of the protocol. We show that an expected level of contamination (∼10 CFU/100 g) could be adequately detected (including key virulence determinants and strain-level specificity) within 8 h of enrichment at a sequencing depth of 10,000,000 reads. We also rationalize the relative benefit of static versus shaking culture conditions and the addition of selected antimicrobial agents, thereby validating the long-standing culture-based parameters behind such protocols. Moreover, the shotgun metagenomic approach was informative regarding the dynamics of microbial communities during the enrichment process, including initial surveys of the microbial loads associated with bagged spinach; the microbes found included key genera such as Pseudomonas, Pantoea, and Exiguobacterium. Collectively, our metagenomic study highlights and considers various parameters required for transitioning to such sequencing-based diagnostics for food safety and the potential to develop better enrichment processes in a high-throughput manner not previously possible. Future studies will investigate new species-specific DNA signature target regimens, rational design of medium components in concert with judicious use of additives, such as antibiotics, and alterations in the sample processing protocol to enhance detection. PMID:26386062

  1. Application of metagenomic sequencing to food safety: detection of Shiga Toxin-producing Escherichia coli on fresh bagged spinach.

    PubMed

    Leonard, Susan R; Mammel, Mark K; Lacher, David W; Elkins, Christopher A

    2015-12-01

    Culture-independent diagnostics reduce the reliance on traditional (and slower) culture-based methodologies. Here we capitalize on advances in next-generation sequencing (NGS) to apply this approach to food pathogen detection utilizing NGS as an analytical tool. In this study, spiking spinach with Shiga toxin-producing Escherichia coli (STEC) following an established FDA culture-based protocol was used in conjunction with shotgun metagenomic sequencing to determine the limits of detection, sensitivity, and specificity levels and to obtain information on the microbiology of the protocol. We show that an expected level of contamination (∼10 CFU/100 g) could be adequately detected (including key virulence determinants and strain-level specificity) within 8 h of enrichment at a sequencing depth of 10,000,000 reads. We also rationalize the relative benefit of static versus shaking culture conditions and the addition of selected antimicrobial agents, thereby validating the long-standing culture-based parameters behind such protocols. Moreover, the shotgun metagenomic approach was informative regarding the dynamics of microbial communities during the enrichment process, including initial surveys of the microbial loads associated with bagged spinach; the microbes found included key genera such as Pseudomonas, Pantoea, and Exiguobacterium. Collectively, our metagenomic study highlights and considers various parameters required for transitioning to such sequencing-based diagnostics for food safety and the potential to develop better enrichment processes in a high-throughput manner not previously possible. Future studies will investigate new species-specific DNA signature target regimens, rational design of medium components in concert with judicious use of additives, such as antibiotics, and alterations in the sample processing protocol to enhance detection. PMID:26386062

  2. Postprandial plasma carotenoid responses following consumption of strawberries, red wine, vitamin C or spinach by elderly women.

    PubMed

    Paiva, S A; Yeum, K J; Cao, G; Prior, R L; Russell, R M

    1998-12-01

    This study investigated the postprandial plasma responses of carotenoids for 24 h after feeding five specific breakfast beverages; four of which had low or no carotenoid content. In seven fasting healthy elderly female subjects a blood sample (baseline) was obtained, after which they were given a breakfast beverage, containing one of the following: 1) strawberries (240 g); 2) ascorbic acid (1250 mg); 3) spinach (294 g); 4) red wine (300 mL); and 5) control (breakfast beverage only). Blood samples were collected at 0.5, 1, 4, 7, 11, 15 and 24 h. Plasma carotenoids were measured using HPLC. No significant differences were found in the levels of the plasma carotenoids measured among the various treatments at baseline. In the spinach treatment, plasma lutein, zeaxanthin and beta-carotene levels at 7, 11, 15 and 24 h were significantly higher than those at baseline, as expected. All of the carotenoids measured in the control and vitamin C treatments, at subsequent sampling times were not significantly different from those at baseline. However, for most carotenoids, strawberry and red wine feeding resulted in significantly lower carotenoids values from baseline at 11 and 15 h. Subjects who received a diet with low levels of carotenoids, but whose postprandial plasma levels of carotenoids remain steady, might be explained by a mechanism that promotes secretion of carotenoids into the circulation. Assuming that plasma carotenoids are being used over time, we hypothesize that strawberries and red wine contain some substances that interfere with the secretion of carotenoids into the circulation. PMID:9868186

  3. Application of metagenomic sequencing to food safety: detection of Shiga Toxin-producing Escherichia coli on fresh bagged spinach.

    PubMed

    Leonard, Susan R; Mammel, Mark K; Lacher, David W; Elkins, Christopher A

    2015-12-01

    Culture-independent diagnostics reduce the reliance on traditional (and slower) culture-based methodologies. Here we capitalize on advances in next-generation sequencing (NGS) to apply this approach to food pathogen detection utilizing NGS as an analytical tool. In this study, spiking spinach with Shiga toxin-producing Escherichia coli (STEC) following an established FDA culture-based protocol was used in conjunction with shotgun metagenomic sequencing to determine the limits of detection, sensitivity, and specificity levels and to obtain information on the microbiology of the protocol. We show that an expected level of contamination (∼10 CFU/100 g) could be adequately detected (including key virulence determinants and strain-level specificity) within 8 h of enrichment at a sequencing depth of 10,000,000 reads. We also rationalize the relative benefit of static versus shaking culture conditions and the addition of selected antimicrobial agents, thereby validating the long-standing culture-based parameters behind such protocols. Moreover, the shotgun metagenomic approach was informative regarding the dynamics of microbial communities during the enrichment process, including initial surveys of the microbial loads associated with bagged spinach; the microbes found included key genera such as Pseudomonas, Pantoea, and Exiguobacterium. Collectively, our metagenomic study highlights and considers various parameters required for transitioning to such sequencing-based diagnostics for food safety and the potential to develop better enrichment processes in a high-throughput manner not previously possible. Future studies will investigate new species-specific DNA signature target regimens, rational design of medium components in concert with judicious use of additives, such as antibiotics, and alterations in the sample processing protocol to enhance detection.

  4. Behavior of Leaf Meristems and Their Modification

    PubMed Central

    Ichihashi, Yasunori; Tsukaya, Hirokazu

    2015-01-01

    A major source of diversity in flowering plant form is the extensive variability of leaf shape and size. Leaf formation is initiated by recruitment of a handful of cells flanking the shoot apical meristem (SAM) to develop into a complex three-dimensional structure. Leaf organogenesis depends on activities of several distinct meristems that are established and spatiotemporally differentiated after the initiation of leaf primordia. Here, we review recent findings in the gene regulatory networks that orchestrate leaf meristem activities in a model plant Arabidopsis thaliana. We then discuss recent key studies investigating the natural variation in leaf morphology to understand how the gene regulatory networks modulate leaf meristems to yield a substantial diversity of leaf forms during the course of evolution. PMID:26648955

  5. Spectral reflectance relationships to leaf water stress

    NASA Technical Reports Server (NTRS)

    Ripple, William J.

    1986-01-01

    Spectral reflectance data were collected from detached snapbean leaves in the laboratory with a multiband radiometer. Four experiments were designed to study the spectral response resulting from changes in leaf cover, relative water content of leaves, and leaf water potential. Spectral regions included in the analysis were red (630-690 nm), NIR (760-900 nm), and mid-IR (2.08-2.35 microns). The red and mid-IR bands showed sensitivity to changes in both leaf cover and relative water content of leaves. The NIR was only highly sensitive to changes in leaf cover. Results provided evidence that mid-IR reflectance was governed primarily by leaf moisture content, although soil reflectance was an important factor when leaf cover was less than 100 percent. High correlations between leaf water potentials and reflectance were attributed to covariances with relative water content of leaves and leaf cover.

  6. An Innovative Way to Monitor Leaf Age

    NASA Astrophysics Data System (ADS)

    Garnello, A.; Paredes, K.; Trinh, U.; Saleska, S. R.; Wu, J.

    2013-12-01

    Anthony John Garnello, Karina Paredes, Uyen Khanh Ho Trinh, Jin Wu, Scott Saleska Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ, USA Abstract: Leaf age is an important characteristic for controlling plant functional performance and is associated with the changes of leaf physical, chemical, and physiological properties. Understanding how plant physiology changes over time will allow more accurate predictions of growth patterns, and a more comprehensive understanding of vegetative life histories. There still lacks an efficient technique in monitoring leaf age, tagging leaves is still the only way to accurately monitor leaf age. The goal of this study is to develop a multi-metric, accurate technique for better monitoring of leaf age. In order to acquire true leaf age records, 10 individual plant species were selected at the University of Arizona campus, and newly flushing leaves were tagged and monitored during the Monsoon season (from early June, 2013, to mid October, 2013). Every 2 weeks, 10 to 15 leaves in relative age order were harvested from each 1-meter branch to measure multiple key leaf metrics, including leaf thickness (via micrometer), fresh and dry weight, fresh and dry area (via ImageJ software), and leaf hyperspectral reflectance (via a handheld ASD Field Pro). Other leaf traits were also derived from our measurements, such as specific leaf area (SLA), leaf density (fresh weight/leaf volume), water percentage, and shrinkage ratio (1-dry area/fresh area). The hyperspectral version of vegetation index (a ratio derived from two spectral channels) was generated for each branch sample, by randomly selecting two channels from within the spectral domain of 350 nm to 2500 nm. The preliminary result documents three types of hyperspectral vegetation index (VI) which are highly related with leaf relative age order (R2>0.9). These include the sensitive spectral domains correlated with (a) leaf pigments (~550nm) and leaf physical

  7. Analysis of Circadian Leaf Movements.

    PubMed

    Müller, Niels A; Jiménez-Gómez, José M

    2016-01-01

    The circadian clock is a molecular timekeeper that controls a wide variety of biological processes. In plants, clock outputs range from the molecular level, with rhythmic gene expression and metabolite content, to physiological processes such as stomatal conductance or leaf movements. Any of these outputs can be used as markers to monitor the state of the circadian clock. In the model plant Arabidopsis thaliana, much of the current knowledge about the clock has been gained from time course experiments profiling expression of endogenous genes or reporter constructs regulated by the circadian clock. Since these methods require labor-intensive sample preparation or transformation, monitoring leaf movements is an interesting alternative, especially in non-model species and for natural variation studies. Technological improvements both in digital photography and image analysis allow cheap and easy monitoring of circadian leaf movements. In this chapter we present a protocol that uses an autonomous point and shoot camera and free software to monitor circadian leaf movements in tomato. PMID:26867616

  8. Studies on the movements of ionic selectivity, compatible solutes, and intracellular ions caused in the leaves of spinach (Spinacia oleracea L.) plants cultured in a nutrient solution with seawater.

    PubMed

    Sun, Jin; Jia, Yongxia; Guo, Shirong; Chen, Lifang

    2010-01-01

    Analyses of ionic selectivity, compatible solutes, and intracellular ions in the leaves of spinach (Spinacia oleracea L.) plants cultured in the Hoagland's nutrient solution with or.without seawater (40%) were carried out using two cultivars--the Helan No.3 (seawater tolerant) and the Yuanye (seawater sensitive). When both cultivars were subjected to seawater stress, the leaves of the Helan No. 3 spinach preferred potassium (K+), calcium (Ca2+), magnesium (Mg2+), and sulfate (SO4(2-)) over sodium (Na+) and chlorine (Cl-) to keep high ratios of K/Na, Mg/Na, Ca/Na, and SO4(2-)/Cl- compared with the Yanye spinach. Moreover, those of the Helan No. 3 spinach under the seawater stress showed high efficiency of accumulation of compatible solutes (sugars and proline), low degradation of proteins, and suppression of free amino acids. However, the activities of plasma membrane H+ -ATPase and tonoplast H+ -ATPase in the leaves of spinach with the stress were enhanced. Taken together, the Helan No. 3 spinach under the seawater stress seems to acquire a high tolerance to the seawater salinity by inducing a high ion uptake, low concentration of Na+ and Cl-, efficient accumulation of compatible solutes, low decomposition of proteins, and suppression of free amino acids in the leaves.

  9. LEAF: A Microcomputer Program for Constructing the Tukey Stem and Leaf Graph.

    ERIC Educational Resources Information Center

    Pascale, Pietro J.; Smith, Joseph

    1986-01-01

    This paper presents a BASIC microcomputer program that constructs the Tukey (1977) stem and leaf graph. Options within the LEAF program include a modified stem and leaf where the stem is split and a parallel stem and leaf graph where two separate sets of data are displayed from a common stem. (Author)

  10. 7 CFR 29.2529 - Leaf scrap.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf scrap. 29.2529 Section 29.2529 Agriculture...-Cured Tobacco (u.s. Types 22, 23, and Foreign Type 96) § 29.2529 Leaf scrap. A byproduct of unstemmed tobacco. Leaf scrap results from handling unstemmed tobacco and consists of loose and tangled whole...

  11. 7 CFR 29.3034 - Leaf scrap.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf scrap. 29.3034 Section 29.3034 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... Leaf scrap. A by-product of unstemmed tobacco. Leaf scrap results from handling unstemmed tobacco...

  12. 7 CFR 29.3526 - Leaf scrap.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf scrap. 29.3526 Section 29.3526 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... Type 95) § 29.3526 Leaf scrap. A byproduct of unstemmed tobacco Leaf scrap results from...

  13. 7 CFR 29.6022 - Leaf scrap.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf scrap. 29.6022 Section 29.6022 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... INSPECTION Standards Definitions § 29.6022 Leaf scrap. A byproduct of unstemmed tobacco Leaf scrap...

  14. 7 CFR 29.3035 - Leaf structure.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Leaf structure. The cell development of a leaf as indicated by its porosity or solidity. (See Elements... 7 Agriculture 2 2011-01-01 2011-01-01 false Leaf structure. 29.3035 Section 29.3035 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  15. 7 CFR 29.3527 - Leaf structure.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Type 95) § 29.3527 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf structure. 29.3527 Section 29.3527 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  16. 7 CFR 29.3035 - Leaf structure.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Leaf structure. The cell development of a leaf as indicated by its porosity or solidity. (See Elements... 7 Agriculture 2 2013-01-01 2013-01-01 false Leaf structure. 29.3035 Section 29.3035 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  17. 7 CFR 29.1030 - Leaf structure.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Type 92) § 29.1030 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2014-01-01 2014-01-01 false Leaf structure. 29.1030 Section 29.1030 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  18. 7 CFR 29.6023 - Leaf structure.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... INSPECTION Standards Definitions § 29.6023 Leaf structure. The cell development of a leaf as indicated by its... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf structure. 29.6023 Section 29.6023 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  19. 7 CFR 29.3527 - Leaf structure.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Type 95) § 29.3527 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2012-01-01 2012-01-01 false Leaf structure. 29.3527 Section 29.3527 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  20. 7 CFR 29.1030 - Leaf structure.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Type 92) § 29.1030 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf structure. 29.1030 Section 29.1030 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  1. 7 CFR 29.3527 - Leaf structure.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Type 95) § 29.3527 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2011-01-01 2011-01-01 false Leaf structure. 29.3527 Section 29.3527 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  2. 7 CFR 29.6023 - Leaf structure.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... INSPECTION Standards Definitions § 29.6023 Leaf structure. The cell development of a leaf as indicated by its... 7 Agriculture 2 2013-01-01 2013-01-01 false Leaf structure. 29.6023 Section 29.6023 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  3. 7 CFR 29.6023 - Leaf structure.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... INSPECTION Standards Definitions § 29.6023 Leaf structure. The cell development of a leaf as indicated by its... 7 Agriculture 2 2014-01-01 2014-01-01 false Leaf structure. 29.6023 Section 29.6023 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  4. 7 CFR 29.1030 - Leaf structure.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Type 92) § 29.1030 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2011-01-01 2011-01-01 false Leaf structure. 29.1030 Section 29.1030 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  5. 7 CFR 29.6023 - Leaf structure.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... INSPECTION Standards Definitions § 29.6023 Leaf structure. The cell development of a leaf as indicated by its... 7 Agriculture 2 2012-01-01 2012-01-01 false Leaf structure. 29.6023 Section 29.6023 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  6. 7 CFR 29.1030 - Leaf structure.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Type 92) § 29.1030 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2012-01-01 2012-01-01 false Leaf structure. 29.1030 Section 29.1030 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  7. 7 CFR 29.3035 - Leaf structure.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Leaf structure. The cell development of a leaf as indicated by its porosity or solidity. (See Elements... 7 Agriculture 2 2012-01-01 2012-01-01 false Leaf structure. 29.3035 Section 29.3035 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  8. 7 CFR 29.3527 - Leaf structure.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Type 95) § 29.3527 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2014-01-01 2014-01-01 false Leaf structure. 29.3527 Section 29.3527 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  9. 7 CFR 29.3035 - Leaf structure.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Leaf structure. The cell development of a leaf as indicated by its porosity or solidity. (See Elements... 7 Agriculture 2 2014-01-01 2014-01-01 false Leaf structure. 29.3035 Section 29.3035 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  10. 7 CFR 29.3035 - Leaf structure.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Leaf structure. The cell development of a leaf as indicated by its porosity or solidity. (See Elements... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf structure. 29.3035 Section 29.3035 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  11. 7 CFR 29.3527 - Leaf structure.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Type 95) § 29.3527 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2013-01-01 2013-01-01 false Leaf structure. 29.3527 Section 29.3527 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  12. 7 CFR 29.6023 - Leaf structure.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... INSPECTION Standards Definitions § 29.6023 Leaf structure. The cell development of a leaf as indicated by its... 7 Agriculture 2 2011-01-01 2011-01-01 false Leaf structure. 29.6023 Section 29.6023 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  13. 7 CFR 29.1030 - Leaf structure.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Type 92) § 29.1030 Leaf structure. The cell development of a leaf as indicated by its porosity. (See... 7 Agriculture 2 2013-01-01 2013-01-01 false Leaf structure. 29.1030 Section 29.1030 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections,...

  14. Influence of different planting seasons of six leaf vegetables on residues of five pesticides.

    PubMed

    Fan, Sufang; Deng, Kailin; Yu, Chuanshan; Zhao, Pengyue; Bai, Aijuan; Li, Yanjie; Pan, Canping; Li, Xuesheng

    2013-09-25

    To investigate the influence of different planting seasons on the dissipation of pesticides, field experiments of thiophanate-methyl, metalaxyl, fluazifop-P-butyl, chlorpyrifos, and λ-cyhalothrin on six crops including pakchoi, rape, crown daisy, amaranth, spinach, and lettuce were designed and conducted. In this study, a high-performance liquid chromatography and electrospray ionization-tandem mass spectrometer with multiple reaction monitoring was used to simultaneously determine thiophanate-methyl and its metabolite carbendazim, metalaxyl, and fluazifop-P-butyl in various samples; gas chromatography with an electron capture detector was used to detect chlorpyrifos and λ-cyhalothrin. The limits of quantitation (LOQs) of these six pesticides were in the range of 0.001-0.01 mg kg(-1) for all samples, and the average recoveries of all pesticides ranged from 60.1 to 119.1% at 0.01 and 0.1 mg kg(-1) spiked levels. The relative standard deviation (RSD) ranged from 1.1 to 13.9%. All maximal concentrations of the six pesticides in six leaf vegetables in autumn were higher than in summer in Beijing. For most pesticides half-lives in autumn were longer than in summer. The results showed that the initial concentration, maximal concentration, and half-lives of pesticides were influenced not only by environmental factors such as light, heat, moisture, and rainy climate but also by plant matrices. PMID:23978278

  15. Comparison of half and full-leaf shape feature extraction for leaf classification

    NASA Astrophysics Data System (ADS)

    Sainin, Mohd Shamrie; Ahmad, Faudziah; Alfred, Rayner

    2016-08-01

    Shape is the main information for leaf feature that most of the current literatures in leaf identification utilize the whole leaf for feature extraction and to be used in the leaf identification process. In this paper, study of half-leaf features extraction for leaf identification is carried out and the results are compared with the results obtained from the leaf identification based on a full-leaf features extraction. Identification and classification is based on shape features that are represented as cosines and sinus angles. Six single classifiers obtained from WEKA and seven ensemble methods are used to compare their performance accuracies over this data. The classifiers were trained using 65 leaves in order to classify 5 different species of preliminary collection of Malaysian medicinal plants. The result shows that half-leaf features extraction can be used for leaf identification without decreasing the predictive accuracy.

  16. 7 CFR 29.1162 - Leaf (B Group).

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Specifications, and Tolerances B1L—Choice Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil... percent. B2L—Fine Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil, deep color.... B3L—Good Quality Lemon Leaf Ripe, firm leaf structure, medium body, oily, strong color...

  17. 7 CFR 29.1162 - Leaf (B Group).

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Specifications, and Tolerances B1L—Choice Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil... percent. B2L—Fine Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil, deep color.... B3L—Good Quality Lemon Leaf Ripe, firm leaf structure, medium body, oily, strong color...

  18. 7 CFR 29.1162 - Leaf (B Group).

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Specifications, and Tolerances B1L—Choice Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil... percent. B2L—Fine Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil, deep color.... B3L—Good Quality Lemon Leaf Ripe, firm leaf structure, medium body, oily, strong color...

  19. 7 CFR 29.1162 - Leaf (B Group).

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Specifications, and Tolerances B1L—Choice Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil... percent. B2L—Fine Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil, deep color.... B3L—Good Quality Lemon Leaf Ripe, firm leaf structure, medium body, oily, strong color...

  20. 7 CFR 29.1162 - Leaf (B Group).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Specifications, and Tolerances B1L—Choice Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil... percent. B2L—Fine Quality Lemon Leaf Ripe, firm leaf structure, medium body, rich in oil, deep color.... B3L—Good Quality Lemon Leaf Ripe, firm leaf structure, medium body, oily, strong color...

  1. Inactivation of ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum and spinach with the new affinity label 2-bromo-1,5-dihydroxy-3-pentanone 1,5-bisphosphate

    SciTech Connect

    Donnelly, M.I.; Hartman, F.C.

    1981-11-16

    In an attempt to identify the active-site base believed to initiate catalysis by ribulosebisphosphate carboxylase, we have synthesized 2-bromo-1, 5-dihydroxy-3-pentanone 1,5-bisphosphate, a reactive analogue of a postulated intermediate of carboxylation. Although highly unstable, this compound can be shown to inactivate the carboxylases from both Rhodospirillum rubrum and spinach rapidly and irreversibly. Inactivation follows pseudo first-order kinetics, shows rate saturation and is greatly reduced by saturating amounts of the competitive inhibitor, 2-carboxyribitol 1,5-bisphosphate. The incorporation of reagent, quantified by reducing the modified carboxylases with (/sup 3/H)NaBH/sub 4/, shows that inactivation results from the modification of approximately one residue per catalytic subunit of the Rhodospirillum rubrum enzyme and less than one residue per protomeric unit of the spinach enzyme.

  2. Leaf physiognomy and climate: A multivariate analysis

    NASA Astrophysics Data System (ADS)

    Davis, J. M.; Taylor, S. E.

    1980-11-01

    Research has demonstrated that leaf physiognomy is representative of the local or microclimate conditions under which plants grow. The physiognomy of leaf samples from Oregon, Michigan, Missouri, Tennessee, and the Panama Canal Zone has been related to the microclimate using Walter diagrams and Thornthwaite water-budget data. A technique to aid paleoclimatologists in identifying the nature of the microclimate from leaf physiognomy utilizes statistical procedures to classify leaf samples into one of six microclimate regimes based on leaf physiognomy information available from fossilized samples.

  3. Nutrient Value of Leaf vs. Seed

    PubMed Central

    Edelman, Marvin; Colt, Monica

    2016-01-01

    Major differences stand out between edible leaves and seeds in protein quality, vitamin, and mineral concentrations and omega 6/omega 3 fatty acid ratios. Data for seeds (wheat, rice, corn, soy, lentil, chick pea) are compared with corresponding data for edible green leaves (kale, spinach, broccoli, duckweed). An x/y representation of data for lysine and methionine content highlights the group differences between grains, pulses, leafy vegetables, and animal foods. Leaves come out with flying colors in all these comparisons. The perspective ends with a discussion on “So why do we eat mainly seeds?” PMID:27493937

  4. Nutrient Value of Leaf vs. Seed.

    PubMed

    Edelman, Marvin; Colt, Monica

    2016-01-01

    Major differences stand out between edible leaves and seeds in protein quality, vitamin, and mineral concentrations and omega 6/omega 3 fatty acid ratios. Data for seeds (wheat, rice, corn, soy, lentil, chick pea) are compared with corresponding data for edible green leaves (kale, spinach, broccoli, duckweed). An x/y representation of data for lysine and methionine content highlights the group differences between grains, pulses, leafy vegetables, and animal foods. Leaves come out with flying colors in all these comparisons. The perspective ends with a discussion on "So why do we eat mainly seeds?" PMID:27493937

  5. Nutrient value of leaf versus seed

    NASA Astrophysics Data System (ADS)

    Edelman, Marvin; Holt, Monica

    2016-07-01

    Major differences stand out between edible leaves and seeds in protein quality, vitamin and mineral concentrations and omega 6 / omega 3 fatty acid ratios. Data for seeds (wheat, rice, corn, soy, lentil, chick pea) are compared with corresponding data for edible green leaves (kale, spinach, broccoli, duckweed). An x/y representation of data for lysine and methionine content highlights the group differences between grains, pulses, leafy vegetables and animal foods. Leaves come out with flying colors in all these comparisons. The perspective ends with a discussion on “So why do we eat mainly seeds?”

  6. Effect of repeated irrigation with water containing varying levels of total organic carbon on the persistence of Escherichia coli O157:H7 on baby spinach.

    PubMed

    Ingram, David T; Patel, Jitu; Sharma, Manan

    2011-05-01

    The California lettuce and leafy greens industry has adopted the Leafy Greens Marketing Agreement (LGMA), which allows for 126 most-probable-number (MPN) Escherichia coli per 100 ml in irrigation water. Repeat irrigation of baby spinach plants with water containing E. coli O157:H7 and different levels of total organic carbon (TOC) was used to determine the epiphytic survival of E. coli O157:H7. Three irrigation treatments (0 ppm of TOC, 12 or 15 ppm of TOC, and 120 or 150 ppm of TOC) were prepared with bovine manure containing E. coli O157:H7 at either low (0 to 1 log CFU/100 ml) or high (5 to 6 log CFU/100 ml) populations, and sprayed onto baby spinach plants in growth chambers by using a fine-mist airbrush. MPN and direct plating techniques were used to determine the E. coli O157:H7 populations on the aerial plant tissue. Plants irrigated with high E. coli O157:H7 populations, regardless of TOC levels, showed a 3-log reduction within the first 24 h. Low levels of E. coli O157:H7 were observed for up to 16 days on all TOC treatments, ranging from 76.4 MPN per plant (day 1) to 0.40 MPN per plant (day 16). No viable cells were detected on spinach tissue 24 h after irrigation with water containing fewer than 126 CFU/100 ml E. coli O157:H7. Under growth chamber conditions in this study, E. coli O157:H7 populations in irrigation water that complies with the LGMA standards will not persist for more than 24 h when applied onto foliar surfaces of spinach plants.

  7. Reduction of Escherichia coli O157:H7 in Fresh Spinach, using lactic acid bacteria and chlorine as a multihurdle intervention.

    PubMed

    Gragg, S E; Brashears, M M

    2010-02-01

    A 12-day shelf life study was conducted at 7 degrees C to determine whether Escherichia coli O157:H7 on spinach can be controlled effectively by selected strains of lactic acid bacteria (LAB) alone or in combination with chlorine as a multihurdle intervention. The multihurdle intervention consisted of both LAB and chlorine and was applied to spinach as a rinse and evaluated in comparison to LAB alone and chlorine and water rinses. Reductions achieved by all treatments also were compared with those observed for an inoculated control. The spinach was inoculated by submersion in a solution containing an E. coli O157:H7 cocktail at 1.0 x 10(6) CFU/ml. LAB were applied postharvest at a concentration of 2.0 x 10(8) CFU/ml, and 200 ppm of chlorine was used for the chlorine rinse. All spinach samples were packaged in commercial packaging, held in a retail display case, and tested for E. coli O157:H7 on days 0, 1, 3, 6, 9, and 12 using the Neo-Grid filtration system and CHROMagar. Survival of LAB throughout the shelf life also was determined. Significant reductions in pathogen populations were achieved by water (P = 0.0008), LAB (P < 0.0001), chlorine (P < 0.0001), and multihurdle (P < 0.0001) treatments when compared with controls. The multihurdle treatment produced the greatest reduction from control populations, a reduction of 1.91 log CFU/ml. This reduction was significantly greater than that achieved with water (P < 0.0001), LAB (P = 0.0025), and chlorine (P < 0.0001) alone, indicating that the application of chlorine and LAB is most effective as a combination treatment. The results obtained from this study indicate that the industry standard chlorine wash may be more effective when applied in combination with LAB.

  8. Reduction of Escherichia coli O157:H7 in Fresh Spinach, using lactic acid bacteria and chlorine as a multihurdle intervention.

    PubMed

    Gragg, S E; Brashears, M M

    2010-02-01

    A 12-day shelf life study was conducted at 7 degrees C to determine whether Escherichia coli O157:H7 on spinach can be controlled effectively by selected strains of lactic acid bacteria (LAB) alone or in combination with chlorine as a multihurdle intervention. The multihurdle intervention consisted of both LAB and chlorine and was applied to spinach as a rinse and evaluated in comparison to LAB alone and chlorine and water rinses. Reductions achieved by all treatments also were compared with those observed for an inoculated control. The spinach was inoculated by submersion in a solution containing an E. coli O157:H7 cocktail at 1.0 x 10(6) CFU/ml. LAB were applied postharvest at a concentration of 2.0 x 10(8) CFU/ml, and 200 ppm of chlorine was used for the chlorine rinse. All spinach samples were packaged in commercial packaging, held in a retail display case, and tested for E. coli O157:H7 on days 0, 1, 3, 6, 9, and 12 using the Neo-Grid filtration system and CHROMagar. Survival of LAB throughout the shelf life also was determined. Significant reductions in pathogen populations were achieved by water (P = 0.0008), LAB (P < 0.0001), chlorine (P < 0.0001), and multihurdle (P < 0.0001) treatments when compared with controls. The multihurdle treatment produced the greatest reduction from control populations, a reduction of 1.91 log CFU/ml. This reduction was significantly greater than that achieved with water (P < 0.0001), LAB (P = 0.0025), and chlorine (P < 0.0001) alone, indicating that the application of chlorine and LAB is most effective as a combination treatment. The results obtained from this study indicate that the industry standard chlorine wash may be more effective when applied in combination with LAB. PMID:20132683

  9. Glutathione reductase: Comparison of steady-state and rapid reaction primary kinetic isotope effects exhibited by the yeast, spinach, and Escherichia coli enzymes

    SciTech Connect

    Vanoni, M.A.; Wong, K.K.; Ballou, D.P.; Blanchard, J.S. )

    1990-06-19

    Kinetic parameters for NADPH and NADH have been determined at pH 8.1 for spinach, yeast, and E. coli glutathione reductases. NADPH exhibited low Km values for all enzymes (3-6 microM), while the Km values for NADH were 100 times higher (approximately 400 microM). Under our experimental conditions, the percentage of maximal velocities with NADH versus those measured with NADPH were 18.4, 3.7, and 0.13% for the spinach, yeast, and E. coli enzymes, respectively. Primary deuterium kinetic isotope effects were independent of GSSG concentration between Km and 15Km levels, supporting a ping-pong kinetic mechanism. For each of the three enzymes, NADPH yielded primary deuterium kinetic isotope effects on Vmax only, while NADH exhibited primary deuterium kinetic isotope effects on both V and V/K. The magnitude of DV/KNADH at pH 8.1 is 4.3 for the spinach enzyme, 2.7 for the yeast enzyme, and 1.6 for the E. coli glutathione reductase. The experimentally determined values of TV/KNADH of 7.4, 4.2, and 2.2 for the spinach, yeast, and E. coli glutathione reductases agree well with those calculated from the corresponding DV/KNADH using the Swain-Schaad expression. This suggests that the intrinsic primary kinetic isotope effect on NADH oxidation is fully expressed. In order to confirm this conclusion, single-turnover experiments have been performed. The measured primary deuterium kinetic isotope effects on the enzyme reduction half-reaction using NADH match those measured in the steady state for each of the three glutathione reductases.

  10. Hormonal regulation of leaf senescence in Lilium.

    PubMed

    Arrom, Laia; Munné-Bosch, Sergi

    2012-10-15

    In addition to floral senescence and longevity, the control of leaf senescence is a major factor determining the quality of several cut flowers, including Lilium, in the commercial market. To better understand the physiological process underlying leaf senescence in this species, we evaluated: (i) endogenous variation in the levels of phytohormones during leaf senescence, (ii) the effects of leaf darkening in senescence and associated changes in phytohormones, and (iii) the effects of spray applications of abscisic acid (ABA) and pyrabactin on leaf senescence. Results showed that while gibberellin 4 (GA(4)) and salicylic acid (SA) contents decreased, that of ABA increased during the progression of leaf senescence. However, dark-induced senescence increased ABA levels, but did not affect GA(4) and SA levels, which appeared to correlate more with changes in air temperature and/or photoperiod than with the induction of leaf senescence. Furthermore, spray applications of pyrabactin delayed the progression of leaf senescence in cut flowers. Thus, we conclude that (i) ABA plays a major role in the regulation of leaf senescence in Lilium, (ii) darkness promotes leaf senescence and increases ABA levels, and (iii) exogenous applications of pyrabactin inhibit leaf senescence in Lilium, therefore suggesting that it acts as an antagonist of ABA in senescing leaves of cut lily flowers. PMID:22854182

  11. Hormonal regulation of leaf senescence in Lilium.

    PubMed

    Arrom, Laia; Munné-Bosch, Sergi

    2012-10-15

    In addition to floral senescence and longevity, the control of leaf senescence is a major factor determining the quality of several cut flowers, including Lilium, in the commercial market. To better understand the physiological process underlying leaf senescence in this species, we evaluated: (i) endogenous variation in the levels of phytohormones during leaf senescence, (ii) the effects of leaf darkening in senescence and associated changes in phytohormones, and (iii) the effects of spray applications of abscisic acid (ABA) and pyrabactin on leaf senescence. Results showed that while gibberellin 4 (GA(4)) and salicylic acid (SA) contents decreased, that of ABA increased during the progression of leaf senescence. However, dark-induced senescence increased ABA levels, but did not affect GA(4) and SA levels, which appeared to correlate more with changes in air temperature and/or photoperiod than with the induction of leaf senescence. Furthermore, spray applications of pyrabactin delayed the progression of leaf senescence in cut flowers. Thus, we conclude that (i) ABA plays a major role in the regulation of leaf senescence in Lilium, (ii) darkness promotes leaf senescence and increases ABA levels, and (iii) exogenous applications of pyrabactin inhibit leaf senescence in Lilium, therefore suggesting that it acts as an antagonist of ABA in senescing leaves of cut lily flowers.

  12. Effects of high voltage nanosecond pulsed plasma and micro DBD plasma on seed germination, growth development and physiological activities in spinach.

    PubMed

    Ji, Sang-Hye; Choi, Ki-Hong; Pengkit, Anchalee; Im, Jun Sup; Kim, Ju Sung; Kim, Yong Hee; Park, Yeunsoo; Hong, Eun Jeong; Jung, Sun Kyung; Choi, Eun-Ha; Park, Gyungsoon

    2016-09-01

    In this study, we analyzed seed germination, seedling growth, and physiological aspects after treatment with high voltage nanosecond pulsed plasma and micro DBD plasma in spinach (Spinacia oleracea L.), a green leafy vegetable known to have low germination rate. Both germination and dry weight of seedlings increased after high voltage pulse shots were applied to spinach seeds. However seeds treated with many shots (10 shots) showed a decrease in germination rate and seedling growth. Seeds treated with air DBD plasma exhibited slightly higher germination and subsequent seedling growth than those treated with N2 plasma. Seed surface was degenerated after treated with high voltage pulsed plasma and micro DBD plasma but no significant difference in the degree of degeneration was observed among micro DBD plasma treatment time. Level of GA3 hormone and mRNA expression of an amylolytic enzyme-related gene in seeds were elevated 1 day after treatment with high voltage pulsed plasma. The relative amount of chlorophyll and total polyphenols in spinach seedlings grown from seeds treated with air DBD plasma was increased in 30 s, 1 min, and 3 min treatments. Taken together, our results suggest a possibility that plasma can enhance seed germination by triggering biochemical processes in seeds. PMID:26944552

  13. Antioxidant assays - consistent findings from FRAP and ORAC reveal a negative impact of organic cultivation on antioxidant potential in spinach but not watercress or rocket leaves.

    PubMed

    Payne, Adrienne C; Mazzer, Alice; Clarkson, Graham J J; Taylor, Gail

    2013-11-01

    Watercress (Rorippa nasturtium-aquaticum), wild rocket (Diplotaxis tenuifolia), and spinach (Spinacia oleracea) are commercial crops reported to have high concentrations of antioxidants, possibly contributing to disease prevention following human consumption. Following analysis of supermarket-purchased salad leaves, we report the antioxidant content potential of these species using two comparable techniques assessing the consistency between the assays - by the ferric reducing antioxidant power (FRAP) assay and the oxygen radical absorbance capacity (ORAC) assay. The leaves were harvested from both conventionally and organically managed crops, to investigate whether organic agriculture results in improved crop quality. Watercress had the highest FRAP and ability to scavenge free radicals, followed by spinach and rocket. For watercress and rocket, there was no significant effect of organic agriculture on FRAP and ORAC, but for spinach, the antioxidant potential was reduced and this was significant at the 5% level of probability for FRAP but not ORAC, although the trend was clear in both tests. We conclude that there is variation in salad crop antioxidant potential and that FRAP and ORAC are useful techniques for measuring antioxidants in these salad crops with similar ranking for each salad crop studied.

  14. Effects of high voltage nanosecond pulsed plasma and micro DBD plasma on seed germination, growth development and physiological activities in spinach.

    PubMed

    Ji, Sang-Hye; Choi, Ki-Hong; Pengkit, Anchalee; Im, Jun Sup; Kim, Ju Sung; Kim, Yong Hee; Park, Yeunsoo; Hong, Eun Jeong; Jung, Sun Kyung; Choi, Eun-Ha; Park, Gyungsoon

    2016-09-01

    In this study, we analyzed seed germination, seedling growth, and physiological aspects after treatment with high voltage nanosecond pulsed plasma and micro DBD plasma in spinach (Spinacia oleracea L.), a green leafy vegetable known to have low germination rate. Both germination and dry weight of seedlings increased after high voltage pulse shots were applied to spinach seeds. However seeds treated with many shots (10 shots) showed a decrease in germination rate and seedling growth. Seeds treated with air DBD plasma exhibited slightly higher germination and subsequent seedling growth than those treated with N2 plasma. Seed surface was degenerated after treated with high voltage pulsed plasma and micro DBD plasma but no significant difference in the degree of degeneration was observed among micro DBD plasma treatment time. Level of GA3 hormone and mRNA expression of an amylolytic enzyme-related gene in seeds were elevated 1 day after treatment with high voltage pulsed plasma. The relative amount of chlorophyll and total polyphenols in spinach seedlings grown from seeds treated with air DBD plasma was increased in 30 s, 1 min, and 3 min treatments. Taken together, our results suggest a possibility that plasma can enhance seed germination by triggering biochemical processes in seeds.

  15. D-Glucosone and L-sorbosone, putative intermediates of L-ascorbic acid biosynthesis in detached bean and spinach leaves. [Phaseolus vulgaris L. ; Spinacia oleracea L

    SciTech Connect

    Saito, Kazumi; Nick, J.A.; Loewus, F.A. )

    1990-11-01

    D-(6-{sup 14}C)Glucosone that had been prepared enzymically from D-(6-{sup 14}C)glucose was used to compare relative efficiencies of these two sugars for L-ascorbic acid (AA) biosynthesis in detached bean (Phaseolus vulgaris L., cv California small white) apices and 4-week-old spinach (Spinacia oleracea L., cv Giant Noble) leaves. At tracer concentration, {sup 14}C from glucosone was utilized by spinach leaves for AA biosynthesis much more effectively than glucose. Carbon-14 from (6-{sup 14}C)glucose underwent considerable redistribution during AA formation, whereas {sup 14}C from (6-{sup 14}C)glucosone remained almost totally in carbon 6 of AA. In other experiments with spinach leaves, L-(U-{sup 14}C)sorbosone was found to be equivalent to (6-{sup 14}C)glucose as a source of {sup 14}C for AA. In the presence of 0.1% D-glucosone, conversion of (6-{sup 14}C) glucose into labeled AA was greatly repressed. In a comparable experiment with L-sorbosone replacing D-glucosone, the effect was much less. The experiments described here give substance to the proposal that D-glucosone and L-sorbosone are putative intermediates in the conversion of D-glucose to AA in higher plants.

  16. The relationship of leaf photosynthetic traits – Vcmax and Jmax – to leaf nitrogen, leaf phosphorus, and specific leaf area: a meta-analysis and modeling study

    PubMed Central

    Walker, Anthony P; Beckerman, Andrew P; Gu, Lianhong; Kattge, Jens; Cernusak, Lucas A; Domingues, Tomas F; Scales, Joanna C; Wohlfahrt, Georg; Wullschleger, Stan D; Woodward, F Ian

    2014-01-01

    Great uncertainty exists in the global exchange of carbon between the atmosphere and the terrestrial biosphere. An important source of this uncertainty lies in the dependency of photosynthesis on the maximum rate of carboxylation (Vcmax) and the maximum rate of electron transport (Jmax). Understanding and making accurate prediction of C fluxes thus requires accurate characterization of these rates and their relationship with plant nutrient status over large geographic scales. Plant nutrient status is indicated by the traits: leaf nitrogen (N), leaf phosphorus (P), and specific leaf area (SLA). Correlations between Vcmax and Jmax and leaf nitrogen (N) are typically derived from local to global scales, while correlations with leaf phosphorus (P) and specific leaf area (SLA) have typically been derived at a local scale. Thus, there is no global-scale relationship between Vcmax and Jmax and P or SLA limiting the ability of global-scale carbon flux models do not account for P or SLA. We gathered published data from 24 studies to reveal global relationships of Vcmax and Jmax with leaf N, P, and SLA. Vcmax was strongly related to leaf N, and increasing leaf P substantially increased the sensitivity of Vcmax to leaf N. Jmax was strongly related to Vcmax, and neither leaf N, P, or SLA had a substantial impact on the relationship. Although more data are needed to expand the applicability of the relationship, we show leaf P is a globally important determinant of photosynthetic rates. In a model of photosynthesis, we showed that at high leaf N (3 gm−2), increasing leaf P from 0.05 to 0.22 gm−2 nearly doubled assimilation rates. Finally, we show that plants may employ a conservative strategy of Jmax to Vcmax coordination that restricts photoinhibition when carboxylation is limiting at the expense of maximizing photosynthetic rates when light is limiting. PMID:25473475

  17. The relationship of leaf photosynthetic traits - V cmax and J max - to leaf nitrogen, leaf phosphorus, and specific leaf area: a meta-analysis and modeling study.

    PubMed

    Walker, Anthony P; Beckerman, Andrew P; Gu, Lianhong; Kattge, Jens; Cernusak, Lucas A; Domingues, Tomas F; Scales, Joanna C; Wohlfahrt, Georg; Wullschleger, Stan D; Woodward, F Ian

    2014-08-01

    Great uncertainty exists in the global exchange of carbon between the atmosphere and the terrestrial biosphere. An important source of this uncertainty lies in the dependency of photosynthesis on the maximum rate of carboxylation (V cmax) and the maximum rate of electron transport (J max). Understanding and making accurate prediction of C fluxes thus requires accurate characterization of these rates and their relationship with plant nutrient status over large geographic scales. Plant nutrient status is indicated by the traits: leaf nitrogen (N), leaf phosphorus (P), and specific leaf area (SLA). Correlations between V cmax and J max and leaf nitrogen (N) are typically derived from local to global scales, while correlations with leaf phosphorus (P) and specific leaf area (SLA) have typically been derived at a local scale. Thus, there is no global-scale relationship between V cmax and J max and P or SLA limiting the ability of global-scale carbon flux models do not account for P or SLA. We gathered published data from 24 studies to reveal global relationships of V cmax and J max with leaf N, P, and SLA. V cmax was strongly related to leaf N, and increasing leaf P substantially increased the sensitivity of V cmax to leaf N. J max was strongly related to V cmax, and neither leaf N, P, or SLA had a substantial impact on the relationship. Although more data are needed to expand the applicability of the relationship, we show leaf P is a globally important determinant of photosynthetic rates. In a model of photosynthesis, we showed that at high leaf N (3 gm(-2)), increasing leaf P from 0.05 to 0.22 gm(-2) nearly doubled assimilation rates. Finally, we show that plants may employ a conservative strategy of J max to V cmax coordination that restricts photoinhibition when carboxylation is limiting at the expense of maximizing photosynthetic rates when light is limiting.

  18. Leaf Senescence by Magnesium Deficiency

    PubMed Central

    Tanoi, Keitaro; Kobayashi, Natsuko I.

    2015-01-01

    Magnesium ions (Mg2+) are the second most abundant cations in living plant cells, and they are involved in various functions, including photosynthesis, enzyme catalysis, and nucleic acid synthesis. Low availability of Mg2+ in an agricultural field leads to a decrease in yield, which follows the appearance of Mg-deficient symptoms such as chlorosis, necrotic spots on the leaves, and droop. During the last decade, a variety of physiological and molecular responses to Mg2+ deficiency that potentially link to leaf senescence have been recognized, allowing us to reconsider the mechanisms of Mg2+ deficiency. This review focuses on the current knowledge about the physiological responses to Mg2+ deficiency including a decline in transpiration, accumulation of sugars and starch in source leaves, change in redox states, increased oxidative stress, metabolite alterations, and a decline in photosynthetic activity. In addition, we refer to the molecular responses that are thought to be related to leaf senescence. With these current data, we give an overview of leaf senescence induced by Mg deficiency. PMID:27135350

  19. Yeasts colonizing the leaf surfaces.

    PubMed

    Sláviková, Elena; Vadkertiová, Renata; Vránová, Dana

    2007-08-01

    The yeasts were isolated from the leaf surfaces of ten species of trees. The study site was a forest park (Zelezná Studnicka) of the Small Carpathians mountain range. One hundred and thirty seven yeast strains belonging to 13 genera were isolated from 320 samples of leaves and needles. Seventeen yeast species were isolated, but only seven occurred regularly: Aureobasidium pullulans, Cryptococcus laurentii, Pichia anomala, Metschnikowia pulcherrima, Saccharomyces sp., Lachancea thermotolerans, and Rhodotorula glutinis. The remaining species were isolated from the leaves and needles of three or less tree species. A. pullulans, Cr. laurentii, and P. anomala were the most frequently found species and they occurred on leaves and needles of all ten tree species. Saccharomyces sp. occurred in leaf samples collected from eight kinds of trees. M. pulcherrima and L. thermotolerans were found in samples collected from six species of trees. Both these species occurred almost always on the leaves of deciduous trees. Rh. glutinis was the most frequently isolated carotenoids producing species. We have found out that the ascomycetous and basidiomycetous species were present in the leaf samples in approximately equal frequency, contrary to the soil samples taken from this forest park, where the ascomycetous species were found rarely.

  20. The Influence of Leaf Angle and Leaf Surface Characteristics on the Process of Rainfall Interception

    NASA Astrophysics Data System (ADS)

    Holder, C.; Ginebra, R.; Webb, R.

    2015-12-01

    Individual choice in plant selection for household landscaping influences differences in runoff from urban watersheds because the variation in plant canopy architecture results in rainfall interception differences. Understanding the variables that influence rainfall interception and understanding the mechanism of rainfall interception are important concepts for sustainable watershed management. The broad objective of this study was to explore the influence of leaf hydrophobicity, water droplet retention, and leaf angle on the mechanism and process of rainfall interception and raindrop impaction on leaf surfaces of common tree species from the semi-arid regions of the western United States. Leaf hydrophobicity is determined by the cohesive forces of the water molecules among themselves and the adhesive forces that result from the molecular interactions between the water droplet and the leaf surface. Water droplet retention is a measure of how easily a water droplet drains off a leaf surface. The specific hypotheses examined were 1) larger raindrops falling on leaf surfaces will deflect the leaf to an angle greater than the water droplet retention angle; 2) an increased leaf angle, whether from natural position or deflection due to droplet impact and retention, reduces interception from raindrop impaction on hydrophobic and hydrophilic leaf surfaces; and 3) increased droplet size and frequency decrease rainfall interception more significantly in the hydrophilic case. These hypotheses were addressed in a laboratory experiment by 1) measuring leaf hydrophobicity and water droplet retention using a goniometer with a tilting base; 2) measuring leaf traits such as leaf area, leaf surface roughness, trichome density, and specific storage capacity; 3) examining raindrop splash on leaf surfaces with varying leaf hydrophobicity, water droplet retention, and leaf angle with a raindrop generator and high-speed video camera; and 4) modeling the impact of raindrop splash on leaf

  1. LeafJ: an ImageJ plugin for semi-automated leaf shape measurement.

    PubMed

    Maloof, Julin N; Nozue, Kazunari; Mumbach, Maxwell R; Palmer, Christine M

    2013-01-21

    High throughput phenotyping (phenomics) is a powerful tool for linking genes to their functions (see review and recent examples). Leaves are the primary photosynthetic organ, and their size and shape vary developmentally and environmentally within a plant. For these reasons studies on leaf morphology require measurement of multiple parameters from numerous leaves, which is best done by semi-automated phenomics tools. Canopy shade is an important environmental cue that affects plant architecture and life history; the suite of responses is collectively called the shade avoidance syndrome (SAS). Among SAS responses, shade induced leaf petiole elongation and changes in blade area are particularly useful as indices. To date, leaf shape programs (e.g. SHAPE, LAMINA, LeafAnalyzer, LEAFPROCESSOR) can measure leaf outlines and categorize leaf shapes, but can not output petiole length. Lack of large-scale measurement systems of leaf petioles has inhibited phenomics approaches to SAS research. In this paper, we describe a newly developed ImageJ plugin, called LeafJ, which can rapidly measure petiole length and leaf blade parameters of the model plant Arabidopsis thaliana. For the occasional leaf that required manual correction of the petiole/leaf blade boundary we used a touch-screen tablet. Further, leaf cell shape and leaf cell numbers are important determinants of leaf size. Separate from LeafJ we also present a protocol for using a touch-screen tablet for measuring cell shape, area, and size. Our leaf trait measurement system is not limited to shade-avoidance research and will accelerate leaf phenotyping of many mutants and screening plants by leaf phenotyping.

  2. Leaf drop affects herbivory in oaks.

    PubMed

    Pearse, Ian S; Karban, Richard

    2013-11-01

    Leaf phenology is important to herbivores, but the timing and extent of leaf drop has not played an important role in our understanding of herbivore interactions with deciduous plants. Using phylogenetic general least squares regression, we compared the phenology of leaves of 55 oak species in a common garden with the abundance of leaf miners on those trees. Mine abundance was highest on trees with an intermediate leaf retention index, i.e. trees that lost most, but not all, of their leaves for 2-3 months. The leaves of more evergreen species were more heavily sclerotized, and sclerotized leaves accumulated fewer mines in the summer. Leaves of more deciduous species also accumulated fewer mines in the summer, and this was consistent with the idea that trees reduce overwintering herbivores by shedding leaves. Trees with a later leaf set and slower leaf maturation accumulated fewer herbivores. We propose that both leaf drop and early leaf phenology strongly affect herbivore abundance and select for differences in plant defense. Leaf drop may allow trees to dispose of their herbivores so that the herbivores must recolonize in spring, but trees with the longest leaf retention also have the greatest direct defenses against herbivores.

  3. Leaf drop affects herbivory in oaks.

    PubMed

    Pearse, Ian S; Karban, Richard

    2013-11-01

    Leaf phenology is important to herbivores, but the timing and extent of leaf drop has not played an important role in our understanding of herbivore interactions with deciduous plants. Using phylogenetic general least squares regression, we compared the phenology of leaves of 55 oak species in a common garden with the abundance of leaf miners on those trees. Mine abundance was highest on trees with an intermediate leaf retention index, i.e. trees that lost most, but not all, of their leaves for 2-3 months. The leaves of more evergreen species were more heavily sclerotized, and sclerotized leaves accumulated fewer mines in the summer. Leaves of more deciduous species also accumulated fewer mines in the summer, and this was consistent with the idea that trees reduce overwintering herbivores by shedding leaves. Trees with a later leaf set and slower leaf maturation accumulated fewer herbivores. We propose that both leaf drop and early leaf phenology strongly affect herbivore abundance and select for differences in plant defense. Leaf drop may allow trees to dispose of their herbivores so that the herbivores must recolonize in spring, but trees with the longest leaf retention also have the greatest direct defenses against herbivores. PMID:23774946

  4. Leaf P increase outpaces leaf N in an Inner Mongolia grassland over 27 years.

    PubMed

    Mi, Zhaorong; Huang, Yuanyuan; Gan, Huijie; Zhou, Wenjia; Flynn, Dan F B; He, Jin-Sheng

    2015-01-01

    The dynamics of leaf nitrogen (N) and phosphorus (P) have been intensively explored in short-term experiments, but rarely at longer timescales. Here, we investigated leaf N : P stoichiometry over a 27-year interval in an Inner Mongolia grassland by comparing leaf N : P concentration of 2006 with that of 1979. Across 80 species, both leaf N and P increased, but the increase in leaf N lagged behind that of leaf P, leading to a significant decrease in the N : P ratio. These changes in leaf N : P stoichiometry varied among functional groups. For leaf N, grasses increased, woody species tended to increase, whereas forbs showed no change. Unlike leaf N, leaf P of grasses and forbs increased, whereas woody species showed no change. Such changes may reflect N deposition and P release induced by soil acidification over the past decades. The interannual effect of precipitation may somewhat have reduced the soil available N, leading to the more modest increase of leaf N than of leaf P. Thus, leaf N : P stoichiometry significantly responded to long-term environmental changes in this temperate steppe, but different functional groups responded differently. Our results indicate that conclusions of plant stoichiometry under short-term N fertilization should be treated with caution when extrapolating to longer timescales.

  5. Erythromycin and 5S rRNA binding properties of the spinach chloroplast ribosomal protein CL22.

    PubMed Central

    Carol, P; Rozier, C; Lazaro, E; Ballesta, J P; Mache, R

    1993-01-01

    The spinach chloroplast ribosomal protein (r-protein) CL22 contains a central region homologous to the Escherichia coli r-protein L22 plus long N- and C-terminal extensions. We show in this study that the CL22 combines two properties which in E. coli ribosome are split between two separate proteins. The CL22 which binds to the 5S rRNA can also be linked to an erythromycin derivative added to the 50S ribosomal subunit. This latter property is similar to that of the E. coli L22 and suggests a similar localization in the 50S subunit. We have overproduced the r-protein CL22 and deleted forms of this protein in E. coli. We show that the overproduced CL22 binds to the chloroplast 5S rRNA and that the deleted protein containing the N- and C-terminal extensions only has lost the 5S rRNA binding property. We suggest that the central homologous regions of the CL22 contains the RNA binding domain. Images PMID:8441674

  6. Comparison of the levels of six endogenous gibberellins in roots and shoots of spinach in relation to photoperiod

    SciTech Connect

    Metzger, J.D.; Zeevaart, J.A.D.

    1980-10-01

    This communication describes the distribution of gibberellins (GAs) in roots and shoots of spinach in relation to photoperiod. From previous work shoots were known to contain GA/sub 53/, GA/sub 44/, GA/sub 19/, GA/sub 17/, GA/sub 20/, and GA/sub 29/. We now show by combined gas chromatography-mass spectrometry that roots contain gas chromatography-selected ion current monitoring. Neither GA/sub 17/ nor GA/sub 20/ were detected in root extracts. Analysis by the d-5 corn bioassay also showed no effect of photoperiodic treatment on the levels of GA-like substances in root extracts. Both phloem and xylem exudates had patterns of GA-like activity similar to those found in shoots and roots, respectively. Moreover, foliar application of (/sup 3/H)GA/sub 20/ resulted in the transport of label from the shoot to the roots. Over half of the label in the roots represented unmetabolized (/sup 3/H)GA/sub 20/, indicating that part of the GA/sub 20/ in the phloem is transported to the roots. Consequently, if GA/sub 20/ is made in, or transported to the roots, it is rapidly metabolized in that organ. This is a clear indication that regulation of GA metabolism is greatly different in roots and shoots.

  7. The Use of Contact Mode Atomic Force Microscopy in Aqueous Medium for Structural Analysis of Spinach Photosynthetic Complexes1[OPEN

    PubMed Central

    Phuthong, Witchukorn; Huang, Zubin; Wittkopp, Tyler M.; Sznee, Kinga; Heinnickel, Mark L.; Dekker, Jan P.; Frese, Raoul N.; Prinz, Fritz B.; Grossman, Arthur R.

    2015-01-01

    To investigate the dynamics of photosynthetic pigment-protein complexes in vascular plants at high resolution in an aqueous environment, membrane-protruding oxygen-evolving complexes (OECs) associated with photosystem II (PSII) on spinach (Spinacia oleracea) grana membranes were examined using contact mode atomic force microscopy. This study represents, to our knowledge, the first use of atomic force microscopy to distinguish the putative large extrinsic loop of Photosystem II CP47 reaction center protein (CP47) from the putative oxygen-evolving enhancer proteins 1, 2, and 3 (PsbO, PsbP, and PsbQ) and large extrinsic loop of Photosystem II CP43 reaction center protein (CP43) in the PSII-OEC extrinsic domains of grana membranes under conditions resulting in the disordered arrangement of PSII-OEC particles. Moreover, we observed uncharacterized membrane particles that, based on their physical characteristics and electrophoretic analysis of the polypeptides associated with the grana samples, are hypothesized to be a domain of photosystem I that protrudes from the stromal face of single thylakoid bilayers. Our results are interpreted in the context of the results of others that were obtained using cryo-electron microscopy (and single particle analysis), negative staining and freeze-fracture electron microscopy, as well as previous atomic force microscopy studies. PMID:26220954

  8. Structure and expression of the nuclear gene coding for the plastid CS1 ribosomal protein from spinach.

    PubMed Central

    Franzetti, B; Zhou, D X; Mache, R

    1992-01-01

    The chloroplast ribosomal protein CS1 is an essential component of the plastids translational machinery involved in translation initiation. Southern analysis suggests that the corresponding nuclear gene is present in one copy in the spinach genome. We have isolated and sequenced the gene (rps1) to study its expression at the transcriptional level. The gene consists of 7 exons and 6 introns including an unusually large intron in the 5' coding region. No canonical TATA-box is found in the 5' upstream region of the gene. rps1 transcripts are detected early during germination and a significant accumulation is observed after the protrusion of the radicle. CS1 mRNAs are present in all organs of young seedlings although there are dramatic differences in the steady state level of the mRNAs between leaves and roots tissues. Transcripts accumulate independently of the presence or absence of light. Band shift analysis shows that the +1, -400 bp region of the gene can bind different sets of proteins isolated from roots and leaves nuclei. We suggest that the expression of the housekeeping plastid-related rps1 gene is regulated in a tissue-specific manner by transcriptional trans-acting factors. Images PMID:1508710

  9. Basella alba rubra spinach pigment-sensitized TiO2 thin film-based solar cells

    NASA Astrophysics Data System (ADS)

    Gokilamani, N.; Muthukumarasamy, N.; Thambidurai, M.; Ranjitha, A.; Velauthapillai, Dhayalan

    2015-03-01

    Nanocrystalline TiO2 thin films have been prepared by sol-gel dip coating method. The X-ray diffraction results showed that TiO2 thin films annealed at 400, 450 and 500 °C are of anatase phase and the peak corresponding to the (101) plane is present in all the samples. The grain size of TiO2 thin films was found to increase with increasing annealing temperature. The grain size is found to be 20, 25 and 33 nm for the films annealed at 400, 450 and 500 °C. The structure of the TiO2 nanocrystalline thin films have been examined by high-resolution transmission electron microscope, Raman spectroscopy and FTIR spectroscopy. TiO2 thin films were sensitized by natural dyes extracted from basella alba rubra spinach. It was found that the absorption peak of basella alba rubra extract is at about 665 nm. The dye-sensitized TiO2-based solar cell sensitized using basella alba rubra exhibited a J sc of 4.35 mA cm-2, V oc of 0.48 V, FF of 0.35 and efficiency of 0.70 %. Natural dyes as sensitizers for dye-sensitized solar cells are promising because of their environmental friendliness, low-cost production and fully biodegradable.

  10. High and low potential forms of the QA quinone electron acceptor in Photosystem II of Thermosynechococcus elongatus and spinach.

    PubMed

    Ido, Kunio; Gross, Christine M; Guerrero, Fernando; Sedoud, Arezki; Lai, Thanh-Lan; Ifuku, Kentaro; Rutherford, A William; Krieger-Liszkay, Anja

    2011-01-01

    The redox potential of Q(A) in Photosystem II (PSII) from Thermosynechococcus elongatus was titrated monitoring chlorophyll fluorescence. A high potential form (E(m)=+60 ± 25 mV) was found in the absence of Mn(4)Ca, the active site for water oxidation. The low potential form (E(m)=-60 ± 48 mV), which is difficult to measure in conventional titration experiments, could be "locked in" by cross-linking the active enzyme. This indicates that the presence of Mn(4)Ca is relayed to the quinone site by significant structural changes in the protein. The presence of high and low potential forms agrees with what has been seen in plants, algae from our lab and in T. elongatus (Shibamoto et al., Biochemistry 48 (2009) 10682-10684). In the latter work, the potentials of Q(A) were shifted to lower potentials compared to other measurements. The redox potential of Q(A) in Mn-depleted PSII from spinach was titrated in the presence of redox mediators and the midpoint potential was shifted by 80 mV towards a more negative value compared to titrations without mediators. The lower values of the midpoint potential of the (Q(A)/Q(A)(-)) redox couple in the literature could be due to a perturbation due to a specific mediator. PMID:21393014

  11. Accumulation of a translation intermediate of D1 protein by light-dark transition in isolated spinach chloroplasts.

    PubMed

    Inagaki, N; Satoh, K

    1992-03-23

    In an in vitro translation experiment using spinach chloroplasts, a novel protein band of about 17.5 kDa appeared by light to dark transition. The protein never accumulated in detectable amounts either in continuous illumination or in continuous darkness. The 17.5 kDa protein accumulated upon light-dark transition, on the other hand, disappeared by the subsequent illumination. Accumulation of the protein in light, however, was observed when stromal level of ATP in chloroplasts was lowered after preillumination by the addition of various chemical compounds which, irrespective of the mode of action, eventually decrease the ATP level, e.g. atrazine, carbonyl-cyanide-m-chlorophenyl hydrazone and glycerate. The dark-accumulated protein was concluded to be a translation intermediate of D1 protein based on the facts that this component precipitates with specific antibodies and is resistant to lysylendopeptidase treatment. The suppression by chloramphenicol of both appearance upon light-dark transition and disappearance by the subsequent illumination of the protein also supported this conclusion. The phenomenon was discussed in terms of pausing in the translation of psbA mRNA.

  12. Low Temperature Electronic Absorption Spectra of Oxidized and Reduced Spinach Ferredoxins. Evidence for Nonequivalent Iron(III) Sites.

    PubMed

    Rawlings, J; Siiman, O; Gray, H B

    1974-01-01

    The electronic absorption spectra of oxidized and reduced spinach ferredoxins have been measured between 1200 and 600 nm at low temperature in D(2)O/ethylene glycol glasses. Relatively weak absorption bands are observed at 720, 820, and 920 nm in oxidized ferredoxin, and at 652, 820, and 920 nm in reduced ferredoxin. The spectral results show that the two Fe(III) centers in oxidized ferredoxin are not equivalent, and that the 820- and 920-nm bands are associated with the nonreducible site. Assignment of the reducible site as tetrahedral Fe(III) is indicated. The 720-nm (13.9 kcm(-1)) band in oxidized ferredoxin is attributed to an intensity-enhanced (6)A(1) --> (4)T(1)d-d transition, whereas the 652-nm (15.3 kcm(-1)) feature of reduced ferredoxin could be due either to (5)E --> (3)T(1) in tetrahedral Fe(II)S(4) or an Fe(II) --> Fe(III) intervalence excitation.

  13. Glyoxylate and Glutamate Effects on Photosynthetic Carbon Metabolism in Isolated Chloroplasts and Mesophyll Cells of Spinach 1

    PubMed Central

    Lawyer, Arthur L.; Cornwell, Karen L.; Gee, Sherry L.; Bassham, James A.

    1983-01-01

    Addition of millimolar sodium glyoxylate to spinach (Spinacia oleracea) chloroplasts was inhibitory to photosynthetic incorporation of 14CO2 under conditions of both low (0.2 millimolar or air levels) and high (9 millimolar) CO2 concentrations. Incorporation of 14C into most metabolites decreased. Labeling of 6-P-gluconate and fructose-1,6-bis-P increased. This suggested that glyoxylate inhibited photosynthetic carbon metabolism indirectly by decreasing the reducing potential of chloroplasts through reduction of glyoxylate to glycolate. This hypothesis was supported by measuring the reduction of [14C]glyoxylate by chloroplasts. Incubation of isolated mesophyll cells with glyoxylate had no effect on net photosynthetic CO2 uptake, but increased labeling was observed in 6-P-gluconate, a key indicator of decreased reducing potential. The possibility that glyoxylate was affecting photosynthetic metabolism by decreasing chloroplast pH cannot be excluded. Increased 14C-labeling of ribulose-1,5-bis-P and decreased 3-P-glyceric acid and glycolate labeling upon addition of glyoxylate to chloroplasts suggested that ribulose-bis-P carboxylase and oxygenase might be inhibited either indirectly or directly by glyoxylate. Glyoxylate addition decreased 14CO2 labeling into glycolate and glycine by isolated mesophyll cells but had no effect on net 14CO2 fixation. Glutamate had little effect on net photosynthetic metabolism in chloroplast preparations but did increase 14CO2 incorporation by 15% in isolated mesophyll cells under air levels of CO2. PMID:16663018

  14. Dynamics of the antioxidant system during seed osmopriming, post-priming germination, and seedling establishment in Spinach (Spinacia oleracea).

    PubMed

    Chen, Keting; Arora, Rajeev

    2011-02-01

    Osmopriming is a pre-sowing treatment that improves seed germination performance and stress tolerance. To understand osmopriming physiology, and its association with post-priming stress tolerance, we investigated the antioxidant system dynamics during three stages: during osmopriming, post-priming germination, and seedling establishment. Spinach seeds (Spinacia oleracea L. cv. Bloomsdale) were primed with -0.6 MPa PEG at 15°C for 8 d, and dried at room temperature for 2 d. Unprimed and primed germinating seeds/seedlings were subjected to a chilling and desiccation stresses. Seed/seedling samples were collected for antioxidant assays and germination performance and stress tolerance were evaluated. Our data indicate that: (1) during osmopriming the transition of seeds from dry to germinating state represses the antioxidant pathways (residing in dry seeds) that involve CAT and SOD enzymes but stimulates another pathway (only detectable in imbibed seeds) involving APX; (2) a renewal of antioxidant system, possibly required by seedling establishment, occurs after roughly 5 d of germination; (3) osmopriming strengthens the antioxidant system and increases seed germination potential, resulting in an increased stress tolerance in germinating seeds. Osmopriming-mediated promotive effect on stress tolerance, however, may diminish in relatively older (e.g. ~5-week) seedlings.

  15. Rapid, enhanced detection of Salmonella Typhimurium on fresh spinach leaves using micron-scale, phage-coated magnetoelastic biosensors

    NASA Astrophysics Data System (ADS)

    Horikawa, Shin; Vaglenov, Kiril A.; Gerken, Dana M.; Chai, Yating; Park, Mi-Kyung; Li, Suiqiong; Petrenko, Valery A.; Chin, Bryan A.

    2012-05-01

    In order to cost-effectively and rapidly detect bacterial food contamination in the field, the potential usefulness of phage-coated magnetoelastic (ME) biosensors has been recently reported. These biosensors are freestanding, mass-sensitive biosensors that can be easily batch-fabricated, thereby reducing the fabrication cost per sensor to a fraction of a cent. In addition, the biosensors can be directly placed on fresh produce surfaces and used to rapidly monitor possible bacterial food contamination without any preceding sample preparation. Previous investigations showed that the limit of detection (LOD) with millimeter-scale ME biosensors was fairly low for fresh produce with smooth surfaces (e.g., tomatoes and shell eggs). However, the LOD is anticipated to be dependent on the size of the biosensors as well as the topography of produce surfaces of interest. This paper presents an investigation into the use of micron-scale, phage-coated ME biosensors for the enhanced detection of Salmonella Typhimurium on fresh spinach leaves.

  16. Partial purification of a spinach thylakoid protein kinase that can phosphorylate light-harvesting chlorophyll a/b proteins

    SciTech Connect

    Clark, R.D.; Hind, G.; Bennett, J.

    1985-01-01

    Protein phosphorylation in plant tissues is particularly marked in chloroplasts, protein kinase activity being associated with the outer envelope, the soluble stromal fraction, and the thylakoid membrane. Furthermore, thylakoid-bound activity probably includes several distinct kinases, as suggested by studies of divalent cation specificity and thermal lability carried out with intact thylakoids and by subfractionation of solubilized membranes. Illumination of thylakoids, particularly with red light, promotes the rapid and extensive phosphorylation of the light-harvesting chlorophyll a/b complex (LHCII) on a threonine residue near the amino terminus of the protein. This phosphorylation is thought to be involved in regulating the distribution of absorbed quanta between photosystems II and I and is modulated by the redox state of the thylakoid plastoquinone pool. Neither of the thylakoid kinases reported to date was capable of phosphorylating purified LHCII in vitro or of incorporating phosphate into threonyl residues of exogenous substrates, that some LHCII phosphorylation was catalyzed by a preliminary fraction led workers to suggest that at least one other kinase remained to be isolated. Here, the authors report the solubilization and partial purification of a protein kinase from spinach thylakoids that is capable of phosphorylating LHCII in vitro, and they show that the specific site of phosphorylation is very nearly the same as, if not identical with, the site phosphorylated in organello.

  17. The Use of Contact Mode Atomic Force Microscopy in Aqueous Medium for Structural Analysis of Spinach Photosynthetic Complexes.

    PubMed

    Phuthong, Witchukorn; Huang, Zubin; Wittkopp, Tyler M; Sznee, Kinga; Heinnickel, Mark L; Dekker, Jan P; Frese, Raoul N; Prinz, Fritz B; Grossman, Arthur R

    2015-10-01

    To investigate the dynamics of photosynthetic pigment-protein complexes in vascular plants at high resolution in an aqueous environment, membrane-protruding oxygen-evolving complexes (OECs) associated with photosystem II (PSII) on spinach (Spinacia oleracea) grana membranes were examined using contact mode atomic force microscopy. This study represents, to our knowledge, the first use of atomic force microscopy to distinguish the putative large extrinsic loop of Photosystem II CP47 reaction center protein (CP47) from the putative oxygen-evolving enhancer proteins 1, 2, and 3 (PsbO, PsbP, and PsbQ) and large extrinsic loop of Photosystem II CP43 reaction center protein (CP43) in the PSII-OEC extrinsic domains of grana membranes under conditions resulting in the disordered arrangement of PSII-OEC particles. Moreover, we observed uncharacterized membrane particles that, based on their physical characteristics and electrophoretic analysis of the polypeptides associated with the grana samples, are hypothesized to be a domain of photosystem I that protrudes from the stromal face of single thylakoid bilayers. Our results are interpreted in the context of the results of others that were obtained using cryo-electron microscopy (and single particle analysis), negative staining and freeze-fracture electron microscopy, as well as previous atomic force microscopy studies.

  18. Molecular cloning and characterization of pathogenesis-related protein family 10 gene from spinach (SoPR10).

    PubMed

    Bai, Xuegui; Long, Juan; He, Xiaozhao; Li, Shun; Xu, Huini

    2014-01-01

    PR10 genes encode small, intracellular proteins that respond to biotic and abiotic stresses. In this study, a cDNA clone (designated as SoPR10, GenBank Accession No. KC142174) encoding a PR10 protein from spinach (Spinacia oleracea L.) was isolated and characterized. SoPR10 encoded a 161-amino acid polypeptide with a predicted molecular mass of 19.76 kDa and a pI of 4.61. Real-time quantitative analysis indicated that SoPR10 was constitutively expressed in root and shoot. The abundance of SoPR10 in salt-resistant cultivar (Chaoji) was generally greater than in salt-sensitive cultivar (Daye) under 160 mM L(-1) NO3(-) treatment for 0.5, 3, and 6 h. The expression of SoPR10 was also induced by other abiotic stresses including polyethylene glycol, NaCl, salicylic acid, and H2O2. Our results indicated that SoPR10 might play important roles under nitrate stress and other abiotic stresses.

  19. Translocation of the potato 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase into isolated spinach chloroplasts

    SciTech Connect

    Zhao, Jianmin; Weaver, L.M.; Herrmann, K.M. )

    1990-05-01

    A cDNA for potato (Solanum tuberosum L.) 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase, the first enzyme of the shikimate pathway, encodes a 56 KD polypeptide whose amino terminus resembles a chloroplast transit sequence. The cDNA was placed downstream of the phage T7 polymerase recognition sequence in plasmid pGEM-3Z. DNA of the resulting plasmid pGEM-DWZ directed T7 polymerase to synthesize potato DAHP synthase mRNA in vitro. The mRNA was used in wheat germ and rabbit reticulocyte lysates for the synthesis of {sup 35}S-labeled pro-DAHP synthase. The predominant translation product is a 59 KD polypeptide that can be immunoprecipitated by rabbit polyclonal antibodies raised against the 53 KD DAHP synthase purified from potato tubers. Isolated spinach chloroplasts process the 59 KD pro-DAHP synthase to a 50 KD polypeptide. The processed polypeptide is protected from protease degradation, suggesting uptake of the enzyme into the cell organelle. Fractionation of reisolated chloroplasts after import of pro-DAHP synthase showed mature enzyme in the stroma. The uptake and processing of DAHP synthase is inhibited by antibodies raised against the mature enzyme. Our results are consistent with the assumption that potato contains a nuclear DNA encoded DAHP synthase that is synthesized as a proenzyme and whose mature form resides in the chloroplasts. Our data provide further evidence that green plants synthesize aromatic amino acids in plastids.

  20. Classification and quantification of leaf curvature

    PubMed Central

    Liu, Zhongyuan; Jia, Liguo; Mao, Yanfei; He, Yuke

    2010-01-01

    Various mutants of Arabidopsis thaliana deficient in polarity, cell division, and auxin response are characterized by certain types of leaf curvature. However, comparison of curvature for clarification of gene function can be difficult without a quantitative measurement of curvature. Here, a novel method for classification and quantification of leaf curvature is reported. Twenty-two mutant alleles from Arabidopsis mutants and transgenic lines deficient in leaf flatness were selected. The mutants were classified according to the direction, axis, position, and extent of leaf curvature. Based on a global measure of whole leaves and a local measure of four regions in the leaves, the curvature index (CI) was proposed to quantify the leaf curvature. The CI values accounted for the direction, axis, position, and extent of leaf curvature in all of the Arabidopsis mutants grown in growth chambers. Comparison of CI values between mutants reveals the spatial and temporal variations of leaf curvature, indicating the strength of the mutant alleles and the activities of the corresponding genes. Using the curvature indices, the extent of curvature in a complicated genetic background becomes quantitative and comparable, thus providing a useful tool for defining the genetic components of leaf development and to breed new varieties with leaf curvature desirable for the efficient capture of sunlight for photosynthesis and high yields. PMID:20400533

  1. Compound leaf development in model plant species.

    PubMed

    Bar, Maya; Ori, Naomi

    2015-02-01

    Plant leaves develop in accordance with a common basic program, which is flexibly adjusted to the species, developmental stage and environment. Two key stages of leaf development are morphogenesis and differentiation. In the case of compound leaves, the morphogenesis stage is prolonged as compared to simple leaves, allowing for the initiation of leaflets. Here, we review recent advances in the understanding of how plant hormones and transcriptional regulators modulate compound leaf development, yielding a substantial diversity of leaf forms, focusing on four model compound leaf organisms: cardamine (Cardamine hirsuta), tomato (Solanum lycopersicum), medicago (Medicago truncatula) and pea (Pisum sativum).

  2. 7 CFR 28.471 - Below Leaf Grade Cotton.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Below Leaf Grade Cotton. 28.471 Section 28.471... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Below Leaf Grade Cotton § 28.471 Below Leaf Grade Cotton. Below leaf grade cotton is American Upland cotton which is lower in leaf grade than...

  3. 7 CFR 28.471 - Below Leaf Grade Cotton.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Below Leaf Grade Cotton. 28.471 Section 28.471... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Below Leaf Grade Cotton § 28.471 Below Leaf Grade Cotton. Below leaf grade cotton is American Upland cotton which is lower in leaf grade than...

  4. 7 CFR 28.471 - Below Leaf Grade Cotton.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Below Leaf Grade Cotton. 28.471 Section 28.471... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Below Leaf Grade Cotton § 28.471 Below Leaf Grade Cotton. Below leaf grade cotton is American Upland cotton which is lower in leaf grade than...

  5. 7 CFR 28.471 - Below Leaf Grade Cotton.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Below Leaf Grade Cotton. 28.471 Section 28.471... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Below Leaf Grade Cotton § 28.471 Below Leaf Grade Cotton. Below leaf grade cotton is American Upland cotton which is lower in leaf grade than...

  6. 7 CFR 28.471 - Below Leaf Grade Cotton.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Below Leaf Grade Cotton. 28.471 Section 28.471... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Below Leaf Grade Cotton § 28.471 Below Leaf Grade Cotton. Below leaf grade cotton is American Upland cotton which is lower in leaf grade than...

  7. 7 CFR 28.464 - Leaf Grade 4.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf Grade 4. 28.464 Section 28.464 Agriculture..., TESTING, AND STANDARDS Standards Official Cotton Standards of the United States for the Leaf Grade of American Upland Cotton § 28.464 Leaf Grade 4. Leaf Grade 4 is leaf which is within the range represented...

  8. 7 CFR 28.461 - Leaf Grade 1.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf Grade 1. 28.461 Section 28.461 Agriculture..., TESTING, AND STANDARDS Standards Official Cotton Standards of the United States for the Leaf Grade of American Upland Cotton § 28.461 Leaf Grade 1. Leaf Grade 1 is leaf which is within the range represented...

  9. 7 CFR 28.466 - Leaf Grade 6.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf Grade 6. 28.466 Section 28.466 Agriculture..., TESTING, AND STANDARDS Standards Official Cotton Standards of the United States for the Leaf Grade of American Upland Cotton § 28.466 Leaf Grade 6. Leaf Grade 6 is leaf which is within the range represented...

  10. 7 CFR 28.465 - Leaf Grade 5.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf Grade 5. 28.465 Section 28.465 Agriculture..., TESTING, AND STANDARDS Standards Official Cotton Standards of the United States for the Leaf Grade of American Upland Cotton § 28.465 Leaf Grade 5. Leaf Grade 5 is leaf which is within the range represented...

  11. 7 CFR 28.462 - Leaf Grade 2.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf Grade 2. 28.462 Section 28.462 Agriculture..., TESTING, AND STANDARDS Standards Official Cotton Standards of the United States for the Leaf Grade of American Upland Cotton § 28.462 Leaf Grade 2. Leaf Grade 2 is leaf which is within the range represented...

  12. 7 CFR 28.463 - Leaf Grade 3.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf Grade 3. 28.463 Section 28.463 Agriculture..., TESTING, AND STANDARDS Standards Official Cotton Standards of the United States for the Leaf Grade of American Upland Cotton § 28.463 Leaf Grade 3. Leaf Grade 3 is leaf which is within the range represented...

  13. 7 CFR 28.467 - Leaf Grade 7.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Leaf Grade 7. 28.467 Section 28.467 Agriculture..., TESTING, AND STANDARDS Standards Official Cotton Standards of the United States for the Leaf Grade of American Upland Cotton § 28.467 Leaf Grade 7. Leaf Grade 7 is leaf which is within the range represented...

  14. Wind-induced leaf transpiration

    NASA Astrophysics Data System (ADS)

    Huang, Cheng-Wei; Chu, Chia-Ren; Hsieh, Cheng-I.; Palmroth, Sari; Katul, Gabriel G.

    2015-12-01

    While the significance of leaf transpiration (fe) on carbon and water cycling is rarely disputed, conflicting evidence has been reported on how increasing mean wind speed (U) impacts fe from leaves. Here, conditions promoting enhancement or suppression of fe with increasing U for a wide range of environmental conditions are explored numerically using leaf-level gas exchange theories that combine a stomatal conductance model based on optimal water use strategies (maximizing the 'net' carbon gain at a given fe), energy balance considerations, and biochemical demand for CO2. The analysis showed monotonic increases in fe with increasing U at low light levels. However, a decline in modeled fe with increasing U were predicted at high light levels but only in certain instances. The dominant mechanism explaining this decline in modeled fe with increasing U is a shift from evaporative cooling to surface heating at high light levels. New and published sap flow measurements for potted Pachira macrocarpa and Messerschmidia argentea plants conducted in a wind tunnel across a wide range of U (2 - 8 m s-1) and two different soil moisture conditions were also employed to assess how fe varies with increasing U. The radiative forcing imposed in the wind tunnel was only restricted to the lower end of expected field conditions. At this low light regime, the findings from the wind tunnel experiments were consistent with the predicted trends.

  15. “Breath figures” on leaf surfaces—formation and effects of microscopic leaf wetness

    PubMed Central

    Burkhardt, Juergen; Hunsche, Mauricio

    2013-01-01

    “Microscopic leaf wetness” means minute amounts of persistent liquid water on leaf surfaces which are invisible to the naked eye. The water is mainly maintained by transpired water vapor condensing onto the leaf surface and to attached leaf surface particles. With an estimated average thickness of less than 1 μm, microscopic leaf wetness is about two orders of magnitude thinner than morning dewfall. The most important physical processes which reduce the saturation vapor pressure and promote condensation are cuticular absorption and the deliquescence of hygroscopic leaf surface particles. Deliquescent salts form highly concentrated solutions. Depending on the type and concentration of the dissolved ions, the physicochemical properties of microscopic leaf wetness can be considerably different from those of pure water. Microscopic leaf wetness can form continuous thin layers on hydrophobic leaf surfaces and in specific cases can act similar to surfactants, enabling a strong potential influence on the foliar exchange of ions. Microscopic leaf wetness can also enhance the dissolution, the emission, and the reaction of specific atmospheric trace gases e.g., ammonia, SO2, or ozone, leading to a strong potential role for microscopic leaf wetness in plant/atmosphere interaction. Due to its difficult detection, there is little knowledge about the occurrence and the properties of microscopic leaf wetness. However, based on the existing evidence and on physicochemical reasoning it can be hypothesized that microscopic leaf wetness occurs on almost any plant worldwide and often permanently, and that it significantly influences the exchange processes of the leaf surface with its neighboring compartments, i.e., the plant interior and the atmosphere. The omission of microscopic water in general leaf wetness concepts has caused far-reaching, misleading conclusions in the past. PMID:24167510

  16. "Breath figures" on leaf surfaces-formation and effects of microscopic leaf wetness.

    PubMed

    Burkhardt, Juergen; Hunsche, Mauricio

    2013-01-01

    "Microscopic leaf wetness" means minute amounts of persistent liquid water on leaf surfaces which are invisible to the naked eye. The water is mainly maintained by transpired water vapor condensing onto the leaf surface and to attached leaf surface particles. With an estimated average thickness of less than 1 μm, microscopic leaf wetness is about two orders of magnitude thinner than morning dewfall. The most important physical processes which reduce the saturation vapor pressure and promote condensation are cuticular absorption and the deliquescence of hygroscopic leaf surface particles. Deliquescent salts form highly concentrated solutions. Depending on the type and concentration of the dissolved ions, the physicochemical properties of microscopic leaf wetness can be considerably different from those of pure water. Microscopic leaf wetness can form continuous thin layers on hydrophobic leaf surfaces and in specific cases can act similar to surfactants, enabling a strong potential influence on the foliar exchange of ions. Microscopic leaf wetness can also enhance the dissolution, the emission, and the reaction of specific atmospheric trace gases e.g., ammonia, SO2, or ozone, leading to a strong potential role for microscopic leaf wetness in plant/atmosphere interaction. Due to its difficult detection, there is little knowledge about the occurrence and the properties of microscopic leaf wetness. However, based on the existing evidence and on physicochemical reasoning it can be hypothesized that microscopic leaf wetness occurs on almost any plant worldwide and often permanently, and that it significantly influences the exchange processes of the leaf surface with its neighboring compartments, i.e., the plant interior and the atmosphere. The omission of microscopic water in general leaf wetness concepts has caused far-reaching, misleading conclusions in the past.

  17. Modeling of the redox state dynamics in photosystem II of Chlorella pyrenoidosa Chick cells and leaves of spinach and Arabidopsis thaliana from single flash-induced fluorescence quantum yield changes on the 100 ns-10 s time scale.

    PubMed

    Belyaeva, N E; Schmitt, F-J; Paschenko, V Z; Riznichenko, G Yu; Rubin, A B

    2015-08-01

    The time courses of the photosystem II (PSII) redox states were analyzed with a model scheme supposing a fraction of 11-25 % semiquinone (with reduced [Formula: see text]) RCs in the dark. Patterns of single flash-induced transient fluorescence yield (SFITFY) measured for leaves (spinach and Arabidopsis (A.) thaliana) and the thermophilic alga Chlorella (C.) pyrenoidosa Chick (Steffen et al. Biochemistry 44:3123-3132, 2005; Belyaeva et al. Photosynth Res 98:105-119, 2008, Plant Physiol Biochem 77:49-59, 2014) were fitted with the PSII model. The simulations show that at high-light conditions the flash generated triplet carotenoid (3)Car(t) population is the main NPQ regulator decaying in the time interval of 6-8 μs. So the SFITFY increase up to the maximum level [Formula: see text]/F 0 (at ~50 μs) depends mainly on the flash energy. Transient electron redistributions on the RC redox cofactors were displayed to explain the SFITFY measured by weak light pulses during the PSII relaxation by electron transfer (ET) steps and coupled proton transfer on both the donor and the acceptor side of the PSII. The contribution of non-radiative charge recombination was taken into account. Analytical expressions for the laser flash, the (3)Car(t) decay and the work of the water-oxidizing complex (WOC) were used to improve the modeled P680(+) reduction by YZ in the state S 1 of the WOC. All parameter values were compared between spinach, A. thaliana leaves and C. pyrenoidosa alga cells and at different laser flash energies. ET from [Formula: see text] slower in alga as compared to leaf samples was elucidated by the dynamics of [Formula: see text] fractions to fit SFITFY data. Low membrane energization after the 10 ns single turnover flash was modeled: the ∆Ψ(t) amplitude (20 mV) is found to be about 5-fold smaller than under the continuous light induction; the time-independent lumen pHL, stroma pHS are fitted close to dark estimates. Depending on the flash energy used at 1

  18. Modeling of the redox state dynamics in photosystem II of Chlorella pyrenoidosa Chick cells and leaves of spinach and Arabidopsis thaliana from single flash-induced fluorescence quantum yield changes on the 100 ns-10 s time scale.

    PubMed

    Belyaeva, N E; Schmitt, F-J; Paschenko, V Z; Riznichenko, G Yu; Rubin, A B

    2015-08-01

    The time courses of the photosystem II (PSII) redox states were analyzed with a model scheme supposing a fraction of 11-25 % semiquinone (with reduced [Formula: see text]) RCs in the dark. Patterns of single flash-induced transient fluorescence yield (SFITFY) measured for leaves (spinach and Arabidopsis (A.) thaliana) and the thermophilic alga Chlorella (C.) pyrenoidosa Chick (Steffen et al. Biochemistry 44:3123-3132, 2005; Belyaeva et al. Photosynth Res 98:105-119, 2008, Plant Physiol Biochem 77:49-59, 2014) were fitted with the PSII model. The simulations show that at high-light conditions the flash generated triplet carotenoid (3)Car(t) population is the main NPQ regulator decaying in the time interval of 6-8 μs. So the SFITFY increase up to the maximum level [Formula: see text]/F 0 (at ~50 μs) depends mainly on the flash energy. Transient electron redistributions on the RC redox cofactors were displayed to explain the SFITFY measured by weak light pulses during the PSII relaxation by electron transfer (ET) steps and coupled proton transfer on both the donor and the acceptor side of the PSII. The contribution of non-radiative charge recombination was taken into account. Analytical expressions for the laser flash, the (3)Car(t) decay and the work of the water-oxidizing complex (WOC) were used to improve the modeled P680(+) reduction by YZ in the state S 1 of the WOC. All parameter values were compared between spinach, A. thaliana leaves and C. pyrenoidosa alga cells and at different laser flash energies. ET from [Formula: see text] slower in alga as compared to leaf samples was elucidated by the dynamics of [Formula: see text] fractions to fit SFITFY data. Low membrane energization after the 10 ns single turnover flash was modeled: the ∆Ψ(t) amplitude (20 mV) is found to be about 5-fold smaller than under the continuous light induction; the time-independent lumen pHL, stroma pHS are fitted close to dark estimates. Depending on the flash energy used at 1

  19. Leaf Histology--Two Modern Methods.

    ERIC Educational Resources Information Center

    Freeman, H. E.

    1984-01-01

    Two methods for examining leaf structure are presented; both methods involve use of "superglue." The first method uses the glue to form a thin, permanent, direct replica of a leaf surface on a microscope slide. The second method uses the glue to examine the three-dimensional structure of spongy mesophyll. (JN)

  20. 7 CFR 30.2 - Leaf tobacco.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Leaf tobacco. 30.2 Section 30.2 Agriculture... Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND STANDARD CONTAINER REGULATIONS TOBACCO STOCKS AND STANDARDS Classification of Leaf Tobacco Covering Classes, Types and Groups of Grades § 30.2...