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Sample records for leaf buds analyses

  1. Use of Stored Carbon Reserves in Growth of Temperate Tree Roots and Leaf Buds: Analyses Using Radiocarbon Measurements and Modeling

    SciTech Connect

    Gaudinski, Julia B.; Torn, Margaret S.; Riley, W. J.; Swanston, Christopher W.; Trumbore, Susan E.; JoslinJr., John D.; Majdi, H; Dawson, Todd E.; Hanson, Paul J

    2009-01-01

    Characterizing the use of C reserves in trees is important for understanding stress responses, impacts of asynchrony between photosynthesis and growth demand, and isotopic exchanges in plant dynamic studies. Using an inadvertent, whole ecosystem radiocarbon (14C) exposure in a temperate deciduous oak forest and numerical modeling, we calculated that the mean age of stored C used to grow leaf buds and new fine root tissue is 0.5-1.0 y. The mean age of stored C used to grow new roots was about 0.7 y across a range of realistic values of 14C inputs to the system. The amount of stored C used on an annual basis to grow fine roots was between 15 and 55% of total root growth, with the range defined by the assumed 14C input profile. We estimate the annually-averaged mean age of C in new root tissues is 1-5 months. Therefore, accounting for storage C use in isotope root models may be unnecessary in all but the fastest cycling root populations (i.e., mean age <1 y). Consistent with the whole ecosystem labeling results, we found, using "bomb-14C," that the mean C age of new root tissues in three additional forest sites (one deciduous, two coniferous) was less than 2 years. We conclude that in many ecosystem types, growth from stored C is insufficient to impact bomb-14C based estimates of long root lifetimes.

  2. The use of stored carbon reserves in growth of temperate tree roots and leaf buds: Analyses using radiocarbon measurements and modeling

    SciTech Connect

    Gaudinski, J.B.; Torn, M.S.; Riley, W.J.; Swanston, C.; Trumbore, S.E.; Joslin, J.D.; Majdi, H.; Dawson, T.E.; Hanson, P.J.

    2009-02-01

    Characterizing the use of carbon (C) reserves in trees is important for understanding regional and global C cycles, stress responses, asynchrony between photosynthetic activity and growth demand, and isotopic exchanges in studies of tree physiology and ecosystem C cycling. Using an inadvertent, whole-ecosystem radiocarbon ({sup 14}C) release in a temperate deciduous oak forest and numerical modeling, we estimated that the mean age of stored C used to grow both leaf buds and new roots is 0.7 years and about 55% of new-root growth annually comes from stored C. Therefore, the calculated mean age of C used to grow new-root tissue is {approx}0.4 years. In short, new roots contain a lot of stored C but it is young in age. Additionally, the type of structure used to model stored C input is important. Model structures that did not include storage, or that assumed stored and new C mixed well (within root or shoot tissues) before being used for root growth, did not fit the data nearly as well as when a distinct storage pool was used. Consistent with these whole-ecosystem labeling results, the mean age of C in new-root tissues determined using 'bomb-{sup 14}C' in three additional forest sites in North America and Europe (one deciduous, two coniferous) was less than 1-2 years. The effect of stored reserves on estimated ages of fine roots is unlikely to be large in most natural abundance isotope studies. However, models of root C dynamics should take stored reserves into account, particularly for pulse-labeling studies and fast-cycling roots (<1 years).

  3. Optical properties of bud scales and protochlorophyll(ide) forms in leaf primordia of closed and opened buds.

    PubMed

    Solymosi, Katalin; Böddi, Béla

    2006-08-01

    The transmission spectra of bud scales of 14 woody species and the 77 K fluorescence emission spectra of the innermost leaf primordia of closed and opened buds of 37 woody species were studied. Pigment concentrations were determined in some species. Bud scales had low transmittance between 400 and 680 nm with a local minimum around 680 nm. Transmittance increased steeply above 680 nm and was > 80% in the 700-800 nm spectral region. Significant protochlorophyllide (Pchlide) accumulation was observed in leaf primordia of tightly packed, closed buds with relatively thick, dark bud scales. In common ash (Fraxinus excelsior L.) and Hungarian ash (Fraxinus angustifolia Vahl.), the innermost leaf primordia of the closed buds contained protochlorophyll (Pchl) and Pchlide (abbreviated as Pchl(ide)), but no chlorophyll. We observed Pchl(ide) forms with emission maxima at 633, 643 and 655 nm in these leaves. Complete transformation of Pchlide(655) (protochlorophyllide form with maximum emission at 655 nm) into Chlide(692) (chlorophyllide form with maximum emission at 692 nm) occurred after irradiation for 10 s. The innermost leaf primordia of the buds of four species (flowering ash (Fraxinus ornus L.), horse chestnut (Aesculus hippocastanum L.), tree of heaven (Ailanthus altissima P. Mill.) and common walnut (Juglans regia L.)) contained Pchl(ide)(633), Pchl(ide)(643) and Pchlide(655) as well as an emission band at 688 nm corresponding to a chlorophyll form. The Pchlide(655) was fully photoactive in these species. The outermost leaf primordia of these four species and the innermost leaf primordia of 28 other species contained all of the above described Pchl(ide) forms in various ratios but in small amounts. In addition, Chl forms were present and the main bands in the fluorescence emission spectra were at 690 or 740 nm, or both. The results indicate that Pchl(ide) accumulation occurs in leaf primordia in near darkness inside the tightly closed buds, where the bud scales and

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

    PubMed

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

    2012-05-01

    To study the formation of the photosynthetic apparatus in nature, the carotenoid and chlorophyllous pigment compositions of differently developed leaf primordia in closed and opening buds of common ash (Fraxinus excelsior L.) and horse chestnut (Aesculus hippocastanum L.) as well as in closed buds of tree of heaven (Ailanthus altissima P. Mill.) were analyzed with HPLC. The native organization of the chlorophyllous pigments was studied using 77 K fluorescence spectroscopy, and plastid ultrastructure was investigated with electron microscopy. Complete etiolation, i.e., accumulation of protochlorophyllide, and absence of chlorophylls occurred in the innermost leaf primordia of common ash buds. The other leaf primordia were partially etiolated in the buds and contained protochlorophyllide (0.5-1 μg g(-1) fresh mass), chlorophyllides (0.2-27 μg g(-1) fresh mass) and chlorophylls (0.9-643 μg g(-1) fresh mass). Etio-chloroplasts with prolamellar bodies and either regular or only low grana were found in leaves having high or low amounts of chlorophyll a and b, respectively. After bud break, etioplast-chloroplast conversion proceeded and the pigment contents increased in the leaves, similarly to the greening processes observed in illuminated etiolated seedlings under laboratory conditions. The pigment contents and the ratio of the different spectral forms had a high biological variability that could be attributed to (i) various light conditions due to light filtering in the buds resulting in differently etiolated leaf primordia, (ii) to differences in the light-exposed and inner regions of the same primordia in opening buds due to various leaf folding, and (iii) to tissue-specific slight variations of plastid ultrastructure. PMID:22160501

  5. Gene expression analysis of bud and leaf color in tea.

    PubMed

    Wei, Kang; Zhang, Yazhen; Wu, Liyun; Li, Hailin; Ruan, Li; Bai, Peixian; Zhang, Chengcai; Zhang, Fen; Xu, Liyi; Wang, Liyuan; Cheng, Hao

    2016-10-01

    Purple shoot tea attributing to the high anthocyanin accumulation is of great interest for its wide health benefits. To better understand potential mechanisms involved in purple buds and leaves formation in tea plants, we performed transcriptome analysis of six green or purple shoot tea individuals from a F1 population using the Illumina sequencing method. Totally 292 million RNA-Seq reads were obtained and assembled into 112,233 unigenes, with an average length of 759 bp and an N50 of 1081 bp. Moreover, totally 2193 unigenes showed significant differences in expression levels between green and purple tea samples, with 1143 up- and 1050 down-regulated in the purple teas. Further real time PCR analysis confirmed RNA-Seq results. Our study identified 28 differentially expressed transcriptional factors and A CsMYB gene was found to be highly similar to AtPAP1 in Arabidopsis. Further analysis of differentially expressed genes involved in anthocyanin biosynthesis and transportation showed that the late biosynthetic genes and genes involved in anthocyanin transportation were largely affected but the early biosynthetic genes were less or none affected. Overall, the identification of a large number of differentially expressed genes offers a global view of the potential mechanisms associated with purple buds and leaves formation, which will facilitate molecular breeding in tea plants. PMID:27362295

  6. Perception of photoperiod in individual buds of mature trees regulates leaf-out.

    PubMed

    Zohner, Constantin M; Renner, Susanne S

    2015-12-01

    Experimental data on the perception of day length and temperature in dormant temperate zone trees are surprisingly scarce. In order to investigate when and where these environmental signals are perceived, we carried out bagging experiments in which buds on branches of Fagus sylvatica, Aesculus hippocastanum and Picea abies trees were exposed to natural light increase or kept at constant 8-h days from December until June. Parallel experiments used twigs cut from the same trees, harvesting treated and control twigs seven times and then exposing them to 8- or 16-h days in a glasshouse. Under 8-h days, budburst in Fagus outdoors was delayed by 41 d and in Aesculus by 4 d; in Picea, day length had no effect. Buds on nearby branches reacted autonomously, and leaf primordia only reacted to light cues in late dormancy after accumulating warm days. Experiments applying different wavelength spectra and high-resolution spectrometry to buds indicate a phytochrome-mediated photoperiod control. By demonstrating local photoperiodic control of buds, revealing the time when these signals are perceived, and showing the interplay between photoperiod and chilling, this study contributes to improved modelling of the impact of climate warming on photosensitive species. PMID:26096967

  7. Volatile compounds from leaf-buds of Populus nigra L (Salicaceae).

    PubMed

    Jerković, Igor; Mastelić, Josip

    2003-05-01

    Volatile components from fresh and air-dried leaf-buds of Populus nigra L. (Salicaceae) were isolated by Likens-Nickerson apparatus and analyzed using GC/MS. Forty-eight components (ca. 95% of the total isolate) were identified among black poplar bud volatiles. Sesquiterpene alcohols beta-eudesmol and alpha-eudesmol represented 26.3-28.7% of the oil. Other major sesquiterpene compounds were gamma-selinene (7.6-8.8%), delta-cadinene (7.8-8.6%), alpha-elemene (3.3-5.2%) and gamma-cadinene (3.9-4.2%). Hemiterpenes were also identified (2.2-7.6%). Monoterpenes were present in low percentages (1.6-5.7%). Aliphatic and aromatic alcohols, carbonyl compounds and aliphatic acids were identified among non-terpene volatiles (9.8-13.5%). The fresh buds contained 0.27% and dried 0.12% essential oil. Air-drying moderately effected the volatiles qualitative and quantitative composition. PMID:12657305

  8. From buds to litter: seasonal changes in leaf wax concentrations and carbon isotopes and implications for the geologic past

    NASA Astrophysics Data System (ADS)

    Suh, Y. J.; Diefendorf, A. F.

    2014-12-01

    The carbon isotope composition (δ13C) of leaf waxes, such as n-alkanes, have extensively been used in paleoenvironmental studies for reconstruction of the past vegetation, climate and carbon cycling. There is however little information available on the seasonal variation of leaf wax concentration and δ13C in modern plants and when the δ13C signal is set. This lack of information confounds interpretations of leaf wax δ13C in sedimentary archives. To address this gap, this study investigates temporal changes in n-alkane and n-alkanoic acid δ13C values in several species (Acer rubrum, Acer saccharum, Ulmus Americana, Sassafras albidum, and Juniperus virginiana) within a single temperate deciduous forest stand in southern Ohio. We sampled atmospheric air, buds, leaves, leaf litter, and surface soil weekly during leaf flush and biweekly thereafter. In A. rubrum, A. saccharum, and U. Americana, buds had one or two dominant n-alkanes, such as C29 and C31. After leaf flush, the concentrations of shorter n-alkanes (C23~C27) significantly increased relative to the longer chain-lengths. We are currently analyzing remaining samples from the growing season and are analyzing bulk leaf and leaf wax (n-alkanes, n-alkanoic acids) δ13C values. This information will be important for identifying environmental and physiological controls on leaf wax δ13C and will improve interpretations of leaf wax δ13C preserved in the geologic record.

  9. Comparison of axillary bud growth and patatin accumulation in potato leaf cuttings as assays for tuber induction

    NASA Technical Reports Server (NTRS)

    Wheeler, R. M.; Hannapel, D. J.; Tibbitts, T. W.

    1988-01-01

    Single-node leaf cuttings from potatoes (Solanum tuberosum L.) cvs. Norland, Superior, Norchip, and Kennebec, were used to assess tuber induction in plants grown under 12, 16, and 20 h daily irradiation (400 micromol s-1 m-2 PPF). Leaf cuttings were taken from plants at four, six and 15 weeks after planting and cultured for 14 d in sand trays in humid environments. Tuber induction was determined by visually rating the type of growth at the attached axillary bud, and by measuring the accumulation of the major tuber protein, patatin, in the base of the petioles. Axillary buds from leaf cuttings of plants grown under the 12 h photoperiod consistently formed round, sessile tubers at the axils for all four cultivars at all harvests. Buds from cuttings of plants grown under the 16 and 20 h photoperiods exhibited mixed tuber, stolon, and leafy shoot growth. Patatin accumulation was highest in petioles of cuttings taken from 12 h plants for all cultivars at all harvests, with levels in 16 and 20 h cuttings approx. one-half that of the 12 h cuttings. Trends, both in visual ratings of axillary buds and in petiole patatin accumulation, followed the harvest index (ratio of tuber to total plant dry matter), suggesting that either method is an acceptable assay for tuber induction in the potato.

  10. CG Methylation Covaries with Differential Gene Expression between Leaf and Floral Bud Tissues of Brachypodium distachyon

    PubMed Central

    Roessler, Kyria; Takuno, Shohei; Gaut, Brandon S.

    2016-01-01

    DNA methylation has the potential to influence plant growth and development through its influence on gene expression. To date, however, the evidence from plant systems is mixed as to whether patterns of DNA methylation vary significantly among tissues and, if so, whether these differences affect tissue-specific gene expression. To address these questions, we analyzed both bisulfite sequence (BSseq) and transcriptomic sequence data from three biological replicates of two tissues (leaf and floral bud) from the model grass species Brachypodium distachyon. Our first goal was to determine whether tissues were more differentiated in DNA methylation than explained by variation among biological replicates. Tissues were more differentiated than biological replicates, but the analysis of replicated data revealed high (>50%) false positive rates for the inference of differentially methylated sites (DMSs) and differentially methylated regions (DMRs). Comparing methylation to gene expression, we found that differential CG methylation consistently covaried negatively with gene expression, regardless as to whether methylation was within genes, within their promoters or even within their closest transposable element. The relationship between gene expression and either CHG or CHH methylation was less consistent. In total, CG methylation in promoters explained 9% of the variation in tissue-specific expression across genes, suggesting that CG methylation is a minor but appreciable factor in tissue differentiation. PMID:26950546

  11. Transient etiolation: protochlorophyll(ide) and chlorophyll forms in differentiating plastids of closed and breaking leaf buds of horse chestnut (Aesculus hippocastanum).

    PubMed

    Solymosi, Katalin; Bóka, Károly; Böddi, Béla

    2006-08-01

    An accompanying paper reports the accumulation of photoactive protochlorophyllide (Pchlide) in the innermost leaf primordia of buds of many tree species. In this paper, we describe plastid differentiation, changes in pigment concentrations and spectral properties of bud scales and leaf primordia of horse chestnut (Aesculus hippocastanum L.) from January until the end of bud break in April. The bud scales contained plastids with grana, stroma thylakoids characteristic of chloroplasts and large dense bodies within the stroma. In January, proplastids and young chloroplasts were present in the leaf primordia, and the fluorescence spectra of the primordia were similar to those of green leaves except for a minor band at 630 nm, indicative of a protochlorophyll(ide). During bud break, the pigment concentrations of the green bud scales and the outermost leaf primordia increased, and Pchlide forms with emission maxima at 633, 644 and 655 nm accumulated in the middle and innermost leaf primordia. Depending on the position of the leaf primordia within the bud, their plastids and their pigment concentrations varied. Etio-chloroplasts with prolamellar bodies (PLBs) and prothylakoids with developing grana were observed in the innermost leaves. Besides the above-mentioned Pchlide forms, the middle and innnermost leaf primordia contained only a Chl band with an emission maximum at 686 nm. The outermost leaf primordia contained etio-chloroplasts with well-developed grana and small, narrow-type PLBs. These outermost leaves contained only chlorophyll forms like the mature green leaves. No Pchlide accumulation was observed after bud break, indicating that etiolation of the innermost and middle leaves is transient. The Pchlide forms and the plastid types of the primordia in buds grown in nature were similar to those of leaves of dark-germinated seedlings and to those of the leaf primordia of dark-forced buds. We conclude that transient etiolation occurs under natural conditions. The

  12. Characterization of bud emergence 46 (BEM46) protein: sequence, structural, phylogenetic and subcellular localization analyses.

    PubMed

    Kumar, Abhishek; Kollath-Leiß, Krisztina; Kempken, Frank

    2013-08-30

    The bud emergence 46 (BEM46) protein from Neurospora crassa belongs to the α/β-hydrolase superfamily. Recently, we have reported that the BEM46 protein is localized in the perinuclear ER and also forms spots close by the plasma membrane. The protein appears to be required for cell type-specific polarity formation in N. crassa. Furthermore, initial studies suggested that the BEM46 amino acid sequence is conserved in eukaryotes and is considered to be one of the widespread conserved "known unknown" eukaryotic genes. This warrants for a comprehensive phylogenetic analysis of this superfamily to unravel origin and molecular evolution of these genes in different eukaryotes. Herein, we observe that all eukaryotes have at least a single copy of a bem46 ortholog. Upon scanning of these proteins in various genomes, we find that there are expansions leading into several paralogs in vertebrates. Usingcomparative genomic analyses, we identified insertion/deletions (indels) in the conserved domain of BEM46 protein, which allow to differentiate fungal classes such as ascomycetes from basidiomycetes. We also find that exonic indels are able to differentiate BEM46 homologs of different eukaryotic lineage. Furthermore, we unravel that BEM46 protein from N. crassa possess a novel endoplasmic-retention signal (PEKK) using GFP-fusion tagging experiments. We propose that three residues namely a serine 188S, a histidine 292H and an aspartic acid 262D are most critical residues, forming a catalytic triad in BEM46 protein from N. crassa. We carried out a comprehensive study on bem46 genes from a molecular evolution perspective with combination of functional analyses. The evolutionary history of BEM46 proteins is characterized by exonic indels in lineage specific manner. PMID:23916612

  13. Proteomic analyses of baculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus budded and occluded virus.

    PubMed

    Braconi, Carla Torres; Ardisson-Araújo, Daniel Mendes Pereira; Paes Leme, Adriana Franco; Oliveira, Juliana Velasco de Castro; Pauletti, Bianca Alves; Garcia-Maruniak, Alejandra; Ribeiro, Bergmann Morais; Maruniak, James E; Zanotto, Paolo Marinho de Andrade

    2014-04-01

    Baculoviruses infect insects, producing two distinct phenotypes during the viral life cycle: the budded virus (BV) and the occlusion-derived virus (ODV) for intra- and inter-host spread, respectively. Since the 1980s, several countries have been using Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) as a biological control agent against the velvet bean caterpillar, A. gemmatalis. The genome of AgMNPV isolate 2D (AgMNPV-2D) carries at least 152 potential genes, with 24 that possibly code for structural proteins. Proteomic studies have been carried out on a few baculoviruses, with six ODV and two BV proteomes completed so far. Moreover, there are limited data on virion proteins carried by AgMNPV-2D. Therefore, structural proteins of AgMNPV-2D were analysed by MALDI- quadrupole-TOF and liquid chromatography MS/MS. A total of 44 proteins were associated with the ODV and 33 with the BV of AgMNPV-2D. Although 38 structural proteins were already known, we found six new proteins in the ODV and seven new proteins carried by the AgMNPV-2D BV. Eleven cellular proteins that were found on several other enveloped viruses were also identified, which are possibly carried with the virion. These findings may provide novel insights into baculovirus biology and their host interaction. Moreover, our data may be helpful in subsequent applied studies aiming to improve AgMNPV use as a biopesticide and a biotechnology tool for gene expression or delivery. PMID:24443474

  14. A Comparative Analysis of Genetic Differentiation across Six Shared Willow Host Species in Leaf- and Bud-Galling Sawflies

    PubMed Central

    Leppänen, Sanna A.; Malm, Tobias; Värri, Kaisa; Nyman, Tommi

    2014-01-01

    Genetic divergence and speciation in plant-feeding insects could be driven by contrasting selection pressures imposed by different plant species and taxa. While numerous examples of host-associated differentiation (HAD) have been found, the overall importance of HAD in insect diversification remains unclear, as few studies have investigated its frequency in relation to all speciation events. One promising way to infer the prevalence and repeatability of HAD is to estimate genetic differentiation in multiple insect taxa that use the same set of hosts. To this end, we measured and compared variation in mitochondrial COI and nuclear ITS2 sequences in population samples of leaf-galling Pontania and bud-galling Euura sawflies (Hymenoptera: Tenthredinidae) collected from six Salix species in two replicate locations in northern Fennoscandia. We found evidence of frequent HAD in both species complexes, as individuals from the same willow species tended to cluster together on both mitochondrial and nuclear phylogenetic trees. Although few fixed differences among the putative species were found, hierarchical AMOVAs showed that most of the genetic variation in the samples was explained by host species rather than by sampling location. Nevertheless, the levels of HAD measured across specific pairs of host species were not correlated in the two focal galler groups. Hence, our results support the hypothesis of HAD as a central force in herbivore speciation, but also indicate that evolutionary trajectories are only weakly repeatable even in temporally overlapping radiations of related insect taxa. PMID:25551608

  15. Characterization of bud emergence 46 (BEM46) protein: Sequence, structural, phylogenetic and subcellular localization analyses

    SciTech Connect

    Kumar, Abhishek; Kollath-Leiß, Krisztina; Kempken, Frank

    2013-08-30

    Highlights: •All eukaryotes have at least a single copy of a bem46 ortholog. •The catalytic triad of BEM46 is illustrated using sequence and structural analysis. •We identified indels in the conserved domain of BEM46 protein. •Localization studies of BEM46 protein were carried out using GFP-fusion tagging. -- Abstract: The bud emergence 46 (BEM46) protein from Neurospora crassa belongs to the α/β-hydrolase superfamily. Recently, we have reported that the BEM46 protein is localized in the perinuclear ER and also forms spots close by the plasma membrane. The protein appears to be required for cell type-specific polarity formation in N. crassa. Furthermore, initial studies suggested that the BEM46 amino acid sequence is conserved in eukaryotes and is considered to be one of the widespread conserved “known unknown” eukaryotic genes. This warrants for a comprehensive phylogenetic analysis of this superfamily to unravel origin and molecular evolution of these genes in different eukaryotes. Herein, we observe that all eukaryotes have at least a single copy of a bem46 ortholog. Upon scanning of these proteins in various genomes, we find that there are expansions leading into several paralogs in vertebrates. Usingcomparative genomic analyses, we identified insertion/deletions (indels) in the conserved domain of BEM46 protein, which allow to differentiate fungal classes such as ascomycetes from basidiomycetes. We also find that exonic indels are able to differentiate BEM46 homologs of different eukaryotic lineage. Furthermore, we unravel that BEM46 protein from N. crassa possess a novel endoplasmic-retention signal (PEKK) using GFP-fusion tagging experiments. We propose that three residues namely a serine 188S, a histidine 292H and an aspartic acid 262D are most critical residues, forming a catalytic triad in BEM46 protein from N. crassa. We carried out a comprehensive study on bem46 genes from a molecular evolution perspective with combination of functional

  16. Proteomics and transcriptomics analyses of Arabidopsis floral buds uncover important functions of ARABIDOPSIS SKP1-LIKE1

    DOE PAGESBeta

    Lu, Dihong; Ni, Weimin; Stanley, Bruce A.; Ma, Hong

    2016-03-03

    The ARABIDOPSIS SKP1-LIKE1 (ASK1) protein functions as a subunit of SKP1-CUL1-F-box (SCF) E3 ubiquitin ligases. Previous genetic studies showed that ASK1 plays important roles in Arabidopsis flower development and male meiosis. However, the molecular impact of ASK1-containing SCF E3 ubiquitin ligases (ASK1-E3s) on the floral proteome and transcriptome is unknown. Here we identified proteins that are potentially regulated by ASK1-E3s by comparing floral bud proteomes of wild-type and the ask1 mutant plants. More than 200 proteins were detected in the ask1 mutant but not in wild-type and >300 were detected at higher levels in the ask1 mutant than in wild-type,more » but their RNA levels were not significantly different between wild-type and ask1 floral buds as shown by transcriptomics analysis, suggesting that they are likely regulated at the protein level by ASK1-E3s. Integrated analyses of floral proteomics and transcriptomics of ask1 and wild-type uncovered several potential aspects of ASK1-E3 functions, including regulation of transcription regulators, kinases, peptidases, and ribosomal proteins, with implications on possible mechanisms of ASK1-E3 functions in floral development. In conclusion, our results suggested that ASK1-E3s play important roles in Arabidopsis protein degradation during flower development. This study opens up new possibilities for further functional studies of these candidate E3 substrates.« less

  17. Species diversity of Fergusonina Malloch gall flies (Diptera: Fergusoninidae) forming leaf bud galls on snow gum (Eucalyptus pauciflora Sieb. ex Spreng. complex), with a description of a new species from Tasmania

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new species of Fergusonina (Diptera: Fergusoninidae) fly is described from terminal leaf bud galls (TLBGs) from the Eucalyptus pauciflora Sieb. ex Spreng. (snow gum) species complex. Fergusonina tasmaniensis Nelson sp.n. is the first species from the genus Fergusonina to be described from Tasmania...

  18. Pear Bud Metabolism: Seasonal Changes in Glucose Utilization

    PubMed Central

    Zimmerman, Richard H.; Faust, Miklos

    1969-01-01

    Utilization of glucose, uracil and valine by flower and leaf buds of seedling pear trees (Pyrus calleryana Decne.) from the time of flower bud initiation to flowering was investigated. A very high rate of glucose utilization through the pentose phosphate pathway was observed throughout the development of buds. There was no difference in the type of glucose metabolism between flower and leaf buds except immediately before flowering, when the metabolism in flower buds was shifted toward the glycolytic pathway. Such a shift did not occur in leaf buds. The incorporation of uracil and valine into the nucleic acid and protein fraction of buds, respectively, was high throughout bud development, perhaps indicating a high rate of turnover in the resting buds. Incorporation of both compounds decreased when buds started to expand prior to flowering. PMID:16657202

  19. Plantecophys--An R Package for Analysing and Modelling Leaf Gas Exchange Data.

    PubMed

    Duursma, Remko A

    2015-01-01

    Here I present the R package 'plantecophys', a toolkit to analyse and model leaf gas exchange data. Measurements of leaf photosynthesis and transpiration are routinely collected with portable gas exchange instruments, and analysed with a few key models. These models include the Farquhar-von Caemmerer-Berry (FvCB) model of leaf photosynthesis, the Ball-Berry models of stomatal conductance, and the coupled leaf gas exchange model which combines the supply and demand functions for CO2 in the leaf. The 'plantecophys' R package includes functions for fitting these models to measurements, as well as simulating from the fitted models to aid in interpreting experimental data. Here I describe the functionality and implementation of the new package, and give some examples of its use. I briefly describe functions for fitting the FvCB model of photosynthesis to measurements of photosynthesis-CO2 response curves ('A-Ci curves'), fitting Ball-Berry type models, modelling C3 photosynthesis with the coupled photosynthesis-stomatal conductance model, modelling C4 photosynthesis, numerical solution of optimal stomatal behaviour, and energy balance calculations using the Penman-Monteith equation. This open-source package makes technically challenging calculations easily accessible for many users and is freely available on CRAN. PMID:26581080

  20. Plantecophys - An R Package for Analysing and Modelling Leaf Gas Exchange Data

    PubMed Central

    Duursma, Remko A.

    2015-01-01

    Here I present the R package 'plantecophys', a toolkit to analyse and model leaf gas exchange data. Measurements of leaf photosynthesis and transpiration are routinely collected with portable gas exchange instruments, and analysed with a few key models. These models include the Farquhar-von Caemmerer-Berry (FvCB) model of leaf photosynthesis, the Ball-Berry models of stomatal conductance, and the coupled leaf gas exchange model which combines the supply and demand functions for CO2 in the leaf. The 'plantecophys' R package includes functions for fitting these models to measurements, as well as simulating from the fitted models to aid in interpreting experimental data. Here I describe the functionality and implementation of the new package, and give some examples of its use. I briefly describe functions for fitting the FvCB model of photosynthesis to measurements of photosynthesis-CO2 response curves ('A-Ci curves'), fitting Ball-Berry type models, modelling C3 photosynthesis with the coupled photosynthesis-stomatal conductance model, modelling C4 photosynthesis, numerical solution of optimal stomatal behaviour, and energy balance calculations using the Penman-Monteith equation. This open-source package makes technically challenging calculations easily accessible for many users and is freely available on CRAN. PMID:26581080

  1. Arenavirus Budding

    PubMed Central

    Urata, Shuzo; de la Torre, Juan Carlos

    2011-01-01

    Several arenaviruses cause hemorrhagic fever disease in humans and pose a significant public health concern in their endemic regions. On the other hand, the prototypic arenavirus LCMV is a superb workhorse for the investigation of virus-host interactions and associated disease. The arenavirus small RING finger protein called Z has been shown to be the main driving force of virus budding. The budding activity of Z is mediated by late (L) domain motifs, PT/SAP, and PPXY, located at the C-terminus of Z. This paper will present the current knowledge on arenavirus budding including the diversity of L domain motifs used by different arenaviruses. We will also discuss how improved knowledge of arenavirus budding may facilitate the development of novel antiviral strategies to combat human pathogenic arenaviruses. PMID:22312335

  2. K-Nutrition, Growth Bud Formation, and Amine and Hydroxycinnamic Acid Amide Contents in Leaf Explants of Nicotiana tabacum Variety Xanthi n.c. Cultivated in Vitro1

    PubMed Central

    Klinguer, Serge; Martin-Tanguy, Josette; Martin, Claude

    1986-01-01

    The effects of K-nutrition on growth (increase of fresh weight), bud formation (time of emergence, number of buds), and amine and hydroxycinnamic acid amide contents in foliar explants of Nicotiana tabacum cv Xanthi n.c. cultivated in vitro were examined. In K-deficient medium and in high K medium growth and bud formation were markedly inhibited. Marked changes of amine content (a diamine, putrescine; a phenolic amine, phenethylamine) were observed after a few days of culture. No apparent relationship was found between these amines and growth or bud differentiation. In contrast, changes in hydroxycinnamic acid levels were shown to correlate well with growth and bud formation. The greatest stimulation of budding and growth was correlated with the greatest accumulation of these amides. The highest contents of hydroxycinnamic acid amides were found during the first 15 days in culture when intensive cell division took place. Then they declined sharply after 26 days in culture as the rate of cell division decreased and differentiation occurred. PMID:16665067

  3. High Throughput Analyses of Budding Yeast ARSs Reveal New DNA Elements Capable of Conferring Centromere-Independent Plasmid Propagation

    PubMed Central

    Hoggard, Timothy; Liachko, Ivan; Burt, Cassaundra; Meikle, Troy; Jiang, Katherine; Craciun, Gheorghe; Dunham, Maitreya J.; Fox, Catherine A.

    2016-01-01

    The ability of plasmids to propagate in Saccharomyces cerevisiae has been instrumental in defining eukaryotic chromosomal control elements. Stable propagation demands both plasmid replication, which requires a chromosomal replication origin (i.e., an ARS), and plasmid distribution to dividing cells, which requires either a chromosomal centromere for segregation or a plasmid-partitioning element. While our knowledge of yeast ARSs and centromeres is relatively advanced, we know less about chromosomal regions that can function as plasmid partitioning elements. The Rap1 protein-binding site (RAP1) present in transcriptional silencers and telomeres of budding yeast is a known plasmid-partitioning element that functions to anchor a plasmid to the inner nuclear membrane (INM), which in turn facilitates plasmid distribution to daughter cells. This Rap1-dependent INM-anchoring also has an important chromosomal role in higher-order chromosomal structures that enhance transcriptional silencing and telomere stability. Thus, plasmid partitioning can reflect fundamental features of chromosome structure and biology, yet a systematic screen for plasmid partitioning elements has not been reported. Here, we couple deep sequencing with competitive growth experiments of a plasmid library containing thousands of short ARS fragments to identify new plasmid partitioning elements. Competitive growth experiments were performed with libraries that differed only in terms of the presence or absence of a centromere. Comparisons of the behavior of ARS fragments in the two experiments allowed us to identify sequences that were likely to drive plasmid partitioning. In addition to the silencer RAP1 site, we identified 74 new putative plasmid-partitioning motifs predicted to act as binding sites for DNA binding proteins enriched for roles in negative regulation of gene expression and G2/M-phase associated biology. These data expand our knowledge of chromosomal elements that may function in plasmid

  4. High Throughput Analyses of Budding Yeast ARSs Reveal New DNA Elements Capable of Conferring Centromere-Independent Plasmid Propagation.

    PubMed

    Hoggard, Timothy; Liachko, Ivan; Burt, Cassaundra; Meikle, Troy; Jiang, Katherine; Craciun, Gheorghe; Dunham, Maitreya J; Fox, Catherine A

    2016-01-01

    The ability of plasmids to propagate in Saccharomyces cerevisiae has been instrumental in defining eukaryotic chromosomal control elements. Stable propagation demands both plasmid replication, which requires a chromosomal replication origin (i.e., an ARS), and plasmid distribution to dividing cells, which requires either a chromosomal centromere for segregation or a plasmid-partitioning element. While our knowledge of yeast ARSs and centromeres is relatively advanced, we know less about chromosomal regions that can function as plasmid partitioning elements. The Rap1 protein-binding site (RAP1) present in transcriptional silencers and telomeres of budding yeast is a known plasmid-partitioning element that functions to anchor a plasmid to the inner nuclear membrane (INM), which in turn facilitates plasmid distribution to daughter cells. This Rap1-dependent INM-anchoring also has an important chromosomal role in higher-order chromosomal structures that enhance transcriptional silencing and telomere stability. Thus, plasmid partitioning can reflect fundamental features of chromosome structure and biology, yet a systematic screen for plasmid partitioning elements has not been reported. Here, we couple deep sequencing with competitive growth experiments of a plasmid library containing thousands of short ARS fragments to identify new plasmid partitioning elements. Competitive growth experiments were performed with libraries that differed only in terms of the presence or absence of a centromere. Comparisons of the behavior of ARS fragments in the two experiments allowed us to identify sequences that were likely to drive plasmid partitioning. In addition to the silencer RAP1 site, we identified 74 new putative plasmid-partitioning motifs predicted to act as binding sites for DNA binding proteins enriched for roles in negative regulation of gene expression and G2/M-phase associated biology. These data expand our knowledge of chromosomal elements that may function in plasmid

  5. Leaf Length Tracker: a novel approach to analyse leaf elongation close to the thermal limit of growth in the field

    PubMed Central

    Kirchgessner, Norbert; Yates, Steven; Hiltpold, Maya; Walter, Achim

    2016-01-01

    Leaf growth in monocot crops such as wheat and barley largely follows the daily temperature course, particularly under cold but humid springtime field conditions. Knowledge of the temperature response of leaf extension, particularly variations close to the thermal limit of growth, helps define physiological growth constraints and breeding-related genotypic differences among cultivars. Here, we present a novel method, called ‘Leaf Length Tracker’ (LLT), suitable for measuring leaf elongation rates (LERs) of cereals and other grasses with high precision and high temporal resolution under field conditions. The method is based on image sequence analysis, using a marker tracking approach to calculate LERs. We applied the LLT to several varieties of winter wheat (Triticum aestivum), summer barley (Hordeum vulgare), and ryegrass (Lolium perenne), grown in the field and in growth cabinets under controlled conditions. LLT is easy to use and we demonstrate its reliability and precision under changing weather conditions that include temperature, wind, and rain. We found that leaf growth stopped at a base temperature of 0°C for all studied species and we detected significant genotype-specific differences in LER with rising temperature. The data obtained were statistically robust and were reproducible in the tested environments. Using LLT, we were able to detect subtle differences (sub-millimeter) in leaf growth patterns. This method will allow the collection of leaf growth data in a wide range of future field experiments on different graminoid species or varieties under varying environmental or treatment conditions. PMID:26818912

  6. Leaf Length Tracker: a novel approach to analyse leaf elongation close to the thermal limit of growth in the field.

    PubMed

    Nagelmüller, Sebastian; Kirchgessner, Norbert; Yates, Steven; Hiltpold, Maya; Walter, Achim

    2016-04-01

    Leaf growth in monocot crops such as wheat and barley largely follows the daily temperature course, particularly under cold but humid springtime field conditions. Knowledge of the temperature response of leaf extension, particularly variations close to the thermal limit of growth, helps define physiological growth constraints and breeding-related genotypic differences among cultivars. Here, we present a novel method, called 'Leaf Length Tracker' (LLT), suitable for measuring leaf elongation rates (LERs) of cereals and other grasses with high precision and high temporal resolution under field conditions. The method is based on image sequence analysis, using a marker tracking approach to calculate LERs. We applied the LLT to several varieties of winter wheat (Triticum aestivum), summer barley (Hordeum vulgare), and ryegrass (Lolium perenne), grown in the field and in growth cabinets under controlled conditions. LLT is easy to use and we demonstrate its reliability and precision under changing weather conditions that include temperature, wind, and rain. We found that leaf growth stopped at a base temperature of 0°C for all studied species and we detected significant genotype-specific differences in LER with rising temperature. The data obtained were statistically robust and were reproducible in the tested environments. Using LLT, we were able to detect subtle differences (sub-millimeter) in leaf growth patterns. This method will allow the collection of leaf growth data in a wide range of future field experiments on different graminoid species or varieties under varying environmental or treatment conditions. PMID:26818912

  7. Dormant buds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dormant vegetative buds from diverse species can be preserved using cryopreservation. Sakai (1960) provided one of the first studies showing that winter twigs of poplar (Populus sieboldi) and willow (Salix koriyanagi) could survive low temperatures if slowly cooled prior to immersion in liquid nitr...

  8. Distinct Domains of Yeast Cortical Tag Proteins Bud8p and Bud9p Confer Polar Localization and Functionality

    PubMed Central

    Krappmann, Anne-Brit; Taheri, Naimeh; Heinrich, Melanie

    2007-01-01

    In Saccharomyces cerevisiae, diploid yeast cells follow a bipolar budding program, which depends on the two transmembrane glycoproteins Bud8p and Bud9p that potentially act as cortical tags to mark the cell poles. Here, we have performed systematic structure-function analyses of Bud8p and Bud9p to identify functional domains. We find that polar transport of Bud8p and Bud9p does not depend on N-terminal sequences but instead on sequences in the median part of the proteins and on the C-terminal parts that contain the transmembrane domains. We show that the guanosine diphosphate (GDP)/guanosine triphosphate (GTP) exchange factor Bud5p, which is essential for bud site selection and physically interacts with Bud8p, also interacts with Bud9p. Regions of Bud8p and Bud9p predicted to reside in the extracellular space are likely to confer interaction with the N-terminal region of Bud5p, implicating indirect interactions between the cortical tags and the GDP/GTP exchange factor. Finally, we have identified regions of Bud8p and Bud9p that are required for interaction with the cortical tag protein Rax1p. In summary, our study suggests that Bud8p and Bud9p carry distinct domains for delivery of the proteins to the cell poles, for interaction with the general budding machinery and for association with other cortical tag proteins. PMID:17581861

  9. Adventitious bud regeneration from leaf expiants of the shrubby ornamental honeysuckle, Lonicera nitida Wils. cv. 'Maigrün': effects of thidiazuron and 2,3,5-triiodobenzoic acid.

    PubMed

    Cambecèdes, J; Duron, M; Decourtye, L

    1991-11-01

    Different combinations of auxins and cytokinins were employed to assess the regeneration capacity from in vitro leaf explants of Lonicera nitida Wils. cv 'Maïgrün'. A high frequency of rhizogenesis was noticed, with 2.3 μM thidiazuron plus 2.9 μM indole-3-acetic acid as the only hormonal combination to support caulogenic responses. Increasing thidiazuron concentration and/or suppressing auxin did not improve caulogenesis. Combining thidiazuron with 2,3,5-triiodobenzoic acid produced a dramatic increase in the percentage of caulogenic explants. A maximum of 74% of adventitious bud forming explants was obtained with 2.3 μM thidiazuron plus 20 μM 2,3,5-triiodobenzoic acid. Buds were often in a rosette form and were vitreous, so that shoot elongation was difficult to obtain. The effect of the duration of the 2,3,5-triiodobenzoic acid treatment on shoot elongation was investigated. PMID:24221854

  10. Testing sustainability by prospective and retrospective demographic analyses: evaluation for palm leaf harvest.

    PubMed

    Zuidema, Pieter A; De Kroon, Hans; Werger, Marinus J A

    2007-01-01

    Harvesting nontimber forest products (NTFPs) is a major economic activity in tropical forests. As many NTFPs are overexploited, sustainability analyses are required to set harvest guidelines. Here we introduce and apply a new approach to evaluating sustainability, which combines prospective (elasticity) and retrospective (Life Table Response Experiments [LTRE]) demographic analyses of matrix population models. We relate the elasticity of vital rates (representing their importance for population growth rate, lamda) to their contribution to harvest-induced change in lamda ("LTRE contribution"). When high-elasticity vital rates have a low LTRE contribution, exploitation is potentially sustainable as negative effects for population growth are buffered. If the reverse is found, there is little scope for sustainability because crucial vital rates are affected. Our approach is less sensitive to chance fluctuations than the commonly used sustainability criterion of lamda = 1.0, as it does not depend on the absolute value of lamda. We applied this analysis to Geonoma deversa, a clustered forest understory palm. We studied three experimentally defoliated and control populations in a Bolivian rainforest during two years. Cutting all leaves of large ramets did not change mortality but strongly affected growth and reproduction. In spite of severe changes in some vital rates, population growth rate was not significantly reduced after defoliation. A literature review revealed that six other understory palms species responded very similarly to defoliation. The combination of LTRE contributions and elasticity analyses showed that low-elasticity vital rates were mainly responsible for the defoliation-induced change in lamda for Geonoma deversa. For two other understory palms (Astrocaryum mexicanum and Chamaedorea radicalis) new demographic analyses yielded very similar results. For Geonoma, the LTRE contribution-elasticity relation strongly changed when we mimicked harvest damage

  11. Late Quaternary climate and environmental changes in a permafrost section near Igarka, Northern Siberia based on leaf wax analyses

    NASA Astrophysics Data System (ADS)

    Schaefer, Imke; Schweri, Lea; Zech, Jana; Tananaev, Nikita; Zech, Roland

    2016-04-01

    Leaf wax biomarkers, such as long chain n-alkanes and n-alkanoic acids, and their carbon isotopic composition are a promising tool for reconstructing past climate and environmental changes and gain more and more attention in paleoresearch. Here we present the results of leaf wax analyses from a permafrost outcrop at the left banks of the Yenisei River near the city of Igarka, Northern Russia. Fluvio-glacial sediments are exposed in the lower part of the outcrop and probably date back to ~60 ka. The upper part consist of aeolian sediments deposited since, overprinted by various pedogenetic processes. First results indicate a continuous contribution of deciduous trees to the vegetation during the last glacial. Compound specific deuterium and radiocarbon analyses are in progress in order to investigate changes in paleoclimate and to establish a robust chronology.

  12. Analyses of the leaf, fruit and seed of Thaumatococcus daniefii (Benth.): exploring potential uses.

    PubMed

    Chinedu, Shalom Nwodo; Oluwadamisi, Adetayo Y; Popoola, Samuel T; David, Bolaji J; Epelle, Tamunotonyesia

    2014-06-01

    Thaumatococcus daniellii is an economic plant with versatile uses in Southern Nigeria. The arils attached to the seeds contain thaumatin, a non-sugar sweetener and taste modifier. This study examined the chemical constituents of the leaf, fruit and seed of T. daniellii. The fresh fruit, on weight basis, consists of 4.8% aril, 22.8% seed and 72.4% fleshy part. The leaf contained (per 100 g): 10.67 g moisture, 8.95 g ash, 17.21 g fat, 21.06 g protein, 24.61 g crude fiber 17.50 g carbohydrate, 0.10 g calcium, 0.08 g magnesium, 0.01 g iron and 0.37 g phosphorus. The fruit (fleshy part) contained 10.04 g moisture, 21.08 g ash, 0.93 g fat, 11.53 g protein, 18.43 g crude fiber, 37.27 g carbohydrate, 0.34 g calcium, 0.30 g magnesium, 0.01 g iron and 0.21 g phosphorus. The seed contained 15.15 g moisture, 11.30 g ash, 0.21 g fat, 10.36 g protein, 20.52 g crude fiber and 42.46 g carbohydrate. Terpenoids, flavonoids, alkaloids and cardiac glycosides were significantly present in both the leaf and fruit whereas phlobatannin, saponin, steroids, anthraquinones and ascorbic acid were absent. Tannin was present only in the leaf. The leaf and fruit of T. daniellii have significant nutritional and medicinal benefits. The leaf is rich in protein and fat. The fruit is a good source of minerals, particularly, calcium and magnesium; the leaf is also rich in phosphorus. PMID:26035959

  13. Coupling between plant leaf water and atmospheric vapor: insights from isotopic analyses

    NASA Astrophysics Data System (ADS)

    Rambo, J. P.; Lai, C.

    2012-12-01

    We measured stable isotopic composition of leaf water and atmospheric water vapor in an old growth forest in the Pacific Northwest of U.S.A. in the summer of 2011. A LGR cavity-enhanced absorption spectroscopy analyzer was used to measure hourly 18O/16O and 2H/1H ratios of atmospheric water vapor (δ18Ov and δDv) at three canopy heights (1m, 10m and 60m aboveground). Modeling studies show that transpiration plays an important role in controlling δ18Ov and δDv variation in this forest. Here we use deuterium excess (d = δD - 8 x δ18O), an isotopic tracer independent of the equilibrium process, to investigate H2O molecular diffusion and exchange between leaf water and atmospheric water vapor. We observed a robust diel pattern in the d values of atmospheric vapor (dv); its daily values typically fluctuate from -20‰ at night to +20‰ at midday. We found a strong correlation between dv and relative humidity (h). In under-saturated conditions, dv and h follow a hyperbolic relationship where values of dv increase asymptotically as h decreases. To investigate how h influences the H2O molecular exchange between leaf water and atmospheric water vapor, bulk leaf waters of dominant understory plant species were sampled every two hours for three consecutive days. Stable isotope analysis of leaf water was used to validate Craig-Gordon model calculations. Model comparisons suggest that retrodiffusion of atmospheric H2O molecules into leaf intercellular space must have intensified beginning in the mid-afternoon and continue throughout the night to counter-balance the evaporative isotopic enrichment in leaf water, which typically occurs after sunrise and peaks at midday. H2O molecular exchange occurring between an evaporating surface and water vapor limits the extent of evaporative enrichment of a finite water reservoir. This idea has been illustrated by laboratory experiments but to our best knowledge, never has been tested in field conditions. Our results demonstrate a

  14. Structure of the Bro1 Domain Protein BROX and Functional Analyses of the ALIX Bro1 Domain in HIV-1 Budding

    SciTech Connect

    Zhai Q.; Robinson H.; Landesman M. B.; Sundquist W. I.; Hill C. P.

    2011-12-01

    Bro1 domains are elongated, banana-shaped domains that were first identified in the yeast ESCRT pathway protein, Bro1p. Humans express three Bro1 domain-containing proteins: ALIX, BROX, and HD-PTP, which function in association with the ESCRT pathway to help mediate intraluminal vesicle formation at multivesicular bodies, the abscission stage of cytokinesis, and/or enveloped virus budding. Human Bro1 domains share the ability to bind the CHMP4 subset of ESCRT-III proteins, associate with the HIV-1 NC{sup Gag} protein, and stimulate the budding of viral Gag proteins. The curved Bro1 domain structure has also been proposed to mediate membrane bending. To date, crystal structures have only been available for the related Bro1 domains from the Bro1p and ALIX proteins, and structures of additional family members should therefore aid in the identification of key structural and functional elements. We report the crystal structure of the human BROX protein, which comprises a single Bro1 domain. The Bro1 domains from BROX, Bro1p and ALIX adopt similar overall structures and share two common exposed hydrophobic surfaces. Surface 1 is located on the concave face and forms the CHMP4 binding site, whereas Surface 2 is located at the narrow end of the domain. The structures differ in that only ALIX has an extended loop that projects away from the convex face to expose the hydrophobic Phe105 side chain at its tip. Functional studies demonstrated that mutations in Surface 1, Surface 2, or Phe105 all impair the ability of ALIX to stimulate HIV-1 budding. Our studies reveal similarities in the overall folds and hydrophobic protein interaction sites of different Bro1 domains, and show that a unique extended loop contributes to the ability of ALIX to function in HIV-1 budding.

  15. Characterization of Xylella fastidiosa pear leaf scorch strain in Taiwan through whole genome sequence analyses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram-negative bacterium causing diseases in many economically important crops mostly in the Americas but also in Asia and Europe. A strain of X. fastidiosa was found to cause pear leaf scorch (PLS) disease in Taiwan in 1992. Because of nutritional fastidiousness, characteriza...

  16. Proteomic and gene expression analyses during bolting-related leaf color change in Brassica rapa.

    PubMed

    Zhang, Y W; Guo, M H; Tang, X B; Jin, D; Fang, Z Y

    2016-01-01

    Bolting and flowering are key processes during the growth and development of Chinese cabbage (Brassica rapa L. ssp pekinensis). Understanding the molecular mechanisms underlying bolting and flowering is of significance for improving production of the vegetable. A leaf-color change from bright green to gray-green has been observed following differentiation of the flowering stem and before bolting in the vegetable, and is considered to be a signal for bolting. Proteomics in meristem tissues of an inbred line (C30) were analyzed by two-dimensional electrophoresis during the transition period. We found that some proteins were specifically expressed while others were differentially expressed. Among these, 17 proteins were specifically expressed before the color change, 18 were specifically expressed after the color change, 21 were downregulated during the color change, and 29 were upregulated. Mass spectrometric analysis (MALDI-TOF-TOF/MS) was used to analyze 17 protein spots, and four proteins (subunit E1 of vacuolar-type H+ transporter ATPase, the large subunit of Rubicon, S-adenosylmethionine synthetase, and tubulin α-2) were identified. qPCR analysis was conducted to quantify the expression of genes encoding these proteins during the transitional period. The expression of BrVHA-E1, BrSAMS, BrrbcL, and BrTUA6 was significantly different before and after the leaf-color change, suggesting that these genes might be involved in regulating flower differentiation and bolting. PMID:27525926

  17. Concepts and Analyses in the CT Scanning of Root Systems and Leaf Canopies: A Timely Summary

    PubMed Central

    Lafond, Jonathan A.; Han, Liwen; Dutilleul, Pierre

    2015-01-01

    Non-medical applications of computed tomography (CT) scanning have flourished in recent years, including in Plant Science. This Perspective article on CT scanning of root systems and leaf canopies is intended to be of interest to three categories of readers: those who have not yet tried plant CT scanning, and should find inspiration for new research objectives; readers who are on the learning curve with applications—here is helpful advice for them; and researchers with greater experience—the field is evolving quickly and it is easy to miss aspects. Our conclusion is that CT scanning of roots and canopies is highly demanding in terms of technology, multidisciplinarity and big-data analysis, to name a few areas of expertise, but eventually, the reward for researchers is directly proportional! PMID:26734022

  18. Concepts and Analyses in the CT Scanning of Root Systems and Leaf Canopies: A Timely Summary.

    PubMed

    Lafond, Jonathan A; Han, Liwen; Dutilleul, Pierre

    2015-01-01

    Non-medical applications of computed tomography (CT) scanning have flourished in recent years, including in Plant Science. This Perspective article on CT scanning of root systems and leaf canopies is intended to be of interest to three categories of readers: those who have not yet tried plant CT scanning, and should find inspiration for new research objectives; readers who are on the learning curve with applications-here is helpful advice for them; and researchers with greater experience-the field is evolving quickly and it is easy to miss aspects. Our conclusion is that CT scanning of roots and canopies is highly demanding in terms of technology, multidisciplinarity and big-data analysis, to name a few areas of expertise, but eventually, the reward for researchers is directly proportional! PMID:26734022

  19. Antiproliferative and phytochemical analyses of leaf extracts of ten Apocynaceae species

    PubMed Central

    Wong, Siu Kuin; Lim, Yau Yan; Abdullah, Noor Rain; Nordin, Fariza Juliana

    2011-01-01

    Background: The anticancer properties of Apocynaceae species are well known in barks and roots but less so in leaves. Materials and Methods: In this study, leaf extracts of 10 Apocynaceae species were assessed for antiproliferative (APF) activities using the sulforhodamine B assay. Their extracts were also analyzed for total alkaloid content (TAC), total phenolic content (TPC), and radical scavenging activity (RSA) using the Dragendorff precipitation, Folin–Ciocalteu, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays, respectively. Results: Leaf extracts of Alstonia angustiloba, Calotropis gigantea, Catharanthus roseus, Nerium oleander, Plumeria obtusa, and Vallaris glabra displayed positive APF activities. Extracts of Allamanda cathartica, Cerbera odollam, Dyera costulata, and Kopsia fruticosa did not show any APF activity. Dichloromethane (DCM) extract of C. gigantea, and DCM and DCM:MeOH extracts of V. glabra showed strong APF activities against all six human cancer cell lines. Against breast cancer cells of MCF-7 and MDA-MB-231, DCM extracts of C. gigantea and N. oleander were stronger than or comparable to standard drugs of xanthorrhizol, curcumin, and tamoxifen. All four extracts of N. oleander were effective against MCF-7 cells. Extracts of Kopsia fruticosa had the highest TAC while those of Dyera costulata had the highest TPC and RSA. Extracts of C. gigantea and V. glabra inhibited the growth of all six cancer cell lines while all extracts of N. oleander were effective against MCF-7 cells. Conclusion: Extracts of C. gigantea, V. glabra, and N. oleander therefore showed great promise as potential candidates for anticancer drugs. The wide-spectrum APF activities of these three species are reported for the first time and their bioactive compounds warrant further investigation. PMID:21772753

  20. Transcriptomic and Proteomic Analyses of Resistant Host Responses in Arachis diogoi Challenged with Late Leaf Spot Pathogen, Phaeoisariopsis personata

    PubMed Central

    Kumar, Dilip; Kirti, Pulugurtha Bharadwaja

    2015-01-01

    Late leaf spot is a serious disease of peanut caused by the imperfect fungus, Phaeoisariopsis personata. Wild diploid species, Arachis diogoi. is reported to be highly resistant to this disease and asymptomatic. The objective of this study is to investigate the molecular responses of the wild peanut challenged with the late leaf spot pathogen using cDNA-AFLP and 2D proteomic study. A total of 233 reliable, differentially expressed genes were identified in Arachis diogoi. About one third of the TDFs exhibit no significant similarity with the known sequences in the data bases. Expressed sequence tag data showed that the characterized genes are involved in conferring resistance in the wild peanut to the pathogen challenge. Several genes for proteins involved in cell wall strengthening, hypersensitive cell death and resistance related proteins have been identified. Genes identified for other proteins appear to function in metabolism, signal transduction and defence. Nineteen TDFs based on the homology analysis of genes associated with defence, signal transduction and metabolism were further validated by quantitative real time PCR (qRT-PCR) analyses in resistant wild species in comparison with a susceptible peanut genotype in time course experiments. The proteins corresponding to six TDFs were differentially expressed at protein level also. Differentially expressed TDFs and proteins in wild peanut indicate its defence mechanism upon pathogen challenge and provide initial breakthrough of genes possibly involved in recognition events and early signalling responses to combat the pathogen through subsequent development of resistivity. This is the first attempt to elucidate the molecular basis of the response of the resistant genotype to the late leaf spot pathogen, and its defence mechanism. PMID:25646800

  1. Microarray analyses for identifying genes conferring resistance to pepper leaf curl virus in chilli pepper (Capsicum spp.).

    PubMed

    Rai, Ved Prakash; Rai, Ashutosh; Kumar, Rajesh; Kumar, Sanjay; Kumar, Sanjeet; Singh, Major; Singh, Sheo Pratap

    2016-09-01

    Pepper leaf curl virus (PepLCV) is a serious threat to pepper (Capsicum spp.) production worldwide. Molecular mechanism underlying pepper plants response to PepLCV infection is key to develop PepLCV resistant varieties. In this study, we generated transcriptome profiles of PepLCV resistant genotype (BS-35) and susceptible genotype (IVPBC-535) after artificial viral inoculation using microarray technology and detail experimental procedures and analyses are described. A total of 319 genes differentially expressed between resistant and susceptible genotypes were identified, out of that 234 unique genes were found to be up-regulated > 2-fold in resistant line BS-35 when compared to susceptible, IVPBC-535. The data set we generated has been analyzed to identify genes that are involved in the regulation of resistance against PepLCV. The raw data have been deposited in the Gene Expression Omnibus (GEO) database under accession number GSE41131. PMID:27556012

  2. Bud Dormancy and Growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nearly all land plants produce ancillary meristems in the form of axillary or adventitious buds in addition to the shoot apical meristem. Outgrowth of these buds has a significant impact on plant architecture and the ability of plants to compete with neighboring plants, as well as to respond to and ...

  3. Tropical Storm Bud

    Atmospheric Science Data Center

    2013-04-19

    article title:  A Strengthening Eastern Pacific Storm     View Larger Image ... Imaging SpectroRadiometer (MISR) show then Tropical Storm Bud as it was intensifying toward hurricane status, which it acquired ...

  4. Influences of polar auxin transport on polarity of adventitious bud formation in hybrid populas

    SciTech Connect

    Kim, Myung Won ); Hackett, W. )

    1989-04-01

    The role of auxin and cytokinin distribution of polar regeneration of adventitious bud has been investigated. Explants from leaf midvein were labelled with {sup 14}C-NAA and {sup 14}C-BA and the radioactivity in proximal, mid, and distal portions was counted after 24h and 48h. Explants showing polar regeneration of buds on the proximal end showed a clear polar distribution of {sup 14}CNAA. Auxin transport inhibitors (NPA, TIBA) eliminated polar distribution of auxin and reduced polarity of bud formation and the total number of buds formed, but did not reduce callus formation. Increased concentration of Ca(NO{sub 3}){sub 2} decreased polarity of bud formation and increased the number of buds formed but did not affect the distribution of auxin of cytokinin. Some factor in addition to polar distribution of auxin or cytokinin-auxin ratio appears to influence the polarity of adventitious bud formation.

  5. Growth profile and SEM analyses of Candida albicans and Escherichia coli with Hymenocallis littoralis (Jacq.) Salisb leaf extract.

    PubMed

    Rosli, N; Sumathy, V; Vikneswaran, M; Sreeramanan, S

    2014-12-01

    Hymenocallis littoralis (Jacq.) Salisb (Melong kecil) commonly known as 'Spider Lily' is an herbaceous plant from the family Amaryllidaceae. Study was carried out to determine the effect of H. littoralis leaf extract on the growth and morphogenesis of two pathogenic microbes, Candida albicans and Escherichia coli. The leaf extract displayed favourable anticandidal and antibacterial activity with a minimum inhibition concentration (MIC) of 6.25 mg/mL. Time kill study showed both microbes were completely killed after treated with leaf extract at 20 h. Both microbes' cell walls were heavily ruptured based on scanning electron microscopy (SEM) analysis. The significant anticandidal and antibacterial activities showed by H. littoralis leaf extract suggested the potential antimicrobial agent against C. albicans and E. coli. PMID:25776614

  6. Dental cell sheet biomimetic tooth bud model.

    PubMed

    Monteiro, Nelson; Smith, Elizabeth E; Angstadt, Shantel; Zhang, Weibo; Khademhosseini, Ali; Yelick, Pamela C

    2016-11-01

    Tissue engineering and regenerative medicine technologies offer promising therapies for both medicine and dentistry. Our long-term goal is to create functional biomimetic tooth buds for eventual tooth replacement in humans. Here, our objective was to create a biomimetic 3D tooth bud model consisting of dental epithelial (DE) - dental mesenchymal (DM) cell sheets (CSs) combined with biomimetic enamel organ and pulp organ layers created using GelMA hydrogels. Pig DE or DM cells seeded on temperature-responsive plates at various cell densities (0.02, 0.114 and 0.228 cells 10(6)/cm(2)) and cultured for 7, 14 and 21 days were used to generate DE and DM cell sheets, respectively. Dental CSs were combined with GelMA encapsulated DE and DM cell layers to form bioengineered 3D tooth buds. Biomimetic 3D tooth bud constructs were cultured in vitro, or implanted in vivo for 3 weeks. Analyses were performed using micro-CT, H&E staining, polarized light (Pol) microscopy, immunofluorescent (IF) and immunohistochemical (IHC) analyses. H&E, IHC and IF analyses showed that in vitro cultured multilayered DE-DM CSs expressed appropriate tooth marker expression patterns including SHH, BMP2, RUNX2, tenascin and syndecan, which normally direct DE-DM interactions, DM cell condensation, and dental cell differentiation. In vivo implanted 3D tooth bud constructs exhibited mineralized tissue formation of specified size and shape, and SHH, BMP2 and RUNX2and dental cell differentiation marker expression. We propose our biomimetic 3D tooth buds as models to study optimized DE-DM cell interactions leading to functional biomimetic replacement tooth formation. PMID:27565550

  7. Berkeley UXO Discriminator (BUD)

    SciTech Connect

    Gasperikova, Erika; Smith, J. Torquil; Morrison, H. Frank; Becker, Alex

    2007-01-01

    The Berkeley UXO Discriminator (BUD) is an optimally designed active electromagnetic system that not only detects but also characterizes UXO. The system incorporates three orthogonal transmitters and eight pairs of differenced receivers. it has two modes of operation: (1) search mode, in which BUD moves along a profile and exclusively detects targets in its vicinity, providing target depth and horizontal location, and (2) discrimination mode, in which BUD, stationary above a target, from a single position, determines three discriminating polarizability responses together with the object location and orientation. The performance of the system is governed by a target size-depth curve. Maximum detection depth is 1.5 m. While UXO objects have a single major polarizability coincident with the long axis of the object and two equal transverse polarizabilities, scrap metal has three different principal polarizabilities. The results clearly show that there are very clear distinctions between symmetric intact UXO and irregular scrap metal, and that BUD can resolve the intrinsic polarizabilities of the target. The field survey at the Yuma Proving Ground in Arizona showed excellent results within the predicted size-depth range.

  8. "Bud, Not Buddy."

    ERIC Educational Resources Information Center

    Brodie, Carolyn S.

    2002-01-01

    Discusses the award-winning book "Bud, Not Buddy" written by Christopher Paul Curtis. Lists different versions of the book; suggests learning activities; lists sources for biographical information and interviews with Curtis, teacher guides, professional articles, and other Depression era novels; and provides a citation for the author's Newberry…

  9. The mode of origin of root buds and root sprouts in the clonal tree Sassafras albidum (Lauraceae).

    PubMed

    Bosela, M; Ewers, F

    1997-11-01

    The developmental anatomy of root buds and root sprouts was examined in the clonal tree Sassafras albidum. Root samples from 13 clones that varied widely in age and vigor were sectioned and two types of buds were found, "additional" buds and "reparative" buds. Additional buds form during the early growth of uninjured roots and they perennate by growing outwards in concert with the vascular cambium such that bud traces are produced in the secondary xylem. Reparative buds form de novo in response to senescence, injuries, or other types of disturbance. Reparative buds were found on the roots of seven of the clones, whereas additional buds were found on the roots of all 13 clones. The reparative buds had originated in the proliferated pericycle, where they were subtended by sphaeroblasts, or spherical nodules of wood. Few of the reparative buds were vascularized and none were connected with the vasculature of their parent roots. In contrast, most of the additional buds were vascularized, and the leaf traces of several of the additional buds appeared to be contiguous with the conducting xylem of their parent roots. To determine whether both bud types were functional, 82 field-collected root sprouts and 44 incubation-induced sprouts were sectioned at the root-sprout junction and examined for evidence relating to their mode of origin. None of the sprouts were subtended by sphaeroblasts, but 98% were subtended by bud traces, which indicated that they had originated from additional buds. Although reparative buds were more common than additional buds on some of the root samples, they appear to be dysfunctional at sprouting. Additional buds, on the other hand, are able to sprout both as a normal part of clonal spread and from root cuttings. PMID:21708553

  10. Molecular Characterization of Tomato Leaf Curl China Virus, Infecting Tomato Plants in China, and Functional Analyses of Its Associated Betasatellite▿†

    PubMed Central

    Yang, Xiuling; Guo, Wei; Ma, Xinying; An, Qianli; Zhou, Xueping

    2011-01-01

    A novel tomato-infecting begomovirus from Guangxi province, China, was identified and characterized, for which the name Tomato leaf curl China virus (ToLCCNV) was proposed. Phylogenetic and recombination analyses of the virus genomic sequences suggested that ToLCCNV may have arisen by recombination among Tomato leaf curl Vietnam virus (ToLCVV), Tomato leaf curl Gujarat virus (ToLCGV), and an unknown virus. A betasatellite molecule was found to be associated with ToLCCNV (ToLCCNB), and its complete nucleotide sequences were determined. Infectious clones of ToLCCNV and ToLCCNB were constructed and then used for agro-inoculation of plants; ToLCCNV alone infected Nicotiana benthamiana, Nicotiana glutinosa, Petunia hybrida, and Solanum lycopersicum plants, but no symptoms were induced. ToLCCNB was required for induction of leaf curl disease in these hosts. The βC1 protein of ToLCCNB was identified as a suppressor of RNA silencing and accumulated primarily in the nucleus. Deletion mutagenesis of βC1 showed that the central part of βC1 (amino acids 44 to 74) was responsible for both the suppressor activity and nuclear localization. PMID:21378048

  11. Leaf waxes, compound-specific D/H and 14C analyses in the Loess Paleosol Sequence Möhlin, Switzerland

    NASA Astrophysics Data System (ADS)

    Wüthrich, Lorenz; Bliedtner, Marcel; Kathrin Schäfer, Imke; Zech, Jana; Gaar, Dorian; Preusser, Frank; Zech, Roland

    2016-04-01

    Leaf waxes, such as long-chain n-alkanes and n-alkanoic acids, and their D/H isotopic composition, are increasingly used for paleoenvironmental and -climate reconstructions. Recent technological innovations now also allow to perform radiocarbon analyses on leaf waxes. For this study, we analyzed leaf waxes and their δD and 14C composition in the 7 m Loess Paleosol Sequence Möhlin, Switzerland. The chain length patterns in the upper part of the profile indicate n-alkane contribution from deciduous trees, while the underlying loess is dominated by inputs from grasses and herbs. Our δD record does not show depleted, glacial values compared to the Holocene, as we had expected in analogy to the Greenland ice core records. Values are most enriched at 1 m depth, i.e. well below the topsoil. Further research is needed to disentangle source effects and evapotranspirative enrichment, before the δD record can be interpreted robustly. Our radiocarbon ages for the leaf waxes are in very good agreement with independent age control based on luminescence ages, corroborating that massive loess accumulation occurred already at 35 ka. Only the uppermost 3 m were deposited during the last glacial maximum.

  12. Leaf waxes and compound-specific δD analyses in a Holocene fluvial sediment-paleosol sequence from the upper Alazani River, SE-Georgia

    NASA Astrophysics Data System (ADS)

    Bliedtner, Marcel; von Suchodoletz, Hans; Schäfer, Imke; Zech, Jana; Zielhofer, Christoph; Zech, Roland

    2016-04-01

    Leaf waxes of terrestrial plants are relatively resistant against degradation and serve as valuable biomarkers preserved in various sedimentary archives. Compound-specific D/H analyses on leaf waxes are increasingly used to reconstruct past climate and environmental conditions. Here, we present a n-alkane and compound-specific δD record from a Holocene fluvial sediment-paleosol sequence along the upper Alazani River in eastern Georgia. Generally, such records from fluvial sedimentary archives must be divided into a catchment signal recorded in the fluvial sediment layers and a local in-situ signal recorded in the intercalated paleosols. The n-alkane homologue pattern shows a clear catchment versus in-situ signal. The paleosols are dominated by n-alkanes derived from the local vegetation, mainly grasses throughout the Holocene. The fluvial sediment layers contain leaf waxes derived from the forested catchment, although with relatively high contributions from grasses between 8 and 5 ka, possibly indicating more arid conditions. Because of the well-known altitude-effect on the isotopic composition of precipitation, we had expected more depleted δD values for the fluvial sediment layers, i.e. the catchment-derived samples, and more enriched δD values for the paleosols, i.e. the low altitude, in-situ signal. This is, however, not the case, and we speculate that the altitude effect might be offset by evapo-transpirative enrichment of tree leaf water and leaf waxes. The catchment and in-situ δD records both show a minor trend to more enriched values during the Holocene, while the recent topsoil is most depleted. Interpretation of these isotope records is not straight-forward and requires disentangling the effects of changing vegetation, source signal and local climate.

  13. The barley Uniculme4 gene encodes a BLADE-ON-PETIOLE-like protein that controls tillering and leaf patterning.

    PubMed

    Tavakol, Elahe; Okagaki, Ron; Verderio, Gabriele; Shariati J, Vahid; Hussien, Ahmed; Bilgic, Hatice; Scanlon, Mike J; Todt, Natalie R; Close, Timothy J; Druka, Arnis; Waugh, Robbie; Steuernagel, Burkhard; Ariyadasa, Ruvini; Himmelbach, Axel; Stein, Nils; Muehlbauer, Gary J; Rossini, Laura

    2015-05-01

    Tillers are vegetative branches that develop from axillary buds located in the leaf axils at the base of many grasses. Genetic manipulation of tillering is a major objective in breeding for improved cereal yields and competition with weeds. Despite this, very little is known about the molecular genetic bases of tiller development in important Triticeae crops such as barley (Hordeum vulgare) and wheat (Triticum aestivum). Recessive mutations at the barley Uniculme4 (Cul4) locus cause reduced tillering, deregulation of the number of axillary buds in an axil, and alterations in leaf proximal-distal patterning. We isolated the Cul4 gene by positional cloning and showed that it encodes a BROAD-COMPLEX, TRAMTRACK, BRIC-À-BRAC-ankyrin protein closely related to Arabidopsis (Arabidopsis thaliana) BLADE-ON-PETIOLE1 (BOP1) and BOP2. Morphological, histological, and in situ RNA expression analyses indicate that Cul4 acts at axil and leaf boundary regions to control axillary bud differentiation as well as the development of the ligule, which separates the distal blade and proximal sheath of the leaf. As, to our knowledge, the first functionally characterized BOP gene in monocots, Cul4 suggests the partial conservation of BOP gene function between dicots and monocots, while phylogenetic analyses highlight distinct evolutionary patterns in the two lineages. PMID:25818702

  14. Histological and Molecular Characterization of Grape Early Ripening Bud Mutant

    PubMed Central

    Yu, Yi-He; Xi, Fei-Fei; Shi, Yan-Yan; Zhang, Guo-Hai

    2016-01-01

    An early ripening bud mutant was analyzed based on the histological, SSR, and methylation-sensitive amplified polymorphism (MSAP) analysis and a layer-specific approach was used to investigate the differentiation between the bud mutant and its parent. The results showed that the thickness of leaf spongy tissue of mutant (MT) is larger than that of wild type (WT) and the differences are significant. The mean size of cell layer L2 was increased in the mutant and the difference is significant. The genetic background of bud mutant revealed by SSR analysis is highly uniform to its parent; just the variations from VVS2 SSR marker were detected in MT. The total methylation ratio of MT is lower than that of the corresponding WT. The outside methylation ratio in MT is much less than that in WT; the average inner methylation ratio in MT is larger than that in WT. The early ripening bud mutant has certain proportion demethylation in cell layer L2. All the results suggested that cell layer L2 of the early ripening bud mutant has changed from the WT. This study provided the basis for a better understanding of the characteristic features of the early ripening bud mutant in grape. PMID:27610363

  15. Histological and Molecular Characterization of Grape Early Ripening Bud Mutant.

    PubMed

    Guo, Da-Long; Yu, Yi-He; Xi, Fei-Fei; Shi, Yan-Yan; Zhang, Guo-Hai

    2016-01-01

    An early ripening bud mutant was analyzed based on the histological, SSR, and methylation-sensitive amplified polymorphism (MSAP) analysis and a layer-specific approach was used to investigate the differentiation between the bud mutant and its parent. The results showed that the thickness of leaf spongy tissue of mutant (MT) is larger than that of wild type (WT) and the differences are significant. The mean size of cell layer L2 was increased in the mutant and the difference is significant. The genetic background of bud mutant revealed by SSR analysis is highly uniform to its parent; just the variations from VVS2 SSR marker were detected in MT. The total methylation ratio of MT is lower than that of the corresponding WT. The outside methylation ratio in MT is much less than that in WT; the average inner methylation ratio in MT is larger than that in WT. The early ripening bud mutant has certain proportion demethylation in cell layer L2. All the results suggested that cell layer L2 of the early ripening bud mutant has changed from the WT. This study provided the basis for a better understanding of the characteristic features of the early ripening bud mutant in grape. PMID:27610363

  16. Xenopus Limb bud morphogenesis.

    PubMed

    Keenan, Samuel R; Beck, Caroline W

    2016-03-01

    Xenopus laevis, the South African clawed frog, is a well-established model organism for the study of developmental biology and regeneration due to its many advantages for both classical and molecular studies of patterning and morphogenesis. While contemporary studies of limb development tend to focus on models developed from the study of chicken and mouse embryos, there are also many classical studies of limb development in frogs. These include both fate and specification maps, that, due to their age, are perhaps not as widely known or cited as they should be. This has led to some inevitable misinterpretations- for example, it is often said that Xenopus limb buds have no apical ectodermal ridge, a morphological signalling centre located at the distal dorsal/ventral epithelial boundary and known to regulate limb bud outgrowth. These studies are valuable both from an evolutionary perspective, because amphibians diverged early from the amniote lineage, and from a developmental perspective, as amphibian limbs are capable of regeneration. Here, we describe Xenopus limb morphogenesis with reference to both classical and molecular studies, to create a clearer picture of what we know, and what is still mysterious, about this process. PMID:26404044

  17. Resistance to Cucurbit Leaf Crumple Virus in Melon

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cucurbit leaf crumple virus (CuLCrV) is a geminivirus common in melons (Cucumis melo L.) planted from July through September in the desert southwest U.S.A. Symptoms include chlorotic leaf spots and terminal buds, leaf curling and crumpling and interveinal yellowing, and plants may be stunted in size...

  18. Phenological synchrony between Scaphoideus titanus (Hemiptera: Cicadellidae) hatchings and grapevine bud break: could this explain the insect's expansion?

    PubMed

    Chuche, J; Desvignes, E; Bonnard, O; Thiéry, D

    2015-02-01

    Scaphoideus titanus is the invasive vector of the phytoplasma causing the Flavescence dorée in European vineyards. This epidemic is a serious threat to viticulture that has been increasing for more than 60 years in Europe. We studied the effect of synchrony with the plant phenology and the effect of plant-sap quality on the individual fitness. Thus, we conducted laboratory experiments to determine if insect hatchings were synchronized with grapevine bud break. We used two natural populations: one from a cold winter vineyard and one from a mild winter vineyard. In both cases, egg hatching was synchronized with bud break and leaf appearance. The phloem quality of the young and old leaves as a food source was analysed by high-performance liquid chromatography, and the effects on S. titanus growth were evaluated. Phloem composition varied with the grapevine cutting's age but also varied between leaves of different ages from the same plant. The older leaves were less nutritious because they had the highest carbon-to-nitrogen ratio and the lowest content of essential amino acids. Despite diverse phloem qualities, no fitness difference was observed. We found that the synchronization of egg hatchings with bud break is well regulated. However, the nymphs are not affected by the phloem-sap quality, suggesting that S. titanus may accept different food qualities and that egg hatching synchrony could contribute to population expansion in vineyards. PMID:25339543

  19. The Barley Uniculme4 Gene Encodes a BLADE-ON-PETIOLE-Like Protein That Controls Tillering and Leaf Patterning1[OPEN

    PubMed Central

    Tavakol, Elahe; Okagaki, Ron; Verderio, Gabriele; Shariati J., Vahid; Hussien, Ahmed; Bilgic, Hatice; Scanlon, Mike J.; Todt, Natalie R.; Close, Timothy J.; Druka, Arnis; Waugh, Robbie; Steuernagel, Burkhard; Ariyadasa, Ruvini; Himmelbach, Axel; Stein, Nils; Muehlbauer, Gary J.

    2015-01-01

    Tillers are vegetative branches that develop from axillary buds located in the leaf axils at the base of many grasses. Genetic manipulation of tillering is a major objective in breeding for improved cereal yields and competition with weeds. Despite this, very little is known about the molecular genetic bases of tiller development in important Triticeae crops such as barley (Hordeum vulgare) and wheat (Triticum aestivum). Recessive mutations at the barley Uniculme4 (Cul4) locus cause reduced tillering, deregulation of the number of axillary buds in an axil, and alterations in leaf proximal-distal patterning. We isolated the Cul4 gene by positional cloning and showed that it encodes a BROAD-COMPLEX, TRAMTRACK, BRIC-À-BRAC-ankyrin protein closely related to Arabidopsis (Arabidopsis thaliana) BLADE-ON-PETIOLE1 (BOP1) and BOP2. Morphological, histological, and in situ RNA expression analyses indicate that Cul4 acts at axil and leaf boundary regions to control axillary bud differentiation as well as the development of the ligule, which separates the distal blade and proximal sheath of the leaf. As, to our knowledge, the first functionally characterized BOP gene in monocots, Cul4 suggests the partial conservation of BOP gene function between dicots and monocots, while phylogenetic analyses highlight distinct evolutionary patterns in the two lineages. PMID:25818702

  20. Seasonal and inter-annual variability of bud development as related to climate in two coexisting Mediterranean Quercus species

    PubMed Central

    Alla, Arben Q.; Camarero, J. Julio; Montserrat-Martí, Gabriel

    2013-01-01

    Background and Aims In trees, bud development is driven by endogenous and exogenous factors such as species and climate, respectively. However, knowledge is scarce on how these factors drive changes in bud size across different time scales. Methods The seasonal patterns of apical bud enlargement are related to primary and secondary growth in two coexisting Mediterranean oaks with contrasting leaf habit (Quercus ilex, evergreen; Quercus faginea, deciduous) over three years. In addition, the climatic factors driving changes in bud size of the two oak species were determined by correlating bud mass with climatic variables at different time scales (from 5 to 30 d) over a 15-year period. Key Results The maximum enlargement rate of buds was reached between late July and mid-August in both species. Moreover, apical bud size increased with minimum air temperatures during the period of maximum bud enlargement rates. Conclusions The forecasted rising minimum air temperatures predicted by climatic models may affect bud size and consequently alter crown architecture differentially in sympatric Mediterranean oaks. However, the involvement of several drivers controlling the final size of buds makes it difficult to predict the changes in bud size as related to ongoing climate warming. PMID:23179859

  1. Proteomic and Metabolomic Analyses of Leaf from Clematis terniflora DC. Exposed to High-Level Ultraviolet-B Irradiation with Dark Treatment.

    PubMed

    Yang, Bingxian; Wang, Xin; Gao, Cuixia; Chen, Meng; Guan, Qijie; Tian, Jingkui; Komatsu, Setsuko

    2016-08-01

    Clematis terniflora DC. has potential pharmaceutical value; on the contrary, high-level UV-B irradiation with dark treatment led to the accumulation of secondary metabolites. Metabolomic and proteomic analyses of leaf of C. terniflora were performed to investigate the systematic response mechanisms to high-level UV-B irradiation with dark treatment. Metabolites related to carbohydrates, fatty acids, and amino acids and/or proteins related to stress, cell wall, and amino acid metabolism were gradually increased in response to high-level UV-B irradiation with dark treatment. On the basis of cluster analysis and mapping of proteins related to amino acid metabolism, the abundances of S-adenosylmethionine synthetase and cysteine synthase as well as 1,1-diphenyl-2-picrylhydrazyl scavenging activity were gradually increased in response to high-level UV-B irradiation with dark treatment. Furthermore, the abundance of dihydrolipoyl dehydrogenase/glutamate dehydrogenase and the content of γ-aminobutyric acid were also increased following high-level UV-B irradiation with dark treatment. Taken together, these results suggest that high-level UV-B irradiation with dark treatment induces the activation of reactive oxygen species scavenging system and γ-aminobutyric acid shunt pathway in leaf of C. terniflora. PMID:27323210

  2. Project BudBurst: Citizen Science for All Seasons

    NASA Astrophysics Data System (ADS)

    Henderson, S.; Brewer, C.; Havens, K.; Meymaris, K.

    2007-12-01

    Project BudBurst is a national citizen science initiative designed to engage the public in observations of phenological (plant life cycle) events that raise awareness of climate change, and create a cadre of informed citizen scientists. Citizen science programs such as Project BudBurst provide the opportunity for students and interested laypersons to actively participate in scientific research. Such programs are important not only from an educational perspective, but because they also enable scientists to broaden the geographic and temporal scale of their observations. Project BudBurst launched a pilot program in the Spring of 2007. The goals of Project BudBurst were to 1) increase awareness of phenology as an area of scientific study; 2) Increase awareness of the impacts of changing climates on plants; and 3) increase science literacy by engaging participants in the scientific process. From April through mid-June 2007, this on-line educational and data-entry program, engaged participants of all ages and walks of life in recording the timing of the leafing and flowering of ~60 easily identifiable, broadly distributed wild and cultivated species found across the continent. We will report on the results of the pilot project and discuss plans to expand Project BudBurst as it becomes a year round event beginning in 2008. A broad consortium of collaborators, representing the Chicago Botanic Garden, Plant Conservation Alliance, ESRI, the USA-National Phenology Network, University Corporation for Atmospheric Research, University of Arizona, University of Montana, University of California-Santa Barbara, University of Wisconsin-Milwaukee and the University of Wisconsin-Madison, came together to design and implement Project BudBurst with seed funding from the U.S. Bureau of Land Management, the National Phenology Network (through a RCN grant from the NSF), and the Plant Conservation Alliance.

  3. The Anillin-Related Region of Bud4 Is the Major Functional Determinant for Bud4's Function in Septin Organization during Bud Growth and Axial Bud Site Selection in Budding Yeast

    PubMed Central

    Wu, Huan; Guo, Jia; Zhou, Ya-Ting

    2015-01-01

    The anillin-related protein Bud4 of Saccharomyces cerevisiae is required for axial bud site selection by linking the axial landmark to the septins, which localize at the mother bud neck. Recent studies indicate that Bud4 plays a role in septin organization during cytokinesis. Here we show that Bud4 is also involved in septin organization during bud growth prior to cytokinesis, as bud4Δ shs1Δ cells displayed an elongated bud morphology and defective septin organization at 18°C. Bud4 overexpression also affected septin organization during bud growth in shs1Δ cells at 30°C. Bud4 was previously thought to associate with the septins via its central region, while the C-terminal anillin-related region was not involved in septin association. Surprisingly, we found that the central region of Bud4 alone targets to the bud neck throughout the cell cycle, unlike full-length Bud4, which localizes to the bud neck only during G2/M phase. We identified the anillin-related region to be a second targeting domain that cooperates with the central region for proper septin association. In addition, the anillin-related region could largely mediate Bud4's function in septin organization during bud growth and bud site selection. We show that this region interacts with the C terminus of Bud3 and the two segments depend on each other for association with the septins. Moreover, like the bud4Δ mutant, the bud3Δ mutant genetically interacts with shs1Δ and cdc12-6 mutants in septin organization, suggesting that Bud4 and Bud3 may cooperate in septin organization during bud growth. These observations provide new insights into the interaction of Bud4 with the septins and Bud3. PMID:25576483

  4. The anillin-related region of Bud4 is the major functional determinant for Bud4's function in septin organization during bud growth and axial bud site selection in budding yeast.

    PubMed

    Wu, Huan; Guo, Jia; Zhou, Ya-Ting; Gao, Xiang-Dong

    2015-03-01

    The anillin-related protein Bud4 of Saccharomyces cerevisiae is required for axial bud site selection by linking the axial landmark to the septins, which localize at the mother bud neck. Recent studies indicate that Bud4 plays a role in septin organization during cytokinesis. Here we show that Bud4 is also involved in septin organization during bud growth prior to cytokinesis, as bud4Δ shs1Δ cells displayed an elongated bud morphology and defective septin organization at 18°C. Bud4 overexpression also affected septin organization during bud growth in shs1Δ cells at 30°C. Bud4 was previously thought to associate with the septins via its central region, while the C-terminal anillin-related region was not involved in septin association. Surprisingly, we found that the central region of Bud4 alone targets to the bud neck throughout the cell cycle, unlike full-length Bud4, which localizes to the bud neck only during G2/M phase. We identified the anillin-related region to be a second targeting domain that cooperates with the central region for proper septin association. In addition, the anillin-related region could largely mediate Bud4's function in septin organization during bud growth and bud site selection. We show that this region interacts with the C terminus of Bud3 and the two segments depend on each other for association with the septins. Moreover, like the bud4Δ mutant, the bud3Δ mutant genetically interacts with shs1Δ and cdc12-6 mutants in septin organization, suggesting that Bud4 and Bud3 may cooperate in septin organization during bud growth. These observations provide new insights into the interaction of Bud4 with the septins and Bud3. PMID:25576483

  5. Spectral and HRTEM analyses of Annona muricata leaf extract mediated silver nanoparticles and its Larvicidal efficacy against three mosquito vectors Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti.

    PubMed

    Santhosh, Shanthi Bhupathi; Ragavendran, Chinnasamy; Natarajan, Devarajan

    2015-12-01

    Mosquitoes transmit various diseases which mainly affect the human beings and every year cause millions of deaths globally. Currently available chemical and synthetic mosquitocidal agents pose severe side effects, pollute the environment vigorously, and become resistance. There is an urgent need to identify and develop the cost effective, compatible and eco-friendly product for mosquito control. The present study was aimed to find out the larvicidal potential of aqueous crude extract and green synthesized silver nanoparticles (AgNPs) from Annona muricata leaves were tested against fourth instar larvae of three important mosquitoes i.e. Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti using different concentrations of AgNPs (10, 20, 30, 40 and 50 ppm) and the aqueous leaf extract (100, 200, 300, 400, and 500 ppm) for 24 and 48 h. The maximum mortality was noticed in AgNPs than aqueous leaf extract of A. muricata against tested mosquitoes with least LC50 values of 37.70, 31.29, and 20.65 ppm (24h) and 546.7, 516.2, and 618.4 ppm (48 h), respectively. All tested concentrations of AgNps exhibited 100% mortality in A. aegypti larvae at 48 hour observation. In addition, the plant mediated AgNPs were characterized by UV-vis spectrum, Fourier transform infrared spectroscopy, particle size analyser, X-ray diffraction, high resonance transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis for confirmation of nanoparticle synthesis. Based on the findings of the study suggests that the use of A. muricata plant mediated AgNPs can act as an alternate insecticidal agents for controlling target mosquitoes. PMID:26410042

  6. Glycoconjugate in rat taste buds.

    PubMed

    Kano, K; Ube, M; Taniguchi, K

    2001-05-01

    The taste buds of the fungiform papillae, circumvallate papilla, foliate papillae, soft palate and epiglottis of the rat oral cavity were examined by lectin histochemistry to elucidate the relationships between expression of glycoconjugates and innervation. Seven out of 21 lectins showed moderate to intense staining in at least more than one taste bud. They were succinylated wheat germ agglutinin (s-WGA). Dolichos biflorus agglutinin (DBA), Bandeiraea simplicifolia lectin-I (BSL-I), Ricinus communis agglutinin-I (RCA-I), peanut agglutinin (PNA), Ulex europaeus agglutinin-I (UEA-I) and Phaseolus vulgaris agglutinin-L (PHA-L). UEA-I and BSL-I showed moderate to intense staining in all of the taste buds examined. They strongly stained the taste buds of the epiglottis, which are innervated by the cranial nerve X. UEA-I intensely stained the taste buds of the fungiform papillae and soft palate, both of which are innervated by the cranial nerve VII. The taste buds of circumvallate papilla and foliate papillae were innervated by the cranial nerve IX and strongly stained by BSL-I. Thus, UEA-I and BSL-I binding glycoconjugates, probably alpha-linked fucose and alpha-D-galactose, respectively, might be specific for taste buds. Although the expression of these glycoconjugates would be related to the innervation of the cranial nerve X, the differential expression of alpha-linked fucose and alpha-D-galactose might be related to the innervation of the cranial nerve VII and IX, respectively. PMID:11411494

  7. Project BudBurst: People, Plants, and Climate Change

    NASA Astrophysics Data System (ADS)

    Henderson, S.; Ward, D.; Havens, K.; Gardiner, L. S.; Alaback, P.

    2010-12-01

    Providing opportunities for individuals to contribute to a better understanding of climate change is the hallmark of Project BudBurst (www.budburst.org). This highly successful, national citizen science program, now in its third year, is bringing climate change education outreach to thousands of individuals. Project BudBurst is a national citizen science initiative designed to engage the public in observations of phenological (plant life cycle) events that raise awareness of climate change, and create a cadre of informed citizen scientists. Citizen science programs such as Project BudBurst provide the opportunity for students and interested laypersons to actively participate in scientific research. Such programs are important not only from an educational perspective, but because they also enable scientists to broaden the geographic and temporal scale of their observations. The goals of Project BudBurst are to 1) increase awareness of phenology as an area of scientific study; 2) Increase awareness of the impacts of changing climates on plants; and 3) increase science literacy by engaging participants in the scientific process. From its 2008 launch in February, this on-line educational and data-entry program, engaged participants of all ages and walks of life in recording the timing of the leafing and flowering of wild and cultivated species found across the continent. Thus far, thousands of participants from all 50 states have submitted data. Project BudBurst has been the subject of almost 200 media outlets including NPR, national and regional television broadcasts, and most of the major national and regional newspapers. This presentation will provide an overview of Project BudBurst and will report on the results of the 2009 field campaign and discuss plans to expand Project BudBurst in 2010 including the use of mobile phones applications for data collection and reporting from the field. Project BudBurst co managed by the National Ecological Observatory Network and

  8. Dentition development and budding morphogenesis.

    PubMed

    Peterková, R; Peterka, M; Viriot, L; Lesot, H

    2000-01-01

    The development of functional teeth in the mouse has been widely used as a model to study general mechanisms of organogenesis. Compared with other mammals, in which three incisors, one canine, four premolars, and three molars may occur even in each dental quadrant, the mouse functional dentition is strongly reduced. It comprises only one incisor separated from three molars by a toothless gap diastema at the location of the missing teeth. However, mouse embryos also develop transient vestigial dental primordia between the incisor and molar germs in both the upper and lower jaws. These rudimental structures regress, and epithelial apoptosis is involved in this process. The existence of the vestigial dental structures allowed a better assessment of the periodicity in the mouse dentition, which extends opportunities for the interpretation of molecular data on tooth development. We compared the dentition development with tentative models of budding morphogenesis in other epithelial appendages lungs and feathers. We suggested how developmental control by signaling molecules, including bone morphogenetic protein (Bmp), sonic hedgehog (Shh), and fibroblast growth factor (Fgf), can be similarly involved during budding morphogenesis of dentition and other epithelial appendages. We propose that epithelial apoptosis plays an important role in achieving specific features of dentition, whose development involves both budding and its more complex variant branching. The failure of segregation of the originating buds supports the participation of the concrescence of several tooth primordia in the evolutionary differentiation of mammalian teeth. PMID:11354512

  9. Bud-Neck Scaffolding as a Possible Driving Force in ESCRT-Induced Membrane Budding

    PubMed Central

    Mercker, Moritz; Marciniak-Czochra, Anna

    2015-01-01

    Membrane budding is essential for processes such as protein sorting and transport. Recent experimental results with ESCRT proteins reveal a novel budding mechanism, with proteins emerging in bud necks but separated from the entire bud surface. Using an elastic model, we show that ESCRT protein shapes are sufficient to spontaneously create experimentally observed structures, with protein-membrane interactions leading to protein scaffolds in bud-neck regions. Furthermore, the model reproduces experimentally observed budding directions and bud sizes. Finally, our results reveal that membrane-mediated sorting has the capability of creating structures more complicated than previously assumed. PMID:25692588

  10. Project BudBurst: Citizen Science for All Seasons

    NASA Astrophysics Data System (ADS)

    Meymaris, K.; Henderson, S.; Alaback, P.; Havens, K.

    2008-12-01

    Providing opportunities for individuals to contribute to a better understanding of climate change is the hallmark of Project BudBurst (www.budburst.org). This highly successful, national citizen science program, now in its second year, is bringing climate change education outreach to thousands of individuals. Project BudBurst is a national citizen science initiative designed to engage the public in observations of phenological (plant life cycle) events that raise awareness of climate change, and create a cadre of informed citizen scientists. Citizen science programs such as Project BudBurst provide the opportunity for students and interested laypersons to actively participate in scientific research. Such programs are important not only from an educational perspective, but because they also enable scientists to broaden the geographic and temporal scale of their observations. The goals of Project BudBurst are to 1) increase awareness of phenology as an area of scientific study; 2) Increase awareness of the impacts of changing climates on plants; and 3) increase science literacy by engaging participants in the scientific process. From its 2008 launch in February, this on-line educational and data-entry program, engaged participants of all ages and walks of life in recording the timing of the leafing and flowering of wild and cultivated species found across the continent. Thus far, participants from 49 states have submitted data that is being submitted to the USA National Phenology Network (www.usanpn.org) database. Project BudBurst has been the subject of almost 200 media outlets including NPR, national and regional television broadcasts, and most of the major national and regional newspapers. This presentation will provide an overview of Project Budburst and will report on the results of the 2008 field campaign and discuss plans to expand Project BudBurst in 2009. Project BudBurst is a Windows to the Universe Citizen Science program managed by the University

  11. Project BudBurst: Continental-scale citizen science for all seasons

    NASA Astrophysics Data System (ADS)

    Henderson, S.; Newman, S. J.; Ward, D.; Havens-Young, K.; Alaback, P.; Meymaris, K.

    2011-12-01

    Project BudBurst's (budburst.org) recent move to the National Ecological Observatory Network (NEON) has benefitted both programs. NEON has been able to use Project BudBurst as a testbed to learn best practices, network with experts in the field, and prototype potential tools for engaging people in continental-scale ecology as NEON develops its citizen science program. Participation in Project BudBurst has grown significantly since the move to NEON. Project BudBurst is a national citizen science initiative designed to engage the public in observations of phenological (plant life cycle) events that raise awareness of climate change, and create a cadre of informed citizen scientists. Citizen science programs such as Project BudBurst provide the opportunity for students and interested laypersons to actively participate in scientific research. Such programs are important not only from an educational perspective, but because they also enable scientists to broaden the geographic and temporal scale of their observations. The goals of Project BudBurst are to 1) increase awareness of phenology as an area of scientific study; 2) Increase awareness of the impacts of changing climates on plants at a continental-scale; and 3) increase science literacy by engaging participants in the scientific process. From its 2008 launch in February, this on-line educational and data-entry program, engaged participants of all ages and walks of life in recording the timing of the leafing and flowering of wild and cultivated species found across the continent. Thus far, thousands of participants from all 50 states have submitted data. This presentation will provide an overview of Project BudBurst and will report on the results of the 2010 field campaign and discuss plans to expand Project BudBurst in 2012 including the use of mobile phones applications for data collection and reporting from the field. Project BudBurst is co-managed by the National Ecological Observatory Network and the Chicago

  12. The Energy of COPI for Budding Membranes

    PubMed Central

    Thiam, Abdou Rachid; Pincet, Frédéric

    2015-01-01

    As a major actor of cellular trafficking, COPI coat proteins assemble on membranes and locally bend them to bud 60 nm-size coated particles. Budding requires the energy of the coat assembly to overcome the one necessary to deform the membrane which primarily depends on the bending modulus and surface tension, γ. Using a COPI-induced oil nanodroplet formation approach, we modulated the budding of nanodroplets using various amounts and types of surfactant. We found a Heaviside-like dependence between the budding efficiency and γ: budding was only dependent on γ and occurred beneath 1.3 mN/m. With the sole contribution of γ to the membrane deformation energy, we assessed that COPI supplies ~1500 kBT for budding particles from membranes, which is consistent with common membrane deformation energies. Our results highlight how a simple remodeling of the composition of membranes could mechanically modulate budding in cells. PMID:26218078

  13. Coevolutionary patterning of teeth and taste buds

    PubMed Central

    Bloomquist, Ryan F.; Parnell, Nicholas F.; Phillips, Kristine A.; Fowler, Teresa E.; Yu, Tian Y.; Sharpe, Paul T.; Streelman, J. Todd

    2015-01-01

    Teeth and taste buds are iteratively patterned structures that line the oro-pharynx of vertebrates. Biologists do not fully understand how teeth and taste buds develop from undifferentiated epithelium or how variation in organ density is regulated. These organs are typically studied independently because of their separate anatomical location in mammals: teeth on the jaw margin and taste buds on the tongue. However, in many aquatic animals like bony fishes, teeth and taste buds are colocalized one next to the other. Using genetic mapping in cichlid fishes, we identified shared loci controlling a positive correlation between tooth and taste bud densities. Genome intervals contained candidate genes expressed in tooth and taste bud fields. sfrp5 and bmper, notable for roles in Wingless (Wnt) and bone morphogenetic protein (BMP) signaling, were differentially expressed across cichlid species with divergent tooth and taste bud density, and were expressed in the development of both organs in mice. Synexpression analysis and chemical manipulation of Wnt, BMP, and Hedgehog (Hh) pathways suggest that a common cichlid oral lamina is competent to form teeth or taste buds. Wnt signaling couples tooth and taste bud density and BMP and Hh mediate distinct organ identity. Synthesizing data from fish and mouse, we suggest that the Wnt-BMP-Hh regulatory hierarchy that configures teeth and taste buds on mammalian jaws and tongues may be an evolutionary remnant inherited from ancestors wherein these organs were copatterned from common epithelium. PMID:26483492

  14. Coevolutionary patterning of teeth and taste buds.

    PubMed

    Bloomquist, Ryan F; Parnell, Nicholas F; Phillips, Kristine A; Fowler, Teresa E; Yu, Tian Y; Sharpe, Paul T; Streelman, J Todd

    2015-11-01

    Teeth and taste buds are iteratively patterned structures that line the oro-pharynx of vertebrates. Biologists do not fully understand how teeth and taste buds develop from undifferentiated epithelium or how variation in organ density is regulated. These organs are typically studied independently because of their separate anatomical location in mammals: teeth on the jaw margin and taste buds on the tongue. However, in many aquatic animals like bony fishes, teeth and taste buds are colocalized one next to the other. Using genetic mapping in cichlid fishes, we identified shared loci controlling a positive correlation between tooth and taste bud densities. Genome intervals contained candidate genes expressed in tooth and taste bud fields. sfrp5 and bmper, notable for roles in Wingless (Wnt) and bone morphogenetic protein (BMP) signaling, were differentially expressed across cichlid species with divergent tooth and taste bud density, and were expressed in the development of both organs in mice. Synexpression analysis and chemical manipulation of Wnt, BMP, and Hedgehog (Hh) pathways suggest that a common cichlid oral lamina is competent to form teeth or taste buds. Wnt signaling couples tooth and taste bud density and BMP and Hh mediate distinct organ identity. Synthesizing data from fish and mouse, we suggest that the Wnt-BMP-Hh regulatory hierarchy that configures teeth and taste buds on mammalian jaws and tongues may be an evolutionary remnant inherited from ancestors wherein these organs were copatterned from common epithelium. PMID:26483492

  15. Cotton buds, momentum, and impulse

    NASA Astrophysics Data System (ADS)

    van den Berg, Ed; Nuñez, Jover; Guirit, Alfredo; van Huis, Cor

    2000-01-01

    Here is a simple experiment demonstrating impulse and momentum that was picked up from a Japanese presenter at a physics teacher conference held in Cebu City. We have not been able to trace the experiment farther and have never seen it in print. After student-author Nuñez demonstrated it during an exam on conducting demonstrations, we converted the qualitative idea into a quanitative experiment and even discovered some possibilities for student research. The lab is also suitable as homework, since it uses universally available "equipment" — cotton buds (swabs), drinking straws, and a ruler.

  16. Experimental evolution in budding yeast

    NASA Astrophysics Data System (ADS)

    Murray, Andrew

    2012-02-01

    I will discuss our progress in analyzing evolution in the budding yeast, Saccharomyces cerevisiae. We take two basic approaches. The first is to try and examine quantitative aspects of evolution, for example by determining how the rate of evolution depends on the mutation rate and the population size or asking whether the rate of mutation is uniform throughout the genome. The second is to try to evolve qualitatively novel, cell biologically interesting phenotypes and track the mutations that are responsible for the phenotype. Our efforts include trying to alter cell morphology, evolve multicellularity, and produce a biological oscillator.

  17. Whole-Transcriptome Analysis of Differentially Expressed Genes in the Vegetative Buds, Floral Buds and Buds of Chrysanthemum morifolium

    PubMed Central

    Liu, Hua; Sun, Ming; Du, Dongliang; Pan, Huitang; Cheng, Tangren; Wang, Jia; Zhang, Qixiang

    2015-01-01

    Background Chrysanthemum morifolium is an important floral crop that is cultivated worldwide. However, due to a lack of genomic resources, very little information is available concerning the molecular mechanisms of flower development in chrysanthemum. Results The transcriptomes of chrysanthemum vegetative buds, floral buds and buds were sequenced using Illumina paired-end sequencing technology. A total of 15.4 Gb of reads were assembled into 91,367 unigenes with an average length of 739 bp. A total of 43,137 unigenes showed similarity to known proteins in the Swissprot or NCBI non-redundant protein databases. Additionally, 25,424, 24,321 and 13,704 unigenes were assigned to 56 gene ontology (GO) categories, 25 EuKaryotic Orthologous Groups (KOG) categories, and 285 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, respectively. A total of 1,876 differentially expressed genes (DEGs) (1,516 up-regulated, 360 down-regulated) were identified between vegetative buds and floral buds, and 3,300 DEGs (1,277 up-regulated, 1,706 down-regulated) were identified between floral buds and buds. Many genes encoding important transcription factors (e.g., AP2, MYB, MYC, WRKY, NAC and CRT) as well as proteins involved in carbohydrate metabolism, protein kinase activity, plant hormone signal transduction, and the defense responses, among others, were considerably up-regulated in floral buds. Genes involved in the photoperiod pathway and flower organ determination were also identified. These genes represent important candidate genes for molecular cloning and functional analysis to study flowering regulation in chrysanthemum. Conclusion This comparative transcriptome analysis revealed significant differences in gene expression and signaling pathway components between the vegetative buds, floral buds and buds of Chrysanthemum morifolium. A wide range of genes was implicated in regulating the phase transition from vegetative to reproductive growth. These results should aid

  18. [Impact of TDZ and NAA on adventitious bud induction and cluster bud multiplication in Tulipa edulis].

    PubMed

    Zhu, Li-Fang; Xu, Chao; Zhu, Zai-Biao; Yang, He-Tong; Guo, Qiao-Sheng; Xu, Hong-jian; Ma, Hong-Jian; Zhao, Gui-Hua

    2014-08-01

    To explore the method of explants directly induced bud and establish the tissue culture system of mutiple shoot by means of direct organogenesis, core bud and daughter bulbs (the top of bud stem expanded to form daughter bulb) of T. edulis were used as explants and treated with thidiazuron (TDZ) and 1-naphthlcetic acid (NAA). The results showed that the optimal medium for bud inducted form core bud and daughter bulb were MS + TDZ 2.0 mg x L(-1) + NAA 4.0 mg x L(-1) and MS +TDZ 2.0 mg x L(-1) + NAA 2.0 mg x L(-1) respectively, both of them had a bud induction rate of 72.92%, 79.22%. The optimal medium for cluster buds multiplication was MS + TDZ 0.2 mg x L(-1) + NAA 0.2 mg x L(-1), and proliferation coefficient was 2.23. After proliferation, cluster buds rooting occurred on MS medium with IBA 1.0 mg x L(-1) and the rooting rate was 52.6%, three to five seedlings in each plant. Using core bud and daughter bulb of T. edulis, the optimum medium for adventitious bud directly inducted from daughter bulb, core bud and cluster bud multiplication were screened out and the tissue culture system of multiple shoot by means of direct organogenesis was established. PMID:25509282

  19. Transcriptome Profiling of Tiller Buds Provides New Insights into PhyB Regulation of Tillering and Indeterminate Growth in Sorghum.

    PubMed

    Kebrom, Tesfamichael H; Mullet, John E

    2016-04-01

    Phytochrome B (phyB) enables plants to modify shoot branching or tillering in response to varying light intensities and ratios of red and far-red light caused by shading and neighbor proximity. Tillering is inhibited in sorghum genotypes that lack phytochrome B (58M, phyB-1) until after floral initiation. The growth of tiller buds in the first leaf axil of wild-type (100M, PHYB) and phyB-1 sorghum genotypes is similar until 6 d after planting when buds of phyB-1 arrest growth, while wild-type buds continue growing and develop into tillers. Transcriptome analysis at this early stage of bud development identified numerous genes that were up to 50-fold differentially expressed in wild-type/phyB-1 buds. Up-regulation of terminal flower1, GA2oxidase, and TPPI could protect axillary meristems in phyB-1 from precocious floral induction and decrease bud sensitivity to sugar signals. After bud growth arrest in phyB-1, expression of dormancy-associated genes such as DRM1, GT1, AF1, and CKX1 increased and ENOD93, ACCoxidase, ARR3/6/9, CGA1, and SHY2 decreased. Continued bud outgrowth in wild-type was correlated with increased expression of genes encoding a SWEET transporter and cell wall invertases. The SWEET transporter may facilitate Suc unloading from the phloem to the apoplast where cell wall invertases generate monosaccharides for uptake and utilization to sustain bud outgrowth. Elevated expression of these genes was correlated with higher levels of cytokinin/sugar signaling in growing buds of wild-type plants. PMID:26893475

  20. Tracing QTLs for Leaf Blast Resistance and Agronomic Performance of Finger Millet (Eleusine coracana (L.) Gaertn.) Genotypes through Association Mapping and in silico Comparative Genomics Analyses.

    PubMed

    Ramakrishnan, M; Antony Ceasar, S; Duraipandiyan, V; Vinod, K K; Kalpana, Krishnan; Al-Dhabi, N A; Ignacimuthu, S

    2016-01-01

    Finger millet is one of the small millets with high nutritive value. This crop is vulnerable to blast disease caused by Pyricularia grisea, which occurs annually during rainy and winter seasons. Leaf blast occurs at early crop stage and is highly damaging. Mapping of resistance genes and other quantitative trait loci (QTLs) for agronomic performance can be of great use for improving finger millet genotypes. Evaluation of one hundred and twenty-eight finger millet genotypes in natural field conditions revealed that leaf blast caused severe setback on agronomic performance for susceptible genotypes, most significant traits being plant height and root length. Plant height was reduced under disease severity while root length was increased. Among the genotypes, IE4795 showed superior response in terms of both disease resistance and better agronomic performance. A total of seven unambiguous QTLs were found to be associated with various agronomic traits including leaf blast resistance by association mapping analysis. The markers, UGEP101 and UGEP95, were strongly associated with blast resistance. UGEP98 was associated with tiller number and UGEP9 was associated with root length and seed yield. Cross species validation of markers revealed that 12 candidate genes were associated with 8 QTLs in the genomes of grass species such as rice, foxtail millet, maize, Brachypodium stacei, B. distachyon, Panicum hallii and switchgrass. Several candidate genes were found proximal to orthologous sequences of the identified QTLs such as 1,4-β-glucanase for leaf blast resistance, cytokinin dehydrogenase (CKX) for tiller production, calmodulin (CaM) binding protein for seed yield and pectin methylesterase inhibitor (PMEI) for root growth and development. Most of these QTLs and their putatively associated candidate genes are reported for first time in finger millet. On validation, these novel QTLs may be utilized in future for marker assisted breeding for the development of fungal

  1. Tracing QTLs for Leaf Blast Resistance and Agronomic Performance of Finger Millet (Eleusine coracana (L.) Gaertn.) Genotypes through Association Mapping and in silico Comparative Genomics Analyses

    PubMed Central

    Ramakrishnan, M.; Antony Ceasar, S.; Duraipandiyan, V.; Vinod, K. K.; Kalpana, Krishnan; Al-Dhabi, N. A.; Ignacimuthu, S.

    2016-01-01

    Finger millet is one of the small millets with high nutritive value. This crop is vulnerable to blast disease caused by Pyricularia grisea, which occurs annually during rainy and winter seasons. Leaf blast occurs at early crop stage and is highly damaging. Mapping of resistance genes and other quantitative trait loci (QTLs) for agronomic performance can be of great use for improving finger millet genotypes. Evaluation of one hundred and twenty-eight finger millet genotypes in natural field conditions revealed that leaf blast caused severe setback on agronomic performance for susceptible genotypes, most significant traits being plant height and root length. Plant height was reduced under disease severity while root length was increased. Among the genotypes, IE4795 showed superior response in terms of both disease resistance and better agronomic performance. A total of seven unambiguous QTLs were found to be associated with various agronomic traits including leaf blast resistance by association mapping analysis. The markers, UGEP101 and UGEP95, were strongly associated with blast resistance. UGEP98 was associated with tiller number and UGEP9 was associated with root length and seed yield. Cross species validation of markers revealed that 12 candidate genes were associated with 8 QTLs in the genomes of grass species such as rice, foxtail millet, maize, Brachypodium stacei, B. distachyon, Panicum hallii and switchgrass. Several candidate genes were found proximal to orthologous sequences of the identified QTLs such as 1,4-β-glucanase for leaf blast resistance, cytokinin dehydrogenase (CKX) for tiller production, calmodulin (CaM) binding protein for seed yield and pectin methylesterase inhibitor (PMEI) for root growth and development. Most of these QTLs and their putatively associated candidate genes are reported for first time in finger millet. On validation, these novel QTLs may be utilized in future for marker assisted breeding for the development of fungal

  2. Fruit load modulates flowering-related gene expression in buds of alternate-bearing ‘Moncada’ mandarin

    PubMed Central

    Muñoz-Fambuena, Natalia; Mesejo, Carlos; González-Mas, M. Carmen; Primo-Millo, Eduardo; Agustí, Manuel; Iglesias, Domingo J.

    2012-01-01

    Background and Aims Gene determination of flowering is the result of complex interactions involving both promoters and inhibitors. In this study, the expression of flowering-related genes at the meristem level in alternate-bearing citrus trees is analysed, together with the interplay between buds and leaves in the determination of flowering. Methods First defruiting experiments were performed to manipulate blossoming intensity in ‘Moncada’ mandarin, Citrus clementina. Further defoliation was performed to elucidate the role leaves play in the flowering process. In both cases, the activity of flowering-related genes was investigated at the flower induction (November) and differentiation (February) stages. Key Results Study of the expression pattern of flowering-genes in buds from on (fully loaded) and off (without fruits) trees revealed that homologues of FLOWERING LOCUS T (CiFT), TWIN SISTER OF FT (TSF), APETALA1 (CsAP1) and LEAFY (CsLFY) were negatively affected by fruit load. CiFT and TSF activities showed a marked increase in buds from off trees through the study period (ten-fold in November). By contrast, expression of the homologues of the flowering inhibitors of TERMINAL FLOWER 1 (CsTFL), TERMINAL FLOWER 2 (TFL2) and FLOWERING LOCUS C (FLC) was generally lower in off trees. Regarding floral identity genes, the increase in CsAP1 expression in off trees was much greater in buds than in leaves, and significant variations in CsLFY expression (approx. 20 %) were found only in February. Defoliation experiments further revealed that the absence of leaves completely abolished blossoming and severely affected the expression of most of the flowering-related genes, particularly decreasing the activity of floral promoters and of CsAP1 at the induction stage. Conclusions These results suggest that the presence of fruit affects flowering by greatly altering gene-expression not only at the leaf but also at the meristem level. Although leaves are required for flowering to

  3. AXR1 Acts after Lateral Bud Formation to Inhibit Lateral Bud Growth in Arabidopsis1

    PubMed Central

    Stirnberg, Petra; Chatfield, Steven P.; Leyser, H.M. Ottoline

    1999-01-01

    The AXR1 gene of Arabidopsis is required for many auxin responses. The highly branched shoot phenotype of mature axr1 mutant plants has been taken as genetic evidence for a role of auxin in the control of shoot branching. We compared the development of lateral shoots in wild-type Columbia and axr1-12 plants. In the wild type, the pattern of lateral shoot development depends on the developmental stage of the plant. During prolonged vegetative growth, axillary shoots arise and develop in a basal-apical sequence. After floral transition, axillary shoots arise rapidly along the primary shoot axis and grow out to form lateral inflorescences in an apical-basal sequence. For both patterns, the axr1 mutation does not affect the timing of axillary meristem formation; however, subsequent lateral shoot development proceeds more rapidly in axr1 plants. The outgrowth of lateral inflorescences from excised cauline nodes of wild-type plants is inhibited by apical auxin. axr1-12 nodes are resistant to this inhibition. These results provide evidence for common control of axillary growth in both patterns, and suggest a role for auxin during the late stages of axillary shoot development following the formation of the axillary bud and several axillary leaf primordia. PMID:10557232

  4. Axillary Meristems and the Development of Epicormic Buds in Wollemi Pine (Wollemia nobilis)

    PubMed Central

    BURROWS, G. E.; OFFORD, C. A.; MEAGHER, P. F.; ASHTON, K.

    2003-01-01

    Intact trees of Wollemia nobilis Jones, Hill and Allen (Araucariaceae) routinely develop multiple coppice shoots as well as orthotropic epicormic shoots that become replacement or additional leaders. As these are unusual architectural features for the Araucariaceae, an investigation was made of the axillary meristems of the main stem and their role in the production of epicormic and possibly coppice shoots. Leaf axils, excised from the apex to the base of 2‐m‐high W. nobilis plants (seedling origin, ex situ grown), were examined anatomically. Small, endogenous, undifferentiated (no leaf primordia, no vascular or provascular connections) meristems were found in the axils from near the shoot apex. In the more proximal positions about half the meristems sampled did not differentiate further, but became tangentially elongated to compensate for increases in stem diameter. In the remaining axils the meristems slowly developed into bud primordia, although these buds usually developed few leaf primordia and their apical ‘domes’ were wide and flat. Associated vascular development was generally restricted to provascular dedifferentiation of the cortical parenchyma, with the procambium usually forming a ‘closed loop’ that did not extend back to the secondary vascular tissues. Development of the meristems was very uneven with adjacent axils often at widely differing stages of development into buds. The study shows that, unlike most conifers, W. nobilis possesses long‐lived meristematic potential in most, if not all, leaf axils. Unlike other araucarias that have been investigated, many of the meristems in the orthotropic main stem will slowly develop into bud primordia beneath the bark in intact plants. It appears likely that this slow but continued development provides a ready source of additional or replacement leaders and thus new branches and leaves. PMID:14612379

  5. Making continental-scale environmental programs relevant locally for educators with Project BudBurst

    NASA Astrophysics Data System (ADS)

    Goehring, L.; Henderson, S.; Wasser, L.; Newman, S. J.; Ward, D.

    2012-12-01

    Project BudBurst is a national citizen science initiative designed to engage non professionals in observations of phenological (plant life cycle) events that raise awareness of climate change, and create a cadre of informed citizen scientists. Citizen science programs such as Project BudBurst provide excellent opportunities for educators and their students to actively participate in scientific research. Such programs are important not only from an educational perspective, but because they also enable scientists to broaden the geographic and temporal scale of their observations. The goals of Project BudBurst are to 1) increase awareness of phenology as an area of scientific study; 2) increase awareness of the impacts of changing climates on plants at a continental-scale; and 3) increase science literacy by engaging participants in the scientific process. From its 2008 launch, this on-line program has engaged participants of all ages and walks of life in recording the timing of the leafing and flowering of wild and cultivated species found across the continent, and in contemplating the meaning of such data in their local environments. Thus far, thousands of participants from all 50 states have submitted data. This presentation will provide an overview of Project BudBurst educational resources and share lessons learned from educators in implementing the program in formal and informal education settings. Lesson plans and tips from educators will be highlighted. Project BudBurst is co-managed by the National Ecological Observatory Network and the Chicago Botanic Garden.

  6. Chances and pitfalls of leaf wax biomarker analyses applied to fluvial sediment sequences - the example of a Holocene fluvial sediment-paleosol sequence from the upper Alazani River, eastern Georgia

    NASA Astrophysics Data System (ADS)

    von Suchodoletz, Hans; Bliedtner, Marcel; Zielhofer, Christoph; Faust, Dominik; Zech, Roland

    2016-04-01

    During the last decades, fluvial sediment sequences in many regions have intensively been studied to reconstruct Late Quaternary palaeoenvironmental and palaeohydrological conditions. However, up to now analyses of leaf wax biomarkers that are increasingly used to reconstruct paleoenvironmental and -climate conditions e.g. from lake sediments or loess-paleosol sequences were not systematically applied to Late Quaternary fluvial sediments. Given the ubiquitous distribution of fluvial sediment sequences on the earth's surface such investigations could potentially strongly enhance the knowledge about former environmental conditions in many regions. For this conceptual study we exemplarily analysed leaf wax biomarker (long-chain n-alkanes, n-alkanoic acids) in a fluvial sediment palaeosol sequence from the upper Alazani River in eastern Georgia to discuss general possibilities and pitfalls: Generally, biomarker records from fluvial archives can be divided into i) a catchment signal recorded in the fluvial sediment layers and ii) a local in-situ signal recorded in the intercalated paleosols. This offers the great chance to reconstruct paleoenvironmental conditions in both the whole catchment and at the sampling site. However, potential pitfalls are, for example, that inherited catchment signals can bias the in-situ signal from paleosols, while intermediate sediment storage in the catchment prior to sediment deposition and postsedimentary processes may alter the original catchment signal in the fluvial sediment layers. Thus, when applying leaf wax biomarker analyses to fluvial sediment sequences one has to be careful: The interpretation of the biomarker record strongly depends on the specific geomorphological and sedimentological conditions of the investigated site and of the catchment area.

  7. Genomic analyses of cherry rusty mottle group and cherry twisted leaf-associated viruses reveal a possible new genus within the family betaflexiviridae.

    PubMed

    Villamor, D E V; Susaimuthu, J; Eastwell, K C

    2015-03-01

    It is demonstrated that closely related viruses within the family Betaflexiviridae are associated with a number of diseases that affect sweet cherry (Prunus avium) and other Prunus spp. Cherry rusty mottle-associated virus (CRMaV) is correlated with the appearance of cherry rusty mottle disease (CRMD), and Cherry twisted leaf-associated virus (CTLaV) is linked to cherry twisted leaf disease (CTLD) and apricot ringpox disease (ARPD). Comprehensive analysis of previously reported full genomic sequences plus those determined in this study representing isolates of CTLaV, CRMaV, Cherry green ring mottle virus, and Cherry necrotic rusty mottle virus revealed segregation of sequences into four clades corresponding to distinct virus species. High-throughput sequencing of RNA from representative source trees for CRMD, CTLD, and ARPD did not reveal additional unique virus sequences that might be associated with these diseases, thereby further substantiating the association of CRMaV and CTLaV with CRMD and CTLD or ARPD, respectively. Based on comparison of the nucleotide and amino acid sequence identity values, phylogenetic relationships with other triple-gene block-coding viruses within the family Betaflexiviridae, genome organization, and natural host range, a new genus (Robigovirus) is suggested. PMID:25496302

  8. The dormant buds of Rhabdopleura compacta (Hemichordata).

    PubMed

    Dilly, P N

    1975-06-13

    Rhabdopleura has an overwintering stage that consists of two layers of cells surrounding a central yolk mass. This cellular part is surrounded by a thick electron dense capsule which is secreted by the bud itself. The capsule is probably impervious and protective to its contents. Blood vessels join the buds to the zooids of the colony. They form the probable route of transfer of yolk from the zooids to the dormant bud. The capsule of the dormant bud has some structural features in common with the black stolon of the adult zooids. The black stolon is probably formed in a manner similar to that which made the fusellar fabric of the periderm of fossil graptolities. PMID:1149105

  9. Structural Characterization of Ginsenosides from Flower Buds of Panax ginseng by RRLC-Q-TOF MS.

    PubMed

    Wu, Wei; Lu, Ziyan; Teng, Yaran; Guo, Yingying; Liu, Shuying

    2016-02-01

    Ginseng flower bud as a part of Panax ginseng has received much attention as a valuable functional food with medicinal potential. A few studies focused on systematic and comprehensive studies on its major ingredients. This study aims to rapidly characterize ginsenosides in ginseng flower buds and provide scientific basis for developing functional food, exploiting pharmaceutical effects and making full use of ginseng resources. A rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-Q-TOF-MS) method was developed for rapid qualitative and quantitative analysis of ginsenosides in ginseng flower buds. The compounds were identified by comparing retention time of the reference standards, accurate mass measurement and the fragment ions obtained from RRLC-Q-TOF-MS/MS analyses. A total of 14 kinds of ginsenosides were identified and 5 kinds of malonyl-ginsenosides were first tentatively identified in ginseng flower buds. Ten kinds of main ginsenosides were quantitatively analyzed. The developed RRLC-Q-TOF-MS method was demonstrated as an effective analytical means for rapid characterization of the ginsenosides in flower buds of P. ginseng. The research result is valuable for quality control, assessment of authenticity and stability evaluation of ginseng flower buds. PMID:26270079

  10. Taste Bud Labeling in Whole Tongue Epithelial Sheet in Adult Mice.

    PubMed

    Venkatesan, Nandakumar; Boggs, Kristin; Liu, Hong-Xiang

    2016-04-01

    Molecular labeling in whole-mount tissues provides an efficient way to obtain general information about the formation, maintenance, degeneration, and regeneration of many organs and tissues. However, labeling of lingual taste buds in whole tongue tissues in adult mice has been problematic because of the strong permeability barrier of the tongue epithelium. In this study, we present a simple method for labeling taste buds in the intact tongue epithelial sheet of an adult mouse. Following intralingual protease injection and incubation, immediate fixation of the tongue on mandible in 4% paraformaldehyde enabled the in situ shape of the tongue epithelium to be well maintained after peeling. The peeled epithelium was accessible to taste bud labeling with a pan-taste cell marker, keratin 8, and a type II taste cell marker, α-gustducin, in all three types of taste papillae, that is, fungiform, foliate, and circumvallate. Overnight incubation of tongue epithelial sheets with primary and secondary antibodies was sufficient for intense labeling of taste buds with both fluorescent and DAB visualizations. Labeled individual taste buds were easy to identify and quantify. This protocol provides an efficient way for phenotypic analyses of taste buds, especially regarding distribution pattern and number. PMID:26701416

  11. Budded baculovirus particle structure revisited.

    PubMed

    Wang, Qiushi; Bosch, Berend-Jan; Vlak, Just M; van Oers, Monique M; Rottier, Peter J; van Lent, Jan W M

    2016-02-01

    Baculoviruses are a group of enveloped, double-stranded DNA insect viruses with budded (BV) and occlusion-derived (ODV) virions produced during their infection cycle. BVs are commonly described as rod shaped particles with a high apical density of protein extensions (spikes) on the lipid envelope surface. However, due to the fragility of BVs the conventional purification and electron microscopy (EM) staining methods considerably distort the native viral structure. Here, we use cryo-EM analysis to reveal the near-native morphology of two intensively studied baculoviruses, Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Spodoptera exigua MNPV (SeMNPV), as models for BVs carrying GP64 and F as envelope fusion protein on the surface. The now well-preserved AcMNPV and SeMNPV BV particles have a remarkable elongated, ovoid shape leaving a large, lateral space between nucleocapsid (NC) and envelope. Consistent with previous findings the NC has a distinctive cap and base structure interacting tightly with the envelope. This tight interaction may explain the partial retaining of the envelope on both ends of the NC and the disappearance of the remainder of the BV envelope in the negative-staining EM images. Cryo-EM also reveals that the viral envelope contains two layers with a total thickness of ≈ 6-7 nm, which is significantly thicker than a usual biological membrane (<4 nm) as measured by X-ray scanning. Most spikes are densely clustered at the two apical ends of the virion although some envelope proteins are also found more sparsely on the lateral regions. The spikes on the surface of AcMNPV BVs appear distinctly different from those of SeMNPV. Based on our observations we propose a new near-native structural model of baculovirus BVs. PMID:26743500

  12. Free Radical Scavenging Activity: Antiproliferative and Proteomics Analyses of the Differential Expression of Apoptotic Proteins in MCF-7 Cells Treated with Acetone Leaf Extract of Diospyros lycioides (Ebenaceae)

    PubMed Central

    Pilane, M. C.; Bagla, V. P.; Mokgotho, M. P.; Mbazima, V.; Matsebatlela, T. M.; Ncube, I.; Mampuru, L.

    2015-01-01

    Breast cancer is the most common cancer in South Africa. The acetone leaf extract of Diospyros lycioides was evaluated qualitatively and quantitatively for its antioxidant potential using DPPH assay and nitric oxide radical scavenging effect, while the viability of MCF-7 cells was evaluated using the MTT. MCF-7 treated cells were stained with Hoechst 335258 dye and annexin-V-FITC to be evaluated for apoptotic effect of the extract, while mRNA expression levels of apoptotic genes were assessed by quantitative real-time PCR and deferential protein expression levels using 2D gel electrophoresis and mass spectrometry. Results revealed presence of antioxidant constituents in the extract. Extract was shown to be cytotoxic in a concentration- and time-dependent manner. Cytotoxicity was demonstrated to be due to apoptosis, with 70% of the extract-treated cells being annexin-V-positive/PI negative at 48 hours. The extract was also shown to upregulate the expression of p53 gene with concomitant downregulation of the Bcl-2 antiapoptotic gene while differentially expressed proteins were identified as enolase, pyruvate kinase, and glyceraldehyde-3-phosphate. The extract in this study was shown to induce apoptosis at an early stage which makes it an ideal source that can be explored for compounds that may be used in the treatment and management of cancer. PMID:26457109

  13. Efficient transmission of Cassava brown streak disease viral pathogens by chip bud grafting

    PubMed Central

    2013-01-01

    Background Techniques to study plant viral diseases under controlled growth conditions are required to fully understand their biology and investigate host resistance. Cassava brown streak disease (CBSD) presents a major threat to cassava production in East Africa. No infectious clones of the causal viruses, Cassava brown streak virus (CBSV) or Ugandan cassava brown streak virus (UCBSV) are available, and mechanical transmission to cassava is not effective. An improved method for transmission of the viruses, both singly and as co-infections has been developed using bud grafts. Findings Axillary buds from CBSD symptomatic plants infected with virulent isolates of CBSV and UCBSV were excised and grafted onto 6–8 week old greenhouse-grown, disease-free cassava plants of cultivars Ebwanateraka, TME204 and 60444. Plants were assessed visually for development of CBSD symptoms and by RT-PCR for presence of the viruses in leaf and storage root tissues. Across replicated experiments, 70-100% of plants inoculated with CBSV developed CBSD leaf and stem symptoms 2–6 weeks after bud grafting. Infected plants showed typical, severe necrotic lesions in storage roots at harvest 12–14 weeks after graft inoculation. Sequential grafting of buds from plants infected with UCBSV followed 10–14 days later by buds carrying CBSV, onto the same test plant, resulted in 100% of the rootstocks becoming co-infected with both pathogens. This dual transmission rate was greater than that achieved by simultaneous grafting with UCBSV and CBSV (67%), or when grafting first with CBSV followed by UCBSV (17%). Conclusions The bud grafting method described presents an improved tool for screening cassava germplasm for resistance to CBSD causal viruses, and for studying pathogenicity of this important disease. Bud grafting provides new opportunities compared to previously reported top and side grafting systems. Test plants can be inoculated as young, uniform plants of a size easily handled in a

  14. Shrinkage of ipsilateral taste buds and hyperplasia of contralateral taste buds following chorda tympani nerve transection

    PubMed Central

    Li, Yi-ke; Yang, Juan-mei; Huang, Yi-bo; Ren, Dong-dong; Chi, Fang-lu

    2015-01-01

    The morphological changes that occur in the taste buds after denervation are not well understood in rats, especially in the contralateral tongue epithelium. In this study, we investigated the time course of morphological changes in the taste buds following unilateral nerve transection. The role of the trigeminal component of the lingual nerve in maintaining the structural integrity of the taste buds was also examined. Twenty-four Sprague-Dawley rats were randomly divided into three groups: control, unilateral chorda tympani nerve transection and unilateral chorda tympani nerve transection + lingual nerve transection. Rats were allowed up to 42 days of recovery before being euthanized. The taste buds were visualized using a cytokeratin 8 antibody. Taste bud counts, volumes and taste receptor cell numbers were quantified and compared among groups. No significant difference was detected between the chorda tympani nerve transection and chorda tympani nerve transection + lingual nerve transection groups. Taste bud counts, volumes and taste receptor cell numbers on the ipsilateral side all decreased significantly compared with control. On the contralateral side, the number of taste buds remained unchanged over time, but they were larger, and taste receptor cells were more numerous postoperatively. There was no evidence for a role of the trigeminal branch of the lingual nerve in maintaining the structural integrity of the anterior taste buds. PMID:26199619

  15. Early Transcriptome Analyses of Z-3-Hexenol-Treated Zea mays Revealed Distinct Transcriptional Networks and Anti-Herbivore Defense Potential of Green Leaf Volatiles

    PubMed Central

    Engelberth, Jurgen; Contreras, Claudia Fabiola; Dalvi, Chinmay; Li, Ting; Engelberth, Marie

    2013-01-01

    Green leaf volatiles (GLV), which are rapidly emitted by plants in response to insect herbivore damage, are now established as volatile defense signals. Receiving plants utilize these molecules to prime their defenses and respond faster and stronger when actually attacked. To further characterize the biological activity of these compounds we performed a microarray analysis of global gene expression. The focus of this project was to identify early transcriptional events elicited by Z-3-hexenol (Z-3-HOL) as our model GLV in maize (Zea mays) seedlings. The microarray results confirmed previous studies on Z-3-HOL -induced gene expression but also provided novel information about the complexity of Z-3-HOL -induced transcriptional networks. Besides identifying a distinct set of genes involved in direct and indirect defenses we also found significant expression of genes involved in transcriptional regulation, Ca2+-and lipid-related signaling, and cell wall reinforcement. By comparing these results with those obtained by treatment of maize seedlings with insect elicitors we found a high degree of correlation between the two expression profiles at this early time point, in particular for those genes related to defense. We further analyzed defense gene expression induced by other volatile defense signals and found Z-3-HOL to be significantly more active than methyl jasmonate, methyl salicylate, and ethylene. The data presented herein provides important information on early genetic networks that are activated by Z-3-HOL and demonstrates the effectiveness of this compound in the regulation of typical plant defenses against insect herbivores in maize. PMID:24155960

  16. Expression of Hex during feather bud development.

    PubMed

    Obinata, Akiko; Akimoto, Yoshihiro

    2005-01-01

    We studied proline-rich divergent homeobox gene Hex/Prh expression in the dorsal skin of chick embryo during feather bud development. Hex mRNA expression was first observed in the dorsolateral ectoderm and mesenchyme at 5 days, then in the epithelium and the dermis of the dorsal skin before placode (primordium of feather bud) formation and then was restricted to the placode and the dermis under the placode. Afterward, Hex expression was seen in the epidermis and the dermis of the posterior region of short bud. In accordance with Hex mRNA expression in the placode, Hex protein was observed in the epidermis as well as in the dermis of the placode. Immunoelectron microscopic study indicated that the protein located both in the nuclei and cytoplasm of the epidermis and the dermis at the short bud stage. The Wnt signaling pathway plays an essential role in the early inductive events in hair (Wnt3a and 7a) and feather (Wnt7a) follicles. The pattern of Hex expression in the epidermis was similar to that of Wnt7a, while little, if any, expression of Wnt7a was detected in the dermis under the placode or the dermis of the short bud compared with that of Hex, suggesting that Hex plays an important role in the initiation of feather morphogenesis. PMID:16172986

  17. Virus Budding and the ESCRT Pathway

    PubMed Central

    Votteler, Jörg; Sundquist, Wesley I.

    2013-01-01

    Enveloped viruses escape infected cells by budding through limiting membranes. In the decade since the discovery that the Human Immunodeficiency Virus (HIV) recruits cellular ESCRT (endosomal sorting complexes required for transport) machinery to facilitate viral budding, this pathway has emerged as the major escape route for enveloped viruses. In cells, the ESCRT pathway catalyzes the analogous membrane fission events required for the abscission stage of cytokinesis and for a series of “reverse topology” vesiculation events. Studies of enveloped virus budding are therefore providing insights into the complex cellular mechanisms of cell division and membrane protein trafficking (and vice versa). Here, we review how viruses mimic cellular recruiting signals to usurp the ESCRT pathway, discuss mechanistic models for ESCRT pathway functions, and highlight important research frontiers. PMID:24034610

  18. Micropropagation of Phalaenopsis and Doritaenopsis by culturing shoot tips of flower stalk buds.

    PubMed

    Tokuhara, K; Mii, M

    1993-11-01

    Green Protocorm-like Bodies (PLB) with high multiplication capacity were induced from shoot tips of flower stalk buds having 1 or 2 leaf primordia using New Dogashima Medium (NDM) containing 0.1 mg l(-1) α-naphthaleneacetic acid (NAA) and 1 mg 1(-1) 6-benzylaminopurine (BAP). These PLB were subcultured on the same medium. More than 10,000 PLBs were obtained from a few buds on a single flower stalk within one year. After transfer onto NDM containing no plant growth regulator (PGR), the PLB developed into plantlets. The micropropagation method formulated in this study was applicable to 12 different genotypes. These results suggest that the methodology could be used on a commercial scale for vegetative propagation of Phalaenopsis and Doritaenopsis. PMID:24196174

  19. 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

  20. Interaction between bud-site selection and polarity-establishment machineries in budding yeast

    PubMed Central

    Wu, Chi-Fang; Savage, Natasha S.; Lew, Daniel J.

    2013-01-01

    Saccharomyces cerevisiae yeast cells polarize in order to form a single bud in each cell cycle. Distinct patterns of bud-site selection are observed in haploid and diploid cells. Genetic approaches have identified the molecular machinery responsible for positioning the bud site: during bud formation, specific locations are marked with immobile landmark proteins. In the next cell cycle, landmarks act through the Ras-family GTPase Rsr1 to promote local activation of the conserved Rho-family GTPase, Cdc42. Additional Cdc42 accumulates by positive feedback, creating a concentrated patch of GTP-Cdc42, which polarizes the cytoskeleton to promote bud emergence. Using time-lapse imaging and mathematical modelling, we examined the process of bud-site establishment. Imaging reveals unexpected effects of the bud-site-selection system on the dynamics of polarity establishment, raising new questions about how that system may operate. We found that polarity factors sometimes accumulate at more than one site among the landmark-specified locations, and we suggest that competition between clusters of polarity factors determines the final location of the Cdc42 cluster. Modelling indicated that temporally constant landmark-localized Rsr1 would weaken or block competition, yielding more than one polarity site. Instead, we suggest that polarity factors recruit Rsr1, effectively sequestering it from other locations and thereby terminating landmark activity. PMID:24062579

  1. Bud development in corydalis (Corydalis bracteata) requires low temperature: a study of developmental and carbohydrate changes

    PubMed Central

    Khodorova, Nadejda V.; Miroslavov, Evgeniy A.; Shavarda, Alexey L.; Laberche, Jean-Claude; Boitel-Conti, Michèle

    2010-01-01

    Background and Aims Spring geophytes require a period of low temperature for proper flower development but the mechanism that underlies the relationship between cold treatment and flowering remains unknown. The present study aims to compare the developmental anatomy and carbohydrate content of the tuberous geophyte Corydalis bracteata growing under natural winter conditions from 10 to −10 °C (field-grown) and under a mild temperature regime of 18 °C (indoor-grown plants). Methods Samples were studied under light and electron microscopy. A histochemical test (periodic acid – Schiff's) was employed to identify starch in sectioned material. Sugars were analysed by capillary gas chromatography. Apoplastic wash fluid was prepared. Key Results Under natural conditions, shoots were elongated, and buds gained in dry mass and developed normally. For indoor-grown plants, these parameters were lower in value and, from December, a progressive necrosis of flower buds was observed. The tuber consisted of the new developing one, which was connected to the bud, and the old tuber with its starch reserve. Due to the absence of plasmodesmata between new and old tuber cells, sugar transport cannot be through the symplast. Thus, a potential apoplastic route is proposed from old tuber phloem parenchyma cells to the adjacent new tuber cells. Sugar content in buds during the autumn months (September–November) was lower for indoor-grown plants than control plants, whereas the sugar content in tubers during the same period was similar for plants from both temperature treatments. However, the amount of apoplastic sugars in tubers of field-grown plants was almost 15-fold higher than in indoor-grown tubers. Conclusions The results suggest that low temperature activates the apoplastic route of sugar transport in C. bracteata tubers and a consequent carbohydrate delivery to the bud. In the absence of cold treatment, the carbohydrate reserve is locked in old tuber cells so the nutrient

  2. Two new species of sympatric Fergusonina flies (Diptera: Fergusoninidae) from bud galls on high elevation snow gums (Eucalyptus pauciflora Sieb. ex Spreng. complex) in the Australian Alps

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two species of Fergusonina Malloch fly, F. daviesae Nelson sp.n. and F. taylori Nelson sp.n. (Diptera: Fergusoninidae), are described from terminal leaf bud galls on high elevation snow gums (Eucalyptus pauciflora complex) in the Australian Alps. These species occur in sympatry at the six locations...

  3. Genome-wide transcriptome profiling provides insights into floral bud development of summer-flowering Camellia azalea

    PubMed Central

    Fan, Zhengqi; Li, Jiyuan; Li, Xinlei; Wu, Bin; Wang, Jiangying; Liu, Zhongchi; Yin, Hengfu

    2015-01-01

    The transition from vegetative to reproductive growth in woody perennials involves pathways controlling flowering timing, bud dormancy and outgrowth in responses to seasonal cues. However little is known about the mechanism governing the adaptation of signaling pathways to environmental conditions in trees. Camellia azalea is a rare species in this genus flowering during summer, which provides a unique resource for floral timing breeding. Here we reported a comprehensive transcriptomics study to capture the global gene profiles during floral bud development in C. azalea. We examined the genome-wide gene expression between three developmental stages including floral bud initiation, floral organ differentiation and bud outgrowth, and identified nine co-expression clusters with distinctive patterns. Further, we identified the differential expressed genes (DEGs) during development and characterized the functional properties of DEGs by Gene Ontology analysis. We showed that transition from floral bud initiation to floral organ differentiation required changes of genes in flowering timing regulation, while transition to floral bud outgrowth was regulated by various pathways such as cold and light signaling, phytohormone pathways and plant metabolisms. Further analyses of dormancy associated MADS-box genes revealed that SVP- and AGL24- like genes displayed distinct expression patterns suggesting divergent roles during floral bud development. PMID:25978548

  4. Competitive canalization of PIN-dependent auxin flow from axillary buds controls pea bud outgrowth.

    PubMed

    Balla, Jozef; Kalousek, Petr; Reinöhl, Vilém; Friml, Jiří; Procházka, Stanislav

    2011-02-01

    Shoot branching is one of the major determinants of plant architecture. Polar auxin transport in stems is necessary for the control of bud outgrowth by a dominant apex. Here, we show that following decapitation in pea (Pisum sativum L.), the axillary buds establish directional auxin export by subcellular polarization of PIN auxin transporters. Apical auxin application on the decapitated stem prevents this PIN polarization and canalization of laterally applied auxin. These results support a model in which the apical and lateral auxin sources compete for primary channels of auxin transport in the stem to control the outgrowth of axillary buds. PMID:21219506

  5. Identification and Quality Assessment of Chrysanthemum Buds by CE Fingerprinting

    PubMed Central

    Xing, Xiaoping; Li, Dan

    2015-01-01

    A simple and efficient fingerprinting method for chrysanthemum buds was developed with the aim of establishing a quality control protocol based on biochemical makeup. Chrysanthemum bud samples were successively extracted by water and alcohol. The fingerprints of the chrysanthemum buds samples were obtained using capillary electrophoresis and electrochemical detection (CE-ED) employing copper and carbon working electrodes to capture all of the chemical information. 10 batches of chrysanthemum buds were collected from different regions and various factories to establish the baseline fingerprint. The experimental data of 10 batches electropherogram buds by CE were analyzed by correlation coefficient and the included angle cosine methods. A standard chrysanthemum bud fingerprint including 24 common peaks was established, 12 from each electrode, which was successfully applied to identify and distinguish between chrysanthemum buds from 2 other chrysanthemum species. These results demonstrate that fingerprint analysis can be used as an important criterion for chrysanthemum buds quality control. PMID:26064777

  6. Signals regulating dormancy in vegetative buds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dormancy in plants involves a temporary suspension of meristem growth, thus insuring bud survival and maintenance of proper shoot system architecture. Dormancy regulation is a complex process involving interactions of various signals through specific and/or overlapping signal transduction pathways. ...

  7. Bilingual Buds: The Evolution of a Program

    ERIC Educational Resources Information Center

    Huang, Sharon

    2009-01-01

    The impetus to begin Bilingual Buds came about six years ago when the author, pregnant with twins and commuting into New York City, was reading about the numerous cognitive benefits for children of acquiring a second language early in their lives. She was surprised to learn that even by the age of six months, children begin to lose the ability to…

  8. Dormant bud preservation for germplasm conservation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The suitability of dormant buds (DB) for cryopreservation of different plant species has been demonstrated in several reports. For the majority of the species, processing DB for long-term liquid nitrogen storage does not involve establishing tissue cultures and the time for growing out post-cryo mat...

  9. Immunohistochemical Analysis of Human Vallate Taste Buds.

    PubMed

    Tizzano, Marco; Grigereit, Laura; Shultz, Nicole; Clary, Matthew S; Finger, Thomas E

    2015-11-01

    The morphology of the vallate papillae from postmortem human samples was investigated with immunohistochemistry. Microscopically, taste buds were present along the inner wall of the papilla, and in some cases in the outer wall as well. The typical taste cell markers PLCβ2, GNAT3 (gustducin) and the T1R3 receptor stain elongated cells in human taste buds consistent with the Type II cells in rodents. In the human tissue, taste bud cells that stain with Type II cell markers, PLCβ2 and GNAT3, also stain with villin antibody. Two typical immunochemical markers for Type III taste cells in rodents, PGP9.5 and SNAP25, fail to stain any taste bud cells in the human postmortem tissue, although these antibodies do stain numerous nerve fibers throughout the specimen. Car4, another Type III cell marker, reacted with only a few taste cells in our samples. Finally, human vallate papillae have a general network of innervation similar to rodents and antibodies directed against SNAP25, PGP9.5, acetylated tubulin and P2X3 all stain free perigemmal nerve endings as well as intragemmal taste fibers. We conclude that with the exception of certain molecular features of Type III cells, human vallate papillae share the structural, morphological, and molecular features observed in rodents. PMID:26400924

  10. Taste Bud-Derived BDNF Is Required to Maintain Normal Amounts of Innervation to Adult Taste Buds123

    PubMed Central

    Meng, Lingbin; Ohman-Gault, Lisa; Ma, Liqun

    2015-01-01

    Abstract Gustatory neurons transmit chemical information from taste receptor cells, which reside in taste buds in the oral cavity, to the brain. As adult taste receptor cells are renewed at a constant rate, nerve fibers must reconnect with new taste receptor cells as they arise. Therefore, the maintenance of gustatory innervation to the taste bud is an active process. Understanding how this process is regulated is a fundamental concern of gustatory system biology. We speculated that because brain-derived neurotrophic factor (BDNF) is required for taste bud innervation during development, it might function to maintain innervation during adulthood. If so, taste buds should lose innervation when Bdnf is deleted in adult mice. To test this idea, we first removed Bdnf from all cells in adulthood using transgenic mice with inducible CreERT2 under the control of the Ubiquitin promoter. When Bdnf was removed, approximately one-half of the innervation to taste buds was lost, and taste buds became smaller because of the loss of taste bud cells. Individual taste buds varied in the amount of innervation each lost, and those that lost the most innervation also lost the most taste bud cells. We then tested the idea that that the taste bud was the source of this BDNF by reducing Bdnf levels specifically in the lingual epithelium and taste buds. Taste buds were confirmed as the source of BDNF regulating innervation. We conclude that BDNF expressed in taste receptor cells is required to maintain normal levels of innervation in adulthood. PMID:26730405

  11. Key Proliferative Activity in the Junction between the Leaf Blade and Leaf Petiole of Arabidopsis1[W][OA

    PubMed Central

    Ichihashi, Yasunori; Kawade, Kensuke; Usami, Takeshi; Horiguchi, Gorou; Takahashi, Taku; Tsukaya, Hirokazu

    2011-01-01

    Leaves are the most important, fundamental units of organogenesis in plants. Although the basic form of a leaf is clearly divided into the leaf blade and leaf petiole, no study has yet revealed how these are differentiated from a leaf primordium. We analyzed the spatiotemporal pattern of mitotic activity in leaf primordia of Arabidopsis (Arabidopsis thaliana) in detail using molecular markers in combination with clonal analysis. We found that the proliferative zone is established after a short interval following the occurrence of a rod-shaped early leaf primordium; it is separated spatially from the shoot apical meristem and seen at the junction region between the leaf blade and leaf petiole and produces both leaf-blade and leaf-petiole cells. This proliferative region in leaf primordia is marked by activity of the ANGUSTIFOLIA3 (AN3) promoter as a whole and seems to be differentiated into several spatial compartments: activities of the CYCLIN D4;2 promoter and SPATULA enhancer mark parts of it specifically. Detailed analyses of the an3 and blade-on-petiole mutations further support the idea that organogenesis of the leaf blade and leaf petiole is critically dependent on the correct spatial regulation of the proliferative region of leaf primordia. Thus, the proliferative zone of leaf primordia is spatially differentiated and supplies both the leaf-blade and leaf-petiole cells. PMID:21880932

  12. HIV-1 Assembly, Budding, and Maturation

    PubMed Central

    Sundquist, Wesley I.; Kräusslich, Hans-Georg

    2012-01-01

    A defining property of retroviruses is their ability to assemble into particles that can leave producer cells and spread infection to susceptible cells and hosts. Virion morphogenesis can be divided into three stages: assembly, wherein the virion is created and essential components are packaged; budding, wherein the virion crosses the plasma membrane and obtains its lipid envelope; and maturation, wherein the virion changes structure and becomes infectious. All of these stages are coordinated by the Gag polyprotein and its proteolytic maturation products, which function as the major structural proteins of the virus. Here, we review our current understanding of the mechanisms of HIV-1 assembly, budding, and maturation, starting with a general overview and then providing detailed descriptions of each of the different stages of virion morphogenesis. PMID:22762019

  13. Synchronization of the Budding Yeast Saccharomyces cerevisiae.

    PubMed

    Foltman, Magdalena; Molist, Iago; Sanchez-Diaz, Alberto

    2016-01-01

    A number of model organisms have provided the basis for our understanding of the eukaryotic cell cycle. These model organisms are generally much easier to manipulate than mammalian cells and as such provide amenable tools for extensive genetic and biochemical analysis. One of the most common model organisms used to study the cell cycle is the budding yeast Saccharomyces cerevisiae. This model provides the ability to synchronise cells efficiently at different stages of the cell cycle, which in turn opens up the possibility for extensive and detailed study of mechanisms regulating the eukaryotic cell cycle. Here, we describe methods in which budding yeast cells are arrested at a particular phase of the cell cycle and then released from the block, permitting the study of molecular mechanisms that drive the progression through the cell cycle. PMID:26519319

  14. Actomyosin ring driven cytokinesis in budding yeast.

    PubMed

    Meitinger, Franz; Palani, Saravanan

    2016-05-01

    Cytokinesis is the final process in the cell cycle that physically divides one cell into two. In budding yeast, cytokinesis is driven by a contractile actomyosin ring (AMR) and the simultaneous formation of a primary septum, which serves as template for cell wall deposition. AMR assembly, constriction, primary septum formation and cell wall deposition are successive processes and tightly coupled to cell cycle progression to ensure the correct distribution of genetic material and cell organelles among the two rising cells prior to cell division. The role of the AMR in cytokinesis and the molecular mechanisms that drive AMR constriction and septation are the focus of current research. This review summarizes the recent progresses in our understanding of how budding yeast cells orchestrate the multitude of molecular mechanisms that control AMR driven cytokinesis in a spatio-temporal manner to achieve an error free cell division. PMID:26845196

  15. Dual Function of CD81 in Influenza Virus Uncoating and Budding

    PubMed Central

    He, Jiang; Sun, Eileen; Bujny, Miriam V.; Kim, Doory; Davidson, Michael W.; Zhuang, Xiaowei

    2013-01-01

    As an obligatory pathogen, influenza virus co-opts host cell machinery to harbor infection and to produce progeny viruses. In order to characterize the virus-host cell interactions, several genome-wide siRNA screens and proteomic analyses have been performed recently to identify host factors involved in influenza virus infection. CD81 has emerged as one of the top candidates in two siRNA screens and one proteomic study. The exact role played by CD81 in influenza infection, however, has not been elucidated thus far. In this work, we examined the effect of CD81 depletion on the major steps of the influenza infection. We found that CD81 primarily affected virus infection at two stages: viral uncoating during entry and virus budding. CD81 marked a specific endosomal population and about half of the fused influenza virus particles underwent fusion within the CD81-positive endosomes. Depletion of CD81 resulted in a substantial defect in viral fusion and infection. During virus assembly, CD81 was recruited to virus budding site on the plasma membrane, and in particular, to specific sub-viral locations. For spherical and slightly elongated influenza virus, CD81 was localized at both the growing tip and the budding neck of the progeny viruses. CD81 knockdown led to a budding defect and resulted in elongated budding virions with a higher propensity to remain attached to the plasma membrane. Progeny virus production was markedly reduced in CD81-knockdown cells even when the uncoating defect was compensated. In filamentous virus, CD81 was distributed at multiple sites along the viral filament. Taken together, these results demonstrate important roles of CD81 in both entry and budding stages of the influenza infection cycle. PMID:24130495

  16. Inter- and intra-tumoral relationships between vasculature characteristics, GLUT1 and budding in colorectal carcinoma.

    PubMed

    Mezheyeuski, Artur; Nerovnya, Alexander; Bich, Tatjana; Tur, Gennadiy; Ostman, Arne; Portyanko, Anna

    2015-10-01

    Vascular characteristics, hypoxia and tumor budding are features that have been implied in the biology and prognosis of colorectal cancer. Internal relationships and the inter- and intra-tumoral variation of these tumor properties remain to be determined. In the current study we have characterized blood vessel status in different areas of CRC and in the peritumoral fibroblastic stroma. Analyses of these characteristics have been supplemented by characterization of budding and hypoxia. Analyses revealed significantly lower values of vessel perimeter (VP) and vessel lumen area (VL) at the invasive front and surrounding stroma as compared to the tumor center. Also, the number of vessels (VN) in the peritumoral stroma was higher than in the center. Thus, tumor center displays larger and fewer vessels as compared to the tumor periphery. GLUT1 expression was correlated directly with VN (r=0.351, p=0.028) and inversely with VL and VP (r=-0.432, p=0.006 and r=-0.484, p=0.002) at the invasive front. Moreover, GLUT1 expression, VP at the invasive front, and VN in the surrounding peritumoral stroma, were associated with budding score (r=0.574, p<0.000, r=-0.340, p=0,034 and r=-0.389, p=0.025 respectively). Furthermore, GLUT1, budding score, vessel number in peritumoral stroma, and vessel size in the invasive front, were significantly different in tumors with or without lymph node metastasis. This study reports previously unrecognized relationships between localization-specific vascular characteristics, hypoxia and tumor budding. The findings suggest potential functional relationships, which should be further explored, and also highlight the inter-tumoral variations in vasculature, which is highly relevant for ongoing efforts to identify vessel-based biomarkers. PMID:25811313

  17. Viral and cellular requirements for the budding of Feline Endogenous Retrovirus RD-114

    PubMed Central

    2011-01-01

    Background RD-114 virus is a feline endogenous retrovirus and produced as infectious viruses in some feline cell lines. Recently, we reported the contamination of an infectious RD-114 virus in a proportion of live attenuated vaccines for dogs and cats. It is very difficult to completely knock out the RD-114 proviruses from cells, as endogenous retroviruses are usually integrated multiply into the host genome. However, it may be possible to reduce the risk of contamination of RD-114 virus by regulating the viral release from cells. Results In this study, to understand the molecular mechanism of RD-114 virus budding, we attempted to identify the viral and cellular requirements for RD-114 virus budding. Analyses of RD-114 L-domain mutants showed that the PPPY sequence in the pp15 region of Gag plays a critical role in RD-114 virus release as viral L-domain. Furthermore, we investigated the cellular factors required for RD-114 virus budding. We demonstrated that RD-114 virus release was inhibited by overexpression of dominant negative mutants of Vps4A, Vps4B, and WWP2. Conclusions These results strongly suggest that RD-114 budding utilizes the cellular multivesicular body sorting pathway similar to many other retroviruses. PMID:22168342

  18. Cell Polarization and Cytokinesis in Budding Yeast

    PubMed Central

    Bi, Erfei; Park, Hay-Oak

    2012-01-01

    Asymmetric cell division, which includes cell polarization and cytokinesis, is essential for generating cell diversity during development. The budding yeast Saccharomyces cerevisiae reproduces by asymmetric cell division, and has thus served as an attractive model for unraveling the general principles of eukaryotic cell polarization and cytokinesis. Polarity development requires G-protein signaling, cytoskeletal polarization, and exocytosis, whereas cytokinesis requires concerted actions of a contractile actomyosin ring and targeted membrane deposition. In this chapter, we discuss the mechanics and spatial control of polarity development and cytokinesis, emphasizing the key concepts, mechanisms, and emerging questions in the field. PMID:22701052

  19. Essential Oil of Betula pendula Roth. Buds

    PubMed Central

    2004-01-01

    The essential oil of Betula pendula Roth. buds was obtained using both hydrodistillation and microdistillation techniques and their chemical compositions were analyzed using both gas chromatography (GC) and gas chromatography–mass spectrometry (GC-MS). Overall, more than 50 compounds were identified representing 80% and 92% for hydrodistillation and microdistillation, respectively. The main components (by hydrodistillation and microdistillation, respectively) found were α-copaene (12% and 10%), germacrene D (11% and 18%) and δ-cadinene (11% and 15%) in the analyzed essential oils. The microdistillation technique proved to be a useful tool and compliant alternative when compared to hydrodistillation. PMID:15841263

  20. Essential Oil of Betula pendula Roth. Buds.

    PubMed

    Demirci, Betül; Paper, Dietrich H; Demirci, Fatih; Can Başer, K Hüsnü; Franz, Gerhard

    2004-12-01

    The essential oil of Betula pendula Roth. buds was obtained using both hydrodistillation and microdistillation techniques and their chemical compositions were analyzed using both gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Overall, more than 50 compounds were identified representing 80% and 92% for hydrodistillation and microdistillation, respectively. The main components (by hydrodistillation and microdistillation, respectively) found were alpha-copaene (12% and 10%), germacrene D (11% and 18%) and delta-cadinene (11% and 15%) in the analyzed essential oils. The microdistillation technique proved to be a useful tool and compliant alternative when compared to hydrodistillation. PMID:15841263

  1. FGF induces new feather buds from developing avian skin.

    PubMed

    Widelitz, R B; Jiang, T X; Noveen, A; Chen, C W; Chuong, C M

    1996-12-01

    Induction of skin appendages involves a cascade of molecular events. The fibroblast growth factor (FGF) family of peptide growth factors is involved in cell proliferation and morphogenesis. We explored the role of the FGFs during skin appendage induction using developing chicken feather buds as a model. FGF-1, FGF-2, or FGF-4 was added directly to the culture medium or was released from pre-soaked Affigel blue beads. Near the midline, FGFs led to fusion of developing feather buds, representing FGFs' ability to expand feather bud domains in developing skin. In lateral regions of the explant where feather placodes have not formed, FGF treatment produces a zone of condensation and a region with an increased number of feather buds. In ventral epidermis that is normally apteric (without feathers), FGFs can also induce new feather buds. Like normal feather buds, the newly induced buds express Shh. The expression of Grb, Ras, Raf, and Erk, intracellular signaling molecules known to be downstream to tyrosine kinase receptors such as the FGF receptor, was enriched in feather bud domains. Genistein, an inhibitor of tyrosine kinase, suppressed feather bud formation and the effect of FGF. These results indicate that there are varied responses to FGFs depending on epithelial competence. All the phenotypic responses, however, show that FGFs facilitate the formation of skin appendage domains. PMID:8941663

  2. Fruit load induces changes in global gene expression and in abscisic acid (ABA) and indole acetic acid (IAA) homeostasis in citrus buds.

    PubMed

    Shalom, Liron; Samuels, Sivan; Zur, Naftali; Shlizerman, Lyudmila; Doron-Faigenboim, Adi; Blumwald, Eduardo; Sadka, Avi

    2014-07-01

    Many fruit trees undergo cycles of heavy fruit load (ON-Crop) in one year, followed by low fruit load (OFF-Crop) the following year, a phenomenon known as alternate bearing (AB). The mechanism by which fruit load affects flowering induction during the following year (return bloom) is still unclear. Although not proven, it is commonly accepted that the fruit or an organ which senses fruit presence generates an inhibitory signal that moves into the bud and inhibits apical meristem transition. Indeed, fruit removal from ON-Crop trees (de-fruiting) induces return bloom. Identification of regulatory or metabolic processes modified in the bud in association with altered fruit load might shed light on the nature of the AB signalling process. The bud transcriptome of de-fruited citrus trees was compared with those of ON- and OFF-Crop trees. Fruit removal resulted in relatively rapid changes in global gene expression, including induction of photosynthetic genes and proteins. Altered regulatory mechanisms included abscisic acid (ABA) metabolism and auxin polar transport. Genes of ABA biosynthesis were induced; however, hormone analyses showed that the ABA level was reduced in OFF-Crop buds and in buds shortly following fruit removal. Additionally, genes associated with Ca(2+)-dependent auxin polar transport were remarkably induced in buds of OFF-Crop and de-fruited trees. Hormone analyses showed that auxin levels were reduced in these buds as compared with ON-Crop buds. In view of the auxin transport autoinhibition theory, the possibility that auxin distribution plays a role in determining bud fate is discussed. PMID:24706719

  3. Fruit load induces changes in global gene expression and in abscisic acid (ABA) and indole acetic acid (IAA) homeostasis in citrus buds

    PubMed Central

    Shalom, Liron; Samuels, Sivan; Zur, Naftali; Shlizerman, Lyudmila; Doron-Faigenboim, Adi; Blumwald, Eduardo; Sadka, Avi

    2014-01-01

    Many fruit trees undergo cycles of heavy fruit load (ON-Crop) in one year, followed by low fruit load (OFF-Crop) the following year, a phenomenon known as alternate bearing (AB). The mechanism by which fruit load affects flowering induction during the following year (return bloom) is still unclear. Although not proven, it is commonly accepted that the fruit or an organ which senses fruit presence generates an inhibitory signal that moves into the bud and inhibits apical meristem transition. Indeed, fruit removal from ON-Crop trees (de-fruiting) induces return bloom. Identification of regulatory or metabolic processes modified in the bud in association with altered fruit load might shed light on the nature of the AB signalling process. The bud transcriptome of de-fruited citrus trees was compared with those of ON- and OFF-Crop trees. Fruit removal resulted in relatively rapid changes in global gene expression, including induction of photosynthetic genes and proteins. Altered regulatory mechanisms included abscisic acid (ABA) metabolism and auxin polar transport. Genes of ABA biosynthesis were induced; however, hormone analyses showed that the ABA level was reduced in OFF-Crop buds and in buds shortly following fruit removal. Additionally, genes associated with Ca2+-dependent auxin polar transport were remarkably induced in buds of OFF-Crop and de-fruited trees. Hormone analyses showed that auxin levels were reduced in these buds as compared with ON-Crop buds. In view of the auxin transport autoinhibition theory, the possibility that auxin distribution plays a role in determining bud fate is discussed. PMID:24706719

  4. Glutamate: Tastant and Neuromodulator in Taste Buds.

    PubMed

    Vandenbeuch, Aurelie; Kinnamon, Sue C

    2016-07-01

    In taste buds, glutamate plays a double role as a gustatory stimulus and neuromodulator. The detection of glutamate as a tastant involves several G protein-coupled receptors, including the heterodimer taste receptor type 1, member 1 and 3 as well as metabotropic glutamate receptors (mGluR1 and mGluR4). Both receptor types participate in the detection of glutamate as shown with knockout animals and selective antagonists. At the basal part of taste buds, ionotropic glutamate receptors [N-methyl-d-aspartate (NMDA) and non-NMDA] are expressed and participate in the modulation of the taste signal before its transmission to the brain. Evidence suggests that glutamate has an efferent function on taste cells and modulates the release of other neurotransmitters such as serotonin and ATP. This short article reviews the recent developments in the field with regard to glutamate receptors involved in both functions as well as the influence of glutamate on the taste signal. PMID:27422519

  5. Socio-cultural practices of deciduous canine tooth bud removal among Maasai children.

    PubMed

    Mutai, J; Muniu, E; Sawe, J; Hassanali, J; Kibet, P; Wanzala, P

    2010-04-01

    The removal of teeth amongst the Maasai is a traditional practice as part of an initiation or to make space for feeding in an event of diseases locking the jaw. Removal of deciduous canine tooth buds (DCB) among infants below 2 years has been reported in several studies to be common mainly amongst communities in East Africa, Ethiopia and Sudan. The main reason for the practice revolves around the belief that tooth buds or 'maggots' are false teeth, nylon or worms and are responsible for diarrhoea, vomiting, fever and growth retardation in children, amongst other illnesses. The main objective of this study was to assess the socio-cultural factors which contribute to this practice. The main methods of data collected included Focus Group Discussions (FGD) with mothers of children in that age group and Traditional Birth Attendants (TBA). In-depth interviews (IDI)were conducted with key informants versed with Maasai traditions whereas observations were done within the manyattas where participants live. Proceedings at both the FGD and the IDI were recorded on paper and were analysed thematically. The study showed that the removal of canine tooth buds amongst children that started initially with calves--that diseases that cause diarrhoea in calves were brought about by the canine tooth buds that were turning reddish in colour--is deeply rooted and practised in the community despite sensitisation interventions mounted jointly by the University of Nairobi, Kenya Medical Research Institute and the Kenya Medical Training College, among others. This study discovered that canine tooth buds are associated with bad spirits that cause diarrhoea and vomiting and the belief that removing them is a sure way of providing a cure for all children's ailments. PMID:20476714

  6. One New Conjugate of a Secoiridoid Glucoside with a Sesquiterpene Glucoside from the Flower Buds of Lonicera japonica.

    PubMed

    Yang, Biao; Meng, Zhaoqing; Ma, Yimin; Wang, Zhenzhong; Ding, Gang; Huang, Wenzhe; Sun, Lin; Hu, Yumei; Liu, Wenjun; Zhang, Chunxiao; Cao, Zeyu; Li, Jiachun; Zhong, Yan; Xiao, Wei

    2015-09-01

    Secosesquside (1), a new secoiridoid glucoside-sesquiterpene conjugate, together with three known secoiridoid derivatives, were isolated from flower buds of Lonicerajaponica. The isolated compounds were elucidated by extensive spectroscopic analyses, especially 2D NMR experiments. The anti-inflammatory activities of the new compound were also evaluated by enzyme-linked immunosorbent assay. PMID:26594743

  7. Specific residues of the GDP/GTP exchange factor Bud5p are involved in establishment of the cell type-specific budding pattern in yeast.

    PubMed

    Kang, Pil Jung; Lee, Bongyong; Park, Hay-Oak

    2004-07-01

    Cells of the budding yeast undergo oriented cell division by choosing a specific site for growth depending on their cell type. Haploid a and alpha cells bud in an axial pattern whereas diploid a/alpha cells bud in a bipolar pattern. The Ras-like GTPase Rsr1p/Bud1p, its GDP-GTP exchange factor Bud5p, and its GTPase-activating protein Bud2p are essential for selecting the proper site for polarized growth in all cell types. Here we showed that specific residues at the N terminus and the C terminus of Bud5p were important for bipolar budding, while some residues were involved in both axial and bipolar budding. These bipolar-specific mutations of BUD5 disrupted proper localization of Bud5p in diploid a/alpha cells without affecting Bud5p localization in haploid alpha cells. In contrast, Bud5p expressed in the bud5 mutants defective in both budding patterns failed to localize in all cell types. Thus, these results identify specific residues of Bud5p that are likely to be involved in direct interaction with spatial landmarks, which recruit Bud5p to the proper bud site. Finally, we found a new start codon of BUD5, which extends the open reading frame to 210 bp upstream of the previously estimated start site, thus encoding a polypeptide of 608 amino acid residues. Bud5p with these additional N-terminal residues interacted with Bud8p, a potential bipolar landmark, suggesting that the N-terminal region is necessary for recognition of the spatial cues. PMID:15136576

  8. Three new species of Fergusonina Malloch fly (Diptera: Fergusoninidae) from bud galls on Eucalyptus L'Her. (E. baxteri (Benth.) Maiden & Blakely complex, E. dalrympleana Maiden and E. pauciflora Sieb. ex Spreng.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three Fergusonina (Diptera: Fergusoninidae) flies are described from terminal leaf bud galls on Eucalyptus L'Hér. from south eastern Australia. Fergusonina omlandi Nelson and Yeates sp. nov. is the third fly from the genus Fergusonina to be described from the Eucalyptus pauciflora Sieb. ex Spreng. (...

  9. An elastic model of partial budding of retroviruses

    NASA Astrophysics Data System (ADS)

    Zhang, Rui; Nguyen, Toan

    2008-03-01

    Retroviruses are characterized by their unique infection strategy of reverse transcription, in which the genetic information flows from RNA back to DNA. The most well known representative is the human immunodeficiency virus (HIV). Unlike budding of traditional enveloped viruses, retrovirus budding happens together with the formation of spherical virus capsids at the cell membrane. Led by this unique budding mechanism, we proposed an elastic model of retrovirus budding in this work. We found that if the lipid molecules of the membrane are supplied fast enough from the cell interior, the budding always proceeds to completion. In the opposite limit, there is an optimal size of partially budded virions. The zenith angle of these partially spherical capsids, α, is given by α˜(2̂/κσ)^1/4, where κ is the bending modulus of the membrane, σ is the surface tension of the membrane, and τ characterizes the strength of capsid protein interaction. If τ is large enough such that α˜π, the budding is complete. Our model explained many features of retrovirus partial budding observed in experiments.

  10. Kinetics of human immunodeficiency virus budding and assembly

    NASA Astrophysics Data System (ADS)

    Zhang, Rui; Nguyen, Toan

    2009-03-01

    Human immunodeficiency virus (HIV) belongs to a large family of RNA viruses, retroviruses. Unlike budding of regular enveloped viruses, retroviruses bud concurrently with the assembly of retroviral capsids on the cell membrane. The kinetics of HIV (and other retroviruses) budding and assembly is therefore strongly affected by the elastic energy of the membrane and fundamentally different from regular viruses. The main result of this work shows that the kinetics is tunable from a fast budding process to a slow and effectively trapped partial budding process, by varying the attractive energy of retroviral proteins (call Gags), relative to the membrane elastic energy. When the Gag-Gag attraction is relatively high, the membrane elastic energy provides a kinetic barrier for the two pieces of the partial capsids to merge. This energy barrier determines the slowest step in the kinetics and the budding time. In the opposite limit, the membrane elastic energy provides not only a kinetic energy barrier, but a free energy barrier. The budding and assembly is effectively trapped at local free energy minimum, corresponding to a partially budded state. The time scale to escape from this metastable state is exponentially large. In both cases, our result fit with experimental measurements pretty well.

  11. A Monitor for Bud Emergence in the Yeast Morphogenesis Checkpoint

    PubMed Central

    Theesfeld, Chandra L.; Zyla, Trevin R.; Bardes, Elaine G.S.; Lew, Daniel J.

    2003-01-01

    Cell cycle transitions are subject to regulation by both external signals and internal checkpoints that monitor satisfactory progression of key cell cycle events. In budding yeast, the morphogenesis checkpoint arrests the cell cycle in response to perturbations that affect the actin cytoskeleton and bud formation. Herein, we identify a step in this checkpoint pathway that seems to be directly responsive to bud emergence. Activation of the kinase Hsl1p is dependent upon its recruitment to a cortical domain organized by the septins, a family of conserved filament-forming proteins. Under conditions that delayed or blocked bud emergence, Hsl1p recruitment to the septin cortex still took place, but hyperphosphorylation of Hsl1p and recruitment of the Hsl1p-binding protein Hsl7p to the septin cortex only occurred after bud emergence. At this time, the septin cortex spread to form a collar between mother and bud, and Hsl1p and Hsl7p were restricted to the bud side of the septin collar. We discuss models for translating cellular geometry (in this case, the emergence of a bud) into biochemical signals regulating cell proliferation. PMID:12925763

  12. Desiccation tolerance of dormant buds from selected Prunus species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dormant buds of woody plant species present a convenient material for backing-up of germplasm in liquid nitrogen. Routinely, this type of material is used in long-term preservation of only a few species (e.g. apple and sour cherry). Cryopreservation procedures of dormant buds are species dependent, ...

  13. beta-catenin signaling can initiate feather bud development.

    PubMed

    Noramly, S; Freeman, A; Morgan, B A

    1999-08-01

    Intercellular signaling by a subset of Wnts is mediated by stabilization of cytoplasmic beta-catenin and its translocation to the nucleus. Immunolocalization of beta-catenin in developing chick skin reveals that this signaling pathway is active in a dynamic pattern from the earliest stages of feather bud development. Forced activation of this pathway by expression of a stabilized beta-catenin in the ectoderm results in the ectopic formation of feather buds. This construct is sufficient to induce bud formation since it does so both within presumptive feather tracts and in normally featherless regions where tract-specific signals are absent. It is also insensitive to the lateral inhibition that mediates the normal spacing of buds and can induce ectopic buds in interfollicular skin. However, additional patterning signals cooperate with this pathway to regulate gene expression within domains of stabilized beta-catenin expression. Localized activation of this pathway within the bud as it develops is required for normal morphogenesis and ectopic activation of the pathway leads to abnormally oriented buds and growths on the feather filaments. These results suggest that activation of the beta-catenin pathway initiates follicle development in embryonic skin and plays important roles in the subsequent morphogenesis of the bud. PMID:10409498

  14. Rabies virus inactivates cofilin to facilitate viral budding and release.

    PubMed

    Zan, Jie; An, Shu-Ting; Mo, Kai-Kun; Zhou, Jian-Wei; Liu, Juan; Wang, Hai-Long; Yan, Yan; Liao, Min; Zhou, Ji-Yong

    2016-09-01

    Cytoplasmic actin and actin-associated proteins have been identified in RABV particles. Although actin is involved in RABV entry into cells, the specific role of actin in RABV budding and release remains unknown. Our study found that RABV M protein-mediated virion budding depends on intact actin filaments. Confocal microscopy demonstrated a block to virions budding, with a number of M protein-mediated budding vesicles detained in the cell cytoplasm. Furthermore, RABV infection resulted in inactivation of cofilin and upregulation of phosphorylated cofilin. Knockdown of cofilin reduced RABV release. These results for the first time indicate that RABV infection resulted in upregulation of phosphorylated cofilin to facililtate actin polymerization for virus budding. PMID:27396619

  15. This bud's for you: mechanisms of cellular nucleocytoplasmic trafficking via nuclear envelope budding.

    PubMed

    Fradkin, Lee G; Budnik, Vivian

    2016-08-01

    The nuclear envelope (NE) physically separates the cytoplasmic and nuclear compartments. While this barrier provides advantages, it also presents a challenge for the nuclear export of large ribonucleoprotein (RNP) complexes. Decades-old dogma holds that all such border-crossing is via the nuclear pore complex (NPC). However, the diameter of the NPC central channel limits the passage of large cargos. Here, we review evidence that such large RNPs employ an endogenous NE-budding pathway, previously thought to be exclusive to the nuclear egress of Herpes viruses. We discuss this and other models proposed, the likelihood that this pathway is conserved, and the consequences of disrupting NE-budding for synapse development, localized translation of synaptic mRNAs, and laminopathies inducing accelerated aging. PMID:27236823

  16. Budding Yeast for Budding Geneticists: A Primer on the Saccharomyces cerevisiae Model System

    PubMed Central

    Duina, Andrea A.; Miller, Mary E.; Keeney, Jill B.

    2014-01-01

    The budding yeast Saccharomyces cerevisiae is a powerful model organism for studying fundamental aspects of eukaryotic cell biology. This Primer article presents a brief historical perspective on the emergence of this organism as a premier experimental system over the course of the past century. An overview of the central features of the S. cerevisiae genome, including the nature of its genetic elements and general organization, is also provided. Some of the most common experimental tools and resources available to yeast geneticists are presented in a way designed to engage and challenge undergraduate and graduate students eager to learn more about the experimental amenability of budding yeast. Finally, a discussion of several major discoveries derived from yeast studies highlights the far-reaching impact that the yeast system has had and will continue to have on our understanding of a variety of cellular processes relevant to all eukaryotes, including humans. PMID:24807111

  17. Calling Card Analysis in Budding Yeast.

    PubMed

    Mayhew, David; Mitra, Robi D

    2016-02-01

    Calling card analysis is a high-throughput method for identifying the genomic binding sites of multiple transcription factors in a single experiment in budding yeast. By tagging a DNA-binding protein with a targeting domain that directs the insertion of the Ty5 retrotransposon, the genomic binding sites for that transcription factor are marked. The transposition locations are then identified en masse by Illumina sequencing. The calling card protocol allows for simultaneous analysis of multiple transcription factors. By cloning barcodes into the Ty5 transposon, it is possible to pair a unique barcode with every transcription factor in the experiment. The method presented here uses expression of transcription factors from their native loci; however, it can also be altered to measure binding sites of transcription factors overexpressed from a plasmid. PMID:26832687

  18. COPI Budding within the Golgi Stack

    PubMed Central

    Popoff, Vincent; Adolf, Frank; Brügger, Britta; Wieland, Felix

    2011-01-01

    The Golgi serves as a hub for intracellular membrane traffic in the eukaryotic cell. Transport within the early secretory pathway, that is within the Golgi and from the Golgi to the endoplasmic reticulum, is mediated by COPI-coated vesicles. The COPI coat shares structural features with the clathrin coat, but differs in the mechanisms of cargo sorting and vesicle formation. The small GTPase Arf1 initiates coating on activation and recruits en bloc the stable heptameric protein complex coatomer that resembles the inner and the outer shells of clathrin-coated vesicles. Different binding sites exist in coatomer for membrane machinery and for the sorting of various classes of cargo proteins. During the budding of a COPI vesicle, lipids are sorted to give a liquid-disordered phase composition. For the release of a COPI-coated vesicle, coatomer and Arf cooperate to mediate membrane separation. PMID:21844168

  19. Measuring mitotic spindle dynamics in budding yeast

    NASA Astrophysics Data System (ADS)

    Plumb, Kemp

    In order to carry out its life cycle and produce viable progeny through cell division, a cell must successfully coordinate and execute a number of complex processes with high fidelity, in an environment dominated by thermal noise. One important example of such a process is the assembly and positioning of the mitotic spindle prior to chromosome segregation. The mitotic spindle is a modular structure composed of two spindle pole bodies, separated in space and spanned by filamentous proteins called microtubules, along which the genetic material of the cell is held. The spindle is responsible for alignment and subsequent segregation of chromosomes into two equal parts; proper spindle positioning and timing ensure that genetic material is appropriately divided amongst mother and daughter cells. In this thesis, I describe fluorescence confocal microscopy and automated image analysis algorithms, which I have used to observe and analyze the real space dynamics of the mitotic spindle in budding yeast. The software can locate structures in three spatial dimensions and track their movement in time. By selecting fluorescent proteins which specifically label the spindle poles and cell periphery, mitotic spindle dynamics have been measured in a coordinate system relevant to the cell division. I describe how I have characterised the accuracy and precision of the algorithms by simulating fluorescence data for both spindle poles and the budding yeast cell surface. In this thesis I also describe the construction of a microfluidic apparatus that allows for the measurement of long time-scale dynamics of individual cells and the development of a cell population. The tools developed in this thesis work will facilitate in-depth quantitative analysis of the non-equilibrium processes in living cells.

  20. Anti-herbivore Structures of Paulownia tomentosa: Morphology, Distribution, Chemical Constituents and Changes During Shoot and Leaf Development

    PubMed Central

    Kobayashi, Sawa; Asai, Teigo; Fujimoto, Yoshinori; Kohshima, Shiro

    2008-01-01

    Background and Aims Recent studies have shown that small structures on plant surfaces serve ecological functions such as resistance against herbivores. The morphology, distribution, chemical composition and changes during shoot and leaf development of such small structures were examined on Paulownia tomentosa. Methods The morphology and distribution of the structures were studied under light microscopy, and their chemical composition was analysed using thin-layer chromatography and high-performance liquid chromatography. To further investigate the function of these structures, several simple field experiments and observations were also conducted. Key Results Three types of small structures on P. tomentosa were investigated: bowl-shaped organs, glandular hairs and dendritic trichomes. The bowl-shaped organs were densely aggregated on the leaves near flower buds and were determined to be extrafloral nectarines (EFNs) that secrete sugar and attract ants. Nectar production of these organs was increased by artificial damage to the leaves, suggesting an anti-herbivore function through symbiosis with ants. Glandular hairs were found on the surfaces of young and/or reproductive organs. Glandular hairs on leaves, stems and flowers secreted mucilage containing glycerides and trapped small insects. Secretions from glandular hairs on flowers and immature fruits contained flavonoids, which may provide protection against some herbivores. Yellow dendritic trichomes on the adaxial side of leaves also contained flavonoids identical to those secreted by the glandular hairs on fruits and flowers. Three special types of leaves, which differed from the standard leaves in shape, size and identity of small structures, developed near young shoot tips or young flower buds. The density of small structures on these leaf types was higher than on standard leaves, suggesting that these leaf types may be specialized to protect young leaves or reproductive organs. Changes in the small structures

  1. [Establishment of high frequency regeneration via leaf explants of 'Red Sun' kiwifruit (Actinidia chinensis)].

    PubMed

    Zhao, Xupeng; Luo, Keming; Zhou, Yue; Wu, Xiuhua; Yang, Li; Tang, Shaohu

    2013-11-01

    A high efficient in vitro regeneration protocol was developed from leaf explants of the female 'Red Sun' kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. This method does not require formation of callus tissues which leads to somaclonal variations. The results show that the adventitious buds developing directly from explants tissue were noticed after 30 d of culture. The maximum regeneration frequency of adventitious buds is 100% and 18.67 shoots was observed in each leaf explants when MS medium was supplemented with 3.0 mg/L BA+1.0 mg/L NAA. The optimal culture medium for bud multiplication is MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3 and the multiplication coefficient reached 8.63. On the rooting medium with 1/2 MS+0.8 mg/L IBA for 15 d, the adventitious plantlets were transferred into matrix perlite supplied with 1/2 MS liquid medium for 15 d and the rooting rate reached 100%. 95 out of 98 plantlets (96.94%) survived acclimatization, producing healthy plants in the greenhouse. Taken together, a highly efficient regeneration method via leaf explants of 'Red Sun' kiwifruit was successfully established. This protocol may be useful for micropropagation and genetic transformation studies of 'Red Sun' kiwifruit. PMID:24701825

  2. Negative feedback regulation of auxin signaling by ATHB8/ACL5-BUD2 transcription module.

    PubMed

    Baima, Simona; Forte, Valentina; Possenti, Marco; Peñalosa, Andrés; Leoni, Guido; Salvi, Sergio; Felici, Barbara; Ruberti, Ida; Morelli, Giorgio

    2014-06-01

    The role of auxin as main regulator of vascular differentiation is well established, and a direct correlation between the rate of xylem differentiation and the amount of auxin reaching the (pro)cambial cells has been proposed. It has been suggested that thermospermine produced by ACAULIS5 (ACL5) and bushy and dwarf2 (BUD2) is one of the factors downstream to auxin contributing to the regulation of this process in Arabidopsis. Here, we provide an in-depth characterization of the mechanism through which ACL5 modulates xylem differentiation. We show that an increased level of ACL5 slows down xylem differentiation by negatively affecting the expression of homeodomain-leucine zipper (HD-ZIP) III and key auxin signaling genes. This mechanism involves the positive regulation of thermospermine biosynthesis by the HD-ZIP III protein Arabidopsis thaliana homeobox8 tightly controlling the expression of ACL5 and BUD2. In addition, we show that the HD-ZIP III protein REVOLUTA contributes to the increased leaf vascularization and long hypocotyl phenotype of acl5 likely by a direct regulation of auxin signaling genes such as like auxin resistant2 (LAX2) and LAX3. We propose that proper formation and differentiation of xylem depend on a balance between positive and negative feedback loops operating through HD-ZIP III genes. PMID:24777988

  3. Characterization of Septin Ultrastructure in Budding Yeast Using Electron Tomography

    PubMed Central

    Bertin, Aurélie; Nogales, Eva

    2015-01-01

    Summary Septins are essential for the completion of cytokinesis. In budding yeast, Saccharomyces cerevisiae, septins are located at the bud neck during mitosis and are closely connected to the inner plasma membrane. In vitro, yeast septins have been shown to self-assemble into a variety of filamentous structures, including rods, paired filaments, bundles and rings [1–3]. Using electron tomography of freeze-substituted section and cryo-electron tomography of frozen sections, we determined the three dimensional organization of the septin cytoskeleton in dividing budding yeast with molecular resolution [4,5]. Here we describe the detailed procedures used for our characterization of the septin cellular ultrastructure. PMID:26519309

  4. Candidate genes associated with bud dormancy release in blackcurrant (Ribes nigrum L.)

    PubMed Central

    2010-01-01

    Background The detrimental effects of mild winter temperatures on the consistency of cropping of blackcurrant (Ribes nigrum L.) in parts of Europe have led to increasing interest in the genetic control of dormancy release in this species. This study examined patterns of gene expression in leaf buds of blackcurrant to identify key differential changes in these profiles around the time of budbreak. Results Using leaf bud tissue of blackcurrant, a cDNA library was generated as a source of blackcurrant ESTs for construction of a custom microarray, which was used to identify differential gene expression during dormancy release. Gene activity was lowest in early stages of dormancy, increasing to reach a maximum around the time of budbreak. Genes with significantly changing expression profiles were clustered and evidence is provided for the transient activity of genes previously associated with dormancy processes in other species. Expression profiling identified candidate genes which were mapped onto a blackcurrant genetic linkage map containing budbreak-related QTL. Three genes, which putatively encode calmodulin-binding protein, beta tubulin and acetyl CoA carboxylase respectively, were found to co-localise with budbreak QTL. Conclusions This study provides insight into the genetic control of dormancy transition in blackcurrant, identifying key changes in gene expression around budbreak. Genetic mapping of ESTs enabled the identification of genes which co-localise with previously-characterised blackcurrant QTL, and it is concluded that these genes have probable roles in release of dormancy and can therefore provide a basis for the development of genetic markers for future breeding deployment. PMID:20840772

  5. Cholesterol modification restricts the spread of Shh gradient in the limb bud.

    PubMed

    Li, Yina; Zhang, Huimin; Litingtung, Ying; Chiang, Chin

    2006-04-25

    Sonic hedgehog (Shh) produced in the zone of polarizing activity is the major determinant of anteroposterior development of the amniote limb. The mature and active Shh protein is cholesterol-modified at its C terminus, and the hydrophobic nature of the modification requires the function of Dispatched (mDispA), a seven-pass transmembrane protein, for Shh release from its source. The current model suggests that the cholesterol moiety promotes the spread of Shh gradient in the limb bud. However, this model is inconsistent with findings in Drosophila and not in line with current thoughts on the role of the cholesterol moiety in Shh multimerization. Therefore, it remains unclear how the cholesterol moiety affects the postrelease extracellular behavior of Shh that relates to the shape of its activity gradient in responsive tissues. Here, we report functional analyses in mice showing that Shh lacking cholesterol modification (ShhN) has an increased propensity to spread long-distance, eliciting ectopic Shh pathway activation consistent with target gene expressions and modulating the level of Gli3 processing in the anterior limb mesoderm. These molecular alterations are reflected in the mispatterning of digits in ShhN mutants. Additionally, we provide direct evidence for the long-distance movement of ShhN across the anteroposterior axis of the limb bud. Our findings suggest that the cholesterol moiety regulates the range and shape of the Shh morphogen gradient by restricting rather than promoting the postrelease spread of Shh across the limb bud during early development. PMID:16611729

  6. Transcriptomic analysis of ‘Suli’ pear (Pyrus pyrifolia white pear group) buds during the dormancy by RNA-Seq

    PubMed Central

    2012-01-01

    Background Bud dormancy is a critical developmental process that allows perennial plants to survive unfavorable environmental conditions. Pear is one of the most important deciduous fruit trees in the world, but the mechanisms regulating bud dormancy in this species are unknown. Because genomic information for pear is currently unavailable, transcriptome and digital gene expression data for this species would be valuable resources to better understand the molecular and biological mechanisms regulating its bud dormancy. Results We performed de novo transcriptome assembly and digital gene expression (DGE) profiling analyses of ‘Suli’ pear (Pyrus pyrifolia white pear group) using the Illumina RNA-seq system. RNA-Seq generated approximately 100 M high-quality reads that were assembled into 69,393 unigenes (mean length = 853 bp), including 14,531 clusters and 34,194 singletons. A total of 51,448 (74.1%) unigenes were annotated using public protein databases with a cut-off E-value above 10-5. We mainly compared gene expression levels at four time-points during bud dormancy. Between Nov. 15 and Dec. 15, Dec. 15 and Jan. 15, and Jan. 15 and Feb. 15, 1,978, 1,024, and 3,468 genes were differentially expressed, respectively. Hierarchical clustering analysis arranged 190 significantly differentially-expressed genes into seven groups. Seven genes were randomly selected to confirm their expression levels using quantitative real-time PCR. Conclusions The new transcriptomes offer comprehensive sequence and DGE profiling data for a dynamic view of transcriptomic variation during bud dormancy in pear. These data provided a basis for future studies of metabolism during bud dormancy in non-model but economically-important perennial species. PMID:23234335

  7. A Comparative Proteomic Analysis of the Buds and the Young Expanding Leaves of the Tea Plant (Camellia sinensis L.)

    PubMed Central

    Li, Qin; Li, Juan; Liu, Shuoqian; Huang, Jianan; Lin, Haiyan; Wang, Kunbo; Cheng, Xiaomei; Liu, Zhonghua

    2015-01-01

    Tea (Camellia sinensis L.) is a perennial woody plant that is widely cultivated to produce a popular non-alcoholic beverage; this beverage has received much attention due to its pleasant flavor and bioactive ingredients, particularly several important secondary metabolites. Due to the significant changes in the metabolite contents of the buds and the young expanding leaves of tea plants, high-performance liquid chromatography (HPLC) analysis and isobaric tags for relative and absolute quantitation (iTRAQ) analysis were performed. A total of 233 differentially expressed proteins were identified. Among these, 116 proteins were up-regulated and 117 proteins were down-regulated in the young expanding leaves compared with the buds. A large array of diverse functions was revealed, including roles in energy and carbohydrate metabolism, secondary metabolite metabolism, nucleic acid and protein metabolism, and photosynthesis- and defense-related processes. These results suggest that polyphenol biosynthesis- and photosynthesis-related proteins regulate the secondary metabolite content of tea plants. The energy and antioxidant metabolism-related proteins may promote tea leaf development. However, reverse transcription quantitative real-time PCR (RT-qPCR) showed that the protein expression levels were not well correlated with the gene expression levels. These findings improve our understanding of the molecular mechanism of the changes in the metabolite content of the buds and the young expanding leaves of tea plants. PMID:26096006

  8. The effect of water stress on some morphological, physiological, and biochemical characteristics and bud success on apple and quince rootstocks.

    PubMed

    Bolat, Ibrahim; Dikilitas, Murat; Ercisli, Sezai; Ikinci, Ali; Tonkaz, Tahsin

    2014-01-01

    The effects of different water stress (control, medium, and severe) on some morphological, physiological, and biochemical characteristics and bud success of M9 apple and MA quince rootstocks were determined. The results showed that water stress significantly affected most morphological, physiological, and biochemical characteristics as well as budding success on the both rootstocks. The increasing water stress decreased the relative shoot length, diameter, and plant total fresh and dry weights. Leaf relative water content and chlorophyll index decreased while electrolyte leakage increased with the increase of water stress in both rootstocks. An increase in water stress also resulted in reduction in budding success in Vista Bella/M9 (79.33% and 46.67%) and Santa Maria/MA (70.33% and 15.33%) combinations. However, the water stress in Santa Maria/MA was more prominent. The increase in water stress resulted in higher peroxidase activities as well as phenol contents in both rootstocks. Although catalase activity, anthocyanin, and proline contents increased with the impact of stress, this was not statistically significant. The results suggest that the impact of stress increased with the increase of water stress; therefore, growers should be careful when using M9 and MA rootstocks in both nursery and orchards where water scarcity is present. PMID:24741357

  9. Mechanical feedback stabilizes budding yeast morphogenesis

    NASA Astrophysics Data System (ADS)

    Banavar, Samhita; Trogdon, Michael; Petzold, Linda; Campas, Otger

    Walled cells have the ability to remodel their shape while sustaining an internal turgor pressure that can reach values up to 10 atmospheres. This requires a tight and simultaneous regulation of cell wall assembly and mechanochemistry, but the underlying mechanisms by which this is achieved remain unclear. Using the growth of mating projections in budding yeast (S. cerevisiae) as a motivating example, we have developed a theoretical description that couples the mechanics of cell wall expansion and assembly via a mechanical feedback. In the absence of a mechanical feedback, cell morphogenesis is inherently unstable. The presence of a mechanical feedback stabilizes changes in cell shape and growth, and provides a mechanism to prevent cell lysis in a wide range of conditions. We solve for the dynamics of the system and obtain the different dynamical regimes. In particular, we show that several parameters affect the stability of growth, including the strength of mechanical feedback in the system. Finally, we compare our results to existing experimental data.

  10. Viral and host proteins that modulate filovirus budding

    PubMed Central

    Liu, Yuliang; Harty, Ronald N

    2010-01-01

    The filoviruses, Ebola and Marburg, utilize a multifaceted mechanism for assembly and budding of infectious virions from mammalian cells. Growing evidence not only demonstrates the importance of multiple viral proteins for efficient assembly and budding, but also the exploitation of various host proteins/pathways by the virus during this late stage of filovirus replication, including endocytic compartments, vacuolar protein sorting pathways, ubiquitination machinery, lipid rafts and cytoskeletal components. Continued elucidation of these complex and orchestrated virus-host interactions will provide a fundamental understanding of the molecular mechanisms of filovirus assembly/budding and ultimately lead to the development of novel viral- and/or host-oriented therapeutics to inhibit filovirus egress and spread. This article will focus on the most recent studies on host interactions and modulation of filovirus budding and summarize the key findings from these investigations. PMID:20730024

  11. Factors influencing axillary shoot proliferation and adventitious budding in cedar.

    PubMed

    Renau-Morata, Begoña; Ollero, Javier; Arrillaga, Isabel; Segura, Juan

    2005-04-01

    We developed procedures for in vitro cloning of Cedrus atlantica Manetti and C. libani A. Rich explants from juvenile and mature plants. Explant size was one determinant of the frequency of axillary bud break in both species. Shoot tips and nodal explants mainly developed calli, whereas bud sprouting occurred in defoliated microcuttings cultured on a modified Murashige and Skoog medium without growth regulators. Isolation and continuous subculture of sprouted buds on the same medium allowed cloning of microcuttings from C. atlantica and C. libani seedlings and bicentennial C. libani trees, thus providing a desirable alternative for multiplying mature trees that have demonstrated superior characteristics. We also report adventitious bud differentiation from isolated embryos of C. atlantica. Neither auxin treatments nor other methods tested, including infection with Agrobacterium rhizogenes, were effective in inducing root initiation. PMID:15687096

  12. Tolerance of budding yeast Saccharomyces cerevisiae to ultra high pressure

    NASA Astrophysics Data System (ADS)

    Shibata, M.; Torigoe, M.; Matsumoto, Y.; Yamamoto, M.; Takizawa, N.; Hada, Y.; Mori, Y.; Takarabe, K.; Ono, F.

    2014-05-01

    Our studies on the tolerance of plants and animals against very high pressure of several GPa have been extended to a smaller sized fungus, the budding yeast Saccharomyces cerevisiae. Several pieces of budding yeast (dry yeast) were sealed in a small teflon capsule with a liquid pressure medium fluorinate, and exposed to 7.5 GPa by using a cubic anvil press. The pressure was kept constant for various duration of time from 2 to 24 h. After the pressure was released, the specimens were brought out from the teflon capsule, and they were cultivated on a potato dextrose agar. It was found that the budding yeast exposed to 7.5 GPa for up to 6 h showed multiplication. However, those exposed to 7.5 GPa for longer than 12 h were found dead. The high pressure tolerance of budding yeast is a little weaker than that of tardigrades.

  13. Studies on Cytokinin-Controlled Bud Formation in Moss Protonemata

    PubMed Central

    Brandes, H.; Kende, H.

    1968-01-01

    Application of cytokinins to moss protonemata of the proper physiological age causes bud formation on specific cells (caulonema). During the early stages of their development, buds revert to protonemal filaments if the cytokinin has been removed by washing the protonemata. This indicates that the hormone is not acting as a “trigger” but has to be present during a critical period of time until differentiation is stabilized. Autoradiographs of protonemata treated with a labeled cytokinin, benzyladenine-benzyl-7-14C, show a striking accumulation of the radioactivity in caulonema cells which are in the stage of bud formation, and in the buds themselves. Cells which did not react to the hormone contained very little radioactivity. The accumulation of benzyladenine in the “target cells” may be due to the presence of binding sites which, in turn, may distinguish responding cells from non-responding ones. Images PMID:16656847

  14. Innervation of the undifferentiated limb bud in rabbit embryo.

    PubMed Central

    Cameron, J; McCredie, J

    1982-01-01

    The concept that there are no nerves in the limb bud of mammalian embryos prior to differentiation has been re-examined. Rabbit embryos were collected at 260 and 290 hours gestation, which is prior to cartilage formation in the forelimb at 320 hours. Forelimb buds and adjacent neural tube were excised, fixed and embedded for light and electron microscopy. The limb buds were sectioned in two planes by serial 1 micrometer sections and inspected by light microscopy. Bundles of nerve fibres were seen within the proximal third of the limb bud, with distal ramification into adjacent zones of condensing mesenchyme. Electron microscopy confirmed the presence of axons and associated immature Schwann cells. These results demonstrate the existence of an anatomical framework through which a neurotrophic influence might be brought to bear upon mesenchyme prior to early differentiation. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 10 PMID:7130041

  15. Leaf waxes in riparian trees: hydrogen isotopes, concentrations, and chain-length patterns

    NASA Astrophysics Data System (ADS)

    Tipple, B. J.; Ehleringer, J.; Doman, C.; Khachaturyan, S.

    2011-12-01

    The stable hydrogen isotope ratios of epicuticular leaf wax n-alkanes record aspects of a plant's ecophysiological conditions. However, it remains unclear as to whether n-alkane hydrogen isotope values (δ2H) directly reflect environmental water (source water or tissue water) or environmental water in combination with a biochemical fractionation. Furthermore, it is uncertain if leaf n-alkane δ2H values reflect a single time interval during leaf expansion or if n-alkane δ2H values record the combination of inputs throughout the entire lifespan of a leaf. These different possibilities will influence how leaf wax biomarkers are interpreted in both ecological and environmental reconstruction contexts. To address these issues, we sampled leaves/buds, stems, and water sources of five common western U.S. riparian species under natural field conditions throughout the growing season. Riparian species were selected because the input water source is most likely to be nearly constant through the growing season. We found that species in this study demonstrated marked and systematic variations in n-alkane concentration, average chain length, and δ2H values. Intraspecific patterns were consistent: average chain lengths and δ2H values increased from bud opening through full leaf expansion with little variation during the remainder of the sampling interval, while leaf-wax concentration as a fraction of total biomass increased throughout the growing season. These data imply that leaf-wax δ2H values reflect multiple periods of wax growth and that the leaf wax is continually produced throughout a leaf's lifespan.

  16. Proteomic analysis of ‘Zaosu’ pear (Pyrus bretschneideri Rehd.) and its red skin bud mutation

    PubMed Central

    2012-01-01

    Background Breeding for strong red skin color is an important objective of the pear breeding program. There are few reports of proteome research in green skin pear and its red skin bud mutation. The manuscript at hand is one of the first studies dealing with 2D-PAGE-based analysis of pear fruits and leaves, establishing a suitable sample preparation and testing different 2D-PAGE protocols. Therefore, it may grant a basis for further studies on the pear proteome being the studies main goal. A proteomic analysis was conducted on leaves and fruits of ‘Zaosu’ pear (Pyrus bretschneideri Rehd.) and its red skin bud mutation in order to reveal their genetic differences in the protein level. Results In the present study, the optimized two-dimensional (2-D) gel electrophoresis system of pear leaf and fruit was set up, and applied to analyze the leaves and fruit protein. The interesting peptide fragments were determined using 4800 Plus MALDI TOF/TOFTM Analyzer mass spectrometer, and the sequence obtained was blasted in NCBInr to identify the differentially-expressed protein. In the 1.5-fold differently-expressed proteins between ‘Zaosu’ pear and its mutant, 10 out of 35 proteins in fruit and 12 out of 24 ones in leaves were identified successfully. Among the 22 identified proteins, 7 protein spots were related to photosynthesis and energy metabolism; 4 were associated with environmental stress; 4 with disease defense; 2 with amino acid metabolism; 2 with cytoskeleton; 1 with antioxidant function; 1 with calcium metabolism; and 1 with unknown function. Moreover, related physiological index, such as chlorophyll content, Rubisco content and polyphone oxidase activity, were different between ‘Zaosu’ pear and its mutant. Conclusion A 2-D gel electrophoresis system of pear leaves and fruits was established, which was suitable for the analysis of proteome comparison. To the best of our knowledge, we have performed the first analysis of the proteomic changes in leaves

  17. Real Life Science with Dandelions and Project BudBurst.

    PubMed

    Johnson, Katherine A

    2016-03-01

    Project BudBurst is a national citizen-science project that tracks bloom times and other phenological data for plants across the country. Data from Project BudBurst are being used to measure the effects of climate change. Students can participate in this project by watching any of the plants on the list, including the common dandelion, which makes the program easy and accessible to everyone. Journal of Microbiology & Biology Education. PMID:27047605

  18. Real Life Science with Dandelions and Project BudBurst

    PubMed Central

    Johnson, Katherine A.

    2016-01-01

    Project BudBurst is a national citizen-science project that tracks bloom times and other phenological data for plants across the country. Data from Project BudBurst are being used to measure the effects of climate change. Students can participate in this project by watching any of the plants on the list, including the common dandelion, which makes the program easy and accessible to everyone. Journal of Microbiology & Biology Education PMID:27047605

  19. Impact of peritumoral and intratumoral budding in esophageal adenocarcinomas.

    PubMed

    Thies, Svenja; Guldener, Lars; Slotta-Huspenina, Julia; Zlobec, Inti; Koelzer, Viktor H; Lugli, Alessandro; Kröll, Dino; Seiler, Christian A; Feith, Marcus; Langer, Rupert

    2016-06-01

    Tumor budding has prognostic significance in many carcinomas and is defined as the presence of detached isolated single cells or small cell clusters up to 5 cells at the invasion front (peritumoral budding [PTB]) or within the tumor (intratumoral budding [ITB]). For esophageal adenocarcinomas (EACs), there are currently only few data about the impact of this morphological feature. We investigated tumor budding in a large collective of 200 primarily resected EACs. Pancytokeratin staining was demonstrated to be superior to hematoxylin and eosin staining for the detection of buds with substantial to excellent interobserver agreement and used for subsequent analysis. PTB and ITB were scored across 10 high-power fields (HPFs). The median count of tumor buds was 130/10 HPFs for PTB (range, 2-593) and 80/10 HPFs for ITB (range, 1-656). PTB and ITB correlated significantly with each other (r = 0.9; P < .001). High PTB and ITB rates were seen in more advanced tumor categories (P < .001 each); tumors with lymph node metastases (P < .001/P = .002); and lymphatic, vascular, and perineural invasion and higher tumor grading (P < .001 each). Survival analysis showed an association with worse survival for high-grade ITB (P = .029) but not PTB (P = .385). However, in multivariate analysis, lymph node and resection status, but not ITB, were independent prognostic parameters. In conclusion, PTB and ITB can be observed in EAC to various degrees. High-grade budding is associated with aggressive tumor phenotype. Assessment of tumor budding, especially ITB, may provide additional prognostic information about tumor behavior and may be useful in specific cases for risk stratification of EAC patients. PMID:26980046

  20. Toward Systems Understanding of Leaf Senescence: An Integrated Multi-Omics Perspective on Leaf Senescence Research.

    PubMed

    Kim, Jeongsik; Woo, Hye Ryun; Nam, Hong Gil

    2016-06-01

    Leaf senescence is a complex but tightly regulated developmental process involving a coordinated sequence of multiple molecular events, which ultimately leads to death of the leaf. Efforts to understand the mechanistic principles underlying leaf senescence have been largely made by transcriptomic, proteomic, and metabolomic studies over the past decade. This review focuses on recent milestones in leaf senescence research obtained using multi-omics technologies, as well as future endeavors toward systems understanding of leaf senescence processes. In particular, we discuss recent advances in understanding molecular events during leaf senescence through genome-wide transcriptome analyses in Arabidopsis. We also describe comparative transcriptome analyses used to unveil the commonality and diversity of regulatory mechanisms governing leaf senescence in the plant kingdom. Finally, we provide current illustrations of epigenomic, proteomic, and metabolomic landscapes of leaf senescence. We envisage that integration of multi-omics leaf senescence data will enable us to address unresolved questions regarding leaf senescence, including determining the molecular principles that coordinate concurrent and ordered changes in biological events during leaf senescence. PMID:27174403

  1. Paraphyly and budding speciation in the hairy snail (Pulmonata, Hygromiidae)

    PubMed Central

    Kruckenhauser, Luise; Duda, Michael; Bartel, Daniela; Sattmann, Helmut; Harl, Josef; Kirchner, Sandra; Haring, Elisabeth

    2014-01-01

    Delimitation of species is often complicated by discordance of morphological and genetic data. This may be caused by the existence of cryptic or polymorphic species. The latter case is particularly true for certain snail species showing an exceptionally high intraspecific genetic diversity. The present investigation deals with the Trochulus hispidus complex, which has a complicated taxonomy. Our analyses of the COI sequence revealed that individuals showing a T. hispidus phenotype are distributed in nine highly differentiated mitochondrial clades (showing p-distances up to 19%). The results of a parallel morphometric investigation did not reveal any differentiation between these clades, although the overall variability is quite high. The phylogenetic analyses based on 12S, 16S and COI sequences show that the T. hispidus complex is paraphyletic with respect to several other morphologically well-defined Trochulus species (T. clandestinus, T. villosus, T. villosulus and T. striolatus) which form well-supported monophyletic groups. The nc marker sequence (5.8S–ITS2–28S) shows only a clear separation of T. o. oreinos and T. o. scheerpeltzi, and a weakly supported separation of T. clandestinus, whereas all other species and the clades of the T. hispidus complex appear within one homogeneous group. The paraphyly of the T. hispidus complex reflects its complicated history, which was probably driven by geographic isolation in different glacial refugia and budding speciation. At our present state of knowledge, it cannot be excluded that several cryptic species are embedded within the T. hispidus complex. However, the lack of morphological differentiation of the T. hispidus mitochondrial clades does not provide any hints in this direction. Thus, we currently do not recommend any taxonomic changes. The results of the current investigation exemplify the limitations of barcoding attempts in highly diverse species such as T. hispidus. PMID:25170185

  2. Early epithelial signaling center governs tooth budding morphogenesis.

    PubMed

    Ahtiainen, Laura; Uski, Isa; Thesleff, Irma; Mikkola, Marja L

    2016-09-12

    During organogenesis, cell fate specification and patterning are regulated by signaling centers, specialized clusters of morphogen-expressing cells. In many organs, initiation of development is marked by bud formation, but the cellular mechanisms involved are ill defined. Here, we use the mouse incisor tooth as a model to study budding morphogenesis. We show that a group of nonproliferative epithelial cells emerges in the early tooth primordium and identify these cells as a signaling center. Confocal live imaging of tissue explants revealed that although these cells reorganize dynamically, they do not reenter the cell cycle or contribute to the growing tooth bud. Instead, budding is driven by proliferation of the neighboring cells. We demonstrate that the activity of the ectodysplasin/Edar/nuclear factor κB pathway is restricted to the signaling center, and its inactivation leads to fewer quiescent cells and a smaller bud. These data functionally link the signaling center size to organ size and imply that the early signaling center is a prerequisite for budding morphogenesis. PMID:27621364

  3. Dielectric modelling of cell division for budding and fission yeast

    NASA Astrophysics Data System (ADS)

    Asami, Koji; Sekine, Katsuhisa

    2007-02-01

    The frequency dependence of complex permittivity or the dielectric spectrum of a system including a cell in cell division has been simulated by a numerical technique based on the three-dimensional finite difference method. Two different types of cell division characteristic of budding and fission yeast were examined. The yeast cells are both regarded as a body of rotation, and thus have anisotropic polarization, i.e. the effective permittivity of the cell depends on the orientation of the cell to the direction of an applied electric field. In the perpendicular orientation, where the rotational axis of the cell is perpendicular to the electric field direction, the dielectric spectra for both yeast cells included one dielectric relaxation and its intensity depended on the cell volume. In the parallel orientation, on the other hand, two dielectric relaxations appeared with bud growth for budding yeast and with septum formation for fission yeast. The low-frequency relaxation was shifted to a lower frequency region by narrowing the neck between the bud and the mother cell for budding yeast and by increasing the degree of septum formation for fission yeast. After cell separation, the low-frequency relaxation disappeared. The simulations well interpreted the oscillation of the relative permittivity of culture broth found for synchronous cell growth of budding yeast.

  4. Cryotolerance of apple tree bud is independent of endodormancy

    PubMed Central

    Bilavcik, Alois; Zamecnik, Jiri; Faltus, Milos

    2015-01-01

    Increasing interest in cryopreservation of dormant buds reveals the need for better understanding of the role of dormancy in cryotolerance. Dormancy stage and low-temperature survival of vegetative apple buds (Malus domestica Borkh.), cultivars ‘Sampion’ and ‘Spartan’, collected from orchard were evaluated during three seasons contrasting in temperature and precipitation throughout the arrested plant growth period. During each season, the cultivars differed either in the onset of the endodormancy or in the length of the endodormant period. A simple relation between endodormancy of the buds and their water content was not detected. The cryosurvival of vegetative apple buds of both cultivars correlated with their cold hardening without direct regard to their particular phase of dormancy. The period of the highest bud cryotolerance after low-temperature exposure overlapped with the endodormant period in some evaluated seasons. Both cultivars had the highest cryosurvival in December and January. The presented data were compared with our previous results from a dormancy study of in vitro apple culture. Endodormancy coincided with the period of successful cryosurvival of apple buds after liquid nitrogen exposure, but as such, it was not decisive for their survival and did not limit their successful cryopreservation. PMID:26442012

  5. Tumor Budding: The Name is EMT. Partial EMT.

    PubMed Central

    Grigore, Alexandru Dan; Jolly, Mohit Kumar; Jia, Dongya; Farach-Carson, Mary C.; Levine, Herbert

    2016-01-01

    Tumor budding is a histological phenomenon encountered in various cancers, whereby individual malignant cells and/or small clusters of malignant cells are seen in the tumor stroma. Postulated to be mirror epithelial-mesenchymal transition, tumor budding has been associated with poor cancer outcomes. However, the vast heterogeneity in its exact definition, methodology of assessment, and patient stratification need to be resolved before it can be routinely used as a standardized prognostic feature. Here, we discuss the heterogeneity in defining and assessing tumor budding, its clinical significance across multiple cancer types, and its prospective implementation in clinical practice. Next, we review the emerging evidence about partial, rather than complete, epithelial-mesenchymal phenotype at the tumor bud level, and its connection with tumor proliferation, quiescence, and stemness. Finally, based on recent literature, indicating a co-expression of epithelial and mesenchymal markers in many tumor buds, we posit tumor budding to be a manifestation of this hybrid epithelial/mesenchymal phenotype displaying collective cell migration. PMID:27136592

  6. Assessing soybean leaf area and leaf biomass by spectral measurements

    NASA Technical Reports Server (NTRS)

    Holben, B. N.; Tucker, C. J.; Fan, C. J.

    1979-01-01

    Red and photographic infrared spectral radiances were correlated with soybean total leaf area index, green leaf area index, chlorotic leaf area index, green leaf biomass, chlorotic leaf biomass, and total biomass. The most significant correlations were found to exist between the IR/red radiance ratio data and green leaf area index and/or green leaf biomass (r squared equals 0.85 and 0.86, respectively). These findings demonstrate that remote sensing data can supply information basic to soybean canopy growth, development, and status by nondestructive determination of the green leaf area or green leaf biomass.

  7. Global Climatic Controls On Leaf Size

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

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

  8. Multiple Pathways Influence Mitochondrial Inheritance in Budding Yeast

    PubMed Central

    Frederick, Rebecca L.; Okamoto, Koji; Shaw, Janet M.

    2008-01-01

    Yeast mitochondria form a branched tubular network. Mitochondrial inheritance is tightly coupled with bud emergence, ensuring that daughter cells receive mitochondria from mother cells during division. Proteins reported to influence mitochondrial inheritance include the mitochondrial rho (Miro) GTPase Gem1p, Mmr1p, and Ypt11p. A synthetic genetic array (SGA) screen revealed interactions between gem1Δ and deletions of genes that affect mitochondrial function or inheritance, including mmr1Δ. Synthetic sickness of gem1Δ mmr1Δ double mutants correlated with defective mitochondrial inheritance by large buds. Additional studies demonstrated that GEM1, MMR1, and YPT11 each contribute to mitochondrial inheritance. Mitochondrial accumulation in buds caused by overexpression of either Mmr1p or Ypt11p did not depend on Gem1p, indicating these three proteins function independently. Physical linkage of mitochondria with the endoplasmic reticulum (ER) has led to speculation that distribution of these two organelles is coordinated. We show that yeast mitochondrial inheritance is not required for inheritance or spreading of cortical ER in the bud. Moreover, Ypt11p overexpression, but not Mmr1p overexpression, caused ER accumulation in the bud, revealing a potential role for Ypt11p in ER distribution. This study demonstrates that multiple pathways influence mitochondrial inheritance in yeast and that Miro GTPases have conserved roles in mitochondrial distribution. PMID:18245340

  9. Are there efferent synapses in fish taste buds?

    PubMed

    Reutter, Klaus; Witt, Martin

    2004-12-01

    In fish, nerve fibers of taste buds are organized within the bud's nerve fiber plexus. It is located between the sensory epithelium consisting of light and dark elongated cells and the basal cells. It comprises the basal parts and processes of light and dark cells that intermingle with nerve fibers, which are the dendritic endings of the taste sensory neurons belonging to the cranial nerves VII, IX or X. Most of the synapses at the plexus are afferent; they have synaptic vesicles on the light (or dark) cells side, which is presynaptic. In contrast, the presumed efferent synapses may be rich in synaptic vesicles on the nerve fibers (presynaptic) side, whereas the cells (postsynaptic) side may contain a subsynaptic cistern; a flat compartment of the smooth endoplasmic reticulum. This structure is regarded as a prerequisite of a typical efferent synapse, as occurring in cochlear and vestibular hair cells. In fish taste buds, efferent synapses are rare and were found only in a few species that belong to different taxa. The significance of efferent synapses in fish taste buds is not well understood, because efferent connections between the gustatory nuclei of the medulla with taste buds are not yet proved. PMID:16217620

  10. Repellence of the red bud borer Resseliella oculiperda from grafted apple trees by impregnation of rubber budding strips with essential oils.

    PubMed

    van Tol, Rob W H M; Swarts, Henk J; van der Linden, Anton; Visser, J H

    2007-05-01

    The red bud borer Resseliella oculiperda (Rübs.) is a pest insect of apple trees when rootstocks are grafted with scion buds by 'shield budding'. The female midges are attracted to the wounds of the grafted buds where they lay their eggs. The larvae feed on the cambium and destroy the buds completely or partially, leading to bad union of the buds with the rootstocks. Budding strips are used very often by growers to bind scion buds to rootstocks. These strips cannot prevent midges from reaching the damaged tissue. Chemical treatments applied to the grafts and other types of strip do not provide better protection against the pest and may cause other risks for growers. In orchard experiments in 2000 and 2001, the authors evaluated the repellent action provided by three essential oils and five compounds of plant origin against the midges by impregnating budding strips with them. The essential oils of lavender, Lavandula angustifolia (P. Mill.), and alpha-terpineol decreased the infestation of buds by more than 95 and 80% respectively. The other potential repellents tested [the essential oil of Juniperus virginiana (L.), citronellal, the essential oil of Cinnamomum camphora (L.) J. Presl, R-carvone, linalool and R-fenchone] decreased infestation by 67, 66, 51, 45, 37 and 25% respectively. The formulation and commercial development of budding strips impregnated with lavender oil is discussed. PMID:17421054

  11. Computational Predictions of Structures of Multichromosomes of Budding Yeast

    PubMed Central

    Gürsoy, Gamze; Xu, Yun; Liang, Jie

    2016-01-01

    Knowledge of the global architecture of the cell nucleus and the spatial organization of genome is critical for understanding gene expression and nuclear function. Single-cell imaging techniques provide a wealth of information on the spatial organization of chromosomes. Computational tools for modelling chromosome structure have broad implications in studying the effect of cell nucleus on higher-order genome organization. Here we describe a multichromosome constrained self-avoiding chromatin model for studying ensembles of genome structural models of budding yeast nucleus. We successfully generated a large number of model genomes of yeast with appropriate chromatin fiber diameter, persistence length, and excluded volume under spatial confinement. By incorporating details of the constraints from single-cell imaging studies, our method can model the budding yeast genome realistically. The model developed here provides a general computational framework for studying the overall architecture of budding yeast genome. PMID:25570855

  12. Light Signaling in Bud Outgrowth and Branching in Plants

    PubMed Central

    Leduc, Nathalie; Roman, Hanaé; Barbier, François; Péron, Thomas; Huché-Thélier, Lydie; Lothier, Jérémy; Demotes-Mainard, Sabine; Sakr, Soulaiman

    2014-01-01

    Branching determines the final shape of plants, which influences adaptation, survival and the visual quality of many species. It is an intricate process that includes bud outgrowth and shoot extension, and these in turn respond to environmental cues and light conditions. Light is a powerful environmental factor that impacts multiple processes throughout plant life. The molecular basis of the perception and transduction of the light signal within buds is poorly understood and undoubtedly requires to be further unravelled. This review is based on current knowledge on bud outgrowth-related mechanisms and light-mediated regulation of many physiological processes. It provides an extensive, though not exhaustive, overview of the findings related to this field. In parallel, it points to issues to be addressed in the near future. PMID:27135502

  13. Psidium guajava and Piper betle Leaf Extracts Prolong Vase Life of Cut Carnation (Dianthus caryophyllus) Flowers

    PubMed Central

    Rahman, M. M.; Ahmad, S. H.; Lgu, K. S.

    2012-01-01

    The effect of leaf extracts of Psidium guajava and Piper betle on prolonging vase life of cut carnation flowers was studied. “Carola” and “Pallas Orange” carnation flowers, at bud stage, were pulsed 24 hours with a floral preservative. Then, flowers were placed in a vase solution containing sprite and a “germicide” (leaf extracts of P. guajava and P. betle, 8-HQC, or a copper coin). Flowers treated with 8-HQC, copper coin, and leaf extracts had longer vase life, larger flower diameter, and higher rate of water uptake compared to control (tap water). The leaf extracts of P. guajava and P. betle showed highest antibacterial and antifungal activities compared to the other treatments. Both showed similar effects on flower quality as the synthetic germicide, 8-HQC. Therefore, these extracts are likely natural germicides to prolong vase life of cut flowers. PMID:22619568

  14. EARLY BUD-BREAK1 (EBB1) defines a conserved mechanism for control of bud-break in woody perennials

    PubMed Central

    Busov, Victor; Carneros, Elena; Yakovlev, Igor

    2016-01-01

    Bud-break is an environmentally and economically important trait in trees, shrubs and vines from temperate latitudes. Poor synchronization of bud-break timing with local climates can lead to frost injuries, susceptibility to pests and pathogens and poor crop yields in fruit trees and vines. The rapid climate changes outpace the adaptive capacities of plants to respond through natural selection. This is particularly true for trees which have long generation cycle and thus the adaptive changes are significantly delayed. Therefore, to devise appropriate breeding and conservation strategies, it is imperative to understand the molecular underpinnings that govern dormancy mechanisms. We have recently identified and characterized the poplar EARLY BUD-BREAK 1 (EBB1) gene. EBB1 is a positive regulator of bud-break and encodes a transcription factor from the AP2/ERF family. Here, using comparative and functional genomics approaches we show that EBB1 function in regulation of bud-break is likely conserved across wide range of woody perennial species with importance to forestry and agriculture. PMID:26317150

  15. Yellow leaf blotch

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yellow leaf blotch occurs worldwide in temperate climates. The disease is reported from countries in Asia, Australasia, Oceania, Europe, North America, Central America, the West Indies, and South America. In the northern Great Plains of North America, it is often the major leaf disease on alfalfa....

  16. [Effects of cold-shock on the growth and flower bud differentiation of tomato seedlings under high temperature stress].

    PubMed

    Li, Sheng-li; Xia, Ya-zhen; Sun, Zhi-qiang

    2016-02-01

    In order to explore the effects of cold-shock on the growth and flower bud differentiation of tomato seedlings under high temperature, tomato seedlings were subjected to cold-shock treat- ments every day with 10 °C for 10 minutes in. an artificial climate chamber. Tomato seedlings were treated with cold-shock at the first true leaf stage and the treatment lasted for 15 days. Tomato seed- lings without cold-shock were used as control. At the fourth true leaf period of tomato seedlings, five plants were randomly sampled and the growth characteristics and the ultrastructure changes of meso- phyll cell of tomato seedlings were examined. The flower bud differentiation process of tomato seed- lings was observed at the periods of the second, fourth and sixth true leaves respectively. Flowering and fruiting of tomato seedlings were also investigated after transplanting. The results showed that the stem diameter and health index of tomato seedlings with cold-shock were enhanced by 7.2% and 55.5% compared with seedlings without cold-shock. Mesophyll cells of the seedlings with cold-shock arranged loosely and various organelles such as chloroplasts and mitochondria were morphologically integrated, while chloroplasts and mitochondria of seedlings mesophyll cells without cold-shock swelled up and thylakoids vacuolized apparently. The flower bud differentiation process of seedlings with cold-shock could be advanced significantly at the early seedling stage compared with the control and the advancement was weakened with the seedling growing. Fruit set number and percentage on the first and second inflorescence of tomato plants transplanted by seedlings with cold-shock were enhanced significantly compared with those of the control. These results indicated that the injury of membrane structure of various organelles, especially chloroplast and mitochondria could be allevia- ted by cold-shock treatment under high temperature tress. Cold-shock treatment could not only im- prove the

  17. Effect of alternating day and night temperature on short day-induced bud set and subsequent bud burst in long days in Norway spruce

    PubMed Central

    Olsen, Jorunn E.; Lee, YeonKyeong; Junttila, Olavi

    2014-01-01

    Young seedlings of the conifer Norway spruce exhibit short day (SD)-induced cessation of apical growth and bud set. Although different, constant temperatures under SD are known to modulate timing of bud set and depth of dormancy with development of deeper dormancy under higher compared to lower temperature, systematic studies of effects of alternating day (DT) and night temperatures (NT) are limited. To shed light on this, seedlings of different provenances of Norway spruce were exposed to a wide range of DT-NT combinations during bud development, followed by transfer to forcing conditions of long days (LD) and 18°C, directly or after different periods of chilling. Although no specific effect of alternating DT/NT was found, the results demonstrate that the effects of DT under SD on bud set and subsequent bud break are significantly modified by NT in a complex way. The effects on bud break persisted after chilling. Since time to bud set correlated with the daily mean temperature under SD at DTs of 18 and 21°C, but not a DT of 15°C, time to bud set apparently also depend on the specific DT, implying that the effect of NT depends on the actual DT. Although higher temperature under SD generally results in later bud break after transfer to forcing conditions, the fastest bud flush was observed at intermediate NTs. This might be due to a bud break-hastening chilling effect of intermediate compared to higher temperatures, and delayed bud development to a stage where bud burst can occur, under lower temperatures. Also, time to bud burst in un-chilled seedlings decreased with increasing SD-duration, suggesting that bud development must reach a certain stage before the processes leading to bud burst are initiated. The present results also indicate that low temperature during bud development had a larger effect on the most southern compared to the most northern provenance studied. Decreasing time to bud burst was observed with increasing northern latitude of origin in un

  18. Electrostatic Levitation of Plant Seeds and Flower Buds

    NASA Astrophysics Data System (ADS)

    Hu, Liang; Wang, Hai-Peng; Li, Liu-Hui; Wei, Bing-Bo

    2012-06-01

    We report the electrostatic levitation of various kinds of seeds and flower buds. Coral berry and pepper near a spherical shape show a stable levitation state. The prolate ellipsoid soybean and flower buds are always “standing" in the free space with satisfactory levitation stability. For the irregular mushroom and wheat grain, the levitation state is characterized as a “top-heavy" posture. These special stable equilibrium states are proved by the analysis of surface charge distribution. The obtained saturation polarization charge of samples presents a good accordance with experimental data. The levitation ability is weighed by the factor m(inr+2)/(inrD2).

  19. AFLP-based genetic mapping of the “bud-flowering” trait in heather (Calluna vulgaris)

    PubMed Central

    2013-01-01

    Background Calluna vulgaris is one of the most important landscaping plants produced in Germany. Its enormous economic success is due to the prolonged flower attractiveness of mutants in flower morphology, the so-called bud-bloomers. In this study, we present the first genetic linkage map of C. vulgaris in which we mapped a locus of the economically highly desired trait “flower type”. Results The map was constructed in JoinMap 4.1. using 535 AFLP markers from a single mapping population. A large fraction (40%) of markers showed distorted segregation. To test the effect of segregation distortion on linkage estimation, these markers were sorted regarding their segregation ratio and added in groups to the data set. The plausibility of group formation was evaluated by comparison of the “two-way pseudo-testcross” and the “integrated” mapping approach. Furthermore, regression mapping was compared to the multipoint-likelihood algorithm. The majority of maps constructed by different combinations of these methods consisted of eight linkage groups corresponding to the chromosome number of C. vulgaris. Conclusions All maps confirmed the independent inheritance of the most important horticultural traits “flower type”, “flower colour”, and “leaf colour”. An AFLP marker for the most important breeding target “flower type” was identified. The presented genetic map of C. vulgaris can now serve as a basis for further molecular marker selection and map-based cloning of the candidate gene encoding the unique flower architecture of C. vulgaris bud-bloomers. PMID:23915059

  20. Molecular Characterization of the Meyer Lemon Isolate of Citrus Tatter Leaf Virus: Complete Genome Sequence and Development of Biologically Active In Vitro Transcripts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tatter leaf virus isolated from Meyer lemon trees (CTLV-ML) from California and Florida induces bud union incompatibility of citrus trees grafted on the widely used trifoliate and trifoliate hybrid rootstocks. The complete genome sequence of CTLV-ML was determined to be 6,495 nucleotides (nts...

  1. Extensive transcriptome changes during natural onset and release of vegetative bud dormancy in Populus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To survive winter conditions, axillary buds of poplar transition from paradormancy to endodormancy. Following sufficient chilling, endodormant axillary buds will transition from endodormancy to ecodormancy. We utilized the near whole genome NimbleGen poplar microarrays to follow transcriptome diff...

  2. Science Shorts: Project BudBurst--Analyzing Data

    ERIC Educational Resources Information Center

    Davis, Kimberly J.; Coskie, Tracy L.

    2008-01-01

    Project BudBurst is a national program intended to get students and other "citizen scientists" to participate in a real study about plants, the environment, and climate change. It also provides an excellent opportunity for students to build data-analysis skills. A collaboration of several agencies and universities, the program began last year and…

  3. Interaction of chill and heat in peach flower bud dormancy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peach bud dormancy requirement is a critical factor in selecting adapted cultivars, but the dormancy process is not well-understood. The Utah model proposes bloom occurs after a cultivar-specific amount of chilling followed by 5000 heat units above 4 °C. This model works well in colder climates, but...

  4. Bud emergence and shoot growth from mature citrus nodal segments

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bud emergence and shoot growth from adult phase citrus nodal cultures were studied using Citrus mitis (calamondin), Citrus paradisi (grapefruit), and Citrus sinensis (sweet orange). The effects of 6-benzylaminopurine (BA), indole 3-acetic acid (IAA), and citrus type on shoot quality and growth fro...

  5. Detection of vegetative bud dormancy QTL in peach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Most temperate tree species cease growth during the winter and enter a state of endodormancy that requires the exposure of vegetative and floral buds to cold temperatures (below 7°C) to initiate normal growth in the spring. The length of the cold treatment required for the resumption of growth is d...

  6. Geographic trend of bud hardiness response in Vitis riparia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A major goal of grapevine breeding efforts for production outside of Mediterranean climates is the production of varieties that have cold tolerance phenotypes. Typically, grapevine breeders use midwinter bud hardiness measures as the descriptive phenotype for cold tolerance. Historical practices of...

  7. Intracellular and extracellular regulation of ureteric bud morphogenesis

    PubMed Central

    DAVIES, JAMIE

    2001-01-01

    The urinary collecting duct system of the permanent kidney develops by growth and branching of an initially unbranched epithelial tubule, the ureteric bud. Formation of the ureteric bud as an outgrowth of the wolffian duct is induced by signalling molecules (such as GDNF) that emanate from the adjacent metanephrogenic mesenchyme. Once it has invaded the mesenchyme, growth and branching of the bud is controlled by a variety of molecules, such as the growth factors GDNF, HGF, TGFβ, activin, BMP-2, BMP-7, and matrix molecules such as heparan sulphate proteoglycans and laminins. These various influences are integrated by signal transduction systems inside ureteric bud cells, with the MAP kinase, protein kinase A and protein kinase C pathways appearing to play major roles. The mechanisms of morphogenetic change that produce branching remain largely obscure, but matrix metalloproteinases are known to be necessary for the process, and there is preliminary evidence for the involvement of the actin/myosin contractile cytoskeleton in creating branch points. PMID:11322719

  8. Extending the dormant bud cryopreservation method to new tree species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In cryopreservation of germplasm, using dormant winter buds (DB) as source plant material is economically favorable over tissue culture options. Although the DB cryopreservation method has been known for many years, the approach is feasible only for cryopreserving a select number of temperate tree s...

  9. 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

  10. Leaf Tissue Senescence

    PubMed Central

    Manos, Peter J.; Goldthwaite, Jonathan

    1975-01-01

    During winter, excised leaf tissue from Rumex obtusifolius degrades chlorophyll at twice the summer rate but the plant hormones, gibberellic acid and zeatin, inhibit the senescence rate by a constant percentage, regardless of season. PMID:16659225

  11. Misexpression of Pknox2 in mouse limb bud mesenchyme perturbs zeugopod development and deltoid crest formation.

    PubMed

    Zhou, Wenrong; Zhu, Huang; Zhao, Jianzhi; Li, Hanjun; Wan, Yong; Cao, Jingjing; Zhao, Haixia; Yu, Jian; Zhou, Rujiang; Yao, Yiyun; Zhang, Lingling; Wang, Lifang; He, Lin; Ma, Gang; Yao, Zhengju; Guo, Xizhi

    2013-01-01

    The TALE (Three Amino acid Loop Extension) family consisting of Meis, Pbx and Pknox proteins is a group of transcriptional co-factors with atypical homeodomains that play pivotal roles in limb development. Compared to the in-depth investigations of Meis and Pbx protein functions, the role of Pknox2 in limb development remains unclear. Here, we showed that Pknox2 was mainly expressed in the zeugopod domain of the murine limb at E10.5 and E11.5. Misexpression of Pknox2 in the limb bud mesenchyme of transgenic mice led to deformities in the zeugopod and forelimb stylopod deltoid crest, but left the autopod and other stylopod skeletons largely intact. These malformations in zeugopod skeletons were recapitulated in mice overexpressing Pknox2 in osteochondroprogenitor cells. Molecular and cellular analyses indicated that the misexpression of Pknox2 in limb bud mesenchyme perturbed the Hox10-11 gene expression profiles, decreased Col2 expression and Bmp/Smad signaling activity in the limb. These results indicated that Pknox2 misexpression affected mesenchymal condensation and early chondrogenic differentiation in the zeugopod skeletons of transgenic embryos, suggesting Pknox2 as a potential regulator of zeugopod and deltoid crest formation. PMID:23717575

  12. A critical three-way junction is conserved in budding yeast and vertebrate telomerase RNAs

    PubMed Central

    Brown, Yogev; Abraham, Mira; Pearl, Sivan; Kabaha, Majdi M.; Elboher, Elhanan; Tzfati, Yehuda

    2007-01-01

    The telomerase ribonucleoprotein copies a short template within its integral RNA moiety onto eukaryotic chromosome ends, compensating for incomplete replication and degradation. Non-template regions of telomerase RNA (TER) are also crucial for telomerase function, yet they are highly divergent in sequence among species and their roles are largely unclear. Using both phylogenetic and mutational analyses, we predicted secondary structures for TERs from Kluyveromyces budding yeast species. A comparison of these secondary structure models with the published model for the Saccharomyces cerevisiae TER reveals a common arrangement into three long arms, a templating domain in the center and several conserved elements in the same positions within the structure. One of them, a three-way junction element, is highly conserved in budding yeast TERs. This element also shows sequence and structure similarity to the critical CR4-CR5 activating domain of vertebrate TERs. Mutational analysis in Kluyveromyces lactis confirmed that this element, and in particular the residues conserved across yeast and vertebrates, is critical for telomerase action both in vivo and in vitro. These findings demonstrate that despite the extreme divergence of TER sequences from different organisms, they do share conserved elements, which presumably carry out common roles in telomerase function. PMID:17855392

  13. Attachment of the Yeast Rhodosporidium toruloides Is Mediated by Adhesives Localized at Sites of Bud Cell Development

    PubMed Central

    Buck, James W.; Andrews, John H.

    1999-01-01

    The basidiomycetous yeast Rhodosporidium toruloides (anamorph, Rhodotorula glutinis) is a common phylloplane epiphyte with biocontrol potential. To understand how R. toruloides adheres to plant surfaces, we obtained nonadherent fungal mutants after chemical mutagenesis with methane-sulfonic acid ethyl ester. Sixteen attachment-minus (Att−) mutants were identified by three methods: (i) screening capsule-minus colonies for loss of adhesive ability; (ii) enrichment for mutants unable to attach to polystyrene; and (iii) selection for reduced fluorescence of fluorescein isothiocyanate-concanavalin A (Con A)-stained cells by fluorescence-activated cell sorting. None of the 16 mutants attached to polystyrene or barley leaves. The lectin Con A eliminated adhesion in all of the wild-type isolates tested. Hapten competition assays indicated that Con A bound to mannose residues on the cell surface. Adhesion of wild-type R. toruloides was transient; nonadhesive cells subsequently became adhesive, with bud development. All Att− mutants and nonattaching wild-type cells lacked polar regions that stained intensely with fluorescein isothiocyanate-Con A and India ink. Lectin, enzyme, and chemical treatments showed that the polar regions consisted of alkali-soluble materials, including mannose residues. Tunicamycin treatment reduced wild-type adhesion, indicating that the mannose residues could be associated with glycoproteins. We concluded that compounds, including mannose residues, that are localized at sites of bud development mediate adhesion of R. toruloides to both polystyrene and barley leaf surfaces. PMID:9925569

  14. 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.

  15. Sensing a bud in the yeast morphogenesis checkpoint: a role for Elm1.

    PubMed

    Kang, Hui; Tsygankov, Denis; Lew, Daniel J

    2016-06-01

    Bud formation by Saccharomyces cerevisiae must be coordinated with the nuclear cycle to enable successful proliferation. Many environmental stresses temporarily disrupt bud formation, and in such circumstances, the morphogenesis checkpoint halts nuclear division until bud formation can resume. Bud emergence is essential for degradation of the mitotic inhibitor, Swe1. Swe1 is localized to the septin cytoskeleton at the bud neck by the Swe1-binding protein Hsl7. Neck localization of Swe1 is required for Swe1 degradation. Although septins form a ring at the presumptive bud site before bud emergence, Hsl7 is not recruited to the septins until after bud emergence, suggesting that septins and/or Hsl7 respond to a "bud sensor." Here we show that recruitment of Hsl7 to the septin ring depends on a combination of two septin-binding kinases: Hsl1 and Elm1. We elucidate which domains of these kinases are needed and show that artificial targeting of those domains suffices to recruit Hsl7 to septin rings even in unbudded cells. Moreover, recruitment of Elm1 is responsive to bud emergence. Our findings suggest that Elm1 plays a key role in sensing bud emergence. PMID:27053666

  16. Sensing a bud in the yeast morphogenesis checkpoint: a role for Elm1

    PubMed Central

    Kang, Hui; Tsygankov, Denis; Lew, Daniel J.

    2016-01-01

    Bud formation by Saccharomyces cerevisiae must be coordinated with the nuclear cycle to enable successful proliferation. Many environmental stresses temporarily disrupt bud formation, and in such circumstances, the morphogenesis checkpoint halts nuclear division until bud formation can resume. Bud emergence is essential for degradation of the mitotic inhibitor, Swe1. Swe1 is localized to the septin cytoskeleton at the bud neck by the Swe1-binding protein Hsl7. Neck localization of Swe1 is required for Swe1 degradation. Although septins form a ring at the presumptive bud site before bud emergence, Hsl7 is not recruited to the septins until after bud emergence, suggesting that septins and/or Hsl7 respond to a “bud sensor.” Here we show that recruitment of Hsl7 to the septin ring depends on a combination of two septin-binding kinases: Hsl1 and Elm1. We elucidate which domains of these kinases are needed and show that artificial targeting of those domains suffices to recruit Hsl7 to septin rings even in unbudded cells. Moreover, recruitment of Elm1 is responsive to bud emergence. Our findings suggest that Elm1 plays a key role in sensing bud emergence. PMID:27053666

  17. Co-ordinated gene expression during phases of dormancy release in raspberry (Rubus idaeus L.) buds.

    PubMed

    Mazzitelli, Luca; Hancock, Robert D; Haupt, Sophie; Walker, Paul G; Pont, Simon D A; McNicol, Jim; Cardle, Linda; Morris, Jenny; Viola, Roberto; Brennan, Rex; Hedley, Peter E; Taylor, Mark A

    2007-01-01

    Bud break in raspberry (Rubus idaeus L.) is often poor and uneven, with many of the subapical buds remaining in a dormant state. In order to determine the dormancy status of raspberry buds, an empirical measure of bud burst in a growth-permissive environment following exposure to chilling (4 degrees C cold storage) was developed. For cv. Glen Ample, percentage bud burst in intact canes and isolated nodes was recorded after 14 d. Isolated nodes (a measure of endodormancy) achieved 100% bud burst after approximately 1500 h chilling whereas buds on intact plants (combined endo- and paradormancy) required an additional 1000 h chilling. A microarray approach was used to follow changes in gene expression that occurred during dormancy transition. The probes for the microarrays were obtained from endodormant and paradormant raspberry bud cDNA libraries. The expression profiles of 5300 clones from these libraries were subjected to principal component analysis to determine the most significant expression patterns. Sequence analysis of these clones, in many cases, enabled their functional categorization and the development of hypotheses concerning the mechanisms of bud dormancy release. Thus a set of novel candidates for key dormancy-related genes from raspberry buds have been identified. Bud dormancy is fundamental to the study of plant developmental processes and, in addition, its regulation is of significant economic importance to fruit and horticultural industries. PMID:17244630

  18. Comparative proteomic and transcriptomic approaches to address the active role of GA4 in Japanese apricot flower bud dormancy release

    PubMed Central

    Zhuang, Weibing; Gao, Zhihong; Zhang, Zhen

    2013-01-01

    Hormones are closely associated with dormancy in deciduous fruit trees, and gibberellins (GAs) are known to be particularly important. In this study, we observed that GA4 treatment led to earlier bud break in Japanese apricot. To understand better the promoting effect of GA4 on the dormancy release of Japanese apricot flower buds, proteomic and transcriptomic approaches were used to analyse the mechanisms of dormancy release following GA4 treatment, based on two-dimensional gel electrophoresis (2-DE) and digital gene expression (DGE) profiling, respectively. More than 600 highly reproducible protein spots (P<0.05) were detected and, following GA4 treatment, 38 protein spots showed more than a 2-fold difference in expression, and 32 protein spots were confidently identified according to the databases. Compared with water treatment, many proteins that were associated with energy metabolism and oxidation–reduction showed significant changes after GA4 treatment, which might promote dormancy release. We observed that genes at the mRNA level associated with energy metabolism and oxidation–reduction also played an important role in this process. Analysis of the functions of the identified proteins and genes and the related metabolic pathways would provide a comprehensive proteomic and transcriptomic view of the coordination of dormancy release after GA4 treatment in Japanese apricot flower buds. PMID:24014872

  19. Cell Biology of Yeast Zygotes, from Genesis to Budding

    PubMed Central

    Tartakoff, Alan M.

    2015-01-01

    The zygote is the essential intermediate that allows interchange of nuclear, mitochondrial and cytosolic determinants between cells. Zygote formation in S. cerevisiae is accomplished by mechanisms that are not characteristic of mitotic cells. These include shifting the axis of growth away from classical cortical landmarks, dramatically reorganizing the cell cortex, remodeling the cell wall in preparation for cell fusion, fusing with an adjacent partner, accomplishing nuclear fusion, orchestrating two steps of septin morphogenesis that account for a delay in fusion of mitochondria, and implementing new norms for bud site selection. This essay emphasizes the sequence of dependent relationships that account for this progression from cell encounters through to zygote budding. It briefly summarizes classical studies of signal transduction and polarity specification and then focuses on downstream events. PMID:25862405

  20. Cell biology of yeast zygotes, from genesis to budding.

    PubMed

    Tartakoff, Alan M

    2015-07-01

    The zygote is the essential intermediate that allows interchange of nuclear, mitochondrial and cytosolic determinants between cells. Zygote formation in Saccharomyces cerevisiae is accomplished by mechanisms that are not characteristic of mitotic cells. These include shifting the axis of growth away from classical cortical landmarks, dramatically reorganizing the cell cortex, remodeling the cell wall in preparation for cell fusion, fusing with an adjacent partner, accomplishing nuclear fusion, orchestrating two steps of septin morphogenesis that account for a delay in fusion of mitochondria, and implementing new norms for bud site selection. This essay emphasizes the sequence of dependent relationships that account for this progression from cell encounters through zygote budding. It briefly summarizes classical studies of signal transduction and polarity specification and then focuses on downstream events. PMID:25862405

  1. 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.

  2. 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

  3. Leaf dynamics in growth and reproduction of Xanthium canadense as influenced by stand density

    PubMed Central

    Ogawa, Takahiro; Oikawa, Shimpei; Hirose, Tadaki

    2015-01-01

    Background and Aims Leaf longevity is controlled by the light gradient in the canopy and also by the nitrogen (N) sink strength in the plant. Stand density may influence leaf dynamics through its effects on light gradient and on plant growth and reproduction. This study tests the hypothesis that the control by the light gradient is manifested more in the vegetative period, whereas the opposite is true when the plant becomes reproductive and develops a strong N sink. Methods Stands of Xanthium canadense were established at two densities. Emergence, growth and death of every leaf on the main stem and branches, and plant growth and N uptake were determined from germination to full senescence. Mean residence time and dry mass productivity were calculated per leaf number, leaf area, leaf mass and leaf N (collectively termed ‘leaf variables’) in order to analyse leaf dynamics and its effect on plant growth. Key Results Branching and reproductive activities were higher at low than at high density. Overall there was no significant difference in mean residence time of leaf variables between the two stands. However, early leaf cohorts on the main stem had a longer retention time at low density, whereas later cohorts had a longer retention time at high density. Branch leaves emerged earlier and tended to live longer at low than at high density. Leaf efficiencies, defined as carbon export per unit investment of leaf variables, were higher at low density in all leaf variables except for leaf number. Conclusions In the vegetative phase of plant growth, the light gradient strongly controls leaf longevity, whereas later the effects of branching and reproductive activities become stronger and over-rule the effect of light environment. As leaf N supports photosynthesis and also works as an N source for plant development, N use is pivotal in linking leaf dynamics with plant growth and reproduction. PMID:26248476

  4. Distinctive expression patterns of heparan sulfate O-sulfotransferases and regional differences in heparan sulfate structure in chick limb buds.

    PubMed

    Nogami, Ken; Suzuki, Hiroaki; Habuchi, Hiroko; Ishiguro, Naoki; Iwata, Hisashi; Kimata, Koji

    2004-02-27

    The skeletal tissue development and patterning in chick limb buds are known to be under the spacio-temporal control of various heparin-binding cell growth factors such as fibroblast growth factors and bone morphogenetic proteins. Different structural regions on heparan sulfate (HS) chains of proteoglycans could be implicated in regional differences in the binding capacities of these cell growth factors, by which they could selectively interact with targeted cells and regulate their signaling in those processes. In this study we first demonstrated by cDNA cloning that one heparan sulfate 2-O-sulfotransferase (HS2ST) and two isoforms of heparan sulfate 6-O-sulfotransferase (HS6ST-1 and -2) occurred in chick embryos and had different substrate specificities each other. We next showed by whole mount in situ hybridization that the HS6ST-1 and HS6ST-2 transcripts were preferentially localized to the anterior proximal region and at the posterior proximal region of the limb bud, respectively, whereas the HS2ST transcript was distributed rather uniformly throughout the bud. Analyses of the structures of HS from different regions of the wing buds have shown variation in that 6-O-sulfated residues are more abundant in the proximal than distal region, whereas iduronosyl 6-O-sulfated residues are abundant in the anterior proximal region and glucuronosyl 6-O-sulfated residues in the posterior proximal region. These results suggest that HS with different sulfation patterns created with multiple sulfotransferase activities provides an appropriate extracellular environment for morphogenetic signal transduction. PMID:14660620

  5. Biomolecules and Natural Medicine Preparations: Analysis of New Sources of Bioactive Compounds from Ribes and Rubus spp. Buds

    PubMed Central

    Donno, Dario; Mellano, Maria Gabriella; Cerutti, Alessandro Kim; Beccaro, Gabriele Loris

    2016-01-01

    It is well known that plants are important sources for the preparation of natural remedies as they contain many biologically active compounds. In particular, polyphenols, terpenic compounds, organic acids, and vitamins are the most widely occurring groups of phytochemicals. Some endemic species may be used for the production of herbal preparations containing phytochemicals with significant bioactivity, as antioxidant activity and anti-inflammatory capacities, and health benefits. Blackberry sprouts and blackcurrant buds are known to contain appreciable levels of bioactive compounds, including flavonols, phenolic acids, monoterpenes, vitamin C, and catechins, with several clinical effects. The aim of this research was to perform an analytical study of blackcurrant and blackberry bud-preparations, in order to identify and quantify the main biomarkers, obtaining a specific phytochemical fingerprint to evaluate the single botanical class contribution to total phytocomplex and relative bioactivity, using a High Performance Liquid Chromatograph−Diode Array Detector; the same analyses were performed both on the University laboratory and commercial preparations. Different chromatographic methods were used to determine concentrations of biomolecules in the preparations, allowing for quantification of statistically significant differences in their bioactive compound content both in the case of Ribes nigrum and Rubus cultivated varieties at different harvest stages. In blackcurrant bud-extracts the most important class was organic acids (50.98%) followed by monoterpenes (14.05%), while in blackberry preparations the main bioactive classes were catechins (50.06%) and organic acids (27.34%). Chemical, pharmaceutical and agronomic-environmental knowledge could be important for obtaining label certifications for the valorization of specific genotypes, with high clinical and pharmaceutical value: this study allowed to develop an effective tool for the natural preparation quality

  6. Biomolecules and Natural Medicine Preparations: Analysis of New Sources of Bioactive Compounds from Ribes and Rubus spp. Buds.

    PubMed

    Donno, Dario; Mellano, Maria Gabriella; Cerutti, Alessandro Kim; Beccaro, Gabriele Loris

    2016-01-01

    It is well known that plants are important sources for the preparation of natural remedies as they contain many biologically active compounds. In particular, polyphenols, terpenic compounds, organic acids, and vitamins are the most widely occurring groups of phytochemicals. Some endemic species may be used for the production of herbal preparations containing phytochemicals with significant bioactivity, as antioxidant activity and anti-inflammatory capacities, and health benefits. Blackberry sprouts and blackcurrant buds are known to contain appreciable levels of bioactive compounds, including flavonols, phenolic acids, monoterpenes, vitamin C, and catechins, with several clinical effects. The aim of this research was to perform an analytical study of blackcurrant and blackberry bud-preparations, in order to identify and quantify the main biomarkers, obtaining a specific phytochemical fingerprint to evaluate the single botanical class contribution to total phytocomplex and relative bioactivity, using a High Performance Liquid Chromatograph-Diode Array Detector; the same analyses were performed both on the University laboratory and commercial preparations. Different chromatographic methods were used to determine concentrations of biomolecules in the preparations, allowing for quantification of statistically significant differences in their bioactive compound content both in the case of Ribes nigrum and Rubus cultivated varieties at different harvest stages. In blackcurrant bud-extracts the most important class was organic acids (50.98%) followed by monoterpenes (14.05%), while in blackberry preparations the main bioactive classes were catechins (50.06%) and organic acids (27.34%). Chemical, pharmaceutical and agronomic-environmental knowledge could be important for obtaining label certifications for the valorization of specific genotypes, with high clinical and pharmaceutical value: this study allowed to develop an effective tool for the natural preparation quality

  7. Elevated CO{sub 2} and leaf shape: Are dandelions getting toothier?

    SciTech Connect

    Thomas, S.C.; Bazzaz, F.A.

    1996-01-01

    Heteroblastic leaf development in Taraxacum officinale is compared between plants grown under ambient (350 ppm) vs. elevated (700 ppm) CO{sub 2} levels. Leaves of elevated CO{sub 2} plants exhibited more deeply incised leaf margins and relatively more slender leaf laminae than leaves of ambient CO{sub 2} plants. These differences were found to be significant in allometric analyses that controlled for differences in leaf size, as well as analyses that controlled for leaf development order. The effects of elevated CO{sub 2} on leaf shape were most pronounced when plants were grown individually, but detectable differences were also found in plants grown at high density. Although less dramatic than in Taraxacum, significant effects of elevated CO{sub 2} on leaf shape were also found in two other weedy rosette species, Plantago major and Rumex crispus. These observations support the long-standing hypothesis that leaf carbohydrate level plays an important role in regulating heteroblastic leaf development, though elevated CO{sub 2} may also affect leaf development through direct hormonal interactions or increased leaf water potential. In Taraxacum, pronounced modifications of leaf shape were found at CO{sub 2} levels predicted to occur within the next century. 33 refs., 5 figs.

  8. Seasonal variability of mercury concentration in soils, buds and leaves of Acer platanoides and Tilia platyphyllos in central Poland.

    PubMed

    Kowalski, Artur; Frankowski, Marcin

    2016-05-01

    In this paper, we present the results of mercury concentration in soils, buds and leaves of maple (Acer platanoides-Ap) and linden (Tilia platyphyllos-Tp) collected in four periods of the growing season of trees, i.e. in April (IV), June (VI), August (VIII) and November (IX) in 2013, from the area of Poznań city (Poland). The highest average concentration of mercury for 88 samples was determined in soils and it equaled 65.8 ± 41.7 ng g(-1) (range 14.5-238.9 ng g(-1)); lower average concentration was found in Ap samples (n = 66): 55.4 ± 18.1 ng g(-1) (range 26.5-106.9 ng g(-1)); in Tp samples 50.4 ± 15.8 ng g(-1) (range 23.1-88.7 ng g(-1)) and in 22 samples of Tp buds 40.8 ± 22.7 ng g(-1) (range 12.4-98.7 ng g(-1)) and Ap buds 28.2 ± 13.6 ng g(-1) (range 8.0-59.5 ng g(-1)). Based on the obtained results, it was observed that the highest concentration of mercury in soils occurred in the centre of Poznań city (95.5 ± 39.1 ng g(-1)), and it was two times higher than the concentration of mercury in other parts of the city. Similar dependencies were not observed for the leaf samples of Ap and Tp. It was found that mercury concentrations in the soil and leaves of maple and linden were different depending on the period of the growing season (April to November). Mercury content in the examined samples was higher in the first two research periods (April IV, June VI), and then, in the following periods, the accumulation of mercury decreased both in soil and leaf samples of the two tree species. There was no correlation found between mercury concentration in leaves and mercury concentration in soils during the four research periods (April-November). When considering the transfer coefficient, it was observed that the main source of mercury in leaves is the mercury coming from the atmosphere. PMID:26846237

  9. Removal of deciduous canine tooth buds in Kenyan rural Maasai.

    PubMed

    Hassanali, J; Amwayi, P; Muriithi, A

    1995-04-01

    The removal of deciduous canine tooth buds in early childhood is a practice that has been documented in Kenya and in neighboring countries. This paper describes the occurrence, rationale and method of this practice amongst rural Kenyan Maasai. In a group of 95 children aged between six months and two years, who were examined in 1991/92, 87% were found to have undergone the removal of one or more deciduous canine tooth buds. In an older age group (3-7 years of age), 72% of the 111 children examined exhibited missing mandibular or maxillary deciduous canines. It was found that the actual removal of a deciduous tooth bud is often performed by middle-aged Maasai women who enucleate the developing tooth using a pointed pen-knife. There exists a strong belief among the Maasai that diarrhoea, vomiting and other febrile illnesses of early childhood are caused by the gingival swelling over the canine region, and which is thought to contain 'worms' or 'nylon' teeth. The immediate and long-term hazards of this practice include profuse bleeding, infection and damage to the developing permanent canines. A multi-disciplinary approach involving social anthropologists in addition to dental and medical personnel, is recommend in order to discourage this harmful operation that appears to be on the increase. PMID:7621751

  10. Leptin's effect on taste bud calcium responses and transmitter secretion.

    PubMed

    Meredith, Tricia L; Corcoran, Alan; Roper, Stephen D

    2015-05-01

    Leptin, a peptide hormone released by adipose tissue, acts on the hypothalamus to control cravings and appetite. Leptin also acts to decrease taste responses to sweet substances, though there is little detailed information regarding where leptin acts in the taste transduction cascade. The present study examined the effects of leptin on sweet-evoked responses and neuro transmitter release from isolated taste buds. Our results indicate that leptin moderately decreased sweet-evoked calcium mobilization in isolated mouse taste buds. We also employed Chinese hamster ovary biosensor cells to examine taste transmitter release from isolated taste buds. Leptin reduced ATP and increased serotonin release in response to sweet stimulation. However, leptin has no effect on bitter-evoked transmitter release, further showing that the action of leptin is sweet specific. Our results support those of previous studies, which state that leptin acts on taste tissue via the leptin receptor, most likely on Type II (Receptor) cells, but also possibly on Type III (Presynaptic) cells. PMID:25537017

  11. Ecological Conditions Favoring Budding in Colonial Organisms under Environmental Disturbance

    PubMed Central

    Nakamaru, Mayuko; Takada, Takenori; Ohtsuki, Akiko; Suzuki, Sayaki U.; Miura, Kanan; Tsuji, Kazuki

    2014-01-01

    Dispersal is a topic of great interest in ecology. Many organisms adopt one of two distinct dispersal tactics at reproduction: the production of small offspring that can disperse over long distances (such as seeds and spawned eggs), or budding. The latter is observed in some colonial organisms, such as clonal plants, corals and ants, in which (super)organisms split their body into components of relatively large size that disperse to a short distance. Contrary to the common dispersal viewpoint, short-dispersal colonial organisms often flourish even in environments with frequent disturbances. In this paper, we investigate the conditions that favor budding over long-distance dispersal of small offspring, focusing on the life history of the colony growth and the colony division ratio. These conditions are the relatively high mortality of very small colonies, logistic growth, the ability of dispersers to peacefully seek and settle unoccupied spaces, and small spatial scale of environmental disturbance. If these conditions hold, budding is advantageous even when environmental disturbance is frequent. These results suggest that the demography or life history of the colony underlies the behaviors of the colonial organisms. PMID:24621824

  12. Paramyxovirus Assembly and Budding: Building Particles that Transmit Infections

    PubMed Central

    Harrison, Megan S.; Sakaguchi, Takemasa; Schmitt, Anthony P.

    2010-01-01

    The paramyxoviruses define a diverse group of enveloped RNA viruses that includes a number of important human and animal pathogens. Examples include human respiratory syncytial virus and the human parainfluenza viruses, which cause respiratory illnesses in young children and the elderly; measles and mumps viruses, which have caused recent resurgences of disease in developed countries; the zoonotic Hendra and Nipah viruses, which have caused several outbreaks of fatal disease in Australia and Asia; and Newcastle disease virus, which infects chickens and other avian species. Like other enveloped viruses, paramyxoviruses form particles that assemble and bud from cellular membranes, allowing the transmission of infections to new cells and hosts. Here, we review recent advances that have improved our understanding of events involved in paramyxovirus particle formation. Contributions of viral matrix proteins, glycoproteins, nucleocapsid proteins, and accessory proteins to particle formation are discussed, as well as the importance of host factor recruitment for efficient virus budding. Trafficking of viral structural components within infected cells is described, together with mechanisms that allow for the selection of specific sites on cellular membranes for the coalescence of viral proteins in preparation of bud formation and virion release. PMID:20398786

  13. Epigenetic regulation of bud dormancy events in perennial plants

    PubMed Central

    Ríos, Gabino; Leida, Carmen; Conejero, Ana; Badenes, María Luisa

    2014-01-01

    Release of bud dormancy in perennial plants resembles vernalization in Arabidopsis thaliana and cereals. In both cases, a certain period of chilling is required for accomplishing the reproductive phase, and several transcription factors with the MADS-box domain perform a central regulatory role in these processes. The expression of DORMANCY-ASSOCIATED MADS-box (DAM)-related genes has been found to be up-regulated in dormant buds of numerous plant species, such as poplar, raspberry, leafy spurge, blackcurrant, Japanese apricot, and peach. Moreover, functional evidence suggests the involvement of DAM genes in the regulation of seasonal dormancy in peach. Recent findings highlight the presence of genome-wide epigenetic modifications related to dormancy events, and more specifically the epigenetic regulation of DAM-related genes in a similar way to FLOWERING LOCUS C, a key integrator of vernalization effectors on flowering initiation in Arabidopsis. We revise the most relevant molecular and genomic contributions in the field of bud dormancy, and discuss the increasing evidence for chromatin modification involvement in the epigenetic regulation of seasonal dormancy cycles in perennial plants. PMID:24917873

  14. Trichomes control flower bud shape by linking together young petals.

    PubMed

    Tan, Jiafu; Walford, Sally-Anne; Dennis, Elizabeth S; Llewellyn, Danny

    2016-01-01

    Trichomes are widespread in plants and develop from surface cells on different tissues(1). They have many forms and functions, from defensive spines to physical barriers that trap layers of air to insulate against desiccation, but there is growing evidence that trichomes can also have developmental roles in regulating flower structure(2,3). We report here that the trichomes on petals of cotton, Gossypium hirsutum L., are essential for correct flower bud shape through a mechanical entanglement of the trichomes on adjacent petals that anchor the edges to counter the opposing force generated by asymmetric expansion of overlapping petals. Silencing a master regulator of petal trichomes, GhMYB-MIXTA-Like10 (GhMYBML10), by RNA interference (RNAi) suppressed petal trichome growth and resulted in flower buds forming into abnormal corkscrew shapes that exposed developing anthers and stigmas to desiccation damage. Artificially gluing petal edges together could partially restore correct bud shape and fertility. Such petal 'Velcro' is present in other Malvaceae and perhaps more broadly in other plant families, although it is not ubiquitous. This mechanism for physical association between separate organs to regulate flower shape and function is different from the usual organ shape control(4) exerted through cell-to-cell communication and differential cell expansion within floral tissues(5,6). PMID:27322517

  15. Topological structure analysis of the protein–protein interaction network in budding yeast

    PubMed Central

    Bu, Dongbo; Zhao, Yi; Cai, Lun; Xue, Hong; Zhu, Xiaopeng; Lu, Hongchao; Zhang, Jingfen; Sun, Shiwei; Ling, Lunjiang; Zhang, Nan; Li, Guojie; Chen, Runsheng

    2003-01-01

    Interaction detection methods have led to the discovery of thousands of interactions between proteins, and discerning relevance within large-scale data sets is important to present-day biology. Here, a spectral method derived from graph theory was introduced to uncover hidden topological structures (i.e. quasi-cliques and quasi-bipartites) of complicated protein–protein interaction networks. Our analyses suggest that these hidden topological structures consist of biologically relevant functional groups. This result motivates a new method to predict the function of uncharacterized proteins based on the classification of known proteins within topological structures. Using this spectral analysis method, 48 quasi-cliques and six quasi-bipartites were isolated from a network involving 11 855 interactions among 2617 proteins in budding yeast, and 76 uncharacterized proteins were assigned functions. PMID:12711690

  16. H2B Ubiquitylation Modulates Spliceosome Assembly and Function in Budding Yeast

    PubMed Central

    Hérissant, Lucas; Moehle, Erica A.; Bertaccini, Diego; Van Dorsselaer, Alain; Schaeffer-Reiss, Christine; Guthrie, Christine; Dargemont, Catherine

    2014-01-01

    Background information Commitment to splicing occurs co-transcriptionally, but a major unanswered question is the extent to which various modifications of chromatin, the template for transcription in vivo, contribute to the regulation of splicing. Results Here we perform genome-wide analyses showing that inhibition of specific marks – H2B ubiquitylation, H3K4 methylation, and H3K36 methylation – perturbs splicing in budding yeast, with each modification exerting gene-specific effects. Furthermore, semi-quantitative mass spectrometry on purified nuclear mRNPs and chromatin immunoprecipitation analysis on intron-containing genes indicated that H2B ubiquitylation, but not Set1-, Set2- or Dot1-dependent H3 methylation, stimulates recruitment of the early splicing factors, namely U1 and U2 snRNPs, onto nascent RNAs. Conclusions These results suggest that histone modifications impact splicing of distinct subsets of genes using distinct pathways. PMID:24476359

  17. Development of automatic image analysis algorithms for protein localization studies in budding yeast

    NASA Astrophysics Data System (ADS)

    Logg, Katarina; Kvarnström, Mats; Diez, Alfredo; Bodvard, Kristofer; Käll, Mikael

    2007-02-01

    Microscopy of fluorescently labeled proteins has become a standard technique for live cell imaging. However, it is still a challenge to systematically extract quantitative data from large sets of images in an unbiased fashion, which is particularly important in high-throughput or time-lapse studies. Here we describe the development of a software package aimed at automatic quantification of abundance and spatio-temporal dynamics of fluorescently tagged proteins in vivo in the budding yeast Saccharomyces cerevisiae, one of the most important model organisms in proteomics. The image analysis methodology is based on first identifying cell contours from bright field images, and then use this information to measure and statistically analyse protein abundance in specific cellular domains from the corresponding fluorescence images. The applicability of the procedure is exemplified for two nuclear localized GFP-tagged proteins, Mcm4p and Nrm1p.

  18. Transcriptomic Analysis for Different Sex Types of Ricinus communis L. during Development from Apical Buds to Inflorescences by Digital Gene Expression Profiling.

    PubMed

    Tan, Meilian; Xue, Jianfeng; Wang, Lei; Huang, Jiaxiang; Fu, Chunling; Yan, Xingchu

    2015-01-01

    The castor plant (Ricinus communis L.) is a versatile industrial oilseed crop with a diversity of sex patterns, its hybrid breeding for improving yield and high purity is still hampered by genetic instability of female and poor knowledge of sex expression mechanisms. To obtain some hints involved in sex expression and provide the basis for further insight into the molecular mechanisms of castor plant sex determination, we performed DGE analysis to investigate differences between the transcriptomes of apices and racemes derived from female (JXBM0705P) and monoecious (JXBM0705M) lines. A total of 18 DGE libraries were constructed from the apices and racemes of a wild monoecious line and its isogenic female derivative at three stages of apex development, in triplicate. Approximately 5.7 million clean tags per library were generated and mapped to the reference castor genome. Transcriptomic analysis showed that identical dynamic changes of gene expression were indicated in monoecious and female apical bud during its development from vegetation to reproduction, with more genes expressed at the raceme formation and infant raceme stages compare to the early leaf bud stage. More than 3000 of differentially expressed genes (DEGs) were detected in Ricinus apices at three developmental stages between two different sex types. A number of DEGs involved in hormone response and biosynthesis, such as auxin response and transport, transcription factors, signal transduction, histone demethylation/methylation, programmed cell death, and pollination, putatively associated with sex expression and reproduction were discovered, and the selected DEGs showed consistent expression between qRT-PCR validation and the DGE patterns. Most of those DEGs were suppressed at the early leaf stage in buds of the mutant, but then activated at the following transition stage (5-7-leaf stage) of buds in the mutant, and ultimately, the number of up-regulated DEGs was equal to that of down-regulation in the

  19. Transcriptomic Analysis for Different Sex Types of Ricinus communis L. during Development from Apical Buds to Inflorescences by Digital Gene Expression Profiling

    PubMed Central

    Tan, Meilian; Xue, Jianfeng; Wang, Lei; Huang, Jiaxiang; Fu, Chunling; Yan, Xingchu

    2016-01-01

    The castor plant (Ricinus communis L.) is a versatile industrial oilseed crop with a diversity of sex patterns, its hybrid breeding for improving yield and high purity is still hampered by genetic instability of female and poor knowledge of sex expression mechanisms. To obtain some hints involved in sex expression and provide the basis for further insight into the molecular mechanisms of castor plant sex determination, we performed DGE analysis to investigate differences between the transcriptomes of apices and racemes derived from female (JXBM0705P) and monoecious (JXBM0705M) lines. A total of 18 DGE libraries were constructed from the apices and racemes of a wild monoecious line and its isogenic female derivative at three stages of apex development, in triplicate. Approximately 5.7 million clean tags per library were generated and mapped to the reference castor genome. Transcriptomic analysis showed that identical dynamic changes of gene expression were indicated in monoecious and female apical bud during its development from vegetation to reproduction, with more genes expressed at the raceme formation and infant raceme stages compare to the early leaf bud stage. More than 3000 of differentially expressed genes (DEGs) were detected in Ricinus apices at three developmental stages between two different sex types. A number of DEGs involved in hormone response and biosynthesis, such as auxin response and transport, transcription factors, signal transduction, histone demethylation/methylation, programmed cell death, and pollination, putatively associated with sex expression and reproduction were discovered, and the selected DEGs showed consistent expression between qRT-PCR validation and the DGE patterns. Most of those DEGs were suppressed at the early leaf stage in buds of the mutant, but then activated at the following transition stage (5-7-leaf stage) of buds in the mutant, and ultimately, the number of up-regulated DEGs was equal to that of down-regulation in the

  20. Automated Sensing of Douglas Fir Bud-Burst

    NASA Astrophysics Data System (ADS)

    Lintz, H. E.; Kruger, A.; Wagner, D. A.; Tenney, I. J.

    2011-12-01

    The timing of plant biological events such as budburst in the spring can have major impacts on plant productivity and ecosystem carbon balance. While research efforts that address the timing of events is gaining considerable momentum, the technology available for sensing and recording the timing of events is limited, especially for trees. Thus, researchers often perform manual measurements, which can be time-consuming and labor-intensive. This has resulted in efforts such as Project BudBurst, a network of professional and volunteer observers across the United States that monitor plants as seasons change. Access to forest trees can be difficult during periods of greatest interest, such as when buds open in the spring. For example, high elevation, snow, and melting snow during the spring hamper access to trees in alpine regions. Researchers at Oregon State University and The University of Iowa are developing instrumentation for automating sensing of budburst in Douglas firs. While the instrumentation targets Douglas firs, it can find application in studying budburst in other species. We present an initial bud-burst sensor that uses optical techniques to sense bud opening. An optical fiber illuminates a target bud with modulated light, a second fiber detects, and guides reflect light to a photodetector and signal processing electronics. Changes in the reflected light indicate the budburst. The instrumentation exploits advances in microelectronics, particularly miniaturization and low power consumption, and uses advanced signal processing techniques such as lock-in detection. The instrumentation records the reflected light every 15 minutes on high-capacity, non-volatile Flash media. Power consumption is very low and sensors have an extrapolated, continuous operating time more than 9 months, suggesting their deployment in the fall, and retrieval in the following spring. We believe the sensor will enable a caliber of research not yet achievable owing to the difficulty of

  1. Differentiation of Apical Bud Cells in a Newly Developed Apical Bud Transplantation Model Using GFP Transgenic Mice as Donor

    PubMed Central

    Sakagami, Ryuji; Yoshinaga, Yasunori; Okamura, Kazuhiko

    2016-01-01

    Rodent mandibular incisors have a unique anatomical structure that allows teeth to grow throughout the lifetime of the rodent. This report presents a novel transplantation technique for studying the apical bud differentiation of rodent mandibular incisors. Incisal apical end tissue with green fluorescent protein from transgenic mouse was transplanted to wild type mice, and the development of the transplanted cells were immunohistologically observed for 12 weeks after the transplantation. Results indicate that the green fluorescent apical end tissue replaced the original tissue, and cells from the apical bud differentiated and extended toward the incisal edge direction. The immunostaining with podoplanin also showed that the characteristics of the green fluorescent tissue were identical to those of the original. The green fluorescent cells were only found in the labial side of the incisor up to 4 weeks. After 12 weeks, however, they were also found in the lingual side. Here the green fluorescent cementocyte-like cells were only present in the cementum close to the dentin surface. This study suggests that some of the cells that form the cellular cementum come from the apical tissue including the apical bud in rodent incisors. PMID:26978064

  2. Strigolactone acts downstream of auxin to regulate bud outgrowth in pea and Arabidopsis.

    PubMed

    Brewer, Philip B; Dun, Elizabeth A; Ferguson, Brett J; Rameau, Catherine; Beveridge, Christine A

    2009-05-01

    During the last century, two key hypotheses have been proposed to explain apical dominance in plants: auxin promotes the production of a second messenger that moves up into buds to repress their outgrowth, and auxin saturation in the stem inhibits auxin transport from buds, thereby inhibiting bud outgrowth. The recent discovery of strigolactone as the novel shoot-branching inhibitor allowed us to test its mode of action in relation to these hypotheses. We found that exogenously applied strigolactone inhibited bud outgrowth in pea (Pisum sativum) even when auxin was depleted after decapitation. We also found that strigolactone application reduced branching in Arabidopsis (Arabidopsis thaliana) auxin response mutants, suggesting that auxin may act through strigolactones to facilitate apical dominance. Moreover, strigolactone application to tiny buds of mutant or decapitated pea plants rapidly stopped outgrowth, in contrast to applying N-1-naphthylphthalamic acid (NPA), an auxin transport inhibitor, which significantly slowed growth only after several days. Whereas strigolactone or NPA applied to growing buds reduced bud length, only NPA blocked auxin transport in the bud. Wild-type and strigolactone biosynthesis mutant pea and Arabidopsis shoots were capable of instantly transporting additional amounts of auxin in excess of endogenous levels, contrary to predictions of auxin transport models. These data suggest that strigolactone does not act primarily by affecting auxin transport from buds. Rather, the primary repressor of bud outgrowth appears to be the auxin-dependent production of strigolactones. PMID:19321710

  3. Ice nucleation activity in various tissues of Rhododendron flower buds: their relevance to extraorgan freezing

    PubMed Central

    Ishikawa, Masaya; Ishikawa, Mikiko; Toyomasu, Takayuki; Aoki, Takayuki; Price, William S.

    2015-01-01

    Wintering flower buds of cold hardy Rhododendron japonicum cooled slowly to subfreezing temperatures are known to undergo extraorgan freezing, whose mechanisms remain obscure. We revisited this material to demonstrate why bud scales freeze first in spite of their lower water content, why florets remain deeply supercooled and how seasonal adaptive responses occur in regard to extraorgan freezing in flower buds. We determined ice nucleation activity (INA) of various flower bud tissues using a test tube-based assay. Irrespective of collection sites, outer and inner bud scales that function as ice sinks in extraorgan freezing had high INA levels whilst florets that remain supercooled and act as a water source lacked INA. The INA level of bud scales was not high in late August when flower bud formation was ending, but increased to reach the highest level in late October just before the first autumnal freeze. The results support the following hypothesis: the high INA in bud scales functions as the subfreezing sensor, ensuring the primary freezing in bud scales at warmer subzero temperatures, which likely allows the migration of floret water to the bud scales and accumulation of icicles within the bud scales. The low INA in the florets helps them remain unfrozen by deep supercooling. The INA in the bud scales was resistant to grinding and autoclaving at 121∘C for 15 min, implying the intrinsic nature of the INA rather than of microbial origin, whilst the INA in stem bark was autoclaving-labile. Anti-nucleation activity (ANA) was implicated in the leachate of autoclaved bud scales, which suppresses the INA at millimolar levels of concentration and likely differs from the colligative effects of the solutes. The tissue INA levels likely contribute to the establishment of freezing behaviors by ensuring the order of freezing in the tissues: from the primary freeze to the last tissue remaining unfrozen. PMID:25859249

  4. Genetic, molecular, and morphological analysis of compound leaf development.

    PubMed

    Goliber, T; Kessler, S; Chen, J J; Bharathan, G; Sinha, N

    1999-01-01

    Leaves, the plant organs responsible for capturing and converting most of the 170 billion metric tons of carbon fixed globally each year, can be broadly grouped into two morphological categories: simple and compound. Although simple-leaved species such as corn and Arabidopsis have traditionally been favored model systems for studying leaf development, recent years have seen an increase in genetic and molecular studies of compound leaf development. Two compound-leaved species in particular have emerged as model systems: tomato and pea. A variety of mutations which alter leaf morphology in these species have been described, and analyses of these mutations have allowed the construction of testable models of leaf development. Also, the knotted-like homeobox (KNOX) genes, which were originally discovered as regulators of meristem function, now appear to have a role in compound leaf development. In addition to the recent genetic and molecular analyses of tomato and pea, insight into the nature of compound leaf development may be gained through the study of (a) heteroblasty and heterophylly, phenomena in which a range of leaf forms can be produced by a single shoot, and (b) the evolutionary origins of compound leaves. PMID:9891889

  5. Effect of CO sub 2 enriched air on the kinetics of leaf expansion. [Pisum sativa; Glycine max

    SciTech Connect

    Potter, J.R. )

    1991-05-01

    Vegetative plants of Pisum sativum (pea) and Glycine max (soybean) were transferred from 350 to 1,200 ppm CO{sub 2} when they had one (pea) or two (soybean) mature leaves and several developing leaves. Controls were kept at 350 ppm. For pea, high CO{sub 2} for 8 days increased dry mass of root, stem, and leaf fractions by 30-50%. Leaf dry mass increase was due primarily to carbohydrate, particularly starch. Dawn levels of starch increased 10-fold within 1 day at high CO{sub 2} and 20-fold at 2 days. At 2 days after transfer leaf starch levels were 1.0 mg cm{sup {minus}2} of leaf area or nearly 30% of leaf dry weight. Soybean data are less complete, but 10 days at high CO{sub 2} increased leaf + stem dry mass by 50% and leaf weight per unit area increased by 14 and 48% at dawn within 1 and 2 days, respectively, at high CO{sub 2}. However 8-10 days at high CO{sub 2} increased total leaf area only slightly (about 15%) for both species, with all the leaf area increase occurring at nodes that were nearly microscopic at the time of transfer. For soybean, most of the increased leaf area due to high CO{sub 2} was from lateral bud break despite a high CO{sub 2} did not stimulated more leaves per plant. Apparently, extra photosynthate had a delayed effect on leaf expansion and did not increase nodes along the main axis. Leaf expansion under high CO{sub 2} was not limited by photosynthate.

  6. Involvement of EARLY BUD-BREAK, an AP2/ERF Transcription Factor Gene, in Bud Break in Japanese Pear (Pyrus pyrifolia Nakai) Lateral Flower Buds: Expression, Histone Modifications and Possible Target Genes.

    PubMed

    Anh Tuan, Pham; Bai, Songling; Saito, Takanori; Imai, Tsuyoshi; Ito, Akiko; Moriguchi, Takaya

    2016-05-01

    In the Japanese pear (Pyrus pyrifolia Nakai) 'Kosui', three developmental stages of lateral flower buds have been proposed to occur during ecodormancy to the flowering phase, i.e. rapid enlargement, sprouting and flowering. Here, we report an APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor gene, named pear EARLY BUD-BREAK (PpEBB), which was highly expressed during the rapid enlargement stage occurring prior to the onset of bud break in flower buds. Gene expression analysis revealed that PpEBB expression was dramatically increased during the rapid enlargement stage in three successive growing seasons. PpEBB transcript levels peaked 1 week prior to onset of bud break in 'Kosui' potted plants treated with hydrogen cyanamide or water under forcing conditions. Chromatin immunoprecipitation-quantitative PCR showed that higher levels of active histone modifications (trimethylation of the histone H3 tail at Lys4) in the 5'-upstream and start codon regions of the PpEBB gene were associated with the induced expression level of PpEBB during the rapid enlargement stage. In addition, we provide evidence that PpEBB may interact with and regulate pear four D-type cyclin (PpCYCD3) genes during bud break in 'Kosui' lateral flower buds. PpEBB significantly increased the promoter activities of four PpCYCD3 genes in a dual-luciferase assay using tobacco leaves. Taken together, our findings uncovered aspects of the bud break regulatory mechanism in the Japanese pear and provided further evidence that the EBB family plays an important role in bud break in perennial plants. PMID:26940832

  7. 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…

  8. Raspberry leaf curl virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Raspberry leaf curl virus (RLCV) is limited to hosts in the genus Rubus and is transmitted persistently by the small raspberry aphid, Aphis rubicola Oestlund. It is found only in North America, principally in the northeastern United States and southeastern Canada and in the Rocky Mountain regions of...

  9. 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...

  10. Model-data assimilation of multiple phenological observations to constrain and predict leaf area index.

    PubMed

    Viskari, Toni; Hardiman, Brady; Desai, Ankur R; Dietze, Michael C

    2015-03-01

    Our limited ability to accurately simulate leaf phenology is a leading source of uncertainty in models of ecosystem carbon cycling. We evaluate if continuously updating canopy state variables with observations is beneficial for predicting phenological events. We employed ensemble adjustment Kalman filter (EAKF) to update predictions of leaf area index (LAI) and leaf extension using tower-based photosynthetically active radiation (PAR) and moderate resolution imaging spectrometer (MODIS) data for 2002-2005 at Willow Creek, Wisconsin, USA, a mature, even-aged, northern hardwood, deciduous forest. The ecosystem demography model version 2 (ED2) was used as the prediction model, forced by offline climate data. EAKF successfully incorporated information from both the observations and model predictions weighted by their respective uncertainties. The resulting. estimate reproduced the observed leaf phenological cycle in the spring and the fall better than a parametric model prediction. These results indicate that during spring the observations contribute most in determining the correct bud-burst date, after which the model performs well, but accurately modeling fall leaf senesce requires continuous model updating from observations. While the predicted net ecosystem exchange (NEE) of CO2 precedes tower observations and unassimilated model predictions in the spring, overall the prediction follows observed NEE better than the model alone. Our results show state data assimilation successfully simulates the evolution of plant leaf phenology and improves model predictions of forest NEE. PMID:26263674

  11. Effects of photoperiod and temperature on the timing of bud burst in Norway spruce (Picea abies).

    PubMed

    Partanen, Jouni; Koski, Veikko; Hänninen, Heikki

    1998-12-01

    We examined the effects of several photoperiod and temperature regimes imposed during the winter-spring period on the timing of bud burst in rooted cuttings of Norway spruce (Picea abies (L.) Karst.) grown in a greenhouse in Finland. The treatments were initiated in November and December after the cuttings had been exposed to natural chilling and freezing events. Irrespective of the treatments applied, time to bud burst decreased with increased duration of previous exposure to natural chilling and freezing events. Fluctuating day/night temperatures and continuous lengthening of the photoperiod hastened bud burst. Shortening the photoperiod delayed bud burst, suggesting that little or no ontogenetic development toward bud burst takes place during mild periods before the winter solstice. In the case of climatic warming, this phenomenon may prevent the premature onset of growth that has been predicted by computer simulations with models that only consider temperature regulation of bud burst. PMID:12651402

  12. Reconstituting ring-rafts in bud-mimicking topography of model membranes

    NASA Astrophysics Data System (ADS)

    Ryu, Yong-Sang; Lee, In-Ho; Suh, Jeng-Hun; Park, Seung Chul; Oh, Soojung; Jordan, Luke R.; Wittenberg, Nathan J.; Oh, Sang-Hyun; Jeon, Noo Li; Lee, Byoungho; Parikh, Atul N.; Lee, Sin-Doo

    2014-07-01

    During vesicular trafficking and release of enveloped viruses, the budding and fission processes dynamically remodel the donor cell membrane in a protein- or a lipid-mediated manner. In all cases, in addition to the generation or relief of the curvature stress, the buds recruit specific lipids and proteins from the donor membrane through restricted diffusion for the development of a ring-type raft domain of closed topology. Here, by reconstituting the bud topography in a model membrane, we demonstrate the preferential localization of cholesterol- and sphingomyelin-enriched microdomains in the collar band of the bud-neck interfaced with the donor membrane. The geometrical approach to the recapitulation of the dynamic membrane reorganization, resulting from the local radii of curvatures from nanometre-to-micrometre scales, offers important clues for understanding the active roles of the bud topography in the sorting and migration machinery of key signalling proteins involved in membrane budding.

  13. Fgf16 is essential for pectoral fin bud formation in zebrafish

    SciTech Connect

    Nomura, Ryohei; Kamei, Eriko; Hotta, Yuuhei; Konishi, Morichika; Miyake, Ayumi; Itoh, Nobuyuki . E-mail: itohnobu@pharm.kyoto-u.ac.jp

    2006-08-18

    Zebrafish pectoral fin bud formation is an excellent model for studying morphogenesis. Fibroblast growth factors (Fgfs) and sonic hedgehog (shh) are essential for pectoral fin bud formation. We found that Fgf16 was expressed in the apical ectodermal ridge (AER) of fin buds. A knockdown of Fgf16 function resulted in no fin bud outgrowth. Fgf16 is required for cell proliferation and differentiation in the mesenchyme and the AER of the fin buds, respectively. Fgf16 functions downstream of Fgf10, a mesenchymal factor, signaling to induce the expression of Fgf4 and Fgf8 in the AER. Fgf16 in the AER and shh in the zone of polarizing activity (ZPA) interact to induce and/or maintain each other's expression. These findings have revealed that Fgf16, a newly identified AER factor, plays a crucial role in pectoral fin bud outgrowth by mediating the interactions of AER-mesenchyme and AER-ZPA.

  14. Reconstituting ring-rafts in bud-mimicking topography of model membranes

    PubMed Central

    Ryu, Yong-Sang; Lee, In-Ho; Suh, Jeng-Hun; Park, Seung Chul; Oh, Soojung; Jordan, Luke R.; Wittenberg, Nathan J.; Oh, Sang-Hyun; Jeon, Noo Li; Lee, Byoungho; Parikh, Atul N.; Lee, Sin-Doo

    2014-01-01

    During vesicular trafficking and release of enveloped viruses, the budding and fission processes dynamically remodel the donor cell membrane in a protein- or a lipid-mediated manner. In all cases, in addition to the generation or relief of the curvature stress, the buds recruit specific lipids and proteins from the donor membrane through restricted diffusion for the development of a ring-type raft domain of closed topology. Here, by reconstituting the bud topography in a model membrane, we demonstrate the preferential localization of cholesterol- and sphingomyelin-enriched microdomains in the collar band of the bud-neck interfaced with the donor membrane. The geometrical approach to the recapitulation of the dynamic membrane reorganization, resulting from the local radii of curvatures from nanometre-to-micrometre scales, offers important clues for understanding the active roles of the bud topography in the sorting and migration machinery of key signalling proteins involved in membrane budding. PMID:25058275

  15. Reconstituting ring-rafts in bud-mimicking topography of model membranes.

    PubMed

    Ryu, Yong-Sang; Lee, In-Ho; Suh, Jeng-Hun; Park, Seung Chul; Oh, Soojung; Jordan, Luke R; Wittenberg, Nathan J; Oh, Sang-Hyun; Jeon, Noo Li; Lee, Byoungho; Parikh, Atul N; Lee, Sin-Doo

    2014-01-01

    During vesicular trafficking and release of enveloped viruses, the budding and fission processes dynamically remodel the donor cell membrane in a protein- or a lipid-mediated manner. In all cases, in addition to the generation or relief of the curvature stress, the buds recruit specific lipids and proteins from the donor membrane through restricted diffusion for the development of a ring-type raft domain of closed topology. Here, by reconstituting the bud topography in a model membrane, we demonstrate the preferential localization of cholesterol- and sphingomyelin-enriched microdomains in the collar band of the bud-neck interfaced with the donor membrane. The geometrical approach to the recapitulation of the dynamic membrane reorganization, resulting from the local radii of curvatures from nanometre-to-micrometre scales, offers important clues for understanding the active roles of the bud topography in the sorting and migration machinery of key signalling proteins involved in membrane budding. PMID:25058275

  16. Sucrose is an early modulator of the key hormonal mechanisms controlling bud outgrowth in Rosa hybrida.

    PubMed

    Barbier, François; Péron, Thomas; Lecerf, Marion; Perez-Garcia, Maria-Dolores; Barrière, Quentin; Rolčík, Jakub; Boutet-Mercey, Stéphanie; Citerne, Sylvie; Lemoine, Remi; Porcheron, Benoît; Roman, Hanaé; Leduc, Nathalie; Le Gourrierec, José; Bertheloot, Jessica; Sakr, Soulaiman

    2015-05-01

    Sugar has only recently been identified as a key player in triggering bud outgrowth, while hormonal control of bud outgrowth is already well established. To get a better understanding of sugar control, the present study investigated how sugar availability modulates the hormonal network during bud outgrowth in Rosa hybrida. Other plant models, for which mutants are available, were used when necessary. Buds were grown in vitro to manipulate available sugars. The temporal patterns of the hormonal regulatory network were assessed in parallel with bud outgrowth dynamics. Sucrose determined bud entrance into sustained growth in a concentration-dependent manner. Sustained growth was accompanied by sustained auxin production in buds, and sustained auxin export in a DR5::GUS-expressing pea line. Several events occurred ahead of sucrose-stimulated bud outgrowth. Sucrose upregulated early auxin synthesis genes (RhTAR1, RhYUC1) and the auxin efflux carrier gene RhPIN1, and promoted PIN1 abundance at the plasma membrane in a pPIN1::PIN1-GFP-expressing tomato line. Sucrose downregulated both RwMAX2, involved in the strigolactone-transduction pathway, and RhBRC1, a repressor of branching, at an early stage. The presence of sucrose also increased stem cytokinin content, but sucrose-promoted bud outgrowth was not related to that pathway. In these processes, several non-metabolizable sucrose analogues induced sustained bud outgrowth in R. hybrida, Pisum sativum, and Arabidopsis thaliana, suggesting that sucrose was involved in a signalling pathway. In conclusion, we identified potential hormonal candidates for bud outgrowth control by sugar. They are central to future investigations aimed at disentangling the processes that underlie regulation of bud outgrowth by sugar. PMID:25873679

  17. Model of human immunodeficiency virus budding and self-assembly: Role of the cell membrane

    NASA Astrophysics Data System (ADS)

    Zhang, Rui; Nguyen, Toan T.

    2008-11-01

    Budding from the plasma membrane of the host cell is an indispensable step in the life cycle of the human immunodeficiency virus (HIV), which belongs to a large family of enveloped RNA viruses, retroviruses. Unlike regular enveloped viruses, retrovirus budding happens concurrently with the self-assembly of the main retrovirus protein subunits (called Gag protein after the name of the genetic material that codes for this protein: Group-specific AntiGen) into spherical virus capsids on the cell membrane. Led by this unique budding and assembly mechanism, we study the free energy profile of retrovirus budding, taking into account the Gag-Gag attraction energy and the membrane elastic energy. We find that if the Gag-Gag attraction is strong, budding always proceeds to completion. During early stage of budding, the zenith angle of partial budded capsids, α , increases with time as α∝t1/2 . However, if the Gag-Gag attraction is weak, a metastable state of partial budding appears. The zenith angle of these partially spherical capsids is given by α0≃(τ2/κσ)1/4 in a linear approximation, where κ and σ are the bending modulus and the surface tension of the membrane, and τ is a line tension of the capsid proportional to the strength of Gag-Gag attraction. Numerically, we find α0<0.3π without any approximations. Using experimental parameters, we show that HIV budding and assembly always proceed to completion in normal biological conditions. On the other hand, by changing Gag-Gag interaction strength or membrane rigidity, it is relatively easy to tune it back and forth between complete budding and partial budding. Our model agrees reasonably well with experiments observing partial budding of retroviruses including HIV.

  18. Identification of budding yeast using a fiber-optic imaging bundle

    NASA Astrophysics Data System (ADS)

    Koschwanez, John; Holl, Mark; Marquardt, Brian; Dragavon, Joe; Burgess, Lloyd; Meldrum, Deirdre

    2004-05-01

    A successful imaging system has been designed and built for yeast pedigree analysis. The system uses a fiber-optic imaging bundle to recognize single yeast cells. Image processing software has been developed to accurately classify the cells as either budding or not budding a daughter cell. This system is intended to replace the body of a microscope for the detection of budding in a microfluidic system.

  19. Measuring Replicative Life Span in the Budding Yeast

    PubMed Central

    Steffen, Kristan K.; Kennedy, Brian K.; Kaeberlein, Matt

    2009-01-01

    Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The budding yeast Saccharomyces cerevisiae has been used extensively to study the biology of aging, and several determinants of yeast longevity have been shown to be conserved in multicellular eukaryotes, including worms, flies, and mice 1. Due to the lack of easily quantified age-associated phenotypes, aging in yeast has been assayed almost exclusively by measuring the life span of cells in different contexts, with two different life span paradigms in common usage 2. Chronological life span refers to the length of time that a mother cell can survive in a non-dividing, quiescence-like state, and is proposed to serve as a model for aging of post-mitotic cells in multicellular eukaryotes. Replicative life span, in contrast, refers the number of daughter cells produced by a mother cell prior to senescence, and is thought to provide a model of aging in mitotically active cells. Here we present a generalized protocol for measuring the replicative life span of budding yeast mother cells. The goal of the replicative life span assay is to determine how many times each mother cell buds. The mother and daughter cells can be easily differentiated by an experienced researcher using a standard light microscope (total magnification 160X), such as the Zeiss Axioscope 40 or another comparable model. Physical separation of daughter cells from mother cells is achieved using a manual micromanipulator equipped with a fiber-optic needle. Typical laboratory yeast strains produce 20-30 daughter cells per mother and one life span experiment requires 2-3 weeks. PMID:19556967

  20. Spaceflight enhances cell aggregation and random budding in Candida albicans.

    PubMed

    Crabbé, Aurélie; Nielsen-Preiss, Sheila M; Woolley, Christine M; Barrila, Jennifer; Buchanan, Kent; McCracken, James; Inglis, Diane O; Searles, Stephen C; Nelman-Gonzalez, Mayra A; Ott, C Mark; Wilson, James W; Pierson, Duane L; Stefanyshyn-Piper, Heidemarie M; Hyman, Linda E; Nickerson, Cheryl A

    2013-01-01

    This study presents the first global transcriptional profiling and phenotypic characterization of the major human opportunistic fungal pathogen, Candida albicans, grown in spaceflight conditions. Microarray analysis revealed that C. albicans subjected to short-term spaceflight culture differentially regulated 452 genes compared to synchronous ground controls, which represented 8.3% of the analyzed ORFs. Spaceflight-cultured C. albicans-induced genes involved in cell aggregation (similar to flocculation), which was validated by microscopic and flow cytometry analysis. We also observed enhanced random budding of spaceflight-cultured cells as opposed to bipolar budding patterns for ground samples, in accordance with the gene expression data. Furthermore, genes involved in antifungal agent and stress resistance were differentially regulated in spaceflight, including induction of ABC transporters and members of the major facilitator family, downregulation of ergosterol-encoding genes, and upregulation of genes involved in oxidative stress resistance. Finally, downregulation of genes involved in actin cytoskeleton was observed. Interestingly, the transcriptional regulator Cap1 and over 30% of the Cap1 regulon was differentially expressed in spaceflight-cultured C. albicans. A potential role for Cap1 in the spaceflight response of C. albicans is suggested, as this regulator is involved in random budding, cell aggregation, and oxidative stress resistance; all related to observed spaceflight-associated changes of C. albicans. While culture of C. albicans in microgravity potentiates a global change in gene expression that could induce a virulence-related phenotype, no increased virulence in a murine intraperitoneal (i.p.) infection model was observed under the conditions of this study. Collectively, our data represent an important basis for the assessment of the risk that commensal flora could play during human spaceflight missions. Furthermore, since the low fluid

  1. Spaceflight Enhances Cell Aggregation and Random Budding in Candida albicans

    PubMed Central

    Woolley, Christine M.; Barrila, Jennifer; Buchanan, Kent; McCracken, James; Inglis, Diane O.; Searles, Stephen C.; Nelman-Gonzalez, Mayra A.; Ott, C. Mark; Wilson, James W.; Pierson, Duane L.; Stefanyshyn-Piper, Heidemarie M.; Hyman, Linda E.; Nickerson, Cheryl A.

    2013-01-01

    This study presents the first global transcriptional profiling and phenotypic characterization of the major human opportunistic fungal pathogen, Candida albicans, grown in spaceflight conditions. Microarray analysis revealed that C. albicans subjected to short-term spaceflight culture differentially regulated 452 genes compared to synchronous ground controls, which represented 8.3% of the analyzed ORFs. Spaceflight-cultured C. albicans–induced genes involved in cell aggregation (similar to flocculation), which was validated by microscopic and flow cytometry analysis. We also observed enhanced random budding of spaceflight-cultured cells as opposed to bipolar budding patterns for ground samples, in accordance with the gene expression data. Furthermore, genes involved in antifungal agent and stress resistance were differentially regulated in spaceflight, including induction of ABC transporters and members of the major facilitator family, downregulation of ergosterol-encoding genes, and upregulation of genes involved in oxidative stress resistance. Finally, downregulation of genes involved in actin cytoskeleton was observed. Interestingly, the transcriptional regulator Cap1 and over 30% of the Cap1 regulon was differentially expressed in spaceflight-cultured C. albicans. A potential role for Cap1 in the spaceflight response of C. albicans is suggested, as this regulator is involved in random budding, cell aggregation, and oxidative stress resistance; all related to observed spaceflight-associated changes of C. albicans. While culture of C. albicans in microgravity potentiates a global change in gene expression that could induce a virulence-related phenotype, no increased virulence in a murine intraperitoneal (i.p.) infection model was observed under the conditions of this study. Collectively, our data represent an important basis for the assessment of the risk that commensal flora could play during human spaceflight missions. Furthermore, since the low fluid

  2. Metabolite changes in conifer buds and needles during forced bud break in Norway spruce (Picea abies) and European silver fir (Abies alba)

    PubMed Central

    Dhuli, Priyanka; Rohloff, Jens; Strimbeck, G. Richard

    2014-01-01

    Environmental changes such as early spring and warm spells induce bud burst and photosynthetic processes in cold-acclimated coniferous trees and consequently, cellular metabolism in overwintering needles and buds. The purpose of the study was to examine metabolism in conifers under forced deacclimation (artificially induced spring) by exposing shoots of Picea abies (boreal species) and Abies alba (temperate species) to a greenhouse environment (22°C, 16/8 h D/N cycle) over a 9 weeks period. Each week, we scored bud opening and collected samples for GC/MS–based metabolite profiling. We detected a total of 169 assigned metabolites and 80 identified metabolites, comprising compounds such as mono- and disaccharides, Krebs cycle acids, amino acids, polyols, phenolics, and phosphorylated structures. Untargeted multivariate statistical analysis based on PCA and cluster analysis segregated samples by species, tissue type, and stage of tissue deacclimations. Similar patterns of metabolic regulation in both species were observed in buds (amino acids, Krebs cycle acids) and needles (hexoses, pentoses, and Krebs cycle acids). Based on correlation of bud opening score with compound levels, distinct metabolites could be associated with bud and shoot development, including amino acids, sugars, and acids with known osmolyte function, and secondary metabolites. This study has shed light on how elevated temperature affects metabolism in buds and needles of conifer species during the deacclimation phase, and contributes to the discussion about how phenological characters in conifers may respond to future global warming. PMID:25566281

  3. γ-Lactam alkaloids from the flower buds of daylily.

    PubMed

    Matsumoto, Takahiro; Nakamura, Seikou; Nakashima, Souichi; Ohta, Tomoe; Yano, Mamiko; Tsujihata, Junichiro; Tsukioka, Junko; Ogawa, Keiko; Fukaya, Masashi; Yoshikawa, Masayuki; Matsuda, Hisashi

    2016-07-01

    Four new alkaloids, hemerocallisamines IV-VII, were isolated from the methanol extract of flower buds of daylily. The chemical structures of the new compounds were elucidated on the basis of chemical and physicochemical evidence. The absolute stereochemistry of the hemerocallisamines IV-VI was elucidated by the application of the modified Mosher's method, HPLC analysis, and optical rotation. In the present study, the isolated alkaloids significantly inhibited the aggregation of Aβ42 in vitro. This is the first report about bioactive alkaloids with a γ-lactam ring from daylily. In addition, isolated nucleosides showed accelerative effects on neurite outgrowth under the non-fasting condition. PMID:26849229

  4. Potato leaf explants as a spaceflight plant test system

    NASA Technical Reports Server (NTRS)

    Wheeler, R. M.

    1986-01-01

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

  5. The Race against Protease Activation Defines the Role of ESCRTs in HIV Budding

    PubMed Central

    Bendjennat, Mourad; Saffarian, Saveez

    2016-01-01

    HIV virions assemble on the plasma membrane and bud out of infected cells using interactions with endosomal sorting complexes required for transport (ESCRTs). HIV protease activation is essential for maturation and infectivity of progeny virions, however, the precise timing of protease activation and its relationship to budding has not been well defined. We show that compromised interactions with ESCRTs result in delayed budding of virions from host cells. Specifically, we show that Gag mutants with compromised interactions with ALIX and Tsg101, two early ESCRT factors, have an average budding delay of ~75 minutes and ~10 hours, respectively. Virions with inactive proteases incorporated the full Gag-Pol and had ~60 minutes delay in budding. We demonstrate that during budding delay, activated proteases release critical HIV enzymes back to host cytosol leading to production of non-infectious progeny virions. To explain the molecular mechanism of the observed budding delay, we modulated the Pol size artificially and show that virion release delays are size-dependent and also show size-dependency in requirements for Tsg101 and ALIX. We highlight the sensitivity of HIV to budding “on-time” and suggest that budding delay is a potent mechanism for inhibition of infectious retroviral release. PMID:27280284

  6. β-Catenin signaling regulates temporally discrete phases of anterior taste bud development.

    PubMed

    Thirumangalathu, Shoba; Barlow, Linda A

    2015-12-15

    The sense of taste is mediated by multicellular taste buds located within taste papillae on the tongue. In mice, individual taste buds reside in fungiform papillae, which develop at mid-gestation as epithelial placodes in the anterior tongue. Taste placodes comprise taste bud precursor cells, which express the secreted factor sonic hedgehog (Shh) and give rise to taste bud cells that differentiate around birth. We showed previously that epithelial activation of β-catenin is the primary inductive signal for taste placode formation, followed by taste papilla morphogenesis and taste bud differentiation, but the degree to which these later elements were direct or indirect consequences of β-catenin signaling was not explored. Here, we define discrete spatiotemporal functions of β-catenin in fungiform taste bud development. Specifically, we show that early epithelial activation of β-catenin, before taste placodes form, diverts lingual epithelial cells from a taste bud fate. By contrast, β-catenin activation a day later within Shh(+) placodes, expands taste bud precursors directly, but enlarges papillae indirectly. Further, placodal activation of β-catenin drives precocious differentiation of Type I glial-like taste cells, but not other taste cell types. Later activation of β-catenin within Shh(+) precursors during papilla morphogenesis also expands taste bud precursors and accelerates Type I cell differentiation, but papilla size is no longer enhanced. Finally, although Shh regulates taste placode patterning, we find that it is dispensable for the accelerated Type I cell differentiation induced by β-catenin. PMID:26525674

  7. Bud protection: a key trait for species sorting in a forest-savanna mosaic.

    PubMed

    Charles-Dominique, Tristan; Beckett, Heath; Midgley, Guy F; Bond, William J

    2015-09-01

    Contrasting fire regimes maintain patch mosaics of savanna, thicket and forest biomes in many African subtropical landscapes. Species dominating each biome are thus expected to display distinct fire-related traits, commonly thought to be bark related. Recent Australian savanna research suggests that bud position, not bark protection alone, determines fire resilience via resprouting. We tested first how bud position influences resprouting ability in 17 tree species. We then compared the effect of both bark-related protection and bud position on the distribution of 63 tree species in 253 transects in all three biomes. Tree species with buds positioned deep under bark had a higher proportion of post-fire aboveground shoot resprouting. Species with low bud protection occurred in fire-prone biomes only if they could root-sucker. The effect of bud protection was supported by a good relationship between species bud protection and distribution across a gradient of fire frequency. Bud protection and high bark production are required to survive frequent fires in savanna. Forests are fire refugia hosting species with little or no bud protection and thin bark. Root-suckering species occur in the three biomes, suggesting that fire is not the only factor filtering this functional type. PMID:25856385

  8. Aip3p/Bud6p, a yeast actin-interacting protein that is involved in morphogenesis and the selection of bipolar budding sites.

    PubMed Central

    Amberg, D C; Zahner, J E; Mulholland, J W; Pringle, J R; Botstein, D

    1997-01-01

    A search for Saccharomyces cerevisiae proteins that interact with actin in the two-hybrid system and a screen for mutants that affect the bipolar budding pattern identified the same gene, AIP3/BUD6. This gene is not essential for mitotic growth but is necessary for normal morphogenesis. MATa/alpha daughter cells lacking Aip3p place their first buds normally at their distal poles but choose random sites for budding in subsequent cell cycles. This suggests that actin and associated proteins are involved in placing the bipolar positional marker at the division site but not at the distal tip of the daughter cell. In addition, although aip3 mutant cells are not obviously defective in the initial polarization of the cytoskeleton at the time of bud emergence, they appear to lose cytoskeletal polarity as the bud enlarges, resulting in the formation of cells that are larger and rounder than normal. aip3 mutant cells also show inefficient nuclear migration and nuclear division, defects in the organization of the secretory system, and abnormal septation, all defects that presumably reflect the involvement of Aip3p in the organization and/or function of the actin cytoskeleton. The sequence of Aip3p is novel but contains a predicted coiled-coil domain near its C terminus that may mediate the observed homo-oligomerization of the protein. Aip3p shows a distinctive localization pattern that correlates well with its likely sites of action: it appears at the presumptive bud site prior to bud emergence, remains near the tips of small bund, and forms a ring (or pair of rings) in the mother-bud neck that is detectable early in the cell cycle but becomes more prominent prior to cytokinesis. Surprisingly, the localization of Aip3p does not appear to require either polarized actin or the septin proteins of the neck filaments. Images PMID:9247651

  9. 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.

  10. Evolution of leaf form correlates with tropical–temperate transitions in Viburnum (Adoxaceae)

    PubMed Central

    Schmerler, Samuel B.; Clement, Wendy L.; Beaulieu, Jeremy M.; Chatelet, David S.; Sack, Lawren; Donoghue, Michael J.; Edwards, Erika J.

    2012-01-01

    Strong latitudinal patterns in leaf form are well documented in floristic comparisons and palaeobotanical studies. However, there is little agreement about their functional significance; in fact, it is still unknown to what degree these patterns were generated by repeated evolutionary adaptation. We analysed leaf form in the woody angiosperm clade Viburnum (Adoxaceae) and document evolutionarily correlated shifts in leafing habit, leaf margin morphology, leaf shape and climate. Multiple independent shifts between tropical and temperate forest habitats have repeatedly been accompanied by a change between evergreen, elliptical leaves with entire margins and deciduous, more rounded leaves with toothed or lobed margins. These consistent shifts in Viburnum support repeated evolutionary adaptation as a major determinant of the global correlation between leaf form and mean annual temperature. Our results provide a new theoretical grounding for the inference of past climates using fossil leaf assemblages. PMID:22810426