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Sample records for barley seed proteomics

  1. Functional proteomics of barley and barley chloroplasts – strategies, methods and perspectives

    PubMed Central

    Petersen, Jørgen; Rogowska-Wrzesinska, Adelina; Jensen, Ole N.

    2013-01-01

    Barley (Hordeum vulgare) is an important cereal grain that is used in a range of products for animal and human consumption. Crop yield and seed quality has been optimized during decades by plant breeding programs supported by biotechnology and molecular biology techniques. The recently completed whole-genome sequencing of barley revealed approximately 26,100 open reading frames, which provides a foundation for detailed molecular studies of barley by functional genomics and proteomics approaches. Such studies will provide further insights into the mechanisms of, for example, drought and stress tolerance, micronutrient utilization, and photosynthesis in barley. In the present review we present the current state of proteomics research for investigations of barley chloroplasts, i.e., the organelle that contain the photosynthetic apparatus in the plant. We describe several different proteomics strategies and discuss their applications in characterization of the barley chloroplast as well as future perspectives for functional proteomics in barley research. PMID:23515231

  2. Identification, cloning and characterization of two thioredoxin h isoforms, HvTrxh1 and HvTrxh2, from the barley seed proteome.

    PubMed

    Maeda, Kenji; Finnie, Christine; ØStergaard, Ole; Svensson, Birte

    2003-06-01

    Two thioredoxin h isoforms, HvTrxh1 and HvTrxh2, were identified in two and one spots, respectively, in a proteome analysis of barley (Hordeum vulgare) seeds based on 2D gel electrophoresis and MS. HvTrxh1 was observed in 2D gel patterns of endosperm, aleurone layer and embryo of mature barley seeds, and HvTrxh2 was present mainly in the embryo. During germination, HvTrxh2 decreased in abundance and HvTrxh1 decreased in the aleurone layer and endosperm but remained at high levels in the embryo. On the basis of MS identification of the two isoforms, expressed sequence tag sequences were identified, and cDNAs encoding HvTrxh1 and HvTrxh2 were cloned by RT-PCR. The sequences were 51% identical, but showed higer similarity to thioredoxin h isoforms from other cereals, e.g. rice Trxh (74% identical with HvTrxh1) and wheat TrxTa (90% identical with HvTrxh2). Recombinant HvTrxh1, HvTrxh2 and TrxTa were produced in Escherichia coli and purified using a three-step procedure. The activity of the purified recombinant thioredoxin h isoforms was demonstrated using insulin and barley alpha-amylase/subtilisin inhibitor as substrates. HvTrxh1 and HvTrxh2 were also efficiently reduced by Arabidopsis thaliana NADP-dependent thioredoxin reductase (NTR). The biochemical properties of HvTrxh2 and TrxTa were similar, whereas HvTrxh1 had higher insulin-reducing activity and was a better substrate for Arabidopsis NTR than HvTrxh2, with a Km of 13 micro m compared with 44 micro m for HvTrxh2. Thus, barley seeds contain two distinct thioredoxin h isoforms which differ in temporal and spatial distribution and kinetic properties, suggesting that they may have different physiological roles. PMID:12787030

  3. Seed proteomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seeds comprise a protective covering, a small embryonic plant, and a nutrient-storage organ. Seeds are protein-rich, and have been the subject of many mass spectrometry-based analyses. Seed storage proteins (SSP), which are transient depots for reduced nitrogen, have been studied for decades by cel...

  4. Seed Proteomics"

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Proteomic analysis of seeds encounters some specific problems that do not impinge on analyses of other plant cells, tissues, or organs. There are anatomic considerations. Seeds comprise the seed coat, the storage organ(s), and the embryonic axis. Are these to be studied individually or as a compo...

  5. Alanine aminotransferase controls seed dormancy in barley.

    PubMed

    Sato, Kazuhiro; Yamane, Miki; Yamaji, Nami; Kanamori, Hiroyuki; Tagiri, Akemi; Schwerdt, Julian G; Fincher, Geoffrey B; Matsumoto, Takashi; Takeda, Kazuyoshi; Komatsuda, Takao

    2016-01-01

    Dormancy allows wild barley grains to survive dry summers in the Near East. After domestication, barley was selected for shorter dormancy periods. Here we isolate the major seed dormancy gene qsd1 from wild barley, which encodes an alanine aminotransferase (AlaAT). The seed dormancy gene is expressed specifically in the embryo. The AlaAT isoenzymes encoded by the long and short dormancy alleles differ in a single amino acid residue. The reduced dormancy allele Qsd1 evolved from barleys that were first domesticated in the southern Levant and had the long dormancy qsd1 allele that can be traced back to wild barleys. The reduced dormancy mutation likely contributed to the enhanced performance of barley in industrial applications such as beer and whisky production, which involve controlled germination. In contrast, the long dormancy allele might be used to control pre-harvest sprouting in higher rainfall areas to enhance global adaptation of barley. PMID:27188711

  6. Alanine aminotransferase controls seed dormancy in barley

    PubMed Central

    Sato, Kazuhiro; Yamane, Miki; Yamaji, Nami; Kanamori, Hiroyuki; Tagiri, Akemi; Schwerdt, Julian G.; Fincher, Geoffrey B.; Matsumoto, Takashi; Takeda, Kazuyoshi; Komatsuda, Takao

    2016-01-01

    Dormancy allows wild barley grains to survive dry summers in the Near East. After domestication, barley was selected for shorter dormancy periods. Here we isolate the major seed dormancy gene qsd1 from wild barley, which encodes an alanine aminotransferase (AlaAT). The seed dormancy gene is expressed specifically in the embryo. The AlaAT isoenzymes encoded by the long and short dormancy alleles differ in a single amino acid residue. The reduced dormancy allele Qsd1 evolved from barleys that were first domesticated in the southern Levant and had the long dormancy qsd1 allele that can be traced back to wild barleys. The reduced dormancy mutation likely contributed to the enhanced performance of barley in industrial applications such as beer and whisky production, which involve controlled germination. In contrast, the long dormancy allele might be used to control pre-harvest sprouting in higher rainfall areas to enhance global adaptation of barley. PMID:27188711

  7. Comparative Proteomic Analysis of Two Barley Cultivars (Hordeum vulgare L.) with Contrasting Grain Protein Content

    PubMed Central

    Guo, Baojian; Luan, Haiye; Lin, Shen; Lv, Chao; Zhang, Xinzhong; Xu, Rugen

    2016-01-01

    Grain protein contents (GPCs) of barley seeds are significantly different between feed and malting barley cultivars. However, there is still no insight into the proteomic analysis of seed proteins between feed and malting barley cultivars. Also, the genetic control of barley GPC is still unclear. GPCs were measured between mature grains of Yangsimai 3 and Naso Nijo. A proteome profiling of differentially expressed protein was established by using a combination of 2-DE and tandem mass spectrometry. In total, 502 reproducible protein spots in barley seed proteome were detected with a pH range of 4–7 and 6–11, among these 41 protein spots (8.17%) were detected differentially expressed between Yangsimai 3 and Naso Nijo. Thirty-four protein spots corresponding to 23 different proteins were identified, which were grouped into eight categories, including stress, protein degradation and post-translational modification, development, cell, signaling, glycolysis, starch metabolism, and other functions. Among the identified proteins, enolase (spot 274) and small subunit of ADP-glucose pyrophosphorylase (spot 271) are exclusively expressed in barley Yangsimai 3, which may be involved in regulating seed protein expression. In addition, malting quality is characterized by an accumulation of serpin protein, Alpha-amylase/trypsin inhibitor CMb and Alpha-amylase inhibitor BDAI-1. Most noticeably, globulin, an important storage protein in barley seed, undergoes post-translational processing in both cultivars, and also displays different expression patterns. PMID:27200019

  8. Search for endophytic diazotrophs in barley seeds

    PubMed Central

    Zawoznik, Myriam S.; Vázquez, Susana C.; Díaz Herrera, Silvana M.; Groppa, María D.

    2014-01-01

    Eight endophytic isolates assigned to Pseudomonas, Azospirillum, and Bacillus genera according to pheno-genotypic features were retrieved from barley seeds under selective pressure for nitrogen-fixers. Genetic relationships among related isolates were investigated through RAPD. Six isolates displayed nitrogen-fixing ability, while all could biosynthesize indolacetic acid in vitro and showed no antibiosis effects against Azospirillum brasilense Az39, a recognized PGPR. PMID:25242949

  9. Isolation and Proteomics Analysis of Barley Centromeric Chromatin Using PICh.

    PubMed

    Zeng, Zixian; Jiang, Jiming

    2016-06-01

    Identification of proteins that are directly or indirectly associated with a specific DNA sequence is often an important goal in molecular biology research. Proteomics of isolated chromatin fragments (PICh) is a technique used to isolate chromatin that contains homologous DNA sequence to a specific nucleic acid probe. All proteins directly and indirectly associated with the DNA sequences that hybridize to the probe are then identified by proteomics.1 We used the PICh technique to isolate chromatin associated with the centromeres of barley (Hordeum vulgare) by using a 2'-deoxy-2'fluoro-ribonucleotides (2'-F RNA) probe that is homologous to the AGGGAG satellite DNA specific to barley centromeres. Proteins associated with the barley centromeric chromatin were then isolated and identified by mass spectrometry. Both alpha-cenH3 and beta-cenH3, the two centromeric histone H3 variants associated with barley centromeres, were positively identified. Interestingly, several different H2A and H2B variants were recovered in the PIChed chromatin. The limitations and future potential of PICh in plant chromatin research are discussed. PMID:27142171

  10. Comparative proteomic analysis of β-aminobutyric acid-mediated alleviation of salt stress in barley.

    PubMed

    Mostek, Agnieszka; Börner, Andreas; Weidner, Stanisław

    2016-02-01

    The non-protein amino acid β-aminobutyric acid (BABA) is known to induce plant resistance to a broad spectrum of biotic and abiotic stresses. This is the first study describing the effect of BABA seed priming on physiological and proteomic changes under salt stress conditions in barley (Hordeum vulgare). The aim of our study was to investigate the changes of fresh weight, dry weight and relative water content (RWC) as well as root proteome changes of two barley lines contrasting in salt tolerance (DH14, DH 187) in response to salt stress after seed priming in water or in 800 μM BABA. Seed priming with BABA significantly increased (p ≤ 0.05) RWC in both barley lines, which indicates considerably lower water loss in BABA-primed plants than in the non-primed control plants. Dry and fresh matter increased significantly in line DH 187, whereas no changes were detected in line DH14. BABA-primed plants of both lines showed different proteomic patterns than the non-primed control plants. The root proteins exhibiting significant abundance changes (1.75-fold change, p ≤ 0.05) were separated by two-dimensional polyacrylamide gel electrophoresis (2D- PAGE). Thirty-one spots, representing 24 proteins, were successfully identified by MALDI TOF/TOF mass spectrometry. The most prominent differences include the up-regulation of antioxidant enzymes (catalase, peroxidase and superoxide dismutase), PR proteins (chitinase, endo-1,3-β-glucosidase), and chaperones (cyclophilin, HSC 70). Our results indicate that BABA induces defence and detoxification processes which may enable faster and more effective responses to salt stress, increasing the chances of survival under adverse environmental conditions. PMID:26760953

  11. Analysis of soybean seed proteins using proteomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This editorial elaborates on investigations consisting of different proteomics technologies and their application to biological sciences. In addition, different classes of soybean seed proteins are discussed. This information will be useful to scientists in obtaining a greater understanding of the...

  12. Involvement of Alternative Splicing in Barley Seed Germination.

    PubMed

    Zhang, Qisen; Zhang, Xiaoqi; Wang, Songbo; Tan, Cong; Zhou, Gaofeng; Li, Chengdao

    2016-01-01

    Seed germination activates many new biological processes including DNA, membrane and mitochondrial repairs and requires active protein synthesis and sufficient energy supply. Alternative splicing (AS) regulates many cellular processes including cell differentiation and environmental adaptations. However, limited information is available on the regulation of seed germination at post-transcriptional levels. We have conducted RNA-sequencing experiments to dissect AS events in barley seed germination. We identified between 552 and 669 common AS transcripts in germinating barley embryos from four barley varieties (Hordeum vulgare L. Bass, Baudin, Harrington and Stirling). Alternative 3' splicing (34%-45%), intron retention (32%-34%) and alternative 5' splicing (16%-21%) were three major AS events in germinating embryos. The AS transcripts were predominantly mapped onto ribosome, RNA transport machineries, spliceosome, plant hormone signal transduction, glycolysis, sugar and carbon metabolism pathways. Transcripts of these genes were also very abundant in the early stage of seed germination. Correlation analysis of gene expression showed that AS hormone responsive transcripts could also be co-expressed with genes responsible for protein biosynthesis and sugar metabolisms. Our RNA-sequencing data revealed that AS could play important roles in barley seed germination. PMID:27031341

  13. Involvement of Alternative Splicing in Barley Seed Germination

    PubMed Central

    Zhang, Qisen; Zhang, Xiaoqi; Wang, Songbo; Tan, Cong; Zhou, Gaofeng; Li, Chengdao

    2016-01-01

    Seed germination activates many new biological processes including DNA, membrane and mitochondrial repairs and requires active protein synthesis and sufficient energy supply. Alternative splicing (AS) regulates many cellular processes including cell differentiation and environmental adaptations. However, limited information is available on the regulation of seed germination at post-transcriptional levels. We have conducted RNA-sequencing experiments to dissect AS events in barley seed germination. We identified between 552 and 669 common AS transcripts in germinating barley embryos from four barley varieties (Hordeum vulgare L. Bass, Baudin, Harrington and Stirling). Alternative 3’ splicing (34%-45%), intron retention (32%-34%) and alternative 5’ splicing (16%-21%) were three major AS events in germinating embryos. The AS transcripts were predominantly mapped onto ribosome, RNA transport machineries, spliceosome, plant hormone signal transduction, glycolysis, sugar and carbon metabolism pathways. Transcripts of these genes were also very abundant in the early stage of seed germination. Correlation analysis of gene expression showed that AS hormone responsive transcripts could also be co-expressed with genes responsible for protein biosynthesis and sugar metabolisms. Our RNA-sequencing data revealed that AS could play important roles in barley seed germination. PMID:27031341

  14. Proteomic insights into seed germination in response to environmental factors.

    PubMed

    Tan, Longyan; Chen, Sixue; Wang, Tai; Dai, Shaojun

    2013-06-01

    Seed germination is a critical process in the life cycle of higher plants. During germination, the imbibed mature seed is highly sensitive to different environmental factors.However, knowledge about the molecular and physiological mechanisms underlying the environmental effects on germination has been lacking. Recent proteomic work has provided invaluable insight into the molecular processes in germinating seeds of Arabidopsis, rice (Oryza sativa), soybean (Glycine max), barley (Hordeum vulgare), maize (Zeamays), tea (Camellia sinensis), European beech (Fagus sylvatica), and Norway maple (Acer platanoides) under different treatments including metal ions (e.g. copper and cadmium), drought, low temperature, hormones, and chemicals (gibberellic acid, abscisic acid, salicylic acid, and α-amanitin), as well as Fusarium graminearum infection. A total of 561 environmental factor-responsive proteins have been identified with various expression patterns in germinating seeds. The data highlight diverse regulatory and metabolic mechanisms upon seed germination, including induction of environmental factor-responsive signaling pathways, seed storage reserve mobilization and utilization, enhancement of DNA repair and modification, regulation of gene expression and protein synthesis, modulation of cell structure, and cell defense. In this review, we summarize the interesting findings and discuss the relevance and significance for our understanding of environmental regulation of seed germination. PMID:23986916

  15. Proteome Analysis of Poplar Seed Vigor.

    PubMed

    Zhang, Hong; Wang, Wei-Qing; Liu, Shu-Jun; Møller, Ian Max; Song, Song-Quan

    2015-01-01

    Seed vigor is a complex property that determines the seed's potential for rapid uniform emergence and subsequent growth. However, the mechanism for change in seed vigor is poorly understood. The seeds of poplar (Populus × Canadensis Moench), which are short-lived, were stored at 30 °C and 75 ± 5% relative humidity for different periods of time (0-90 days) to obtain different vigor seeds (from 95 to 0% germination). With decreasing seed vigor, the temperature range of seed germination became narrower; the respiration rate of the seeds decreased markedly, while the relative electrolyte leakage increased markedly, both levelling off after 45 days. A total of 81 protein spots showed a significant change in abundance (≥ 1.5-fold, P < 0.05) when comparing the proteomes among seeds with different vigor. Of the identified 65 proteins, most belonged to the groups involved in metabolism (23%), protein synthesis and destination (22%), energy (18%), cell defense and rescue (17%), and storage protein (15%). These proteins accounted for 95% of all the identified proteins. During seed aging, 53 and 6 identified proteins consistently increased and decreased in abundance, respectively, and they were associated with metabolism (22%), protein synthesis and destination (22%), energy (19%), cell defense and rescue (19%), storage proteins (15%), and cell growth and structure (3%). These data show that the decrease in seed vigor (aging) is an energy-dependent process, which requires protein synthesis and degradation as well as cellular defense and rescue. PMID:26172265

  16. Identification of Microbial Metabolites Elevating Vitamin Contents in Barley Seeds.

    PubMed

    Yousaf, Anam; Qadir, Abdul; Anjum, Tehmina; Ahmad, Aqeel

    2015-08-19

    The current investigation analyzes metabolites of Acetobacter aceti to explore chemical compounds responsible for the induction of vitamins in barley seeds. A bioactivity guided assay of bacterial extracts and chromatographic analyses of barley produce revealed 13 chemical compounds, which were subjected to principal component analysis (PCA). PCA determined four chemical compounds (i.e., quinolinic acid, pyridoxic acid, p-aminobenzoate, and α-oxobutanoic acid) highly associated with increased quantities of vitamins. Further experimentations confirmed that quinolinic acid and p-aminobenzoate were the most efficient vitamin inducers. The results indicated chloroform/ethanol (4:1) as the best solvent system for the extraction of active compounds from crude metabolites of A. aceti. Significant quantities of mevalonic acid were detected in the extracted fraction, indicating the possible induction of the isoprenoid pathway. Altogether, the current investigation broadens the frontiers in plant-microbe interaction. PMID:26173019

  17. A Low Phytic Acid Barley Mutation Alters Gene Expression in Early Seed Development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley (Hordeum vulgare L.) low phytic acid (lpa) mutants have reduced levels of seed phytate, the most abundant form of phosphorus in seeds, and increases in seed inorganic phosphorus. To understand how lpa mutations affect metabolic and developmental processes during seed growth, gene expression ...

  18. Proteomic analysis of mature Lagenaria siceraria seed.

    PubMed

    Kumari, Neha; Tajmul, Md; Yadav, Savita

    2015-04-01

    Lagenaria siceraria (bottle gourd) class belongs to Magnoliopsida family curcurbitaceae that is a traditionally used medicinal plant. Fruit of this plant are widely used as a therapeutic vegetable in various diseases, all over the Asia and Africa. Various parts of this plant like fruit, seed, leaf and root are used as alternative medicine. In the present study, primarily, we have focused on proteomic analysis of L. siceraria seed using phenol extraction method for protein isolation. Twenty-four colloidal coomassie blue stained protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) after resolving on two-dimensional gel electrophoresis. Out of 24 identified protein spots, four were grouped as unidentified proteins which clearly suggest that less work has been done in the direction of plant seed proteomics. These proteins have been found to implicate in various functions such as biosynthesis of plant cell wall polysaccharides and glycoproteins, serine/threonine kinase activity, plant disease resistance and transferase activity against insects by means of insecticidal and larval growth inhibitory, anti-HIV, antihelmintic and antimicrobial properties. By Blast2GO annotation analysis, amongst the identified proteins of L. siceraria, molecular function for majority of proteins has indispensable role in catalytic activity, few in binding activity and antioxidant activity; it is mostly distributed in cell, organelle, membrane and macromolecular complex. Most of them involved in biological process such as metabolic process, cellular process, response to stimulus, single organism process, signalling, biological recognition, cellular component organization or biogenesis and localization. PMID:25672325

  19. Viability of barley seeds after long-term exposure to outer side of international space station

    NASA Astrophysics Data System (ADS)

    Sugimoto, Manabu; Ishii, Makoto; Mori, Izumi C.; Elena, Shagimardanova; Gusev, Oleg A.; Kihara, Makoto; Hoki, Takehiro; Sychev, Vladimir N.; Levinskikh, Margarita A.; Novikova, Natalia D.; Grigoriev, Anatoly I.

    2011-09-01

    Barley seeds were exposed to outer space for 13 months in a vented metal container without a climate control system to assess the risk of physiological and genetic mutation during long-term storage in space. The space-stored seeds (S0 generation), with an 82% germination rate in 50 seeds, lost about 20% of their weight after the exposure. The germinated seeds showed normal growth, heading, and ripening. The harvested seeds (S1 generation) also germinated and reproduced (S2 generation) as did the ground-stored seeds. The culm length, ear length, number of seed, grain weight, and fertility of the plants from the space-stored seeds were not significantly different from those of the ground-stored seeds in each of the S0 and S1 generation. Furthermore, the S1 and S2 space-stored seeds respectively showed similar β-glucan content to those of the ground-stored seeds. Amplified fragment length polymorphism analysis with 16 primer combinations showed no specific fragment that appears or disappears significantly in the DNA isolated from the barley grown from the space-stored seeds. Though these data are derived from nine S0 space-stored seeds in a single exposure experiment, the results demonstrate the preservation of barley seeds in outer space for 13 months without phenotypic or genotypic changes and with healthy and vigorous growth in space.

  20. Proteomic analysis of the testa from developing soybean seeds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Soybean (Glycine max (L.) Merr. cv Jack) seed development was separated into nine defined stages (S1 to S9). Testa (seed coats) were removed from developing seeds at stages S2, 4, 6, 8, and 9, and subjected to shotgun proteomic profiling. For each stage "total proteins” were isolated from 150 mg dry...

  1. Barley

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This book chapter is part of a series reviewing advances in transgenic crop plants. The chapter covers advances in barley transformation. Conventional and biotechnological approaches to barley improvement are discussed. Experiments conducted around the world to improve barley food, feed and malting ...

  2. Comparative proteomic analysis of aluminum tolerance in tibetan wild and cultivated barleys.

    PubMed

    Dai, Huaxin; Cao, Fangbin; Chen, Xianhong; Zhang, Mian; Ahmed, Imrul Mosaddek; Chen, Zhong-Hua; Li, Chengdao; Zhang, Guoping; Wu, Feibo

    2013-01-01

    Aluminum (Al) toxicity is a major limiting factor for plant production in acid soils. Wild barley germplasm is rich in genetic diversity and may provide elite genes for crop Al tolerance improvement. The hydroponic-experiments were performed to compare proteomic and transcriptional characteristics of two contrasting Tibetan wild barley genotypes Al- resistant/tolerant XZ16 and Al-sensitive XZ61 as well as Al-resistant cv. Dayton. Results showed that XZ16 had less Al uptake and translocation than XZ61 and Dayton under Al stress. Thirty-five Al-tolerance/resistance-associated proteins were identified and categorized mainly in metabolism, energy, cell growth/division, protein biosynthesis, protein destination/storage, transporter, signal transduction, disease/defense, etc. Among them, 30 were mapped on barley genome, with 16 proteins being exclusively up-regulated by Al stress in XZ16, including 4 proteins (S-adenosylmethionine-synthase 3, ATP synthase beta subunit, triosephosphate isomerase, Bp2A) specifically expressed in XZ16 but not Dayton. The findings highlighted the significance of specific-proteins associated with Al tolerance, and verified Tibetan wild barley as a novel genetic resource for Al tolerance. PMID:23691047

  3. Viability and Biological Properties of Barley Seeds Expose to Outside of International Space Station

    NASA Astrophysics Data System (ADS)

    Sugimoto, Manabu; Ishii, Makoto; Mori, Izumi; Shagimardanova, Elena; Gusev, Oleg; Sychev, Vladimir; Levinskikh, Margarita; Novikova, Nataliya; Grigoriev, Anatoly

    Plants play an important role in supplying nutrients and oxygen to human under material recycle system in space as well as on earth, therefore, seed storage in space should be necessary to self-supply foods when number of astronauts would stay and investigate for a long-term habitation of orbit and the bases of the Moon and Mars. In order to understand the effect of real space environment on the preservation of seeds, the seeds of malting barley, Haruna Nijo, were exposed to outside of the Pier docking station of International Space Station in the framework of the Biorisk-MSN program. After exposure to outside of International Space Station for 13 months, the seeds (SP) were transported to Earth, soaked in water, and germinated on the filter paper filled with water. The germination ratio of SP was 82%, while that of the ground control was 96%, showing that the barley seeds survived cosmic radiation, vacuum, and temperature excursion in space. The germinated seeds of SP and ground control were transplanted to the Wagner pots filled with soil and grown for 5 months in the greenhouse. The agronomic character, such as number of main stem leaf and ear, straw weight, culm length, ear length, thousand kernel weight, and percentage of ripening, were not different significantly between SP and ground control. The germination ratio of the harvested SP was 96% as same as that of the harvested ground control. Genomic DNA and protein were extracted from leaves of the barleys and analyzed by AFLP and 2-DE, respectively. The results demonstrated no significant difference in genetic polymorphism and protein production in these barleys. From our results, barley seeds could survive real space environment for the long-term habitation without phenotypic and genotypic damages.

  4. Barley (Hordeum vulgare L.) low phytic acid 1-1: an endosperm-specific filial determinant of seed total phosphorus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In cultivated cereal and legume seed crops, inositol hexaphosphate (Ins P6 or “phytic acid”) typically accounts for 75% (±10%) of seed total phosphorus (P). Genetic blocks in seed Ins P6 accumulation in some cases can also alter the distribution or total amount of seed P. In non-mutant barley (Horde...

  5. Mitogen-Activated Protein Kinase Kinase 3 Regulates Seed Dormancy in Barley.

    PubMed

    Nakamura, Shingo; Pourkheirandish, Mohammad; Morishige, Hiromi; Kubo, Yuta; Nakamura, Masako; Ichimura, Kazuya; Seo, Shigemi; Kanamori, Hiroyuki; Wu, Jianzhong; Ando, Tsuyu; Hensel, Goetz; Sameri, Mohammad; Stein, Nils; Sato, Kazuhiro; Matsumoto, Takashi; Yano, Masahiro; Komatsuda, Takao

    2016-03-21

    Seed dormancy has fundamental importance in plant survival and crop production; however, the mechanisms regulating dormancy remain unclear [1-3]. Seed dormancy levels generally decrease during domestication to ensure that crops successfully germinate in the field. However, reduction of seed dormancy can cause devastating losses in cereals like wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) due to pre-harvest sprouting, the germination of mature seed (grain) on the mother plant when rain occurs before harvest. Understanding the mechanisms of dormancy can facilitate breeding of crop varieties with the appropriate levels of seed dormancy [4-8]. Barley is a model crop [9, 10] and has two major seed dormancy quantitative trait loci (QTLs), SD1 and SD2, on chromosome 5H [11-19]. We detected a QTL designated Qsd2-AK at SD2 as the single major determinant explaining the difference in seed dormancy between the dormant cultivar "Azumamugi" (Az) and the non-dormant cultivar "Kanto Nakate Gold" (KNG). Using map-based cloning, we identified the causal gene for Qsd2-AK as Mitogen-activated Protein Kinase Kinase 3 (MKK3). The dormant Az allele of MKK3 is recessive; the N260T substitution in this allele decreases MKK3 kinase activity and appears to be causal for Qsd2-AK. The N260T substitution occurred in the immediate ancestor allele of the dormant allele, and the established dormant allele became prevalent in barley cultivars grown in East Asia, where the rainy season and harvest season often overlap. Our findings show fine-tuning of seed dormancy during domestication and provide key information for improving pre-harvest sprouting tolerance in barley and wheat. PMID:26948880

  6. Comparative Proteomic Analysis of Peanut Seed and Seed Coat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanut (Arachis hypogaea L.) is grown extensively worldwide for its edible seed and oil. In a peanut, within the hull and encasing the cotyledon is the seed coat, which is commonly referred to within the peanut industry as the skin. The seed coat is a distinct plant structure critical for seed deve...

  7. Quantitative analysis of proteome extracted from barley crowns grown under different drought conditions

    PubMed Central

    Vítámvás, Pavel; Urban, Milan O.; Škodáček, Zbynek; Kosová, Klára; Pitelková, Iva; Vítámvás, Jan; Renaut, Jenny; Prášil, Ilja T.

    2015-01-01

    Barley cultivar Amulet was used to study the quantitative proteome changes through different drought conditions utilizing two-dimensional difference gel electrophoresis (2D-DIGE). Plants were cultivated for 10 days under different drought conditions. To obtain control and differentially drought-treated plants, the soil water content was kept at 65, 35, and 30% of soil water capacity (SWC), respectively. Osmotic potential, water saturation deficit, 13C discrimination, and dehydrin accumulation were monitored during sampling of the crowns for proteome analysis. Analysis of the 2D-DIGE gels revealed 105 differentially abundant spots; most were differentially abundant between the controls and drought-treated plants, and 25 spots displayed changes between both drought conditions. Seventy-six protein spots were successfully identified by tandem mass spectrometry. The most frequent functional categories of the identified proteins can be put into the groups of: stress-associated proteins, amino acid metabolism, carbohydrate metabolism, as well as DNA and RNA regulation and processing. Their possible role in the response of barley to drought stress is discussed. Our study has shown that under drought conditions barley cv. Amulet decreased its growth and developmental rates, displayed a shift from aerobic to anaerobic metabolism, and exhibited increased levels of several protective proteins. Comparison of the two drought treatments revealed plant acclimation to milder drought (35% SWC); but plant damage under more severe drought treatment (30% SWC). The results obtained revealed that cv. Amulet is sensitive to drought stress. Additionally, four spots revealing a continuous and significant increase with decreasing SWC (UDP-glucose 6-dehydrogenase, glutathione peroxidase, and two non-identified) could be good candidates for testing of their protein phenotyping capacity together with proteins that were significantly distinguished in both drought treatments. PMID:26175745

  8. Molecular Clustering Interrelationships and Carbohydrate Conformation in Hull and Seeds Among Barley Cultivars

    SciTech Connect

    N Liu; P Yu

    2011-12-31

    The objective of this study was to use molecular spectral analyses with the diffuse reflectance Fourier transform infrared spectroscopy (DRIFT) bioanlytical technique to study carbohydrate conformation features, molecular clustering and interrelationships in hull and seed among six barley cultivars (AC Metcalfe, CDC Dolly, McLeod, CDC Helgason, CDC Trey, CDC Cowboy), which had different degradation kinetics in rumen. The molecular structure spectral analyses in both hull and seed involved the fingerprint regions of ca. 1536-1484 cm{sup -1} (attributed mainly to aromatic lignin semicircle ring stretch), ca. 1293-1212 cm{sup -1} (attributed mainly to cellulosic compounds in the hull), ca. 1269-1217 cm{sup -1} (attributed mainly to cellulosic compound in the seeds), and ca. 1180-800 cm{sup -1} (attributed mainly to total CHO C-O stretching vibrations) together with an agglomerative hierarchical cluster (AHCA) and principal component spectral analyses (PCA). The results showed that the DRIFT technique plus AHCA and PCA molecular analyses were able to reveal carbohydrate conformation features and identify carbohydrate molecular structure differences in both hull and seeds among the barley varieties. The carbohydrate molecular spectral analyses at the region of ca. 1185-800 cm{sup -1} together with the AHCA and PCA were able to show that the barley seed inherent structures exhibited distinguishable differences among the barley varieties. CDC Helgason had differences from AC Metcalfe, MeLeod, CDC Cowboy and CDC Dolly in carbohydrate conformation in the seed. Clear molecular cluster classes could be distinguished and identified in AHCA analysis and the separate ellipses could be grouped in PCA analysis. But CDC Helgason had no distinguished differences from CDC Trey in carbohydrate conformation. These carbohydrate conformation/structure difference could partially explain why the varieties were different in digestive behaviors in animals. The molecular spectroscopy

  9. Physiological and proteomic analyses on artificially aged Brassica napus seed

    PubMed Central

    Yin, Xiaojian; He, Dongli; Gupta, Ravi; Yang, Pingfang

    2015-01-01

    Plant seeds lose their viability when they are exposed to long term storage or controlled deterioration treatments, by a process known as seed aging. Based on previous studies, artificially aging treatments have been developed to accelerate the process of seed aging in order to understand its underlying mechanisms. In this study, we used Brassica napus seeds to investigate the mechanisms of aging initiation. B. napus seeds were exposed to artificially aging treatment (40°C and 90% relative humidity) and their physio-biochemical characteristics were analyzed. Although the treatment delayed germination, it did not increase the concentration of cellular reactive oxygen species (ROS). Comparative proteomic analysis was conducted among the control and treated seeds at different stages of germination. The proteins responded to the treatment were mainly involved in metabolism, protein modification and destination, stress response, development, and miscellaneous enzymes. Except for peroxiredoxin, no changes were observed in the accumulation of other antioxidant enzymes in the artificially aged seeds. Increased content of abscisic acid (ABA) was observed in the artificially treated seeds which might be involved in the inhibition of germination. Taken together, our results highlight the involvement of ABA in the initiation of seed aging in addition to the ROS which was previously reported to mediate the seed aging process. PMID:25763006

  10. Epigenetic chromatin modifiers in barley: IV. The study of barley Polycomb group (PcG) genes during seed development and in response to external ABA

    PubMed Central

    2010-01-01

    Background Epigenetic phenomena have been associated with the regulation of active and silent chromatin states achieved by modifications of chromatin structure through DNA methylation, and histone post-translational modifications. The latter is accomplished, in part, through the action of PcG (Polycomb group) protein complexes which methylate nucleosomal histone tails at specific sites, ultimately leading to chromatin compaction and gene silencing. Different PcG complex variants operating during different developmental stages have been described in plants. In particular, the so-called FIE/MEA/FIS2 complex governs the expression of genes important in embryo and endosperm development in Arabidopsis. In our effort to understand the epigenetic mechanisms regulating seed development in barley (Hordeum vulgare), an agronomically important monocot plant cultivated for its endosperm, we set out to characterize the genes encoding barley PcG proteins. Results Four barley PcG gene homologues, named HvFIE, HvE(Z), HvSu(z)12a, and HvSu(z)12b were identified and structurally and phylogenetically characterized. The corresponding genes HvFIE, HvE(Z), HvSu(z)12a, and HvSu(z)12b were mapped onto barley chromosomes 7H, 4H, 2H and 5H, respectively. Expression analysis of the PcG genes revealed significant differences in gene expression among tissues and seed developmental stages and between barley cultivars with varying seed size. Furthermore, HvFIE and HvE(Z) gene expression was responsive to the abiotic stress-related hormone abscisic acid (ABA) known to be involved in seed maturation, dormancy and germination. Conclusion This study reports the first characterization of the PcG homologues, HvFIE, HvE(Z), HvSu(z)12a and HvSu(z)12b in barley. All genes co-localized with known chromosomal regions responsible for malting quality related traits, suggesting that they might be used for developing molecular markers to be applied in marker assisted selection. The PcG differential expression

  11. Response to temperature stress of reactive oxygen species scavenging enzymes in the cross-tolerance of barley seed germination*

    PubMed Central

    Mei, Yu-qin; Song, Song-quan

    2010-01-01

    A number of studies have shown the existence of cross-tolerance in plants, but the physiological mechanism is poorly understood. In this study, we used the germination of barley seeds as a system to investigate the cross-tolerance of low-temperature pretreatment to high-temperature stress and the possible involvement of reactive oxygen species (ROS) scavenging enzymes in the cross-tolerance. After pretreatment at 0 °C for different periods of time, barley seeds were germinated at 35 °C, and the content of malondialdehyde (MDA) and the activities of ROS scavenging enzymes were measured by a spectrophotometer analysis. The results showed that barley seed germinated very poorly at 35 °C, and this inhibitive effect could be overcome by pretreatment at 0 °C. The MDA content varied, depending on the temperature at which seeds germinated, while barley seeds pretreated at 0 °C did not change the MDA content. Compared with seeds germinated directly at 35 °C, the seeds pretreated first at 0 °C and then germinated at 35 °C had markedly increased activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), and glutathione reductase (GR). The SOD and APX activities of seeds germinated at 35 °C after 0 °C-pretreatment were even substantially higher than those at 25 °C, and GR activity was similar to that at 25 °C, at which the highest germination performance of barley seeds was achieved. These results indicate that low-temperature pretreatment can markedly increase the tolerance of barley seed to high temperature during germination, this being related to the increase in ROS scavenging enzyme activity. This may provide a new method for increasing seed germination under stress environments, and may be an excellent model system for the study of cross-tolerance. PMID:21121076

  12. [Influence of γ-Irradiated Seeds on the Enzyme Activity in Barley Seedlings].

    PubMed

    Volkova, P Yu; Churyukin, R S; Geras'kin, S A

    2016-01-01

    Influence of γ-irradiation of barley seeds (Nur variety) at the doses of 8-50 Gy on catalase, pyruvate kinase, glucose-6-phosphate dehydrogenase, and guaiacol peroxidase activities was studied in the seedlings on the 3, 5 and 7 days after germination. It has been shown that activities of the studied enzymes increase in the dose range that causes the growth stimulation in the seedlings (16-20 Gy). PMID:27534070

  13. Proteomic analysis of endoplasmic reticulum stress responses in rice seeds

    PubMed Central

    Qian, Dandan; Tian, Lihong; Qu, Leqing

    2015-01-01

    The defects in storage proteins secretion in the endosperm of transgenic rice seeds often leads to endoplasmic reticulum (ER) stress, which produces floury and shrunken seeds, but the mechanism of this response remains unclear. We used an iTRAQ-based proteomics analysis of ER-stressed rice seeds due to the endosperm-specific suppression of OsSar1 to identify changes in the protein levels in response to ER stress. ER stress changed the expression of 405 proteins in rice seed by >2.0- fold compared with the wild-type control. Of these proteins, 140 were upregulated and 265 were downregulated. The upregulated proteins were mainly involved in protein modification, transport and degradation, and the downregulated proteins were mainly involved in metabolism and stress/defense responses. A KOBAS analysis revealed that protein-processing in the ER and degradation-related proteasome were the predominant upregulated pathways in the rice endosperm in response to ER stress. Trans-Golgi protein transport was also involved in the ER stress response. Combined with bioinformatic and molecular biology analyses, our proteomic data will facilitate our understanding of the systemic responses to ER stress in rice seeds. PMID:26395408

  14. Proteomic analysis of endoplasmic reticulum stress responses in rice seeds.

    PubMed

    Qian, Dandan; Tian, Lihong; Qu, Leqing

    2015-01-01

    The defects in storage proteins secretion in the endosperm of transgenic rice seeds often leads to endoplasmic reticulum (ER) stress, which produces floury and shrunken seeds, but the mechanism of this response remains unclear. We used an iTRAQ-based proteomics analysis of ER-stressed rice seeds due to the endosperm-specific suppression of OsSar1 to identify changes in the protein levels in response to ER stress. ER stress changed the expression of 405 proteins in rice seed by >2.0- fold compared with the wild-type control. Of these proteins, 140 were upregulated and 265 were downregulated. The upregulated proteins were mainly involved in protein modification, transport and degradation, and the downregulated proteins were mainly involved in metabolism and stress/defense responses. A KOBAS analysis revealed that protein-processing in the ER and degradation-related proteasome were the predominant upregulated pathways in the rice endosperm in response to ER stress. Trans-Golgi protein transport was also involved in the ER stress response. Combined with bioinformatic and molecular biology analyses, our proteomic data will facilitate our understanding of the systemic responses to ER stress in rice seeds. PMID:26395408

  15. Proteomics of seed development, desiccation tolerance, germination and vigor.

    PubMed

    Wang, Wei-Qing; Liu, Shu-Jun; Song, Song-Quan; Møller, Ian Max

    2015-01-01

    Proteomics, the large-scale study of the total complement of proteins in a given sample, has been applied to all aspects of seed biology mainly using model species such as Arabidopsis or important agricultural crops such as corn and rice. Proteins extracted from the sample have typically been separated and quantified by 2-dimensional polyacrylamide gel electrophoresis followed by liquid chromatography and mass spectrometry to identify the proteins in the gel spots. In this way, qualitative and quantitative changes in the proteome during seed development, desiccation tolerance, germination, dormancy release, vigor alteration and responses to environmental factors have all been studied. Many proteins or biological processes potentially important for each seed process have been highlighted by these studies, which greatly expands our knowledge of seed biology. Proteins that have been identified to be particularly important for at least two of the seed processes are involved in detoxification of reactive oxygen species, the cytoskeleton, glycolysis, protein biosynthesis, post-translational modifications, methionine metabolism, and late embryogenesis-abundant (LEA) proteins. It will be useful for molecular biologists and molecular plant breeders to identify and study genes encoding particularly interesting target proteins with the aim to improve the yield, stress tolerance or other critical properties of our crop species. PMID:25461695

  16. Metabolite fingerprinting of barley whole seeds, endosperms, and embryos during industrial malting.

    PubMed

    Gorzolka, K; Lissel, M; Kessler, N; Loch-Ahring, S; Niehaus, K

    2012-06-15

    Samples of whole seeds, isolated endosperms including the aleurone layer and isolated embryos with attached scutellum from an industrial scale barley malting process (variety Braemar) were analysed for their water soluble metabolites by gas chromatography-mass spectrometry (GC-MS). 73 known metabolites and about 350 unknown signals were detected. Principal component analysis (PCA) showed a time dependent shift of sample profiles. Whole seeds and endosperm samples showed very similar patterns with nearly all compounds rising until the end of germination. In the embryos a maximum concentration of compounds was reached after 72-96 h of malting. Most concentrations decreased afterwards. The kilning step, namely the drying and roasting of germinated seeds, induced variable effects of increases, stability or decreases of metabolites and thereby separated kilned samples from germinated seeds in the PCA. A second barley cultivar (Quench) underwent the same malting and analysis procedures and gave nearly identical results. Fructose, malate, myo-inositol and raffinose exhibited the potential to serve as markers for specific developmental stages of seeds in both varieties. Biological markers represent targets for industrial process control. Their potential application would meet the maltsters' demand to flatten variances in germination properties and to produce equal composed malt by directed malting management. PMID:22465293

  17. Barley Seed Aging: Genetics behind the Dry Elevated Pressure of Oxygen Aging and Moist Controlled Deterioration.

    PubMed

    Nagel, Manuela; Kodde, Jan; Pistrick, Sibylle; Mascher, Martin; Börner, Andreas; Groot, Steven P C

    2016-01-01

    Experimental seed aging approaches intend to mimic seed deterioration processes to achieve a storage interval reduction. Common methods apply higher seed moisture levels and temperatures. In contrast, the "elevated partial pressure of oxygen" (EPPO) approach treats dry seed stored at ambient temperatures with high oxygen pressure. To analyse the genetic background of seed longevity and the effects of seed aging under dry conditions, the EPPO approach was applied to the progeny of the Oregon Wolfe Barley (OWB) mapping population. In comparison to a non-treated control and a control high-pressure nitrogen treatment, EPPO stored seeds showed typical symptoms of aging with a significant reduction of normal seedlings, slower germination, and less total germination. Thereby, the parent Dom ("OWB-D"), carrying dominant alleles, is more sensitive to aging in comparison to the population mean and in most cases to the parent Rec ("OWB-R"), carrying recessive alleles. Quantitative trait locus (QTL) analyses using 2832 markers revealed 65 QTLs, including two major loci for seed vigor on 2H and 7H. QTLs for EPPO tolerance were detected on 3H, 4H, and 5H. An applied controlled deterioration (CD) treatment (aged at higher moisture level and temperature) revealed a tolerance QTL on 5H, indicating that the mechanism of seed deterioration differs in part between EPPO or CD conditions. PMID:27066038

  18. Barley Seed Aging: Genetics behind the Dry Elevated Pressure of Oxygen Aging and Moist Controlled Deterioration

    PubMed Central

    Nagel, Manuela; Kodde, Jan; Pistrick, Sibylle; Mascher, Martin; Börner, Andreas; Groot, Steven P. C.

    2016-01-01

    Experimental seed aging approaches intend to mimic seed deterioration processes to achieve a storage interval reduction. Common methods apply higher seed moisture levels and temperatures. In contrast, the “elevated partial pressure of oxygen” (EPPO) approach treats dry seed stored at ambient temperatures with high oxygen pressure. To analyse the genetic background of seed longevity and the effects of seed aging under dry conditions, the EPPO approach was applied to the progeny of the Oregon Wolfe Barley (OWB) mapping population. In comparison to a non-treated control and a control high-pressure nitrogen treatment, EPPO stored seeds showed typical symptoms of aging with a significant reduction of normal seedlings, slower germination, and less total germination. Thereby, the parent Dom (“OWB-D”), carrying dominant alleles, is more sensitive to aging in comparison to the population mean and in most cases to the parent Rec (“OWB-R”), carrying recessive alleles. Quantitative trait locus (QTL) analyses using 2832 markers revealed 65 QTLs, including two major loci for seed vigor on 2H and 7H. QTLs for EPPO tolerance were detected on 3H, 4H, and 5H. An applied controlled deterioration (CD) treatment (aged at higher moisture level and temperature) revealed a tolerance QTL on 5H, indicating that the mechanism of seed deterioration differs in part between EPPO or CD conditions. PMID:27066038

  19. In Planta Proteomics and Proteogenomics of the Biotrophic Barley Fungal Pathogen Blumeria graminis f. sp. hordei*

    PubMed Central

    Bindschedler, Laurence V.; Burgis, Timothy A.; Mills, Davinia J. S.; Ho, Jenny T. C.; Cramer, Rainer; Spanu, Pietro D.

    2009-01-01

    To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity. PMID:19602707

  20. The study of a barley epigenetic regulator, HvDME, in seed development and under drought

    PubMed Central

    2013-01-01

    Background Epigenetic factors such as DNA methylation and histone modifications regulate a wide range of processes in plant development. Cytosine methylation and demethylation exist in a dynamic balance and have been associated with gene silencing or activation, respectively. In Arabidopsis, cytosine demethylation is achieved by specific DNA glycosylases, including AtDME (DEMETER) and AtROS1 (REPRESSOR OF SILENCING1), which have been shown to play important roles in seed development. Nevertheless, studies on monocot DNA glycosylases are limited. Here we present the study of a DME homologue from barley (HvDME), an agronomically important cereal crop, during seed development and in response to conditions of drought. Results An HvDME gene, identified in GenBank, was found to encode a protein with all the characteristic modules of DME-family DNA glycosylase proteins. Phylogenetic analysis revealed a high degree of homology to other monocot DME glycosylases, and sequence divergence from the ROS1, DML2 and DML3 orthologues. The HvDME gene contains the 5′ and 3′ Long Terminal Repeats (LTR) of a Copia retrotransposon element within the 3′ downstream region. HvDME transcripts were shown to be present both in vegetative and reproductive tissues and accumulated differentially in different seed developmental stages and in two different cultivars with varying seed size. Additionally, remarkable induction of HvDME was evidenced in response to drought treatment in a drought-tolerant barley cultivar. Moreover, variable degrees of DNA methylation in specific regions of the HvDME promoter and gene body were detected in two different cultivars. Conclusion A gene encoding a DNA glycosylase closely related to cereal DME glycosylases was characterized in barley. Expression analysis during seed development and under dehydration conditions suggested a role for HvDME in endosperm development, seed maturation, and in response to drought. Furthermore, differential DNA methylation

  1. A proteomic analysis of seed development in Brassica campestri L.

    PubMed

    Li, Wenlan; Gao, Yi; Xu, Hong; Zhang, Yu; Wang, Jianbo

    2012-01-01

    To gain insights into the protein dynamics during seed development, a proteomic study on the developing Brassica campestri L. seeds with embryos in different embryogenesis stages was carried out. The seed proteins at 10, 16, 20, 25 and 35 DAP (days after pollination), respectively, were separated using two-dimensional gel electrophoresis and identities of 209 spots with altered abundance were determined by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS). These proteins were classified into 16 groups according to their functions. The most abundant proteins were related to primary metabolism, indicating the heavy demand of materials for rapid embryo growth. Besides, the high amount of proteins involved in protein processing and destination indicated importance of protein renewal during seed development. The remaining were those participated in oxidation/detoxification, energy, defense, transcription, protein synthesis, transporter, cell structure, signal transduction, secondary metabolism, transposition, DNA repair, storage and so on. Protein abundance profiles of each functional class were generated and hierarchical cluster analysis established 8 groups of dynamic patterns. Our results revealed novel characters of protein dynamics in seed development in Brassica campestri L. and provided valuable information about the complex process of seed development in plants. PMID:23189193

  2. A Proteomic Analysis of Seed Development in Brassica campestri L

    PubMed Central

    Li, Wenlan; Gao, Yi; Xu, Hong; Zhang, Yu; Wang, Jianbo

    2012-01-01

    To gain insights into the protein dynamics during seed development, a proteomic study on the developing Brassica campestri L. seeds with embryos in different embryogenesis stages was carried out. The seed proteins at 10, 16, 20, 25 and 35 DAP (days after pollination), respectively, were separated using two-dimensional gel electrophoresis and identities of 209 spots with altered abundance were determined by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS). These proteins were classified into 16 groups according to their functions. The most abundant proteins were related to primary metabolism, indicating the heavy demand of materials for rapid embryo growth. Besides, the high amount of proteins involved in protein processing and destination indicated importance of protein renewal during seed development. The remaining were those participated in oxidation/detoxification, energy, defense, transcription, protein synthesis, transporter, cell structure, signal transduction, secondary metabolism, transposition, DNA repair, storage and so on. Protein abundance profiles of each functional class were generated and hierarchical cluster analysis established 8 groups of dynamic patterns. Our results revealed novel characters of protein dynamics in seed development in Brassica campestri L. and provided valuable information about the complex process of seed development in plants. PMID:23189193

  3. A Role for Reactive Oxygen Species Produced by NADPH Oxidases in the Embryo and Aleurone Cells in Barley Seed Germination

    PubMed Central

    Ishibashi, Yushi; Kasa, Shinsuke; Sakamoto, Masatsugu; Aoki, Nozomi; Kai, Kyohei; Yuasa, Takashi; Hanada, Atsushi; Yamaguchi, Shinjiro; Iwaya-Inoue, Mari

    2015-01-01

    Reactive oxygen species (ROS) promote the germination of several seeds, and antioxidants suppress it. However, questions remain regarding the role and production mechanism of ROS in seed germination. Here, we focused on NADPH oxidases, which produce ROS. After imbibition, NADPH oxidase mRNAs were expressed in the embryo and in aleurone cells of barley seed; these expression sites were consistent with the sites of ROS production in the seed after imbibition. To clarify the role of NADPH oxidases in barley seed germination, we examined gibberellic acid (GA) / abscisic acid (ABA) metabolism and signaling in barley seeds treated with diphenylene iodonium chloride (DPI), an NADPH oxidase inhibitor. DPI significantly suppressed germination, and suppressed GA biosynthesis and ABA catabolism in embryos. GA, but not ABA, induced NADPH oxidase activity in aleurone cells. Additionally, DPI suppressed the early induction of α-amylase by GA in aleurone cells. These results suggest that ROS produced by NADPH oxidases promote GA biosynthesis in embryos, that GA induces and activates NADPH oxidases in aleurone cells, and that ROS produced by NADPH oxidases induce α-amylase in aleurone cells. We conclude that the ROS generated by NADPH oxidases regulate barley seed germination through GA / ABA metabolism and signaling in embryo and aleurone cells. PMID:26579718

  4. A Role for Reactive Oxygen Species Produced by NADPH Oxidases in the Embryo and Aleurone Cells in Barley Seed Germination.

    PubMed

    Ishibashi, Yushi; Kasa, Shinsuke; Sakamoto, Masatsugu; Aoki, Nozomi; Kai, Kyohei; Yuasa, Takashi; Hanada, Atsushi; Yamaguchi, Shinjiro; Iwaya-Inoue, Mari

    2015-01-01

    Reactive oxygen species (ROS) promote the germination of several seeds, and antioxidants suppress it. However, questions remain regarding the role and production mechanism of ROS in seed germination. Here, we focused on NADPH oxidases, which produce ROS. After imbibition, NADPH oxidase mRNAs were expressed in the embryo and in aleurone cells of barley seed; these expression sites were consistent with the sites of ROS production in the seed after imbibition. To clarify the role of NADPH oxidases in barley seed germination, we examined gibberellic acid (GA) / abscisic acid (ABA) metabolism and signaling in barley seeds treated with diphenylene iodonium chloride (DPI), an NADPH oxidase inhibitor. DPI significantly suppressed germination, and suppressed GA biosynthesis and ABA catabolism in embryos. GA, but not ABA, induced NADPH oxidase activity in aleurone cells. Additionally, DPI suppressed the early induction of α-amylase by GA in aleurone cells. These results suggest that ROS produced by NADPH oxidases promote GA biosynthesis in embryos, that GA induces and activates NADPH oxidases in aleurone cells, and that ROS produced by NADPH oxidases induce α-amylase in aleurone cells. We conclude that the ROS generated by NADPH oxidases regulate barley seed germination through GA / ABA metabolism and signaling in embryo and aleurone cells. PMID:26579718

  5. [Modification of Barley Development at Early Stages after Exposure of Seeds to γ-Irradiation].

    PubMed

    Geras'kin, S A; Churukin, R S; Kazakova, E A

    2015-01-01

    The reaction of barley seeds (Nur and Grace varieties) to γ-irradiation in the dose range of 2-50 Gy was studied. The length and weight of a root, the length of a seedling and germination rate were investigated. The dose range in which we observed stimulation of plant development was evaluated. It was shown that the increase of root and seedling sizes after irradiation of seeds at stimulating doses is associated with the rise in the developmental speed, rather than with their earlier germination. Also the effects of a dose rate, a quality of seeds, humidity and a period of storage on the manifestation of radiation exposure were studied. PMID:26964346

  6. Genetic changes induced by space flight factors in barley seeds on Soyuz-5 and Soyuz-9 craft

    NASA Technical Reports Server (NTRS)

    Nuzhdin, N. I.; Dozortseva, R. L.

    1980-01-01

    Air-dry seeds of the barley Zimujuschij moscowskyi of the 1969 harvest were taken into space onboard the spaceships Soyuz-5 and Soyuz-9. A cytological study of the mitoses in meristemic cells in rootlet terminals revealed that space flight factors (SFF) in nonirradiated seeds induced about 3% of aberrant cells. After irradiation the effect of SFF increased over two-fold. Although the radio protectors ensured the seeds against from the SFF-induced damage either in irradiated or nonirradiated seed cells which is inconsistent with the previously obtained data.

  7. Protein repair L-isoaspartyl methyltransferase in plants. Phylogenetic distribution and the accumulation of substrate proteins in aged barley seeds.

    PubMed Central

    Mudgett, M B; Lowenson, J D; Clarke, S

    1997-01-01

    Protein L-isoaspartate (D-aspartate) O-methyltransferases (MTs; EC 2.1.1.77) can initiate the conversion of detrimental L-isoaspartyl residues in spontaneously damaged proteins to normal L-aspartyl residues. We detected this enzyme in 45 species from 23 families representing most of the divisions of the plant kingdom. MT activity is often localized in seeds, suggesting that it has a role in their maturation, quiescence, and germination. The relationship among MT activity, the accumulation of abnormal protein L-isoaspartyl residues, and seed viability was explored in barley (Hordeum vulgare cultivar Himalaya) seeds, which contain high levels of MT. Natural aging of barley seeds for 17 years resulted in a significant reduction in MT activity and in seed viability, coupled with increased levels of "unrepaired" L-isoaspartyl residues. In seeds heated to accelerate aging, we found no reduction of MT activity, but we did observe decreased seed viability and the accumulation of isoaspartyl residues. Among populations of accelerated aged seed, those possessing the highest levels of L-isoaspartyl-containing proteins had the lowest germination percentages. These results suggest that the MT present in seeds cannot efficiently repair all spontaneously damaged proteins containing altered aspartyl residues, and their accumulation during aging may contribute to the loss of seed viability. PMID:9414558

  8. Measurement of Wheat Hardness by Seed Scarifier and Barley Pearler And Comparison with Single-Kernel Characterization System

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new procedure based on a seed scarifier (SS) for measuring wheat hardness was described and investigated along with methods of barley pearler (BP) and single kernel characterization system (SKCS). Hardness measured by SS and BP was expressed as percentage of kernel weight remained after abrading ...

  9. Extraction of high-quality RNA from germinating barley (Hordeum vulgare L.) seeds containing high levels of starch.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Comparative evaluation of gene expression levels can lead to improved understanding of the gene networks underlying traits of economic importance. Extraction of high-quality RNA from germinating barley seeds that contain high levels of starch is of vital importance for analysing the expression of ca...

  10. Proteomic studies on soybean seed quality as affected by high temperature

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Long-term high temperature stresses can occur during soybean seed development through maturity and negatively impact seed quality. We investigated soybean seed quality as affected by high temperature using a proteomic approach. The effects of a prolonged high temperature treatment (37/30ºC day/nigh...

  11. Stimulation of Phenolics, Antioxidant and α-Glucosidase Inhibitory Activities During Barley (Hordeum vulgare L.) Seed Germination.

    PubMed

    Ha, K-S; Jo, S-H; Mannam, V; Kwon, Y-I; Apostolidis, E

    2016-06-01

    The rationale of this study was to enhance the nutritional quality of dry barley seeds. In this study we are evaluating the effect of germination on barley seeds relevant to total phenolic contents, antioxidant activity (in terms of DPPH free-radical scavenging) and the in vitro α-glucosidase inhibitory activities. Barley seeds were germinated for 18.5, 24, 30, 48, and 67 h and then extracted in water. The total phenolic contents, antioxidant activities and α-glucosidase inhibitory activities changed with germination time. More specifically, within the first 48 h of germination the total phenolic content increased from 1.1 mg/g fresh weight (0 h) to 3.4 mg/g fresh weight (48 h) and then slightly reduced by 67 h. Similarly, α-glucosidase inhibitory activity was significantly increased from an IC50 128.82 mg/mL (0 h) to an IC50 18.88 mg/mL (48 h) and then slightly reduced by 67 h. Significant maltase inhibitory activity was observed only with 48 h-germinated extract. Antioxidant activities increased continuously from an IC50 15.72 mg/mL at 0 h to and IC50 5.72 mg/mL after 48 h of germination. Based on our observations, barley seed germination was over after 48 h. During the progress of germination phenolic compounds are becoming available and are more easily extracted. After 48 h, lignification is initiated resulting to the decreased total phenolic content and observed antioxidant and carbohydrate hydrolyzing enzyme inhibition activities. The above results indicate the positive effect of germination in barley seeds for enhanced antioxidant and α-glucosidase inhibitory activities. PMID:27188780

  12. Salt stress-induced alterations in the root proteome of barley genotypes with contrasting response towards salinity.

    PubMed

    Witzel, Katja; Weidner, Annette; Surabhi, Giridara-Kumar; Börner, Andreas; Mock, Hans-Peter

    2009-01-01

    In addition to drought and extreme temperatures, soil salinity represents a growing threat to crop productivity. Among the cereal crops, barley is considered as notably salt tolerant, and cultivars show considerable variation for tolerance towards salinity stress. In order to unravel the molecular mechanisms underlying salt stress tolerance and to utilize the natural genetic variation of barley accessions, a series of hydroponics-based salinity stress experiments was conducted using two genetic mapping parents, cvs Steptoe and Morex, which display contrasting levels of salinity tolerance. The proteome of roots from both genotypes was investigated as displayed by two-dimensional gel electrophoresis, and comparisons were made between plants grown under non-saline and saline conditions. Multivariate analysis of the resulting protein patterns revealed cultivar-specific and salt stress-responsive protein expression. Mass spectrometry-based identification was successful for 26 out of 39 selected protein spots. Hierarchical clustering was applied to detect similar protein expression patterns. Among those, two proteins involved in the glutathione-based detoxification of reactive oxygen species (ROS) were more abundant in the tolerant genotype, while proteins involved in iron uptake were expressed at a higher level in the sensitive one. This study emphasizes the role of proteins involved in ROS detoxification during salinity stress, and identified potential candidates for increasing salt tolerance in barley. PMID:19671579

  13. Abnormal etioplast development in barley seedlings infected with BSMV by seed transmission.

    PubMed

    Harsányi, Anett; Böddi, Béla; Bóka, Károly; Almási, Asztéria; Gáborjányi, Richard

    2002-01-01

    The effect of barley stripe mosaic hordeivirus (BSMV) was studied on the ultrastructure of etioplasts, protochlorophyllide forms and the greening process of barley (Hordeum vulgare cv. Pannónia) plants infected by seed transmission. The leaves of 7- to 11-day-old etiolated seedlings were examined by transmission electron microscopy, fluorescence and absorption spectroscopy. The etioplasts of infected seedlings contained smaller prolamellar bodies with less regular membrane structure, while prothylakoid content was higher than in the control. The protochlorophyllide content of virus-infected seedlings was reduced to 74% of the control. In the 77 K fluorescence spectra the relative amount of 655 nm emitting photoactive protochlorophyllide form decreased, and the amount of the 645 and 633 nm emitting forms increased in the infected leaves. A characteristic effect was observed in the process of the Shibata-shift: 40 min delay was observed in the infected leaves. The results of this work proved that BSMV infection delays or inhibits plastid development and the formation of photosynthetic apparatus. PMID:11982946

  14. Expression, purification and preliminary crystallographic studies of alpha-amylase isozyme 1 from barley seeds.

    PubMed

    Robert, Xavier; Gottschalk, Tine E; Haser, Richard; Svensson, Birte; Aghajari, Nushin

    2002-04-01

    The germinating barley seed contains two major alpha-amylase isozyme families, AMY1 and AMY2, involved in starch degradation to provide energy used by the plant embryo for growth. Many years of difficulty in growing three-dimensional crystals of natural AMY1 have now been overcome by a nonapeptide truncation of the enzyme C-terminus. The truncated enzyme was overexpressed in Pichia pastoris, purified and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 8000 as precipitant and 2-propanol as an additive. Crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 88.36, b = 72.82, c = 61.74 A and one molecule per asymmetric unit. PMID:11914496

  15. Glyoxylate cycle and metabolism of organic acids in the scutellum of barley seeds during germination.

    PubMed

    Ma, Zhenguo; Marsolais, Frédéric; Bernards, Mark A; Sumarah, Mark W; Bykova, Natalia V; Igamberdiev, Abir U

    2016-07-01

    During the developmental processes from dry seeds to seedling establishment, the glyoxylate cycle becomes active in the mobilization of stored oils in the scutellum of barley (Hordeum vulgare L.) seeds, as indicated by the activities of isocitrate lyase and malate synthase. The succinate produced is converted to carbohydrates via phosphoenolpyruvate carboxykinase and to amino acids via aminotransferases, while free organic acids may participate in acidifying the endosperm tissue, releasing stored starch into metabolism. The abundant organic acid in the scutellum was citrate, while malate concentration declined during the first three days of germination, and succinate concentration was low both in scutellum and endosperm. Malate was more abundant in endosperm tissue during the first three days of germination; before citrate became predominant, indicating that malate may be the main acid acidifying the endosperm. The operation of the glyoxylate cycle coincided with an increase in the ATP/ADP ratio, a buildup of H2O2 and changes in the redox state of ascorbate and glutathione. It is concluded that operation of the glyoxylate cycle in the scutellum of cereals may be important not only for conversion of fatty acids to carbohydrates, but also for the acidification of endosperm and amino acid synthesis. PMID:27181945

  16. Carbon nanofibers suppress fungal inhibition of seed germination of maize (Zea mays) and barley (Hordeum vulgare L.) crop

    NASA Astrophysics Data System (ADS)

    Joshi, Anjali; Sharma, Arti; Nayyar, Harsh; Verma, Gaurav; Dharamvir, Keya

    2015-08-01

    Carbon nanofibers (CNFs) are one of allotropes of carbon, consists of graphene layers arrangement in the form of stacked cones or like a cup diameter in nanometer and several millimeters in length. Their extraordinary mechanical, chemical and electronic properties are due to their small size. CNFs have been successfully applied in field of medicine in variety of diagnostic methods. They proven to be an excellent system for drug delivery, tissue regeneration, biosensor etc. This research focuses the applications of CNFs in all fields of Agriculture. In the we treated some fungal disease seed of maize and barley using functionalised CNFs. We find that the tested seeds grow just as well as the healthy seeds whereas the untreated fungal disease seeds, by themselves show very poor germination and seedling growth. This simple experiment shows the extraordinary ability of Carbon nanofibers in carrying effectively inside the germinated seeds.

  17. Carbon nanofibers suppress fungal inhibition of seed germination of maize (Zea mays) and barley (Hordeum vulgare L.) crop

    SciTech Connect

    Joshi, Anjali Sharma, Arti; Nayyar, Harsh; Verma, Gaurav; Dharamvir, Keya

    2015-08-28

    Carbon nanofibers (CNFs) are one of allotropes of carbon, consists of graphene layers arrangement in the form of stacked cones or like a cup diameter in nanometer and several millimeters in length. Their extraordinary mechanical, chemical and electronic properties are due to their small size. CNFs have been successfully applied in field of medicine in variety of diagnostic methods. They proven to be an excellent system for drug delivery, tissue regeneration, biosensor etc. This research focuses the applications of CNFs in all fields of Agriculture. In the we treated some fungal disease seed of maize and barley using functionalised CNFs. We find that the tested seeds grow just as well as the healthy seeds whereas the untreated fungal disease seeds, by themselves show very poor germination and seedling growth. This simple experiment shows the extraordinary ability of Carbon nanofibers in carrying effectively inside the germinated seeds.

  18. Subcellular analysis of starch metabolism in developing barley seeds using a non-aqueous fractionation method

    PubMed Central

    Tiessen, Axel; Nerlich, Annika; Faix, Benjamin; Hümmer, Christine; Fox, Simon; Trafford, Kay; Weber, Hans; Weschke, Winfriede; Geigenberger, Peter

    2012-01-01

    Compartmentation of metabolism in developing seeds is poorly understood due to the lack of data on metabolite distributions at the subcellular level. In this report, a non-aqueous fractionation method is described that allows subcellular concentrations of metabolites in developing barley endosperm to be calculated. (i) Analysis of subcellular volumes in developing endosperm using micrographs shows that plastids and cytosol occupy 50.5% and 49.9% of the total cell volume, respectively, while vacuoles and mitochondria can be neglected. (ii) By using non-aqueous fractionation, subcellular distribution between the cytosol and plastid of the levels of metabolites involved in sucrose degradation, starch synthesis, and respiration were determined. With the exception of ADP and AMP which were mainly located in the plastid, most other metabolites of carbon and energy metabolism were mainly located outside the plastid in the cytosolic compartment. (iii) In developing barley endosperm, the ultimate precursor of starch, ADPglucose (ADPGlc), was mainly located in the cytosol (80–90%), which was opposite to the situation in growing potato tubers where ADPGlc was almost exclusively located in the plastid (98%). This reflects the different subcellular distribution of ADPGlc pyrophosphorylase (AGPase) in these tissues. (iv) Cytosolic concentrations of ADPGlc were found to be close to the published Km values of AGPase and the ADPGlc/ADP transporter at the plastid envelope. Also the concentrations of the reaction partners glucose-1-phosphate, ATP, and inorganic pyrophosphate were close to the respective Km values of AGPase. (v) Knock-out of cytosolic AGPase in Riso16 mutants led to a strong decrease in ADPGlc level, in both the cytosol and plastid, whereas knock-down of the ADPGlc/ADP transporter led to a large shift in the intracellular distribution of ADPGlc. (v) The thermodynamic structure of the pathway of sucrose to starch was determined by calculating the mass–action ratios

  19. Subcellular analysis of starch metabolism in developing barley seeds using a non-aqueous fractionation method.

    PubMed

    Tiessen, Axel; Nerlich, Annika; Faix, Benjamin; Hümmer, Christine; Fox, Simon; Trafford, Kay; Weber, Hans; Weschke, Winfriede; Geigenberger, Peter

    2012-03-01

    Compartmentation of metabolism in developing seeds is poorly understood due to the lack of data on metabolite distributions at the subcellular level. In this report, a non-aqueous fractionation method is described that allows subcellular concentrations of metabolites in developing barley endosperm to be calculated. (i) Analysis of subcellular volumes in developing endosperm using micrographs shows that plastids and cytosol occupy 50.5% and 49.9% of the total cell volume, respectively, while vacuoles and mitochondria can be neglected. (ii) By using non-aqueous fractionation, subcellular distribution between the cytosol and plastid of the levels of metabolites involved in sucrose degradation, starch synthesis, and respiration were determined. With the exception of ADP and AMP which were mainly located in the plastid, most other metabolites of carbon and energy metabolism were mainly located outside the plastid in the cytosolic compartment. (iii) In developing barley endosperm, the ultimate precursor of starch, ADPglucose (ADPGlc), was mainly located in the cytosol (80-90%), which was opposite to the situation in growing potato tubers where ADPGlc was almost exclusively located in the plastid (98%). This reflects the different subcellular distribution of ADPGlc pyrophosphorylase (AGPase) in these tissues. (iv) Cytosolic concentrations of ADPGlc were found to be close to the published K(m) values of AGPase and the ADPGlc/ADP transporter at the plastid envelope. Also the concentrations of the reaction partners glucose-1-phosphate, ATP, and inorganic pyrophosphate were close to the respective K(m) values of AGPase. (v) Knock-out of cytosolic AGPase in Riso16 mutants led to a strong decrease in ADPGlc level, in both the cytosol and plastid, whereas knock-down of the ADPGlc/ADP transporter led to a large shift in the intracellular distribution of ADPGlc. (v) The thermodynamic structure of the pathway of sucrose to starch was determined by calculating the mass-action ratios

  20. The Optical Absorption Coefficient of Barley Seeds Investigated by Photoacoustic Spectroscopy and Their Effects by Laser Biostimulation

    NASA Astrophysics Data System (ADS)

    Pérez Reyes, Ma. C.; Hernandez-Aguilar, C.; Dominguez-Pacheco, A.; Cruz-Orea, A.; Moreno Martínez, E.

    2015-09-01

    Laser light as a biostimulator has been applied in agriculture, and some scientific reports evidence its usefulness. A knowledge about seed optical parameters is of great relevance in the biostimulation process, because information can be provided about the light absorption of seeds. Thus, the objective of the present study was to determine the optical absorption coefficient (β ) of barley ( Hordeum vulgare L.) seeds by means of photoacoustic spectroscopy; these seeds were studied in two conditions: seeds in their natural color and seeds dyed with methylene blue. The seeds were biostimulated by a laser beam (650 nm wavelength) to evaluate the effects of pre-sowing biostimulation in natural mycobiota associated with different laser irradiation times (0 s, 60 s, 120 s, 240 s, and 480 s). The results of this research demonstrated changes in the optical parameters (absorption and penetration) that occur in the seeds by changing the natural condition to a dyed condition. The dyed seeds, by the methylene blue photosensitizer, become optically opaque, producing greater optical absorption at 650 nm which causes an increase in the effect of laser stimulation. The experimental results showed that the biggest mycobiota reduction (52 %) corresponded to dyed seeds irradiated with a laser for 120 s.

  1. Differences in Grain Ultrastructure, Phytochemical and Proteomic Profiles between the Two Contrasting Grain Cd-Accumulation Barley Genotypes

    PubMed Central

    Sun, Hongyan; Cao, Fangbin; Wang, Nanbo; Zhang, Mian; Mosaddek Ahmed, Imrul; Zhang, Guoping; Wu, Feibo

    2013-01-01

    To reveal grain physio-chemical and proteomic differences between two barley genotypes, Zhenong8 and W6nk2 of high- and low- grain-Cd-accumulation, grain profiles of ultrastructure, amino acid and proteins were compared. Results showed that W6nk2 possesses significantly lower protein content, with hordein depicting the greatest genotypic difference, compared with Zhenong8, and lower amino acid contents with especially lower proportion of Glu, Tyr, Phe and Pro. Both scanning and transmission electron microscopy observation declared that the size of A-type starch molecule in W6nk2 was considerably larger than that of Zhenong8. Grains of Zhenong8 exhibited more protein-rich deposits around starch granules, with some A-type granules having surface pits. Seventeen proteins were identified in grains, using 2-DE coupled with mass spectrometry, with higher expression in Zhenong8 than that in W6nk2; including z-type serpin, serpin-Z7 and alpha-amylase/trypsin inhibitor CM, carbohydrate metabolism, protein synthesis and signal transduction related proteins. Twelve proteins were less expressed in Zhenong8 than that in W6nk2; including barley trypsin inhibitor chloroform/methanol-soluble protein (BTI-CMe2.1, BTI-CMe2.2), trypsin inhibitor, dehydroascorbate reductase (DHAR), pericentrin, dynein heavy chain and some antiviral related proteins. The data extend our understanding of mechanisms underlying Cd accumulation/tolerance and provides possible utilization of elite genetic resources in developing low-grain-Cd barley cultivars. PMID:24260165

  2. In situ study of water uptake by the seeds, endosperm and husk of barley using infrared spectroscopy.

    PubMed

    Cozzolino, D; Degner, S; Eglinton, J K

    2015-11-01

    Variations in the amount and rates of water uptake influence the seed hydration as well as the modification of the endosperm for industrial uses (e.g., malting). The aim of this study was to investigate and interpret absorption frequencies in the mid infrared (MIR) region associated with water uptake in whole seeds, husk and endosperm of barley seeds during the initial period of soaking in water. Partial least squares (PLS) regression models for the prediction of water uptake in the set of samples yield a coefficient of determination (R(2)) and a standard error in cross validation of 0.75 and 2.57 (% w/w), respectively. The biological implications of this study are that the first stages of germination can be monitored using the information derived from the MIR spectra. These results also demonstrated that whole seeds, endosperm and husk derived from the same variety or genotype have different patterns in the MIR region. PMID:26048560

  3. A major cysteine proteinase, EPB, in germinating barley seeds: structure of two intronless genes and regulation of expression.

    PubMed

    Mikkonen, A; Porali, I; Cercos, M; Ho, T H

    1996-05-01

    The barley cysteine proteinase B (EPB) is the main protease responsible for the degradation of endosperm storage proteins providing nitrogenous nutrients to support the growth of young seedlings. The expression of this enzyme is induced in the germinating seeds by the phytohormone, gibberellin, and suppressed by another phytohormone, abscisic acid. In situ hybridization experiments indicate that EPB is expressed in the scutellar epithelium within 24 h of seed germination, but the aleurone tissue surrounding the starchy endosperm eventually becomes the main tissue expressing this enzyme. The EPB gene family of barley consists of two very similar genes, EPB1 and EPB2, both of which have been mapped to chromosome 3. The sequences of EPB1 and EPB2 match with the two previously published cDNA clones indicating that both genes are expressed. Interestingly, neither of these genes contain any introns, a rare phenomenon in which all members of a small gene family are active intronless genes. Sequence comparison indicates that the barley EPB family can be classified as cathepsin L-like endopeptidases and is most closely related to two legume cysteine proteinases (Phaseolus vulgaris EP-C1 and Vigna mungo SHEP) which are also involved in seed storage protein degradation. The promoters of EPB1 and EPB2 have been linked to the coding sequence of a reporter gene, GUS, encoding beta-glucuronidase, and introduced into barley aleurone cells using the particle bombardment method. Transient expression studies indicate that EPB promoters are sufficient to confer the hormonal regulation of these genes. PMID:8756590

  4. Nitric Oxide and Reactive Oxygen Species Mediate Metabolic Changes in Barley Seed Embryo during Germination.

    PubMed

    Ma, Zhenguo; Marsolais, Frédéric; Bykova, Natalia V; Igamberdiev, Abir U

    2016-01-01

    The levels of nitric oxide (NO) and reactive oxygen species (ROS), ATP/ADP ratios, reduction levels of ascorbate and glutathione, expression of the genes encoding proteins involved in metabolism of NO and activities of the enzymes involved in fermentation and in metabolism of NO and ROS were studied in the embryos of germinating seeds of two barley (Hordeum vulgare L.) cultivars differing in dormancy level. The level of NO production continuously increased after imbibition while the level of nitrosylated SH-groups in proteins increased. This corresponded to the decrease of free SH-groups in proteins. At early stage of germination (0-48 h post imbibition) the genes encoding class 1 phytoglobin (the protein scavenging NO) and S-nitrosoglutathione reductase (scavenging S-nitrosoglutathione) were markedly expressed. More dormant cultivar exhibited lower ATP/ADP and ascorbate/dehydroascorbate ratios and lower lactate and alcohol dehydrogenase activities, while the production of NO and nitrosylation of proteins was higher as compared to the non-dormant cultivar. The obtained data indicate that at the onset of germination NO is actively generated causing nitrosylation of SH-groups and a switch from respiration to fermentation. After radicle protrusion the metabolism changes in a more reducing type as recorded by ratio of reduced and oxidized glutathione and ascorbate. The turnover of NO by the scavenging systems (phytoglobin, S-nitrosoglutathione reductase and interaction with ROS) might contribute to the maintenance of redox and energy balance of germinating seeds and lead to alleviation of dormancy. PMID:26909088

  5. Nitric Oxide and Reactive Oxygen Species Mediate Metabolic Changes in Barley Seed Embryo during Germination

    PubMed Central

    Ma, Zhenguo; Marsolais, Frédéric; Bykova, Natalia V.; Igamberdiev, Abir U.

    2016-01-01

    The levels of nitric oxide (NO) and reactive oxygen species (ROS), ATP/ADP ratios, reduction levels of ascorbate and glutathione, expression of the genes encoding proteins involved in metabolism of NO and activities of the enzymes involved in fermentation and in metabolism of NO and ROS were studied in the embryos of germinating seeds of two barley (Hordeum vulgare L.) cultivars differing in dormancy level. The level of NO production continuously increased after imbibition while the level of nitrosylated SH-groups in proteins increased. This corresponded to the decrease of free SH-groups in proteins. At early stage of germination (0–48 h post imbibition) the genes encoding class 1 phytoglobin (the protein scavenging NO) and S-nitrosoglutathione reductase (scavenging S-nitrosoglutathione) were markedly expressed. More dormant cultivar exhibited lower ATP/ADP and ascorbate/dehydroascorbate ratios and lower lactate and alcohol dehydrogenase activities, while the production of NO and nitrosylation of proteins was higher as compared to the non-dormant cultivar. The obtained data indicate that at the onset of germination NO is actively generated causing nitrosylation of SH-groups and a switch from respiration to fermentation. After radicle protrusion the metabolism changes in a more reducing type as recorded by ratio of reduced and oxidized glutathione and ascorbate. The turnover of NO by the scavenging systems (phytoglobin, S-nitrosoglutathione reductase and interaction with ROS) might contribute to the maintenance of redox and energy balance of germinating seeds and lead to alleviation of dormancy. PMID:26909088

  6. Comparative Transcriptional Profiling Established the Awn as the Major Photosynthetic Organ of the Barley Spike while the Lemma and the Palea Primarily Protect the Seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The lemma, palea, and awn of barley are photosynthetic organs and supply the developing seed with carbohydrates. In addition, the lemma and the palea cover the seed and protect it from pathogens and insects. In spite of the important role they play, there is very little information about gene expr...

  7. Exogenous melatonin improves corn (Zea mays L.) embryo proteome in seeds subjected to chilling stress.

    PubMed

    Kołodziejczyk, Izabela; Dzitko, Katarzyna; Szewczyk, Rafał; Posmyk, Małgorzata M

    2016-04-01

    Melatonin (MEL; N-acetyl-5-methoxytryptamine) plays an important role in plant stress defense. Various plant species rich in this indoleamine have shown a higher capacity for stress tolerance. Moreover, it has great potential for plant biostimulation, is biodegradable and non-toxic for the environment. All this indicates that our concept of seed enrichment with exogenous MEL is justified. This work concerns the effects of corn (Zea mays L.) seed pre-sowing treatments supplemented with MEL. Non-treated seeds (nt), and those hydroprimed with water (H) or with MEL solutions 50 and 500 μM (HMel50, HMel500) were compared. Positive effects of seed priming are particularly apparent during germination under suboptimal conditions. The impact of MEL applied by priming on seed protein profiles during imbibition/germination at low temperature has not been investigated to date. In order to identify changes in the corn seed proteome after applying hydropriming techniques, purified protein extracts of chilling stressed seed embryos (14 days, 5°C) were separated by two-dimensional electrophoresis. Then proteome maps were graphically and statistically compared and selected protein spots were qualitatively analyzed using mass spectrometry techniques and identified. This study aimed to analyze the priming-induced changes in maize embryo proteome and at identifying priming-associated and MEL-associated proteins in maize seeds subjected to chilling. We attempt to explain how MEL expands plant capacity for stress tolerance. PMID:26945210

  8. The study of two barley Type I-like MADS-box genes as potential targets of epigenetic regulation during seed development

    PubMed Central

    2012-01-01

    Background MADS-box genes constitute a large family of transcription factors functioning as key regulators of many processes during plant vegetative and reproductive development. Type II MADS-box genes have been intensively investigated and are mostly involved in vegetative and flowering development. A growing number of studies of Type I MADS-box genes in Arabidopsis, have assigned crucial roles for these genes in gamete and seed development and have demonstrated that a number of Type I MADS-box genes are epigenetically regulated by DNA methylation and histone modifications. However, reports on agronomically important cereals such as barley and wheat are scarce. Results Here we report the identification and characterization of two Type I-like MADS-box genes, from barley (Hordeum vulgare), a monocot cereal crop of high agronomic importance. Protein sequence and phylogenetic analysis showed that the putative proteins are related to Type I MADS-box proteins, and classified them in a distinct cereal clade. Significant differences in gene expression among seed developmental stages and between barley cultivars with varying seed size were revealed for both genes. One of these genes was shown to be induced by the seed development- and stress-related hormones ABA and JA whereas in situ hybridizations localized the other gene to specific endosperm sub-compartments. The genomic organization of the latter has high conservation with the cereal Type I-like MADS-box homologues and the chromosomal position of both genes is close to markers associated with seed quality traits. DNA methylation differences are present in the upstream and downstream regulatory regions of the barley Type I-like MADS-box genes in two different developmental stages and in response to ABA treatment which may be associated with gene expression differences. Conclusions Two barley MADS-box genes were studied that are related to Type I MADS-box genes. Differential expression in different seed developmental

  9. Detection of Pyrenophora teres in conidia and barley seed by a rapid PCR technique

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pyrenophora teres, the causal agent of net blotch of barley survives as seedborne mycelium or pseudothesia in infested host residues. Besides the common netlike and occasional spot form symptoms on barley leaves, diffused dark or pale symptoms which are difficult to distinguish from other fungi are ...

  10. Proteomic analysis of rice after different seed space flights by two-dimensional difference electrophoresis

    NASA Astrophysics Data System (ADS)

    Wang, Wei; Liang, Shujian; Sun, Yeqing

    To investigate the biological effects of space environment in rice plants, proteomic profiles of six rice cultivars growing after twice different seed space flights were analyzed by two-dimensional difference gel electrophoresis (2-D DIGE) coupled with mass spectrometry (MS). Over 1500 protein spots were detected in each paired space/ground-control comparison and more than 800 protein spots were reproducible across all the samples. Six proteins including peroxiredoxin and rubisco were found significantly changed in most of the six cultivars after both of the seed space flights, indicating they might be associated with the responses of rice cells to the space environment. Cluster analyses were also applied using the quantitative protein expression data: cultivar hierarchical clustering and principal component analysis both indicated that the rice proteome changed its expression profiles after seed space environment exposures while protein hierarchical clustering revealed that there might be a decrease of protein expression in rice plants after seed space flights.

  11. Quantitative Proteomics Analysis of Camelina sativa Seeds Overexpressing the AGG3 Gene to Identify the Proteomic Basis of Increased Yield and Stress Tolerance.

    PubMed

    Alvarez, Sophie; Roy Choudhury, Swarup; Sivagnanam, Kumaran; Hicks, Leslie M; Pandey, Sona

    2015-06-01

    Camelina sativa, a close relative of Arabidopsis, is an oilseed plant that is emerging as an important biofuel resource. The genome and transcriptome maps of Camelina have become available recently, but its proteome composition remained unexplored. A labeling LC-based quantitative proteomics approach was applied to decipher the Camelina seed proteome, which led to the identification of 1532 proteins. In addition, the effect of overexpression of the Arabidopsis G-protein γ subunit 3 (AGG3) on the Camelina seed proteome was elucidated to identify the proteomic basis of its increased seed size and improved stress tolerance. The comparative analysis showed a significantly higher expression of proteins involved in primary and secondary metabolism, nucleic acid and protein metabolism, and abscisic acid related responses, corroborating the physiological effects of AGG3 overexpression. More importantly, the proteomic data suggested involvement of the AGG3 protein in the regulation of oxidative stress and heavy metal stress tolerance. These observations were confirmed by the physiological and biochemical characterization of AGG3-overexpressing seeds, which exhibit a higher tolerance to exogenous cadmium in a glutathione-dependent manner. The activity of multiple redox-regulating enzymes is higher in seeds expressing enhanced levels of AGG3. Overall, these data provide critical evidence for the role of redox regulation by the AGG3 protein in mediating important seed-related traits. PMID:25944359

  12. Control of the Water Transport Activity of Barley HvTIP3;1 Specifically Expressed in Seeds.

    PubMed

    Utsugi, Shigeko; Shibasaka, Mineo; Maekawa, Masahiko; Katsuhara, Maki

    2015-09-01

    Tonoplast intrinsic proteins (TIPs) are involved in the transport and storage of water, and control intracellular osmotic pressure by transporting material related to the water potential of cells. In the present study, we focused on HvTIP3;1 during the periods of seed development and desiccation in barley. HvTIP3;1 was specifically expressed in seeds. An immunochemical analysis showed that HvTIP3;1 strongly accumulated in the aleurone layers and outer layers of barley seeds. The water transport activities of HvTIP3;1 and HvTIP1;2, which also accumulated in seeds, were measured in the heterologous expression system of Xenopus oocytes. When they were expressed individually, HvTIP1;2 transported water, whereas HvTIP3;1 did not. However, HvTIP3;1 exhibited water transport activity when co-expressed with HvTIP1;2 in oocytes, and this activity was higher than when HvTIP1;2 was expressed alone. This is the first report to demonstrate that the water permeability of a TIP aquaporin was activated when co-expressed with another TIP. The split-yellow fluorescent protein (YFP) system in onion cells revealed that HvTIP3;1 interacted with HvTIP1;2 to form a heterotetramer in plants. These results suggest that HvTIP3;1 functions as an active water channel to regulate water movement through tissues during the periods of seed development and desiccation. PMID:26177807

  13. Comparative Proteomic Analysis and IgE Binding Properties of Peanut Seed and Testa (Skin)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To investigate the protein composition and potential allergenicity of peanut testae or skins, proteome analysis was conducted using nanoLC-MS/MS sequencing. Initial amino acid analysis suggested differences in protein compositions between the blanched seed (skins removed) and skin. Phenolic compou...

  14. Assessment of the natural variation of low abundant metabolic proteins in soybean seeds using proteomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry, we investigated the distribution of the low abundant proteins that are involved in soybean seed development in four wild and twelve cultivated soybean genotypes. We found proteomic variation of these proteins within and...

  15. Proteomic Analysis of the Protein Expression Profile in the Mature Nigella sativa (Black Seed).

    PubMed

    Alanazi, Ibrahim O; Benabdelkamel, Hicham; Alfadda, Assim A; AlYahya, Sami A; Alghamdi, Waleed M; Aljohi, Hasan A; Almalik, Abdulaziz; Masood, Afshan

    2016-08-01

    Nigella sativa (N. sativa) seed has been used as an important nutritional flavoring agent and in traditional medicine for treating many illnesses since ancient times. Understanding the proteomic component of the seed may lead to enhance the understanding of its structural and biological functional complexity. In this study, we have analyzed its proteome profile based on gel-based proteome mapping technique that includes one-dimensional gel electrophoresis followed by liquid chromatography and tandem mass spectrometry strategy. We have not come across any such studies that have been performed in N. sativa seeds up to date. A total of 277 proteins were identified, and their functional, metabolic, and location-wise annotations were carried out using the UniProt database. The majority of proteins identified in the proteome dataset based on their function were those involved in enzyme catalytic activity, nucleotide binding, and protein binding while the major cellular processes included regulation of biological process followed by regulation of secondary biological process, cell organization and biogenesis, protein metabolism, and transport. The identified proteome was localized mainly to the nucleus then to the cytoplasm, plasma membrane, mitochondria, plastid, and others. A majority of the proteins were involved in biochemical pathways involving carbohydrate metabolism, amino acid and shikimate pathway, lipid metabolism, nucleotide, cell organization and biogenesis, transport, and defense processes. The identified proteins in the dataset help to improve our understanding of the pathways involved in N. sativa seed metabolism and its biochemical features and detail out useful information that may help to utilize these proteins. This study could thus pave a way for future further high-throughput studies using a more targeted proteomic approach. PMID:27020565

  16. Characterization of quinoa seed proteome combining different protein precipitation techniques: Improvement of knowledge of nonmodel plant proteomics.

    PubMed

    Capriotti, Anna Laura; Cavaliere, Chiara; Piovesana, Susy; Stampachiacchiere, Serena; Ventura, Salvatore; Zenezini Chiozzi, Riccardo; Laganà, Aldo

    2015-03-01

    A shotgun proteomics approach was used to characterize the quinoa seed proteome. To obtain comprehensive proteomic data from quinoa seeds three different precipitation procedures were employed: MeOH/CHCl3 /double-distilled H2 O, acetone either alone or with trichloroacetic acid; the isolated proteins were then in-solution digested and the resulting peptides were analyzed by nano-liquid chromatography coupled to tandem mass spectrometry. However, since quinoa is a nonmodel plant species, only a few protein sequences are included in the most widely known protein sequence databases. To improve the data reliability a UniProt subdatabase, containing only proteins of Caryophillales order, was used. A total of 352 proteins were identified and evaluated both from a qualitative and quantitative point of view. This combined approach is certainly useful to increase the final number of identifications, but no particular class of proteins was extracted and identified in spite of the different chemistries and the different precipitation protocols. However, with respect to the other two procedures, from the relative quantitative analysis, based on the number of spectral counts, the trichloroacetic acid/acetone protocol was the best procedure for sample handling and quantitative protein extraction. This study could pave the way to further high-throughput studies on Chenopodium Quinoa. PMID:25580831

  17. Evaluation of phytotoxicity effect of olive mill wastewater treated by different technologies on seed germination of barley (Hordeum vulgare L.).

    PubMed

    Rusan, Munir J M; Albalasmeh, Ammar A; Zuraiqi, Said; Bashabsheh, Mohammad

    2015-06-01

    Olive-mill wastewater (OMW) is a by-product effluent of olive oil extraction process that is produced in large amount in the Mediterranean region. OMW is believed to induce phytotoxic effect on organisms including seed germination and plant growth. The objective of this study was to evaluate the impact of untreated and treated OMW with different techniques on seed germination of barley (Hordeum vulgare L.). The following treatments were investigated: (1) tap water (control); (2) OMW treated by aerobic biological technology in a Jacto Reactor (JR); (3) OMW treated by solar fenton oxidation (SFO); (4) OMW treated by microfiltration followed by nanofiltration (MF+NF); (5) OMW treated by microfiltration followed by reverse osmosis (MF+RO) process; (6) diluted OMW with tap water (25 % OMW); (7) diluted OMW with tap water (50 % OMW); (8) diluted OMW with tap water (75 % OMW); and (9) untreated OMW (100 % OMW). A germination test was conducted in an incubator at temperature of 23 (∘)C. In each petri dish, a filter paper was mounted and ten seeds of barley were placed on the filter paper. Five milliliter of water were added to each petri dish. The seed germination was determined by counting the number of germinated seeds to calculate the percentage of germination (G %). Germination rate index (GRI), seed vigor index (SVI), and phytotoxicity index (PI) were also calculated. Then, the dry weights and lengths of the shoots and the roots of the germinated seeds were measured. The results show that 100, 75, and 50 %OMW were very phytotoxic and completely prohibited seed germination. However, phytotoxicity decreased significantly following treatments of OMW with all techniques investigated and by the 25 % OMW dilution, as results of removing the phenols and other phytotoxic organic compounds from the OMW or by diluting it. This was evidenced by relative enhancement of the dry weights and lengths of shoot and root as well as the G %, GRI, SVG, and PI. It was concluded that if

  18. Expect the Unexpected Enrichment of "Hidden Proteome" of Seeds and Tubers by Depletion of Storage Proteins.

    PubMed

    Gupta, Ravi; Min, Cheol W; Wang, Yiming; Kim, Yong C; Agrawal, Ganesh K; Rakwal, Randeep; Kim, Sun T

    2016-01-01

    Dynamic resolution of seed and tuber protein samples is highly limited due to the presence of high-abundance storage proteins (SPs). These proteins inevitably obscure the low-abundance proteins (LAPs) impeding their identification and characterization. To facilitate the detection of LAPs, several methods have been developed during the past decade, enriching the proteome with extreme proteins. Most of these methods, if not all, are based on the specific removal of SPs which ultimately magnify the proteome coverage. In this mini-review, we summarize the available methods that have been developed over the years for the enrichment of LAPs either from seeds or tubers. Incorporation of these methods during the protein extraction step will be helpful in understanding the seed/tuber biology in greater detail. PMID:27313590

  19. Proteomic Analysis Reveals Key Proteins and Phosphoproteins upon Seed Germination of Wheat (Triticum aestivum L.).

    PubMed

    Dong, Kun; Zhen, Shoumin; Cheng, Zhiwei; Cao, Hui; Ge, Pei; Yan, Yueming

    2015-01-01

    Wheat (Triticum aestivum L.) is one of the oldest cultivated crops and the second most important food crop in the world. Seed germination is the key developmental process in plant growth and development, and poor germination directly affects plant growth and subsequent grain yield. In this study, we performed the first dynamic proteome analysis of wheat seed germination using a two-dimensional differential gel electrophoresis (2D-DIGE)-based proteomic approach. A total of 166 differentially expressed protein (DEP) spots representing 73 unique proteins were identified, which are mainly involved in storage, stress/defense/detoxification, carbohydrate metabolism, photosynthesis, cell metabolism, and transcription/translation/transposition. The identified DEPs and their dynamic expression profiles generally correspond to three distinct seed germination phases after imbibition: storage degradation, physiological processes/morphogenesis, and photosynthesis. Some key DEPs involved in storage substance degradation and plant defense mechanisms, such as globulin 3, sucrose synthase type I, serpin, beta-amylase, and plastid ADP-glucose pyrophosphorylase (AGPase) small subunit, were found to be phosphorylated during seed germination. Particularly, the phosphorylation site Ser(355) was found to be located in the enzyme active region of beta-amylase, which promotes substrate binding. Phosphorylated modification of several proteins could promote storage substance degradation and environmental stress defense during seed germination. The central metabolic pathways involved in wheat seed germination are proposed herein, providing new insights into the molecular mechanisms of cereal seed germination. PMID:26635843

  20. Proteomic Analysis Reveals Key Proteins and Phosphoproteins upon Seed Germination of Wheat (Triticum aestivum L.)

    PubMed Central

    Dong, Kun; Zhen, Shoumin; Cheng, Zhiwei; Cao, Hui; Ge, Pei; Yan, Yueming

    2015-01-01

    Wheat (Triticum aestivum L.) is one of the oldest cultivated crops and the second most important food crop in the world. Seed germination is the key developmental process in plant growth and development, and poor germination directly affects plant growth and subsequent grain yield. In this study, we performed the first dynamic proteome analysis of wheat seed germination using a two-dimensional differential gel electrophoresis (2D-DIGE)-based proteomic approach. A total of 166 differentially expressed protein (DEP) spots representing 73 unique proteins were identified, which are mainly involved in storage, stress/defense/detoxification, carbohydrate metabolism, photosynthesis, cell metabolism, and transcription/translation/transposition. The identified DEPs and their dynamic expression profiles generally correspond to three distinct seed germination phases after imbibition: storage degradation, physiological processes/morphogenesis, and photosynthesis. Some key DEPs involved in storage substance degradation and plant defense mechanisms, such as globulin 3, sucrose synthase type I, serpin, beta-amylase, and plastid ADP-glucose pyrophosphorylase (AGPase) small subunit, were found to be phosphorylated during seed germination. Particularly, the phosphorylation site Ser355 was found to be located in the enzyme active region of beta-amylase, which promotes substrate binding. Phosphorylated modification of several proteins could promote storage substance degradation and environmental stress defense during seed germination. The central metabolic pathways involved in wheat seed germination are proposed herein, providing new insights into the molecular mechanisms of cereal seed germination. PMID:26635843

  1. Role of reactive oxygen species produced by NADPH oxidase in gibberellin biosynthesis during barley seed germination.

    PubMed

    Kai, Kyohei; Kasa, Shinsuke; Sakamoto, Masatsugu; Aoki, Nozomi; Watabe, Gaku; Yuasa, Takashi; Iwaya-Inoue, Mari; Ishibashi, Yushi

    2016-05-01

    NADPH oxidase catalyzes the production of the superoxide anion (O2(-)), a reactive oxygen species (ROS), and regulates the germination of barley (Hordeum vulgare L.). Diphenyleneiodonium (DPI) chloride, an NADPH oxidase inhibitor, delayed barley germination, and exogenous H2O2 (an ROS) partially rescued it. Six enzymes, ent-copalyl diphosphate synthase (CPS), ent-kaurene synthase (KS), ent-kaurene oxidase (KO), ent-kaurenoic acid oxidase (KAO), GA20-oxidase (GA20ox) and GA3-oxidase (GA3ox), catalyze the transformation of trans-geranylgeranyl diphosphate to active gibberellin, which promotes germination. Exogenous H2O2 promoted the expressions of HvKAO1 and HvGA3ox1 in barley embryos. These results suggest that ROS produced by NADPH oxidase are involved in gibberellin biosynthesis through the regulation of HvKAO1 and HvGA3ox1. PMID:27110861

  2. Differential RNA Expression of Bmy1 During Late Seed Development in Wild and Cultivated Barley and the Association With ß-Amylase Activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four genotypes carrying different ß-amylase 1 (Bmy1) intron III alleles (Bmy1.a, Bmy1.b, Bmy1.c, and Bmy1.d) were analyzed for differences in Bmy1 DNA sequence, Bmy1 RNA expression, ß-amylase activity and protein, and total protein during late seed development. Wild barleys Ashqelon (Bmy1.c) and PI...

  3. Leaf proteome alterations in the context of physiological and morphological responses to drought and heat stress in barley (Hordeum vulgare L.).

    PubMed

    Rollins, J A; Habte, E; Templer, S E; Colby, T; Schmidt, J; von Korff, M

    2013-08-01

    The objective of this study was to identify barley leaf proteins differentially regulated in response to drought and heat and the combined stresses in context of the morphological and physiological changes that also occur. The Syrian landrace Arta and the Australian cultivar Keel were subjected to drought, high temperature, or a combination of both treatments starting at heading. Changes in the leaf proteome were identified using differential gel electrophoresis and mass spectrometry. The drought treatment caused strong reductions of biomass and yield, while photosynthetic performance and the proteome were not significantly changed. In contrast, the heat treatment and the combination of heat and drought reduced photosynthetic performance and caused changes of the leaf proteome. The proteomic analysis identified 99 protein spots differentially regulated in response to heat treatment, 14 of which were regulated in a genotype-specific manner. Differentially regulated proteins predominantly had functions in photosynthesis, but also in detoxification, energy metabolism, and protein biosynthesis. The analysis indicated that de novo protein biosynthesis, protein quality control mediated by chaperones and proteases, and the use of alternative energy resources, i.e. glycolysis, play important roles in adaptation to heat stress. In addition, genetic variation identified in the proteome, in plant growth and photosynthetic performance in response to drought and heat represent stress adaption mechanisms to be exploited in future crop breeding efforts. PMID:23918963

  4. Leaf proteome alterations in the context of physiological and morphological responses to drought and heat stress in barley (Hordeum vulgare L.)

    PubMed Central

    von Korff, M.

    2013-01-01

    The objective of this study was to identify barley leaf proteins differentially regulated in response to drought and heat and the combined stresses in context of the morphological and physiological changes that also occur. The Syrian landrace Arta and the Australian cultivar Keel were subjected to drought, high temperature, or a combination of both treatments starting at heading. Changes in the leaf proteome were identified using differential gel electrophoresis and mass spectrometry. The drought treatment caused strong reductions of biomass and yield, while photosynthetic performance and the proteome were not significantly changed. In contrast, the heat treatment and the combination of heat and drought reduced photosynthetic performance and caused changes of the leaf proteome. The proteomic analysis identified 99 protein spots differentially regulated in response to heat treatment, 14 of which were regulated in a genotype-specific manner. Differentially regulated proteins predominantly had functions in photosynthesis, but also in detoxification, energy metabolism, and protein biosynthesis. The analysis indicated that de novo protein biosynthesis, protein quality control mediated by chaperones and proteases, and the use of alternative energy resources, i.e. glycolysis, play important roles in adaptation to heat stress. In addition, genetic variation identified in the proteome, in plant growth and photosynthetic performance in response to drought and heat represent stress adaption mechanisms to be exploited in future crop breeding efforts. PMID:23918963

  5. A Simple Procedure for Depletion of Storage Proteins From Soybean (Glycine max) Seeds to Aid Proteome Analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two-dimensional electrophoretic analysis of plant proteomes containing thousands of proteins, has limited dynamic resolution because only abundant proteins can be detected. Proteomic assessment of the low abundance proteins within seeds is difficult when 60 – 80% is storage proteins. Resolution can ...

  6. Modification of the radiosensitivity of barley seed by post-treatment with caffein. IV. Effect of the moisture content of seed and storage temperature after irradiation.

    PubMed

    Kesavan, P C; Nadkarni, S

    1977-02-01

    The oxygen-dependent damage which develops in barley seeds with approximately 7-8 per cent moisture content disappears after post-irradiation storage in vacuo for 48 hours at 40 degrees C and for 24 hours at 50 degrees C. When the diration of storage at 40 degrees C is extended to 384 hours, oxygen-independent damage becomes potentiated. There is oxygen-dependent damage in seeds of approximately 13.3 per cent moisture content and after the seeds have been stored in vacuo at 50 degrees C, the oxygen-dependent damage begins to increase by 168 hours, and it is very significantly potentiated by 192 hours. Under these circumstances, caffeine acts as a radioprotector only as long as the precursors of oxic damage are present in the seeds. Once these sites are lost, caffeine acts only as a radiosensitizer. The oxygen-independent damage which increases with storage at high temperature is further potentiated by caffeine. PMID:300722

  7. Proteome balancing of the maize seed for higher nutritional value.

    PubMed

    Wu, Yongrui; Messing, Joachim

    2014-01-01

    Most flowering plant seeds are composed of the embryo and endosperm, which are surrounded by maternal tissue, in particular the seed coat. Whereas the embryo is the dormant progeny, the endosperm is a terminal organ for storage of sugars and amino acids in proteins and carbohydrates, respectively. Produced in maternal leaves during photosynthesis, sugars, and amino acids are transported to developing seeds after flowering, and during germination they nourish early seedlings growth. Maize endosperm usually contains around 10% protein and 70% starch, and their composition ratio is rather stable, because it is strictly regulated through a pre-set genetic program that is woven by networks of many interacting or counteracting genes and pathways. Endosperm protein, however, is of low nutritional value due mainly to the high expression of the α-zein gene family, which encodes lysine-free proteins. Reduced levels of these proteins in the opaque 2 (o2) mutant and α-zein RNAi (RNA interference) transgenic seed is compensated by an increase of non-zein proteins, leading to the rebalancing of the nitrogen sink and producing more or less constant levels of total proteins in the seed. The same rebalancing of zeins and non-zeins has been observed for maize seeds bred for 30% protein. In contrast to the nitrogen sink, storage of sulfur is controlled through the accumulation of specialized sulfur-rich proteins in maize endosperm. Silencing the synthesis of α-zeins through RNAi fails to raise sulfur-rich proteins. Although overexpression of the methionine-rich δ-zein can increase the methionine level in seeds, it occurs at least in part at the expense of the cysteine-rich β- and γ-zeins, demonstrating a balance between cysteine and methionine in sulfur storage. Therefore, we propose that the throttle for the flow of sulfur is placed before the synthesis of sulfur amino acids when sulfur is taken up and reduced during photosynthesis. PMID:24910639

  8. Isolation and purification of a papain inhibitor from Egyptian genotypes of barley seeds and its in vitro and in vivo effects on the cowpea bruchid, Callosobruchus maculatus (F.).

    PubMed

    Abd El-Latif, Ashraf Oukasha

    2015-02-01

    The cysteine inhibitors that are known as cystatin have been identified and characterized from several plant species. In the current study, 44 barley (Hordeum vulgare) genotypes including 3 varieties and 41 promising lines were screened for their potential as protease inhibitors. The barley genotypes showed low inhibitory activity against trypsin and chymotrypsin enzymes with a mean of 4.15 TIU/mg protein and 4.40 CIU/mg protein. The barley variety, Giza 123, showed strong papain inhibitory activity of 97.09 PIU/mg proteins and was subjected for further purification studies using ammonium sulfate fractionation and DEAE-Sephadex A-25 column. Barley purified proteins showed two bands on SDS-PAGE corresponding to a molecular mass of 12.4-54.8 kDa. The purified barley PI was found to be stable at a temperature below 80 °C and at a wide range of pH from 2 to 12. Barley PI was found to have higher potential inhibitory activity against papain enzyme compared to the standard papain inhibitor, E-64 with an IC50 value of 21.04 µg/ml and 25.62 µg/ml for barley PI and E-64, respectively. The kinetic analysis revealed a non-competitive type of inhibition with a Ki value of 1.95 × 10(-3 )µM. The antimetabolic effect of barley PI was evaluated against C. maculatus by incorporating the F30-60 protein of the purified inhibitor into the artificial diet using artificial seeds. Barley PI significantly prolonged the development of C. maculatus in proportion to PI concentration. Barley PI significantly increased the mortality of C. maculatus and caused a significant reduction in its fecundity. On the other hand, barley PI seemed to have non-significant effects on the adult longevity and the adult dry weight. The in vitro and in vivo results proved the efficiency of the papain inhibitory protein isolated from barley as a tool for managing the cowpea bruchid, C. maculatus. PMID:25752426

  9. Metabolic profiling of early F. graminearum infection in barley seed spike tissues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several studies have utilized microarray analysis to characterize gene expression occurring during infection of barley and wheat by the fungal pathogen Fusarium graminearum (F.g.). We have analyzed this plant-microbe interaction from the other extreme - the changes in the metabolome arising from th...

  10. Drought Response in the Spikes of Barley: Gene Expression in the Lemma, Palea, Awn, and Seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The photosynthetic organs of the barley spike (lemma, palea, and awn) are considered resistant to drought. This is a beneficial trait because they can sustain grain-filling when drought occurs at the reproductive stage. However, there is little information about gene expression in the spike organs u...

  11. Comparative proteomic analysis of green malts between barley (Hordeum vulgare) cultivars.

    PubMed

    Jin, Zhao; Cai, Guo-Lin; Li, Xiao-Min; Gao, Fei; Yang, Jing-Jing; Lu, Jian; Dong, Jian-jun

    2014-05-15

    The variety of metabolic proteins theoretically reaches a peak at the end of germination in large-scale malting. In the present study, comparative proteomics based on two-dimensional fluorescent difference gel electrophoresis (2D-DIGE) was employed to quantitatively analyse the low-salt soluble proteins in green malts from cultivars of Dan'er and Metcalfe. Fifty-nine metabolic proteins with significant differences between cultivars were successfully identified using MALDI-TOF/TOF. The roles of differential proteins in malt quality discrimination were elucidated according to their functions. Among them, 18 proteins exhibited differences in the green malts but not in the malts between the two cultivars. They could be considered as supplementary contributors to the quality defects of Dan'er malt, and new markers for malt quality improvement. PMID:24423531

  12. Comparative proteomic analysis of seed embryo proteins associated with seed storability in rice (Oryza sativa L) during natural aging.

    PubMed

    Gao, Jiadong; Fu, Hua; Zhou, Xinqiao; Chen, Zhongjian; Luo, Yi; Cui, Baiyuan; Chen, Guanghui; Liu, Jun

    2016-06-01

    Seed storability is considered an important trait in rice breeding; however, the underlying regulating mechanisms remain largely unknown. Here, we carried out a physiological and proteomic study to identify proteins possibly related to seed storability under natural conditions. Two hybrid cultivars, IIYou998 (IIY998) and BoYou998 (BY998), were analyzed in parallel because they share the same restorer line but have significant differences in seed storability. After a 2-year storage period, the germination percentage of IIY998 was significantly lower than that of BY998, whereas the level of malondialdehyde was reversed, indicating that IIY998 seeds may suffer from more severe damage than BY998 during storage. However, we did not find correlation between activities of antioxidant enzymes of superoxide dismutase, peroxidase, and catalase and seed storability. We identified 78 embryo proteins in embryo whose abundance varied more than 3-fold different during storage or between IIY998 and BY998. More proteins changed in abundance in IIY998 embryo (67 proteins) during storage than in BY998 (10 proteins). Several redox regulation proteins, mainly glutathione-related proteins, exhibited different degree of change during storage between BY998 and IIY998 and might play an important role protecting embryo proteins from oxidation. In addition, some disease/defense proteins, including DNA-damage-repair/toleration proteins, and a putative late embryogenesis abundant protein were significantly downregulated in IIY998, whereas their levels did not change in BY998, indicating that they might be correlated with seed storability. Further studies on these candidate seed storage proteins might help improve our understanding of seed aging. PMID:26950923

  13. Proteomic Analysis of the Endosperm Ontogeny of Jatropha curcas L. Seeds.

    PubMed

    Shah, Mohibullah; Soares, Emanoella L; Carvalho, Paulo C; Soares, Arlete A; Domont, Gilberto B; Nogueira, Fábio C S; Campos, Francisco A P

    2015-06-01

    Seeds of Jatropha curcas L. represent a potential source of raw material for the production of biodiesel. However, this use is hampered by the lack of basic information on the biosynthetic pathways associated with synthesis of toxic diterpenes, fatty acids, and triacylglycerols, as well as the pattern of deposition of storage proteins during seed development. In this study, we performed an in-depth proteome analysis of the endosperm isolated from five developmental stages which resulted in the identification of 1517, 1256, 1033, 752, and 307 proteins, respectively, summing up 1760 different proteins. Proteins with similar label free quantitation expression pattern were grouped into five clusters. The biological significance of these identifications is discussed with special focus on the analysis of seed storage proteins, proteins involved in the metabolism of fatty acids, carbohydrates, toxic components and proteolytic processing. Although several enzymes belonging to the biosynthesis of diterpenoid precursors were identified, we were unable to find any terpene synthase/cyclase, indicating that the synthesis of phorbol esters, the main toxic diterpenes, does not occur in seeds. The strategy used enabled us to provide a first in depth proteome analysis of the developing endosperm of this biodiesel plant, providing an important glimpse into the enzymatic machinery devoted to the production of C and N sources to sustain seed development. PMID:25920442

  14. Proteome analysis of metabolic proteins (pI 4-7) in barley (Hordeum vulgare) malts and initial application in malt quality discrimination.

    PubMed

    Jin, Zhao; Mu, Yu-Wen; Sun, Jun-Yong; Li, Xiao-Min; Gao, Xian-Li; Lu, Jian

    2013-01-16

    Barley malt is essential for beer production. In the present study, the nonprolamin fractions including proteins with structural functions or metabolic activities were extracted from barley malts of the widely used cultivars Gangpi and Baudin in China. The metabolic proteomes (pI 4-7) were constructed and compared using two-dimensional electrophoresis (2DE) followed by matrix-assisted laser desorption/ionization-tandem time-of-flight mass spectrometry (MALDI-TOF/TOF) identification. There were 333 and 354 spots detected in the 2DE gels of Gangpi and Baudin malts, respectively, and about 90% of these spots were shared by the two malts. For all, 377 were successfully identified to 192 proteins, most of which were enzymes and enzyme inhibitors, suggesting important roles in barley malting and the mashing stage of brewing. The Baudin malt was found to contain more spots representing amylases, pathogen-related proteins, and chaperones than the Gangpi malt. In addition, enzymes involved in glycolysis and redox pathways showed significantly different profiles between the two malts, permitting a more in-depth elucidation of the relationship between differential proteins and malt qualities. PMID:23198685

  15. Comparative nutritional compositions and proteomics analysis of transgenic Xa21 rice seeds compared to conventional rice.

    PubMed

    Gayen, Dipak; Paul, Soumitra; Sarkar, Sailendra Nath; Datta, Swapan K; Datta, Karabi

    2016-07-15

    Transgenic rice expressing the Xa21 gene have enhanced resistant to most devastating bacterial blight diseases caused by Xanthomonas oryzae pv. oryzae (Xoo). However, identification of unintended modifications, owing to the genetic modification, is an important aspect of transgenic crop safety assessment. In this study, the nutritional compositions of seeds from transgenic rice plants expressing the Xa21 gene were compared against non-transgenic rice seeds. In addition, to detect any changes in protein translation levels as a result of Xa21 gene expression, rice seed proteome analyses were also performed by two-dimensional gel electrophoresis. No significant differences were found in the nutritional compositions (proximate components, amino acids, minerals, vitamins and anti-nutrients) of the transgenic and non-transgenic rice seeds. Although gel electrophoresis identified 11 proteins that were differentially expressed between the transgenic and non-transgenic seed, only one of these (with a 20-fold up-regulation in the transgenic seed) shows nutrient reservoir activity. No new toxins or allergens were detected in the transgenic seeds. PMID:26948618

  16. Analysis of Drought-Induced Proteomic and Metabolomic Changes in Barley (Hordeum vulgare L.) Leaves and Roots Unravels Some Aspects of Biochemical Mechanisms Involved in Drought Tolerance

    PubMed Central

    Chmielewska, Klaudia; Rodziewicz, Paweł; Swarcewicz, Barbara; Sawikowska, Aneta; Krajewski, Paweł; Marczak, Łukasz; Ciesiołka, Danuta; Kuczyńska, Anetta; Mikołajczak, Krzysztof; Ogrodowicz, Piotr; Krystkowiak, Karolina; Surma, Maria; Adamski, Tadeusz; Bednarek, Paweł; Stobiecki, Maciej

    2016-01-01

    In this study, proteomic and metabolomic changes in leaves and roots of two barley (Hordeum vulgare L.) genotypes, with contrasting drought tolerance, subjected to water deficit were investigated. Our two-dimensional electrophoresis (2D-PAGE) combined with matrix-assisted laser desorption time of flight mass spectrometry (MALDI-TOF and MALDI-TOF/TOF) analyses revealed 121 drought-responsive proteins in leaves and 182 in roots of both genotypes. Many of the identified drought-responsive proteins were associated with processes that are typically severely affected during water deficit, including photosynthesis and carbon metabolism. However, the highest number of identified leaf and root proteins represented general defense mechanisms. In addition, changes in the accumulation of proteins that represent processes formerly unassociated with drought response, e.g., phenylpropanoid metabolism, were also identified. Our tandem gas chromatography – time of flight mass spectrometry (GC/MS TOF) analyses revealed approximately 100 drought-affected low molecular weight compounds representing various metabolite types with amino acids being the most affected metabolite class. We compared the results from proteomic and metabolomic analyses to search for existing relationship between these two levels of molecular organization. We also uncovered organ specificity of the observed changes and revealed differences in the response to water deficit of drought susceptible and tolerant barley lines. Particularly, our results indicated that several of identified proteins and metabolites whose accumulation levels were increased with drought in the analyzed susceptible barley variety revealed elevated constitutive accumulation levels in the drought-resistant line. This may suggest that constitutive biochemical predisposition represents a better drought tolerance mechanism than inducible responses. PMID:27512399

  17. Analysis of Drought-Induced Proteomic and Metabolomic Changes in Barley (Hordeum vulgare L.) Leaves and Roots Unravels Some Aspects of Biochemical Mechanisms Involved in Drought Tolerance.

    PubMed

    Chmielewska, Klaudia; Rodziewicz, Paweł; Swarcewicz, Barbara; Sawikowska, Aneta; Krajewski, Paweł; Marczak, Łukasz; Ciesiołka, Danuta; Kuczyńska, Anetta; Mikołajczak, Krzysztof; Ogrodowicz, Piotr; Krystkowiak, Karolina; Surma, Maria; Adamski, Tadeusz; Bednarek, Paweł; Stobiecki, Maciej

    2016-01-01

    In this study, proteomic and metabolomic changes in leaves and roots of two barley (Hordeum vulgare L.) genotypes, with contrasting drought tolerance, subjected to water deficit were investigated. Our two-dimensional electrophoresis (2D-PAGE) combined with matrix-assisted laser desorption time of flight mass spectrometry (MALDI-TOF and MALDI-TOF/TOF) analyses revealed 121 drought-responsive proteins in leaves and 182 in roots of both genotypes. Many of the identified drought-responsive proteins were associated with processes that are typically severely affected during water deficit, including photosynthesis and carbon metabolism. However, the highest number of identified leaf and root proteins represented general defense mechanisms. In addition, changes in the accumulation of proteins that represent processes formerly unassociated with drought response, e.g., phenylpropanoid metabolism, were also identified. Our tandem gas chromatography - time of flight mass spectrometry (GC/MS TOF) analyses revealed approximately 100 drought-affected low molecular weight compounds representing various metabolite types with amino acids being the most affected metabolite class. We compared the results from proteomic and metabolomic analyses to search for existing relationship between these two levels of molecular organization. We also uncovered organ specificity of the observed changes and revealed differences in the response to water deficit of drought susceptible and tolerant barley lines. Particularly, our results indicated that several of identified proteins and metabolites whose accumulation levels were increased with drought in the analyzed susceptible barley variety revealed elevated constitutive accumulation levels in the drought-resistant line. This may suggest that constitutive biochemical predisposition represents a better drought tolerance mechanism than inducible responses. PMID:27512399

  18. Proteomic Analysis of Embryogenesis and the Acquisition of Seed Dormancy in Norway Maple (Acer platanoides L.)

    PubMed Central

    Staszak, Aleksandra Maria; Pawłowski, Tomasz Andrzej

    2014-01-01

    The proteome of zygotic embryos of Acer platanoides L. was analyzed via high-resolution 2D-SDS-PAGE and MS/MS in order to: (1) identify significant physiological processes associated with embryo development; and (2) identify changes in the proteome of the embryo associated with the acquisition of seed dormancy. Seventeen spots were identified as associated with morphogenesis at 10 to 13 weeks after flowering (WAF). Thirty-three spots were associated with maturation of the embryo at 14 to 22 WAF. The greatest changes in protein abundance occurred at 22 WAF, when seeds become fully mature. Overall, the stage of morphogenesis was characterized by changes in the abundance of proteins (tubulins and actin) associated with the growth and development of the embryo. Enzymes related to energy supply were especially elevated, most likely due to the energy demand associated with rapid growth and cell division. The stage of maturation is crucial to the establishment of seed dormancy and is associated with a higher abundance of proteins involved in genetic information processing, energy and carbon metabolism and cellular and antioxidant processes. Results indicated that a glycine-rich RNA-binding protein and proteasome proteins may be directly involved in dormancy acquisition control, and future studies are warranted to verify this association. PMID:24941250

  19. Proteomic and Physiological Analysis of the Response of Oat (Avena sativa) Seeds to Heat Stress under Different Moisture Conditions

    PubMed Central

    Chen, Lingling; Chen, Quanzhu; Kong, Lingqi; Xia, Fangshan; Yan, Huifang; Zhu, Yanqiao; Mao, Peisheng

    2016-01-01

    Seeds lose their viability when they are exposed to high temperature and moisture content (MC) during storage. The expression and metabolism of proteins plays a critical role in seed resistance to heat stress. However, the proteome response to heat stress in oat (Avena sativa) seeds during storage has not been revealed. To understand mechanisms of heat stress acclimation and tolerance in oat seeds, an integrated physiological and comparative proteomic analysis was performed on oat seeds with different MC during heat stress. Oat seeds with 10% and 16% MC were subjected to high temperatures (35, 45, and 50°C) for 24 and 2 days, respectively, and changes in physiological and biochemical characteristics were analyzed. The results showed that seed vigor decreased significantly with temperature increase from 35 to 50°C. Also, the proline content in 10% MC seeds decreased significantly (p < 0.05) whereas that in 16% MC seeds increased significantly (p < 0.05) during heat treatment from 35 to 50°C. There were no significant differences in malondialdehyde content in 10% MC seeds with temperature from 35 to 50°C, but a significant (p < 0.05) decline occurred in 16% MC seeds at 45°C. Proteome analysis revealed 21 significantly different proteins, including 19 down-regulated and two up-regulated proteins. The down-regulated proteins, notably six heat shock proteins and two ATP synthases, have important roles in the mobilization of carbohydrates and energy, and in the balance between synthesis and degradation of other proteins during seed deterioration. The up-regulation of argininosuccinate synthase participated in proline biosynthesis at 16% MC, which is important for maintaining reactive oxygen species homeostasis for the resistance of heat stress. In summary, heat-responsive protein species and mitochondrial respiratory metabolism were sensitive to high temperature and MC treatment. These studies provide a new insight into acclimation and tolerance to heat stress in

  20. Proteomic and Physiological Analysis of the Response of Oat (Avena sativa) Seeds to Heat Stress under Different Moisture Conditions.

    PubMed

    Chen, Lingling; Chen, Quanzhu; Kong, Lingqi; Xia, Fangshan; Yan, Huifang; Zhu, Yanqiao; Mao, Peisheng

    2016-01-01

    Seeds lose their viability when they are exposed to high temperature and moisture content (MC) during storage. The expression and metabolism of proteins plays a critical role in seed resistance to heat stress. However, the proteome response to heat stress in oat (Avena sativa) seeds during storage has not been revealed. To understand mechanisms of heat stress acclimation and tolerance in oat seeds, an integrated physiological and comparative proteomic analysis was performed on oat seeds with different MC during heat stress. Oat seeds with 10% and 16% MC were subjected to high temperatures (35, 45, and 50°C) for 24 and 2 days, respectively, and changes in physiological and biochemical characteristics were analyzed. The results showed that seed vigor decreased significantly with temperature increase from 35 to 50°C. Also, the proline content in 10% MC seeds decreased significantly (p < 0.05) whereas that in 16% MC seeds increased significantly (p < 0.05) during heat treatment from 35 to 50°C. There were no significant differences in malondialdehyde content in 10% MC seeds with temperature from 35 to 50°C, but a significant (p < 0.05) decline occurred in 16% MC seeds at 45°C. Proteome analysis revealed 21 significantly different proteins, including 19 down-regulated and two up-regulated proteins. The down-regulated proteins, notably six heat shock proteins and two ATP synthases, have important roles in the mobilization of carbohydrates and energy, and in the balance between synthesis and degradation of other proteins during seed deterioration. The up-regulation of argininosuccinate synthase participated in proline biosynthesis at 16% MC, which is important for maintaining reactive oxygen species homeostasis for the resistance of heat stress. In summary, heat-responsive protein species and mitochondrial respiratory metabolism were sensitive to high temperature and MC treatment. These studies provide a new insight into acclimation and tolerance to heat stress in

  1. A systematic proteomic analysis of NaCl-stressed germinating maize seeds.

    PubMed

    Meng, Ling-Bo; Chen, Yi-Bo; Lu, Tian-Cong; Wang, Yue-Feng; Qian, Chun-Rong; Yu, Yang; Ge, Xuan-Liang; Li, Xiao-Hui; Wang, Bai-Chen

    2014-05-01

    Salt (NaCl) is a common physiological stressor of plants. To better understand how germinating seeds respond to salt stress, we examined the changes that occurred in the proteome of maize seeds during NaCl-treated germination. Phenotypically, salt concentrations less than 0.2 M appear to delay germination, while higher concentrations disrupt development completely, leading to seed death. The identities of 96 proteins with expression levels altered by NaCl-incubation were established using 2-DE-MALDI-TOF-MS and 2-DE-MALDI-TOF-MS/MS. Of these 96 proteins, 79 were altered greater than twofold when incubated with a 0.2 M salt solution, while 51 were altered when incubated with a 0.1 M salt solution. According to their functional annotations in the Swiss-Prot protein-sequence databases, these proteins are mainly involved in seed storage, energy metabolism, stress response, and protein metabolism. Notably, the expression of proteins that respond to abscisic acid signals increased in response to salt stress. The results of this study provide important clues as to how NaCl stresses the physiology of germinating maize seeds. PMID:24700167

  2. Environmental Effects of Nanoceria on Seed Production of Common Bean (Phaseolus vulgaris): A Proteomic Analysis.

    PubMed

    Majumdar, Sanghamitra; Almeida, Igor C; Arigi, Emma A; Choi, Hyungwon; VerBerkmoes, Nathan C; Trujillo-Reyes, Jesica; Flores-Margez, Juan P; White, Jason C; Peralta-Videa, Jose R; Gardea-Torresdey, Jorge L

    2015-11-17

    The rapidly growing literature on the response of edible plants to nanoceria has provided evidence of its uptake and bioaccumulation, which delineates a possible route of entry into the food chain. However, little is known about how the residing organic matter in soil may affect the bioavailability and resulting impacts of nanoceria on plants. Here, we examined the effect of nanoceria exposure (62.5-500 mg/kg) on kidney bean (Phaseolus vulgaris) productivity and seed quality as a function of soil organic matter content. Cerium accumulation in the seeds produced from plants in organic matter enriched soil showed a dose-dependent increase, unlike in low organic matter soil treatments. Seeds obtained upon nanoceria exposure in soils with higher organic matter were more susceptible to changes in nutrient quality. A quantitative proteomic analysis of the seeds produced upon nanoceria exposure provided evidence for upregulation of stress-related proteins at 62.5 and 125 mg/kg nanoceria treatments. Although the plants did not exhibit overt toxicity, the major seed proteins primarily associated with nutrient storage (phaseolin) and carbohydrate metabolism (lectins) were significantly down-regulated in a dose dependent manner upon nanoceria exposure. This study thus suggests that nanoceria exposures may negatively affect the nutritional quality of kidney beans at the cellular and molecular level. More confirmatory studies with nanoceria along different species using alternative and orthogonal "omic" tools are currently under active investigation, which will enable the identification of biomarkers of exposure and susceptibility. PMID:26488752

  3. A proteomic analysis of rice seed germination as affected by high temperature and ABA treatment.

    PubMed

    Liu, Shu-Jun; Xu, Heng-Heng; Wang, Wei-Qing; Li, Ni; Wang, Wei-Ping; Møller, Ian Max; Song, Song-Quan

    2015-05-01

    Seed germination is a critical phase in the plant life cycle, but the specific events associated with seed germination are still not fully understood. In this study, we used two-dimensional gel electrophoresis followed by mass spectrometry to investigate the changes in the proteome during imbibition of Oryza sativa seeds at optimal temperature with or without abscisic acid (ABA) and high temperature (germination thermoinhibition) to further identify and quantify key proteins required for seed germination. A total of 121 protein spots showed a significant change in abundance (1.5-fold increase/decrease) during germination under all conditions. Among these proteins, we found seven proteins specifically associated with seed germination including glycosyl hydrolases family 38 protein, granule-bound starch synthase 1, Os03g0842900 (putative steroleosin-B), N-carbamoylputrescine amidase, spermidine synthase 1, tubulin α-1 chain and glutelin type-A; and a total of 20 imbibition response proteins involved in energy metabolism, cell growth, cell defense and storage proteins. High temperature inhibited seed germination by decreasing the abundance of proteins involved in methionine metabolism, amino acid biosynthesis, energy metabolism, reserve degradation, protein folding and stress responses. ABA treatment inhibited germination and decreased the abundance of proteins associated with methionine metabolism, energy production and cell division. Our results show that changes in many biological processes including energy metabolism, protein synthesis and cell defense and rescue occurred as a result of all treatments, while enzymes involved in methionine metabolism and weakening of cell wall specifically accumulated when the seeds germinated at the optimal temperature. PMID:25270993

  4. Genome-wide association mapping and biochemical markers reveal that seed ageing and longevity are intricately affected by genetic background and developmental and environmental conditions in barley.

    PubMed

    Nagel, Manuela; Kranner, Ilse; Neumann, Kerstin; Rolletschek, Hardy; Seal, Charlotte E; Colville, Louise; Fernández-Marín, Beatriz; Börner, Andreas

    2015-06-01

    Globally, over 7.4 million accessions of crop seeds are stored in gene banks, and conservation of genotypic variation is pivotal for breeding. We combined genetic and biochemical approaches to obtain a broad overview of factors that influence seed storability and ageing in barley (Hordeum vulgare). Seeds from a germplasm collection of 175 genotypes from four continents grown in field plots with different nutrient supply were subjected to two artificial ageing regimes. Genome-wide association mapping revealed 107 marker trait associations, and hence, genotypic effects on seed ageing. Abiotic and biotic stresses were found to affect seed longevity. To address aspects of abiotic, including oxidative, stress, two major antioxidant groups were analysed. No correlation was found between seed deterioration and the lipid-soluble tocochromanols, nor with oil, starch and protein contents. Conversely, the water-soluble glutathione and related thiols were converted to disulphides, indicating a strong shift towards more oxidizing intracellular conditions, in seeds subjected to long-term dry storage at two temperatures or to two artificial ageing treatments. The data suggest that intracellular pH and (bio)chemical processes leading to seed deterioration were influenced by the type of ageing or storage. Moreover, seed response to ageing or storage treatment appears to be significantly influenced by both maternal environment and genetic background. PMID:25328120

  5. The Red Queen and the seed bank: pathogen resistance of ex situ and in situ conserved barley

    PubMed Central

    Jensen, Helen R; Dreiseitl, Antonín; Sadiki, Mohammed; Schoen, Daniel J

    2012-01-01

    Plant geneticists have proposed that the dynamic conservation of crop plants in farm environments (in situ conservation) is complementary to static conservation in seed banks (ex situ conservation) because it may help to ensure adaptation to changing conditions. Here, we test whether collections of a traditional variety of Moroccan barley (Hordeum vulgare ssp. vulgare) conserved ex situ showed differences in qualitative and quantitative resistance to the endemic fungal pathogen, Blumeria graminis f.sp. hordei, compared to collections that were continuously cultivated in situ. In detached-leaf assays for qualitative resistance, there were some significant differences between in situ and ex situ conserved collections from the same localities. Some ex situ conserved collections showed lower resistance levels, while others showed higher resistance levels than their in situ conserved counterparts. In field trials for quantitative resistance, similar results were observed, with the highest resistance observed in situ. Overall, this study identifies some cases where the Red Queen appears to drive the evolution of increased resistance in situ. However, in situ conservation does not always result in improved adaptation to pathogen virulence, suggesting a more complex evolutionary scenario, consistent with several published examples of plant–pathogen co-evolution in wild systems. PMID:25568056

  6. Comparative proteomic analysis and IgE binding properties of peanut seed and testa (skin).

    PubMed

    White, Brittany L; Gökce, Emine; Nepomuceno, Angelito I; Muddiman, David C; Sanders, Timothy H; Davis, Jack P

    2013-04-24

    To investigate the protein composition and potential allergenicity of peanut testae or skins, proteome analysis was conducted using nanoLC-MS/MS sequencing. Initial amino acid analysis suggested differences in protein compositions between the blanched seed (skins removed) and skin. Phenolic compounds hindered analysis of proteins in skins when the conventional extraction method was used; therefore, phenol extraction of proteins was necessary. A total of 123 proteins were identified in blanched seed and skins, and 83 of the proteins were common between the two structures. The skins contained all of the known peanut allergens in addition to 38 proteins not identified in the seed. Multiple defense proteins with antifungal activity were identified in the skins. Western blotting using sera from peanut-allergic patients revealed that proteins extracted from both the blanched seed and skin bound significant levels of IgE. However, when phenolic compounds were present in the skin protein extract, no IgE binding was observed. These findings indicate that peanut skins contain potentially allergenic proteins; however, the presence of phenolic compounds may attenuate this effect. PMID:23534881

  7. Proteome analysis of the inner integument from developing Jatropha curcas L. seeds.

    PubMed

    Soares, Emanoella L; Shah, Mohibullah; Soares, Arlete A; Costa, José H; Carvalho, Paulo; Domont, Gilberto B; Nogueira, Fábio C S; Campos, Francisco A P

    2014-08-01

    In this study, we performed a systematic proteomic analysis of the inner integument from developing seeds of Jatropha curcas and further explored the protein machinery responsible for generating the carbon and nitrogen sources to feed the growing embryo and endosperm. The inner integument of developing seeds was dissected into two sections called distal and proximal, and proteins were extracted from these sections and from the whole integument and analyzed using an EASY-nanoLC system coupled to an ESI-LTQ-Orbitrap Velos mass spectrometer. We identified 1526, 1192, and 1062 proteins from the proximal, distal, and whole inner integuments, respectively. The identifications include those of peptidases and other hydrolytic enzymes that play a key role in developmental programmed cell death and proteins associated with the cell-wall architecture and modification. Because many of these proteins are differentially expressed within the integument cell layers, these findings suggest that the cells mobilize an array of hydrolases to produce carbon and nitrogen sources from proteins, carbohydrates, and lipids available within the cells. Not least, the identification of several classes of seed storage proteins in the inner integument provides additional evidence of the role of the seed coat as a transient source of reserves for the growing embryo and endosperm. PMID:25010673

  8. Dynamic proteomics emphasizes the importance of selective mRNA translation and protein turnover during Arabidopsis seed germination.

    PubMed

    Galland, Marc; Huguet, Romain; Arc, Erwann; Cueff, Gwendal; Job, Dominique; Rajjou, Loïc

    2014-01-01

    During seed germination, the transition from a quiescent metabolic state in a dry mature seed to a proliferative metabolic state in a vigorous seedling is crucial for plant propagation as well as for optimizing crop yield. This work provides a detailed description of the dynamics of protein synthesis during the time course of germination, demonstrating that mRNA translation is both sequential and selective during this process. The complete inhibition of the germination process in the presence of the translation inhibitor cycloheximide established that mRNA translation is critical for Arabidopsis seed germination. However, the dynamics of protein turnover and the selectivity of protein synthesis (mRNA translation) during Arabidopsis seed germination have not been addressed yet. Based on our detailed knowledge of the Arabidopsis seed proteome, we have deepened our understanding of seed mRNA translation during germination by combining two-dimensional gel-based proteomics with dynamic radiolabeled proteomics using a radiolabeled amino acid precursor, namely [(35)S]-methionine, in order to highlight de novo protein synthesis, stability, and turnover. Our data confirm that during early imbibition, the Arabidopsis translatome keeps reflecting an embryonic maturation program until a certain developmental checkpoint. Furthermore, by dividing the seed germination time lapse into discrete time windows, we highlight precise and specific patterns of protein synthesis. These data refine and deepen our knowledge of the three classical phases of seed germination based on seed water uptake during imbibition and reveal that selective mRNA translation is a key feature of seed germination. Beyond the quantitative control of translational activity, both the selectivity of mRNA translation and protein turnover appear as specific regulatory systems, critical for timing the molecular events leading to successful germination and seedling establishment. PMID:24198433

  9. Dynamic Proteomics Emphasizes the Importance of Selective mRNA Translation and Protein Turnover during Arabidopsis Seed Germination*

    PubMed Central

    Galland, Marc; Huguet, Romain; Arc, Erwann; Cueff, Gwendal; Job, Dominique; Rajjou, Loïc

    2014-01-01

    During seed germination, the transition from a quiescent metabolic state in a dry mature seed to a proliferative metabolic state in a vigorous seedling is crucial for plant propagation as well as for optimizing crop yield. This work provides a detailed description of the dynamics of protein synthesis during the time course of germination, demonstrating that mRNA translation is both sequential and selective during this process. The complete inhibition of the germination process in the presence of the translation inhibitor cycloheximide established that mRNA translation is critical for Arabidopsis seed germination. However, the dynamics of protein turnover and the selectivity of protein synthesis (mRNA translation) during Arabidopsis seed germination have not been addressed yet. Based on our detailed knowledge of the Arabidopsis seed proteome, we have deepened our understanding of seed mRNA translation during germination by combining two-dimensional gel-based proteomics with dynamic radiolabeled proteomics using a radiolabeled amino acid precursor, namely [35S]-methionine, in order to highlight de novo protein synthesis, stability, and turnover. Our data confirm that during early imbibition, the Arabidopsis translatome keeps reflecting an embryonic maturation program until a certain developmental checkpoint. Furthermore, by dividing the seed germination time lapse into discrete time windows, we highlight precise and specific patterns of protein synthesis. These data refine and deepen our knowledge of the three classical phases of seed germination based on seed water uptake during imbibition and reveal that selective mRNA translation is a key feature of seed germination. Beyond the quantitative control of translational activity, both the selectivity of mRNA translation and protein turnover appear as specific regulatory systems, critical for timing the molecular events leading to successful germination and seedling establishment. PMID:24198433

  10. Proteome analysis of plastids from developing seeds of Jatropha curcas L.

    PubMed

    Pinheiro, Camila B; Shah, Mohibullah; Soares, Emanoella L; Nogueira, Fábio C S; Carvalho, Paulo C; Junqueira, Magno; Araújo, Gabriel D T; Soares, Arlete A; Domont, Gilberto B; Campos, Francisco A P

    2013-11-01

    In this study, we performed a proteomic analysis of plastids isolated from the endosperm of developing Jatropha curcas seeds that were in the initial stage of deposition of protein and lipid reserves. Proteins extracted from the plastids were digested with trypsin, and the peptides were applied to an EASY-nano LC system coupled inline to an ESI-LTQ-Orbitrap Velos mass spectrometer, and this led to the identification of 1103 proteins representing 804 protein groups, of which 923 proteins were considered as true identifications, and this considerably expands the repertoire of J. curcas proteins identified so far. Of the identified proteins, only five are encoded in the plastid genome, and none of them are involved in photosynthesis, evidentiating the nonphotosynthetic nature of the isolated plastids. Homologues for 824 out of 923 identified proteins were present in PPDB, SUBA, or PlProt databases while homologues for 13 proteins were not found in any of the three plastid proteins databases but were marked as plastidial by at least one of the three prediction programs used. Functional classification showed that proteins belonging to amino acids metabolism comprise the main functional class, followed by carbohydrate, energy, and lipid metabolisms. The small and large subunits of Rubisco were identified, and their presence in the plastids is considered to be an adaptive feature counterbalancing for the loss of one-third of the carbon as CO2 as a result of the conversion of carbohydrate to oil through glycolysis. While several enzymes involved in the biosynthesis of several precursors of diterpenoids were identified, we were unable to identify any terpene synthase/cyclase, which suggests that the plastids isolated from the endosperm of developing seeds do not synthesize phorbol esters. In conclusion, our study provides insights into the major biosynthetic pathways and certain unique features of the plastids from the endosperm of developing seeds at the whole proteome

  11. 2015 nationwide survey revealed Barley stripe mosaic virus in Korean barley fields

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A seed-transmitted virus has consistently caused significant economic damage to barley crops in Korea in recent years, and may be increasing because many farmers save seed for replanting. Because some barley seed is imported, there is the potential for introduction of new seed-transmitted viruses, c...

  12. Deciphering Mineral Homeostasis in Barley Seed Transfer Cells at Transcriptional Level

    PubMed Central

    Borg, Søren

    2015-01-01

    In addition to the micronutrient inadequacy of staple crops for optimal human nutrition, a global downtrend in crop-quality has emerged from intensive breeding for yield. This trend will be aggravated by elevated levels of the greenhouse gas carbon dioxide. Therefore, crop biofortification is inevitable to ensure a sustainable supply of minerals to the large part of human population who is dietary dependent on staple crops. This requires a thorough understanding of plant-mineral interactions due to the complexity of mineral homeostasis. Employing RNA sequencing, we here communicate transfer cell specific effects of excess iron and zinc during grain filling in our model crop plant barley. Responding to alterations in mineral contents, we found a long range of different genes and transcripts. Among them, it is worth to highlight the auxin and ethylene signaling factors Arfs, Abcbs, Cand1, Hps4, Hac1, Ecr1, and Ctr1, diurnal fluctuation components Sdg2, Imb1, Lip1, and PhyC, retroelements, sulfur homeostasis components Amp1, Hmt3, Eil3, and Vip1, mineral trafficking components Med16, Cnnm4, Aha2, Clpc1, and Pcbps, and vacuole organization factors Ymr155W, RabG3F, Vps4, and Cbl3. Our analysis introduces new interactors and signifies a broad spectrum of regulatory levels from chromatin remodeling to intracellular protein sorting mechanisms active in the plant mineral homeostasis. The results highlight the importance of storage proteins in metal ion toxicity-resistance and chelation. Interestingly, the protein sorting and recycling factors Exoc7, Cdc1, Sec23A, and Rab11A contributed to the response as well as the polar distributors of metal-transporters ensuring the directional flow of minerals. Alternative isoform switching was found important for plant adaptation and occurred among transcripts coding for identical proteins as well as transcripts coding for protein isoforms. We also identified differences in the alternative-isoform preference between the treatments

  13. Radioactive Chernobyl environment has produced high-oil flax seeds that show proteome alterations related to carbon metabolism during seed development.

    PubMed

    Klubicová, Katarína; Danchenko, Maksym; Skultety, Ludovit; Berezhna, Valentyna V; Rashydov, Namik M; Hajduch, Martin

    2013-11-01

    Starting in 2007, we have grown soybean (Glycine max [L.] Merr. variety Soniachna) and flax (Linum usitatissimum, L. variety Kyivskyi) in the radio-contaminated Chernobyl area and analyzed the seed proteomes. In the second-generation flax seeds, we detected a 12% increase in oil content. To characterize the bases for this increase, seed development has been studied. Flax seeds were harvested in biological triplicate at 2, 4, and 6 weeks after flowering and at maturity from plants grown in nonradioactive and radio-contaminated plots in the Chernobyl area for two generations. Quantitative proteomic analyses based on 2-D gel electrophoresis (2-DE) allowed us to establish developmental profiles for 199 2-DE spots in both plots, out of which 79 were reliably identified by tandem mass spectrometry. The data suggest a statistically significant increased abundance of proteins associated with pyruvate biosynthesis via cytoplasmic glycolysis, L-malate decarboxylation, isocitrate dehydrogenation, and ethanol oxidation to acetaldehyde in early stages of seed development. This was followed by statistically significant increased abundance of ketoacyl-[acylcarrier protein] synthase I related to condensation of malonyl-ACP with elongating fatty acid chains. On the basis of these and previous data, we propose a preliminary model for plant adaptation to growth in a radio-contaminated environment. One aspect of the model suggests that changes in carbon assimilation and fatty acid biosynthesis are an integral part of plant adaptation. PMID:24111740

  14. Characterization, crystallization and preliminary X-ray crystallographic analysis of the complex between barley alpha-amylase and the bifunctional alpha-amylase/subtilisin inhibitor from barley seeds.

    PubMed

    Vallée, F; Kadziola, A; Bourne, Y; Abe, J; Svensson, B; Haser, R

    1994-02-11

    The complex between a member of the barley malt alpha-amylase isozyme 2 family (AMY2-2), and the endogenous bifunctional alpha-amylase/subtilisin inhibitor, BASI, has been crystallized by the hanging drop vapour diffusion technique at a AMY2-2: BASI molar ratio of 1:1. Crystals have been grown within 4 days from solutions containing polyethylene glycol and calcium chloride. Analysis of single crystals by gel electrophoresis showed the presence of both proteins in the crystal lattice. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit cell dimensions a = 74.5 A, b = 96.9 A, c = 171.3 A and they diffract to 2.0 A resolution. The presence of two molecules of the 1:1 complex in the asymmetric unit gives a solvent content of 45% by volume. The 1:1 stoichiometry of the complex was confirmed by the molecular replacement method, using as a search model the recently determined three-dimensional structure of the barley alpha-amylase. PMID:8107117

  15. Proteomic and Epigenetic Analysis of Rice after Seed Spaceflight and Ground-Base Ion Radiations

    NASA Astrophysics Data System (ADS)

    Wang, Wei; Sun, Yeqing; Peng, Yuming; Zhao, Qian; Wen, Bin; Yang, Jun

    Highly ionizing radiation (HZE) in space is considered as main factor causing biological effects to plant seeds. In previous work, we compared the proteomic profiles of rice plants growing after seed spaceflights to ground controls by two-dimensional difference gel electrophoresis (2-D DIGE) with mass spectrometry and found that the protein expression profiles were changed and differentially expressed proteins participated in most of the biological processes of rice. To further evaluate the dosage effects of space radiation and compare between low- and high-dose ion effects, we carried out three independent ground-base ionizing radiation experiments with different cumulative doses (low-dose range: 2~1000mGy, high-dose range: 2000~20000mGy) to rice seeds and performed proteomic analysis of seedlings. We found that protein expression profiles showed obvious boundaries between low- and high-dose radiation groups. Rates of differentially expressed proteins presented a dose-dependent effect, it reached the highest value at 2000mGy dosage point in all three radiation experiments coincidently; while proteins responded to low-dose radiations preferred to change their expressions at the minimum dosage (2mGy). Proteins participating in rice biological processes also responded differently between low- and high-dose radiations: proteins involved in energy metabolism and photosynthesis tended to be regulated after low-dose radiations while stress responding, protein folding and cell redox homeostasis related proteins preferred to change their expressions after high-dose radiations. By comparing the proteomic profiles between ground-base radiations and spaceflights, it was worth noting that ground-base low-dose ion radiation effects shared similar biological effects as space environment. In addition, we discovered that protein nucleoside diphosphate kinase 1 (NDPK1) showed obvious increased regulation after spaceflights and ion radiations. NDPK1 catalyzes nucleotide metabolism

  16. Leaf proteomic analysis of three rice heritable mutants after seed space flight

    NASA Astrophysics Data System (ADS)

    Wang, W.; Gu, D. P.; Zheng, Q.; Sun, Y. Q.

    2008-09-01

    To explore the proteomic changes of heritable variant rice plants induced by space environment, three mutants were selected after seed space flight by comparing the phenotypes with their on-ground controls. R955 grew more tillers and became dwarf, 971-5 acquired higher grain yield and better stress resistance, 974-5 matured earlier. Leaf proteins were extracted during the tiller development and analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). More than 300 proteins were detected as reproducible Coomassie Brilliant Blue stained spots with p I values from around 4.0 to 7.0. Five proteins that changed significantly over the controls were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). The main functions of these proteins were photosynthesis, stress defense and metabolism including RuBisCO activase, glycine rich RNA binding protein, peroxidase, triosephosphate isomerase and phosphoenolpyruvate carboxylase, which might be probably associated with the altered phenotypes. Quantitative analyses were also applied: less total protein spots and more down-regulated protein spots were detected in the mutants, indicating there might be a major loss of protein in heritable variant rice plants after seed space flight. These results may provide new insights to understand the biological effects of space environment to rice.

  17. Two traditional maize inbred lines of contrasting technological abilities are discriminated by the seed flour proteome.

    PubMed

    Pinheiro, Carla; Sergeant, Kjell; Machado, Cátia M; Renaut, Jenny; Ricardo, Cândido P

    2013-07-01

    The seed proteome of two traditional maize inbred lines (pb269 and pb369) contrasting in grain hardness and in preferable use for bread-making was evaluated. The pb269 seeds, of flint type (i.e., hard endosperm), are preferably used by manufacturers, while pb369 (dent, soft endosperm) is rejected. The hypothesis that the content and relative amounts of specific proteins in the maize flour are relevant for such discrimination of the inbred lines was tested. The flour proteins were sequentially extracted following the Osborne fractionation (selective solubilization), and the four Osborne fractions were submitted to two-dimensional electrophoresis (2DE). The total amount of protein extracted from the seeds was not significantly different, but pb369 flour exhibited significantly higher proportions of salt-extracted proteins (globulins) and ethanol-extracted proteins (alcohol-soluble prolamins). The proteome analysis allowed discrimination between the two inbred lines, with pb269 demonstrating higher heterogeneity than pb369. From the 967 spots (358 common to both lines, 208 specific to pb269, and 401 specific to pb369), 588 were submitted to mass spectrometry (MS). Through the combined use of trypsin and chymotrypsin it was possible to identify proteins in 436 spots. The functional categorization in combination with multivariate analysis highlighted the most discriminant biological processes (carbohydrate metabolic process, response to stress, chitin catabolic process, oxidation-reduction process) and molecular function (nutrient reservoir activity). The inbred lines exhibited quantitative and qualitative differences in these categories. Differences were also revealed in the amounts, proportions, and distribution of several groups of storage proteins, which can have an impact on the organization of the protein body and endosperm hardness. For some proteins (granule-bound starch synthase-1, cyclophilin, zeamatin), a change in the protein solubility rather than in the

  18. Proteomic analysis of lettuce seed germination and thermoinhibition by sampling of individual seeds at germination and removal of storage proteins by polyethylene glycol fractionation.

    PubMed

    Wang, Wei-Qing; Song, Bin-Yan; Deng, Zhi-Jun; Wang, Yue; Liu, Shu-Jun; Møller, Ian Max; Song, Song-Quan

    2015-04-01

    Germination and thermoinhibition in lettuce (Lactuca sativa 'Jianyexianfeng No. 1') seeds were investigated by a proteomic comparison among dry seeds, germinated seeds at 15°C, at 15°C after imbibition at 25°C for 48 h, or at 25°C in KNO3 (all sampled individually at germination), and ungerminated seeds at 25°C, a thermoinhibitory temperature. Before two-dimensional gel electrophoresis analysis, storage proteins (greater than 50% of total extractable protein) were removed by polyethylene glycol precipitation, which significantly improved the detection of less abundant proteins on two-dimensional gels. A total of 108 protein spots were identified to change more than 2-fold (P<0.05) in abundance in at least one germination treatment. Nineteen proteins increasing and one protein decreasing in abundance during germination had higher abundance in germinated 15°C, 15°C after imbibition at 25°C for 48 h, and 25°C in KNO3 seeds than in ungerminated 25°C seeds. Gene expression of 12 of those proteins correlated well with the protein accumulation. Methionine metabolism, ethylene production, lipid mobilization, cell elongation, and detoxification of aldehydes were revealed to be potentially related to lettuce seed germination and thermoinhibition. Accumulation of three proteins and expression of five genes participating in the mevalonate (MVA) pathway of isoprenoid biosynthesis correlated positively with seed germinability. Inhibition of this pathway by lovastatin delayed seed germination and increased the sensitivity of germination to abscisic acid. MVA pathway-derived products, cytokinins, partially reversed the lovastatin inhibition of germination and released seed thermoinhibition at 25°C. We conclude that the MVA pathway for isoprenoid biosynthesis is involved in lettuce seed germination and thermoinhibition. PMID:25736209

  19. Proteomic Analysis of Lettuce Seed Germination and Thermoinhibition by Sampling of Individual Seeds at Germination and Removal of Storage Proteins by Polyethylene Glycol Fractionation1

    PubMed Central

    Song, Bin-Yan; Deng, Zhi-Jun; Wang, Yue; Liu, Shu-Jun; Møller, Ian Max; Song, Song-Quan

    2015-01-01

    Germination and thermoinhibition in lettuce (Lactuca sativa ‘Jianyexianfeng No. 1’) seeds were investigated by a proteomic comparison among dry seeds, germinated seeds at 15°C, at 15°C after imbibition at 25°C for 48 h, or at 25°C in KNO3 (all sampled individually at germination), and ungerminated seeds at 25°C, a thermoinhibitory temperature. Before two-dimensional gel electrophoresis analysis, storage proteins (greater than 50% of total extractable protein) were removed by polyethylene glycol precipitation, which significantly improved the detection of less abundant proteins on two-dimensional gels. A total of 108 protein spots were identified to change more than 2-fold (P < 0.05) in abundance in at least one germination treatment. Nineteen proteins increasing and one protein decreasing in abundance during germination had higher abundance in germinated 15°C, 15°C after imbibition at 25°C for 48 h, and 25°C in KNO3 seeds than in ungerminated 25°C seeds. Gene expression of 12 of those proteins correlated well with the protein accumulation. Methionine metabolism, ethylene production, lipid mobilization, cell elongation, and detoxification of aldehydes were revealed to be potentially related to lettuce seed germination and thermoinhibition. Accumulation of three proteins and expression of five genes participating in the mevalonate (MVA) pathway of isoprenoid biosynthesis correlated positively with seed germinability. Inhibition of this pathway by lovastatin delayed seed germination and increased the sensitivity of germination to abscisic acid. MVA pathway-derived products, cytokinins, partially reversed the lovastatin inhibition of germination and released seed thermoinhibition at 25°C. We conclude that the MVA pathway for isoprenoid biosynthesis is involved in lettuce seed germination and thermoinhibition. PMID:25736209

  20. NanoUPLC-MSE proteomic data assessment of soybean seeds using the Uniprot database

    PubMed Central

    2012-01-01

    detection range of typical NanoUPLC-MSE experiments, i.e., 500 to 5000 Da, 64 proteins will not be identified. Conclusions NanoUPLC-MSE experiments provide good protein coverage within a peptide error of 5 ppm and a wide MW detection range from 500 to 5000 Da. A second digestion enzyme should be used depending on the tissue or proteins to be analyzed. In the case of seed tissue, trypsin protein digestion results offer good databank coverage. The Uniprot database has many duplicate entries that may result in false protein homolog associations when using NanoUPLC-MSE analysis. The proteomic profile of the EMBRAPA BR-16 seed lacks certain described proteins relative to the profiles of transgenic soybeans reported in other works. PMID:23126227

  1. Proteomics analysis reveals distinct involvement of embryo and endosperm proteins during seed germination in dormant and non-dormant rice seeds.

    PubMed

    Xu, Heng-Heng; Liu, Shu-Jun; Song, Shun-Hua; Wang, Rui-Xia; Wang, Wei-Qing; Song, Song-Quan

    2016-06-01

    Seed germination is a complex trait which is influenced by many genetic, endogenous and environmental factors, but the key event(s) associated with seed germination are still poorly understood. In present study, the non-dormant cultivated rice Yannong S and the dormant Dongxiang wild rice seeds were used as experimental materials, we comparatively investigated the water uptake, germination time course, and the differential proteome of the effect of embryo and endosperm on germination of these two types of seeds. A total of 231 and 180 protein spots in embryo and endosperm, respectively, showed a significant change in abundance during germination. We observed that the important proteins associated with seed germination included those involved in metabolism, energy production, protein synthesis and destination, storage protein, cell growth and division, signal transduction, cell defense and rescue. The contribution of embryo and endosperm to seed germination is different. In embryo, the proteins involved in amino acid activation, sucrose cleavage, glycolysis, fermentation and protein synthesis increased; in endosperm, the proteins involved in sucrose cleavage and glycolysis decreased, and those with ATP and CoQ synthesis and proteolysis increased. Our results provide some new knowledge to understand further the mechanism of seed germination. PMID:27035683

  2. Proteome Analysis of Dormancy-Released Seeds of Fraxinus mandshurica Rupr. in Response to Re-Dehydration under Different Conditions

    PubMed Central

    Zhang, Peng; Liu, Di; Shen, Hailong; Li, Yuhua; Nie, Yuzhe

    2015-01-01

    Desiccation tolerance is the ability of orthodox seeds to achieve equilibrium with atmospheric relative humidity and to survive in this state. Understanding how orthodox seeds respond to dehydration is important for improving quality and long-term storage of seeds under low temperature and drought stress conditions. Long-term storage of seeds is an artificial situation, because in most natural situations a seed that has been shed may not remain in a desiccated state for very long, and if dormant it may undergo repeated cycles of hydration. Different types of seeds are differentially sensitive to desiccation and this directly affects long-term storage. For these reasons, many researchers are investigating loss of desiccation tolerance during orthodox seed development to understand how it is acquired. In this study, the orthodox seed proteome response of Fraxinus mandshurica Rupr. to dehydration (to a relative water content of 10%, which mimics seed dehydration) was investigated under four different conditions viz. 20 °C; 20 °C with silica gel; 1 °C; and 1 °C after pretreatment with Ca2+. Proteins from seeds dehydrated under different conditions were extracted and separated by two-dimensional difference gel electrophoresis (2D-DIGE). A total of 2919 protein spots were detected, and high-resolution 2D-DIGE indicated there were 27 differentially expressed. Seven of these were identified using MALDI TOF/TOF mass spectrometry. Inferences from bioinformatics annotations of these proteins established the possible involvement of detoxifying enzymes, transport proteins, and nucleotide metabolism enzymes in response to dehydration. Of the seven differentially abundant proteins, the amounts of six were down-regulated and one was up-regulated. Also, a putative acyl-coenzyme A oxidase of the glyoxylate cycle increased in abundance. In particular, the presence of kinesin-1, a protein important for regulation and cargo interaction, was up-regulated in seeds exposed to low

  3. Back to Osborne. Sequential protein extraction and LC-MS analysis for the characterization of the Holm oak seed proteome.

    PubMed

    Romero-Rodríguez, M Cristina; Maldonado-Alconada, Ana M; Valledor, Luis; Jorrin-Novo, Jesus V

    2014-01-01

    It is impossible to capture in just one experiment all or most of the total set of protein species that constitute the cell's proteome. Thus, according to our results, and even considering that they depend on the experimental system carried out (plant, yeast, fungi, or bacteria), the best protein extraction protocol yielded less than 20 % of the total amount of proteins, as determined by the Kjeldahl method. For this reason, protein cataloguing and the whole proteome characterization require the use of firstly, fractionation techniques at the cellular, subcellular, protein, or peptide level, and secondly, the use of complementary approaches.Within our current research on Holm oak (Quercus ilex subsp. ballota), we aim to characterize its seed proteome. For that we have optimized an experimental workflow in which the Osborne sequential protein extraction (Osborne, Science 28:417-427, 1908) is combined with downstream electrophoretic protein separation or shotgun MS analysis. In general, it can be used to study any plant seed, as well as to investigate on seed maturation and germination, genotype characterization, allergens identification, food traceability, and substantial equivalence, among others. PMID:24136536

  4. Differential RNA Expression of ßm1 during Late Seed Development in Cultivated and Wild Barleys Carrying Different ßmy1 Intron III Alleles and the Association with Beta-Amylase Activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four genotypes carrying different beta-amylase 1 (Bmy1) intron III alleles (Bmy1.a, Bmy1.b, Bmy1.c, and Bmy1.d) were analyzed for differences in Bmy1 mRNA accumulation, beta-amylase activity and protein, and total protein during late seed development. Wild barleys (Hordeum vulgare ssp. spontaneum) ...

  5. A Developmental Switch of Gene Expression in the Barley Seed Mediated by HvVP1 (Viviparous-1) and HvGAMYB Interactions.

    PubMed

    Abraham, Zamira; Iglesias-Fernández, Raquel; Martínez, Manuel; Rubio-Somoza, Ignacio; Díaz, Isabel; Carbonero, Pilar; Vicente-Carbajosa, Jesús

    2016-04-01

    The accumulation of storage compounds in the starchy endosperm of developing cereal seeds is highly regulated at the transcriptional level. These compounds, mainly starch and proteins, are hydrolyzed upon germination to allow seedling growth. The transcription factor HvGAMYB is a master activator both in the maturation phase of seed development and upon germination, acting in combination with other transcription factors. However, the precise mechanism controlling the switch from maturation to germination programs remains unclear. We report here the identification and molecular characterization of Hordeum vulgare VIVIPAROUS1 (HvVP1), orthologous to ABA-INSENSITIVE3 from Arabidopsis thaliana HvVP1 transcripts accumulate in the endosperm and the embryo of developing seeds at early stages and in the embryo and aleurone of germinating seeds up to 24 h of imbibition. In transient expression assays, HvVP1 controls the activation of Hor2 and Amy6.4 promoters exerted by HvGAMYB. HvVP1 interacts with HvGAMYB in Saccharomyces cerevisiae and in the plant nuclei, hindering its interaction with other transcription factors involved in seed gene expression programs, like BPBF. Similarly, this interaction leads to a decrease in the DNA binding of HvGAMYB and the Barley Prolamine-Box binding Factor (BPBF) to their target sequences. Our results indicate that the HvVP1 expression pattern controls the full Hor2 expression activated by GAMYB and BPBF in the developing endosperm and the Amy6.4 activation in postgerminative reserve mobilization mediated by GAMYB. All these data demonstrate the participation of HvVP1 in antagonistic gene expression programs and support its central role as a gene expression switch during seed maturation and germination. PMID:26858366

  6. Integrative proteomics to understand the transmission mechanism of Barley yellow dwarf virus-GPV by its insect vector Rhopalosiphum padi

    PubMed Central

    Wang, Hui; Wu, Keke; Liu, Yan; Wu, Yunfeng; Wang, Xifeng

    2015-01-01

    Barley yellow dwarf virus-GPV (BYDV-GPV) is transmitted by Rhopalosiphum padi and Schizaphis graminum in a persistent nonpropagative manner. To improve our understanding of its transmission mechanism by aphid vectors, we used two approaches, isobaric tags for relative and absolute quantitation (iTRAQ) and yeast two-hybrid (YTH) system, to identify proteins in R. padi that may interact with or direct the spread of BYDV-GPV along the circulative transmission pathway. Thirty-three differential aphid proteins in viruliferous and nonviruliferous insects were identified using iTRAQ coupled to 2DLC-MS/MS. With the yeast two-hybrid system, 25 prey proteins were identified as interacting with the readthrough protein (RTP) and eight with the coat protein (CP), which are encoded by BYDV-GPV. Among the aphid proteins identified, most were involved in primary energy metabolism, synaptic vesicle cycle, the proteasome pathway and the cell cytoskeleton organization pathway. In a systematic comparison of the two methods, we found that the information generated by the two methods was complementary. Taken together, our findings provide useful information on the interactions between BYDV-GPV and its vector R. padi to further our understanding of the mechanisms regulating circulative transmission in aphid vectors. PMID:26161807

  7. Comparative proteomic analysis of rice after seed ground simulated radiation and spaceflight explains the radiation effects of space environment

    NASA Astrophysics Data System (ADS)

    Wang, Wei; Shi, Jinming; Liang, Shujian; Lei, Huang; Shenyi, Zhang; Sun, Yeqing

    In previous work, we compared the proteomic profiles of rice plants growing after seed space-flights with ground controls by two-dimensional difference gel electrophoresis (2-D DIGE) and found that the protein expression profiles were changed after seed space environment exposures. Spaceflight represents a complex environmental condition in which several interacting factors such as cosmic radiation, microgravity and space magnetic fields are involved. Rice seed is in the process of dormant of plant development, showing high resistance against stresses, so the highly ionizing radiation (HZE) in space is considered as main factor causing biological effects to seeds. To further investigate the radiation effects of space environment, we performed on-ground simulated HZE particle radiation and compared between the proteomes of seed irra-diated plants and seed spaceflight (20th recoverable satellite) plants from the same rice variety. Space ionization shows low-dose but high energy particle effects, for searching the particle effects, ground radiations with the same low-dose (2mGy) but different liner energy transfer (LET) values (13.3KeV/µm-C, 30KeV/µm-C, 31KeV/µm-Ne, 62.2KeV/µm-C, 500Kev/µm-Fe) were performed; using 2-D DIGE coupled with clustering and principle component analysis (PCA) for data process and comparison, we found that the holistic protein expression patterns of plants irradiated by LET-62.2KeV/µm carbon particles were most similar to spaceflight. In addition, although space environment presents a low-dose radiation (0.177 mGy/day on the satellite), the equivalent simulated radiation dose effects should still be evaluated: radiations of LET-62.2KeV/µm carbon particles with different cumulative doses (2mGy, 20mGy, 200mGy, 2000mGy) were further carried out and resulted that the 2mGy radiation still shared most similar proteomic profiles with spaceflight, confirming the low-dose effects of space radiation. Therefore, in the protein expression level

  8. Proteome of Soybean Seed Exudates Contains Plant Defense-Related Proteins Active against the Root-Knot Nematode Meloidogyne incognita.

    PubMed

    Rocha, Raquel O; Morais, Janne K S; Oliveira, Jose T A; Oliveira, Hermogenes D; Sousa, Daniele O B; Souza, Carlos Eduardo A; Moreno, Frederico B; Monteiro-Moreira, Ana Cristina O; de Souza Júnior, José Dijair Antonino; de Sá, Maria F Grossi; Vasconcelos, Ilka M

    2015-06-10

    Several studies have described the effects of seed exudates against microorganisms, but only few of them have investigated the proteins that have defensive activity particularly against nematode parasites. This study focused on the proteins released in the exudates of soybean seeds and evaluated their nematicidal properties against Meloidogyne incognita. A proteomic approach indicated the existence of 63 exuded proteins, including β-1,3-glucanase, chitinase, lectin, trypsin inhibitor, and lipoxygenase, all of which are related to plant defense. The presence of some of these proteins was confirmed by their in vitro activity. The soybean exudates were able to reduce the hatching of nematode eggs and to cause 100% mortality of second-stage juveniles (J2). The pretreatment of J2 with these exudates resulted in a 90% reduction of the gall number in tobacco plants. These findings suggest that the exuded proteins are directly involved in plant defense against soil pathogens, including nematodes, during seed germination. PMID:26034922

  9. Mapping field spatial distribution patterns of isoproturon-mineralizing activity over a three-year winter wheat/rape seed/barley rotation.

    PubMed

    Hussain, S; Devers-Lamrani, M; Spor, A; Rouard, N; Porcherot, M; Beguet, J; Martin-Laurent, F

    2013-03-01

    The temporal and spatial variability of the activity of soil microorganisms able to mineralize the herbicide isoproturon (IPU) pesticide was investigated over a three-year long crop rotation between 2008 and 2010. Isoproturon mineralization was higher in 2008, when winter wheat was treated with this herbicide, than in 2009 and 2010, when rape seed and barley were treated with different herbicides. Under laboratory conditions, we showed that isoproturon mineralization was not promoted by sulfonylurea herbicide applied on barley crop in 2010. IPU mineralization was shown to be highly variable at the field scale in years 2009 and 2010. Principal component analyses and analyses of similarities revealed that soil pH and equivalent humidity, and to a lesser extent soil organic matter content and cation exchange capacity (CEC) were the main drivers of isoproturon-mineralizing activity variance. Using a rather simple model that yields the rate of isoproturon mineralization as a function of soil pH and equivalent humidity, we explained up to 85% of the variance observed. Mapping field-scale distribution of isoproturon mineralization over the three-year survey indicated higher variability in 2009 and in 2010 as compared to 2008, suggesting that isoproturon treatment applied to winter wheat promoted isoproturon mineralization activity and reduced its spatial variability. Field-scale distribution of isoproturon mineralization showed important similarity to the distribution of soil pH, equivalent humidity and to a lesser extent to soil organic matter and cation exchange capacity (CEC) thereby confirming our model. PMID:23246724

  10. Characterization of the Microchemical Structure of Seed Endosperm within a Cellular Dimension among Six Barley Varieties with Distinct Degradation Kinetics, Using Ultraspatially Resolved Synchrotron-Based Infrared Synchrotron-Based Infrared

    SciTech Connect

    Liu, N.; Yu, P

    2010-01-01

    Barley varieties have similar chemical composition but exhibit different rumen degradation kinetics and nutrient availability. These biological differences may be related to molecular, structural, and chemical makeup among the seed endosperm tissue. No detailed study was carried out. The objectives of this study were: (1) to use a molecular spectroscopy technique, synchrotron-based Fourier transform infrared microspectroscopy (SFTIRM), to determine the microchemical-structural features in seed endosperm tissue of six developed barley varieties; (2) to study the relationship among molecular-structural characteristics, degradation kinetics, and nutrient availability in six genotypes of barley. The results showed that inherent microchemical-structural differences in the endosperm among the six barley varieties were detected by the synchrotron-based analytical technique, SFTIRM, with the univariate molecular spectral analysis. The SFTIRM spectral profiles differed (P < 0.05) among the barley samples in terms of the peak ratio and peak area and height intensities of amides I (ca. 1650 cm{sup -1}) and II (ca. 1550 cm{sup -1}), cellulosic compounds (ca. 1240 cm{sup -1}), CHO component peaks (the first peak at the region ca. 1184-1132 cm{sup -1}, the second peak at ca. 1132-1066 cm{sup -1}, and the third peak at ca. 1066-950 cm{sup -1}). With the SFTIRM technique, the structural characteristics of the cereal seeds were illuminated among different cultivars at an ultraspatial resolution. The structural differences of barley seeds may be one reason for the various digestive behaviors and nutritive values in ruminants. The results show weak correlations between the functional groups spectral data (peak area, height intensities, and ratios) and rumen biodegradation kinetics (rate and extent of nutrient degradation). Weak correlations may indicate that limited variations of these six barley varieties might not be sufficient to interpret the relationship between spectroscopic

  11. The DOF protein, SAD, interacts with GAMYB in plant nuclei and activates transcription of endosperm-specific genes during barley seed development.

    PubMed

    Diaz, Isabel; Martinez, Manuel; Isabel-LaMoneda, Ines; Rubio-Somoza, Ignacio; Carbonero, Pilar

    2005-06-01

    The DOF protein, SAD, previously shown to be a transcriptional activator in barley aleurone cells upon seed germination, also has an important role in gene regulation during endosperm development. mRNA was detected in early (10 days after flowering) developing barley seeds where it accumulated in the starchy endosperm, aleurone cells, nucellar projection, vascular tissues and the immature embryo, as shown by RT-PCR and in situ hybridization analyses. The SAD protein, expressed in bacteria, binds to oligonucleotides containing the prolamine box, 5'-A/TAAAG-3'sequence, derived from the promoter regions of the endosperm-specific genes Hor2 and Itr1, encoding a B-hordein and trypsin-inhibitor BTI-CMe, respectively. SAD competed for the same binding sites with another endosperm-expressed DOF protein, BPBF. Transient expression experiments in co-bombarded developing endosperms demonstrated that SAD trans-activated transcription from Hor2 and Itr1 promoters through binding to the intact DOF motifs. When the two DOF factors are co-bombarded together an additive effect was observed upon the expression of the Itr1 gene. In-frame fusion of the Sad ORF to the reporter green fluorescent protein gene directs the fluorescence expression to the nucleus in transiently transformed onion epidermal layers. The visualization of fluorescence in the nucleus of onion cells, using the bimolecular fluorescent complex (BiFC) approach, has shown the in vivo interaction between SAD and the R2R3MYB protein GAMYB. The interaction in plant cells has also been documented for the DOF protein BPBF and GAMYB, but nuclear interaction could not be detected between BPBF and SAD by this procedure. PMID:15918880

  12. Analysis of Proteome Profile in Germinating Soybean Seed, and Its Comparison with Rice Showing the Styles of Reserves Mobilization in Different Crops

    PubMed Central

    Han, Chao; Yin, Xiaojian; He, Dongli; Yang, Pingfang

    2013-01-01

    Background Seed germination is a complex physiological process during which mobilization of nutrient reserves happens. In different crops, this event might be mediated by different regulatory and metabolic pathways. Proteome profiling has been proved to be an efficient way that can help us to construct these pathways. However, no such studies have been performed in soybean germinating seeds up to date. Results Proteome profiling was conducted through one-dimensional gel electrophoresis followed by liquid chromatography and tandem mass spectrometry strategy in the germinating seeds of soybean (glycine max). Comprehensive comparisons were also carried out between rice and soybean germinating seeds. 764 proteins belonging to 14 functional groups were identified and metabolism related proteins were the largest group. Deep analyses of the proteins and pathways showed that lipids were degraded through lipoxygenase dependent pathway and proteins were degraded through both protease and 26S proteosome system, and the lipoxygenase could also help to remove the reactive oxygen species during the rapid mobilization of reserves of soybean germinating seeds. The differences between rice and soybean germinating seeds proteome profiles indicate that each crop species has distinct mechanism for reserves mobilization during germination. Different reserves could be converted into starches before they are totally utilized during the germination in different crops seeds. Conclusions This study is the first comprehensive analysis of proteome profile in germinating soybean seeds to date. The data presented in this paper will improve our understanding of the physiological and biochemical status in the imbibed soybean seeds just prior to germination. Comparison of the protein profile with that of germinating rice seeds gives us new insights on mobilization of nutrient reserves during the germination of crops seeds. PMID:23460823

  13. Barley Oil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley (Hordeum vulgare) is an ancient grain that has was domesticated for use as a food. Currently only about 2% is used for food, about two thirds is used for animal feed and one third for malting. Because the oil content of most barley cultivars is low (<2%), obtaining oil from whole barley gra...

  14. A proteomic analysis of seeds from Bt-transgenic Brassica napus and hybrids with wild B. juncea

    PubMed Central

    Liu, Yongbo; Zhang, Ying-Xue; Song, Song-Quan; Li, Junsheng; Neal Stewart Jr., C.; Wei, Wei; Zhao, Yujie; Wang, Wei-Qing

    2015-01-01

    Transgene insertions might have unintended side effects on the transgenic host, both crop and hybrids with wild relatives that harbor transgenes. We employed proteomic approaches to assess protein abundance changes in seeds from Bt-transgenic oilseed rape (Brassica napus) and its hybrids with wild mustard (B. juncea). A total of 24, 15 and 34 protein spots matching to 23, 13 and 31 unique genes were identified that changed at least 1.5 fold (p < 0.05, Student’s t-test) in abundance between transgenic (tBN) and non-transgenic (BN) oilseed rape, between hybrids of B. juncea (BJ) × tBN (BJtBN) and BJ × BN (BJBN) and between BJBN and BJ, respectively. Eight proteins had higher abundance in tBN than in BN. None of these proteins was toxic or nutritionally harmful to human health, which is not surprising since the seeds are not known to produce toxic proteins. Protein spots varying in abundance between BJtBN and BJBN seeds were the same or homologous to those in the respective parents. None of the differentially-accumulated proteins between BJtBN and BJBN were identical to those between tBN and BN. Results indicated that unintended effects resulted from transgene flow fell within the range of natural variability of hybridization and those found in the native host proteomes. PMID:26486652

  15. A proteomic analysis of seeds from Bt-transgenic Brassica napus and hybrids with wild B. juncea.

    PubMed

    Liu, Yongbo; Zhang, Ying-Xue; Song, Song-Quan; Li, Junsheng; Neal Stewart, C; Wei, Wei; Zhao, Yujie; Wang, Wei-Qing

    2015-01-01

    Transgene insertions might have unintended side effects on the transgenic host, both crop and hybrids with wild relatives that harbor transgenes. We employed proteomic approaches to assess protein abundance changes in seeds from Bt-transgenic oilseed rape (Brassica napus) and its hybrids with wild mustard (B. juncea). A total of 24, 15 and 34 protein spots matching to 23, 13 and 31 unique genes were identified that changed at least 1.5 fold (p < 0.05, Student's t-test) in abundance between transgenic (tBN) and non-transgenic (BN) oilseed rape, between hybrids of B. juncea (BJ) × tBN (BJtBN) and BJ × BN (BJBN) and between BJBN and BJ, respectively. Eight proteins had higher abundance in tBN than in BN. None of these proteins was toxic or nutritionally harmful to human health, which is not surprising since the seeds are not known to produce toxic proteins. Protein spots varying in abundance between BJtBN and BJBN seeds were the same or homologous to those in the respective parents. None of the differentially-accumulated proteins between BJtBN and BJBN were identical to those between tBN and BN. Results indicated that unintended effects resulted from transgene flow fell within the range of natural variability of hybridization and those found in the native host proteomes. PMID:26486652

  16. Regulation of Phosphoenolpyruvate Carboxylase Phosphorylation by Metabolites and Abscisic Acid during the Development and Germination of Barley Seeds1[C][W

    PubMed Central

    Feria, Ana-Belén; Alvarez, Rosario; Cochereau, Ludivine; Vidal, Jean; García-Mauriño, Sofía; Echevarría, Cristina

    2008-01-01

    During barley (Hordeum vulgare) seed development, phosphoenolpyruvate carboxylase (PEPC) activity increased and PEPC-specific antibodies revealed housekeeping (103-kD) and inducible (108-kD) subunits. Bacterial-type PEPC fragments were immunologically detected in denatured protein extracts from dry and imbibed conditions; however, on nondenaturing gels, the activity of the recently reported octameric PEPC (in castor [Ricinus communis] oil seeds) was not detected. The phosphorylation state of the PEPC, as judged by l-malate 50% inhibition of initial activity values, phosphoprotein chromatography, and immunodetection of the phosphorylated N terminus, was found to be high between 8 and 18 d postanthesis (DPA) and during imbibition. In contrast, the enzyme appeared to be in a low phosphorylation state from 20 DPA up to dry seed. The time course of 32/36-kD, Ca2+-independent PEPC kinase activity exhibited a substantial increase after 30 DPA that did not coincide with the PEPC phosphorylation profile. This kinase was found to be inhibited by l-malate and not by putative protein inhibitors, and the PEPC phosphorylation status correlated with high glucose-6-phosphate to malate ratios, thereby suggesting an in vivo metabolic control of the kinase. PEPC phosphorylation was also regulated by photosynthate supply at 11 DPA. In addition, when fed exogenously to imbibing seeds, abscisic acid significantly increased PEPC kinase activity. This was further enhanced by the cytosolic protein synthesis inhibitor cycloheximide but blocked by protease inhibitors, thereby suggesting that the phytohormone acts on the stability of the kinase. We propose that a similar abscisic acid-dependent effect may contribute to produce the increase in PEPC kinase activity during desiccation stages. PMID:18753284

  17. A redox-dependent dimerization switch regulates activity and tolerance for reactive oxygen species of barley seed glutathione peroxidase.

    PubMed

    Navrot, Nicolas; Skjoldager, Nicklas; Bunkenborg, Jakob; Svensson, Birte; Hägglund, Per

    2015-05-01

    Monomeric and dimeric forms of recombinant barley (Hordeum vulgare subsp. vulgare) glutathione peroxidase 2 (HvGpx2) are demonstrated to display distinctly different functional properties in vitro. Monomeric HvGpx2 thus has five fold higher catalytic efficiency than the dimer towards tert-butyl hydroperoxide, but is more sensitive to inactivation by hydrogen peroxide. Treatment of the monomer with hydrogen peroxide results in dimer formation. This observed new behavior of a plant glutathione peroxidase suggests a mechanism involving a switch from a highly catalytically competent monomer to a less active, but more oxidation-resistant dimer. PMID:25796076

  18. Laboratory Methods to Remove Surface Layers from Cereal Grains Using a Seed Scarifier and comparison with a barley pearler.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The distribution of nutrients and undesirable components within a cereal seed is heterogeneous, and many of the valuable nutrients are located in the outer layers. Efficient laboratory methods to remove these layers will allow for the experimental production of value-added products. A seed scarife...

  19. Proteomic analysis reveals differential accumulation of small heat shock proteins and late embryogenesis abundant proteins between ABA-deficient mutant vp5 seeds and wild-type Vp5 seeds in maize.

    PubMed

    Wu, Xiaolin; Gong, Fangping; Yang, Le; Hu, Xiuli; Tai, Fuju; Wang, Wei

    2014-01-01

    ABA is a major plant hormone that plays important roles during many phases of plant life cycle, including seed development, maturity and dormancy, and especially the acquisition of desiccation tolerance. Understanding of the molecular basis of ABA-mediated plant response to stress is of interest not only in basic research on plant adaptation but also in applied research on plant productivity. Maize mutant viviparous-5 (vp5), deficient in ABA biosynthesis in seeds, is a useful material for studying ABA-mediated response in maize. Due to carotenoid deficiency, vp5 endosperm is white, compared to yellow Vp5 endosperm. However, the background difference at proteome level between vp5 and Vp5 seeds is unclear. This study aimed to characterize proteome alterations of maize vp5 seeds and to identify ABA-dependent proteins during seed maturation. We compared the embryo and endosperm proteomes of vp5 and Vp5 seeds by gel-based proteomics. Up to 46 protein spots, most in embryos, were found to be differentially accumulated between vp5 and Vp5. The identified proteins included small heat shock proteins (sHSPs), late embryogenesis abundant (LEA) proteins, stress proteins, storage proteins and enzymes among others. However, EMB564, the most abundant LEA protein in maize embryo, accumulated in comparable levels between vp5 and Vp5 embryos, which contrasted to previously characterized, greatly lowered expression of emb564 mRNA in vp5 embryos. Moreover, LEA proteins and sHSPs displayed differential accumulations in vp5 embryos: six out of eight identified LEA proteins decreased while nine sHSPs increased in abundance. Finally, we discussed the possible causes of global proteome alterations, especially the observed differential accumulation of identified LEA proteins and sHSPs in vp5 embryos. The data derived from this study provides new insight into ABA-dependent proteins and ABA-mediated response during maize seed maturation. PMID:25653661

  20. ALPHA-GLUCOSIDASES FROM THE GLYCOSIDE HYDROLASE FAMILY 31 IN GERMINATING SEEDS AND SEEDLING LEAVES OF BARLEY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Of the four starch degrading enzymes in plants, only alpha-amylase and alpha-glucosidase have been extensively studied. Both alpha-amylase and alpha-glucosidase are important in germinating seeds in direct initiation of attack on starch grains. Five different alpha-glucosidases have been found in ...

  1. A Proteomics-Based Platform for Systems Biology Analysis of Soybean Seed Development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A system based on fresh weight and color was used to define eight stages of soybean (Glycine max (L.) Merrill, cv. Jack) seed development. Storage protein, oil, and starch were quantified from each stage, and used along with the morphological characteristics to establish a first-stage model of seed...

  2. Grain sorghum proteomics: An integrated approach towards characterization of seed storage proteins in kafirin allelic variants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seed protein composition determines quality traits, such as value for food, feedstock and biomaterials uses. Sorghum seed proteins are predominantly prolamins known as kafirins. Located primarily on the periphery of storage protein bodies, cysteine-rich ß- and gama-kafirins are thought to prevent en...

  3. Shotgun label-free quantitative proteomics of developing peanut (Arachis hypogaea L.) seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Legume seeds and peanuts, in particular, are an inexpensive source of plant proteins and edible oil. Owing to their importance in global food security, it is necessary to understand the genetic, biochemical, and physiological mechanisms controlling seed quality and nutritive attributes. A comprehens...

  4. Analysis of the embryo proteome of sycamore (Acer pseudoplatanus L.) seeds reveals a distinct class of proteins regulating dormancy release.

    PubMed

    Pawłowski, Tomasz Andrzej; Staszak, Aleksandra Maria

    2016-05-20

    Acer pseudoplatanus seeds are characterized by a deep physiological embryo dormancy that requires a few weeks of cold stratification in order to promote germination. Understanding the function of proteins and their related metabolic pathways, in conjunction with the plant hormones implicated in the breaking of seed dormancy, would expand our knowledge pertaining to this process. In this study, a proteomic approach was used to analyze the changes occurring in seeds in response to cold stratification, which leads to dormancy release. In addition, the involvement of abscisic (ABA) and gibberellic acids (GA) was also examined. Fifty-three proteins showing significant changes were identified by mass spectrometry. An effect of ABA on protein variation was observed at the beginning of stratification, while the influence of GA on protein abundance was observed during the middle phase of stratification. The majority of proteins associated with dormancy breaking in the presence of only water, and also ABA or GA, were classified as being involved in metabolism and genetic information processing. For metabolic-related proteins, the effect of ABA on protein abundance was stimulatory for half of the proteins and inhibitory for half of the proteins. On the other hand, the effect on genetic information processing related proteins was stimulatory. GA was found to upregulate both metabolic-related and genetic information processing-related proteins. While seed dormancy breaking depends on proteins involved in a variety of processes, proteins associated with methionine metabolism (adenosine kinase, methionine synthase) and glycine-rich RNA binding proteins appear to be of particular importance. PMID:26970688

  5. Crystal structure of an essential enzyme in seed starch degradation: barley limit dextrinase in complex with cyclodextrins.

    PubMed

    Vester-Christensen, Malene Bech; Abou Hachem, Maher; Svensson, Birte; Henriksen, Anette

    2010-11-12

    Barley limit dextrinase [Hordeum vulgare limit dextrinase (HvLD)] catalyzes the hydrolysis of α-1,6 glucosidic linkages in limit dextrins. This activity plays a role in starch degradation during germination and presumably in starch biosynthesis during grain filling. The crystal structures of HvLD in complex with the competitive inhibitors α-cyclodextrin (CD) and β-CD are solved and refined to 2.5 Å and 2.1 Å, respectively, and are the first structures of a limit dextrinase. HvLD belongs to glycoside hydrolase 13 family and is composed of four domains: an immunoglobulin-like N-terminal eight-stranded β-sandwich domain, a six-stranded β-sandwich domain belonging to the carbohydrate binding module 48 family, a catalytic (β/α)(8)-like barrel domain that lacks α-helix 5, and a C-terminal eight-stranded β-sandwich domain of unknown function. The CDs are bound at the active site occupying carbohydrate binding subsites +1 and +2. A glycerol and three water molecules mimic a glucose residue at subsite -1, thereby identifying residues involved in catalysis. The bulky Met440, a unique residue at its position among α-1,6 acting enzymes, obstructs subsite -4. The steric hindrance observed is proposed to affect substrate specificity and to cause a low activity of HvLD towards amylopectin. An extended loop (Asp513-Asn520) between β5 and β6 of the catalytic domain also seems to influence substrate specificity and to give HvLD a higher affinity for α-CD than pullulanases. The crystal structures additionally provide new insight into cation sites and the concerted action of the battery of hydrolytic enzymes in starch degradation. PMID:20863834

  6. Low Phytic Acid Barley Responses to Phosphorus Rates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Low phytic acid (LPA) barley (Hordeum vulgare L.) cultivars partition phosphorus in seed tissue differently than conventional barley cultivars through a reduction in seed phytic acid (myo-inositol-1,2,3,4,5,6-hexkisphosphate) coupled with an increase in inorganic phosphorus. The response of the LPA...

  7. Barley Genomics: An Overview

    PubMed Central

    Sreenivasulu, Nese; Graner, Andreas; Wobus, Ulrich

    2008-01-01

    Barley (Hordeum vulgare), first domesticated in the Near East, is a well-studied crop in terms of genetics, genomics, and breeding and qualifies as a model plant for Triticeae research. Recent advances made in barley genomics mainly include the following: (i) rapid accumulation of EST sequence data, (ii) growing number of studies on transcriptome, proteome, and metabolome, (iii) new modeling techniques, (iv) availability of genome-wide knockout collections as well as efficient transformation techniques, and (v) the recently started genome sequencing effort. These developments pave the way for a comprehensive functional analysis and understanding of gene expression networks linked to agronomically important traits. Here, we selectively review important technological developments in barley genomics and related fields and discuss the relevance for understanding genotype-phenotype relationships by using approaches such as genetical genomics and association studies. High-throughput genotyping platforms that have recently become available will allow the construction of high-density genetic maps that will further promote marker-assisted selection as well as physical map construction. Systems biology approaches will further enhance our knowledge and largely increase our abilities to design refined breeding strategies on the basis of detailed molecular physiological knowledge. PMID:18382615

  8. Proteomic identification of allergenic seed proteins, napin and cruciferin, from cold-pressed rapeseed oils.

    PubMed

    Puumalainen, T J; Puustinen, A; Poikonen, S; Turjanmaa, K; Palosuo, T; Vaali, K

    2015-05-15

    In Finland and France atopic children commonly react to seeds of oilseed rape and turnip rape in skin prick tests (SPT) and open food challenges. These seeds are not as such in dietary use and therefore the routes of sensitization are unknown. Possible allergens were extracted from commercial cold-pressed and refined rapeseed oils and identified by gel-based tandem nanoflow liquid chromatography mass spectrometry (LC-MS/MS). Napin (a 2S albumin), earlier identified as a major allergen in the seeds of oilseed rape and turnip rape, and cruciferin (an 11S globulin), a new potential seed allergen, were detected in cold-pressed oils, but not in refined oils. Pooled sera from five children sensitized or allergic to oilseed rape and turnip rape seeds reacted to these proteins from cold-pressed oil preparations and individual sera from five children reacted to these proteins extracted from the seeds when examined with IgE immunoblotting. Hence cold-pressed rapeseed oil might be one possible route of sensitization for these allergens. PMID:25577095

  9. Proteins with High Turnover Rate in Barley Leaves Estimated by Proteome Analysis Combined with in Planta Isotope Labeling1[W][OPEN

    PubMed Central

    Nelson, Clark J.; Alexova, Ralitza; Jacoby, Richard P.; Millar, A. Harvey

    2014-01-01

    Protein turnover is a key component in cellular homeostasis; however, there is little quantitative information on degradation kinetics for individual plant proteins. We have used 15N labeling of barley (Hordeum vulgare) plants and gas chromatography-mass spectrometry analysis of free amino acids and liquid chromatography-mass spectrometry analysis of proteins to track the enrichment of 15N into the amino acid pools in barley leaves and then into tryptic peptides derived from newly synthesized proteins. Using information on the rate of growth of barley leaves combined with the rate of degradation of 14N-labeled proteins, we calculate the turnover rates of 508 different proteins in barley and show that they vary by more than 100-fold. There was approximately a 9-h lag from label application until 15N incorporation could be reliably quantified in extracted peptides. Using this information and assuming constant translation rates for proteins during the time course, we were able to quantify degradation rates for several proteins that exhibit half-lives on the order of hours. Our workflow, involving a stringent series of mass spectrometry filtering steps, demonstrates that 15N labeling can be used for large-scale liquid chromatography-mass spectrometry studies of protein turnover in plants. We identify a series of abundant proteins in photosynthesis, photorespiration, and specific subunits of chlorophyll biosynthesis that turn over significantly more rapidly than the average protein involved in these processes. We also highlight a series of proteins that turn over as rapidly as the well-known D1 subunit of photosystem II. While these proteins need further verification for rapid degradation in vivo, they cluster in chlorophyll and thiamine biosynthesis. PMID:25082890

  10. Digging deeper into the seed proteome: Pre-fractionation of total proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seeds are a common experimental system for many reasons. Among these they occupy a major niche in agriculture and human nutrition, they are a rich source of critical genetic information, and they are a near-ideal system for the study of phytohormone action or the transition from either dormancy or ...

  11. Unraveling the seed endosperm proteome of the lotus (Nelumbo nucifera Gaertn.) utilizing 1DE and 2DE separation in conjunction with tandem mass spectrometry.

    PubMed

    Moro, Carlo F; Fukao, Yoichiro; Shibato, Junko; Rakwal, Randeep; Timperio, Anna Maria; Zolla, Lello; Agrawal, Ganesh Kumar; Shioda, Seiji; Kouzuma, Yoshiaki; Yonekura, Masami

    2015-05-01

    Nelumbo nucifera (Gaertn.) or lotus, is an aquatic plant native to India, and presently consumed as food mainly in China and Japan. Lotus is also widely used in Indian and Chinese traditional medicine. Extracts from different parts of the lotus plant have been reported to show diverse biological activities-antioxidant, free radical scavenging, anti-inflammatory, and immunomodulatory. Despite this, little work has been done in isolating and identifying proteins responsible for these activities, or yet importantly to establish a lotus proteome. The aim of our group is to develop a proteome catalog of the lotus plant, starting with its seed, the nutrient rich food source. In this present study, the seed endosperm-most abundant in proteins, and main nutrient storage tissue-was targeted for protein extraction by testing five different extraction protocols, followed by their proteomic analyses using complementary 1DE and 2DE approaches in conjunction with MS/MS. The inventory of 66 nonredundant proteins obtained by 1DE-MS and the 30 obtained by 2DE-MS provides the first catalog of the lotus seed endosperm, where the most abundant protein functions were in categories of metabolic activities related to carbohydrate metabolism and nutrient storage. PMID:25545995

  12. Proteomic Analysis of Phosphoproteins in the Rice Nucleus During the Early Stage of Seed Germination.

    PubMed

    Li, Ming; Yin, Xiaojian; Sakata, Katsumi; Yang, Pingfang; Komatsu, Setsuko

    2015-07-01

    The early stage of seed germination is the first step in the plant life cycle without visible morphological change. To investigate the mechanism controlling the early stage of rice seed germination, we performed gel-and label-free nuclear phosphoproteomics. A total of 3467 phosphopeptides belonging to 102 nuclear phosphoproteins from rice embryos were identified. Protein-synthesis-related proteins were mainly phosphorylated. During the first 24 h following imbibition, 115 nuclear phosphoproteins were identified, and significant changes in the phosphorylation level over time were observed in 29 phosphoproteins. Cluster analysis indicated that nucleotide-binding proteins and zinc finger CCCH- and BED-type proteins increased in abundance during the first 12 h of imbibition and then decreased. The in silico protein-protein interactions for 29 nuclear phosphoproteins indicated that the Sas10/Utp3 protein, which functions in snoRNA binding and gene silencing, was the center of the phosphoprotein network in nuclei. The germination rate of seeds was significantly slowed with phosphatase inhibitor treatment. The mRNA expression of the zinc finger CCCH-type protein did not change, and the zinc finger BED-type protein was upregulated in rice embryos during the early stage of germination with phosphatase inhibitor treatment. These results suggest that the phosphorylation and dephosphorylation of nuclear proteins are involved in rice seed germination. Furthermore, transcription factors such as zinc finger CCCH- and BED-type proteins might play a key role through nuclear phosphoproteins, and Sas10/Utp3 protein might interact with nuclear phosphoproteins in rice embryos to mediate the early stage of seed germination. PMID:26035336

  13. Investigation of relationships between barley stress peptides and beer gushing using SDS-PAGE and MS screening.

    PubMed

    Hégrová, Blanka; Farková, Marta; Macuchová, Simona; Havel, Josef; Preisler, Jan

    2009-12-01

    The relationship between gushing and antifungal peptides in barley and malt kernels was examined for five barley varieties produced in the Czech Republic with four conditions of infection and treatment. Proteome changes during pathogen-seed interaction were observed with SDS-PAGE and MALDI-TOF MS. These methods were applied as a fast screening for observing the relationship between gushing and peptides/proteins. It was found that the presence of basic peptides, presumably hordothionins and non-specific lipid transfer protein type 1, did not correlate with the degree of gushing for malt (/r/ in <0.07, 0.34>), (/r/ in <0.01, 0.49>), respectively, as detected by both methods. PMID:19950350

  14. Proteomic analysis of mature soybean seeds from the Chernobyl area suggests plant adaptation to the contaminated environment.

    PubMed

    Danchenko, Maksym; Skultety, Ludovit; Rashydov, Namik M; Berezhna, Valentyna V; Mátel, L'ubomír; Salaj, Terézia; Pret'ová, Anna; Hajduch, Martin

    2009-06-01

    The explosion in one of the four reactors of the Chernobyl Nuclear Power Plant (CNPP, Chernobyl) caused the worst nuclear environmental disaster ever seen. Currently, 23 years after the accident, the soil in the close vicinity of CNPP is still significantly contaminated with long-living radioisotopes, such as (137)Cs. Despite this contamination, the plants growing in Chernobyl area were able to adapt to the radioactivity, and survive. The aim of this study was to investigate plant adaptation mechanisms toward permanently increased level of radiation using a quantitative high-throughput proteomics approach. Soybeans of a local variety (Soniachna) were sown in contaminated and control fields in the Chernobyl region. Mature seeds were harvested and the extracted proteins were subjected to two-dimensional gel electrophoresis (2-DE). In total, 9.2% of 698 quantified protein spots on 2-D gel were found to be differentially expressed with a p-value seed storage proteins are involved in plant adaptation mechanism to radioactivity in the Chernobyl region. PMID:19320472

  15. Comparative proteomic analysis of embryos between a maize hybrid and its parental lines during early stages of seed germination.

    PubMed

    Guo, Baojian; Chen, Yanhong; Zhang, Guiping; Xing, Jiewen; Hu, Zhaorong; Feng, Wanjun; Yao, Yingyin; Peng, Huiru; Du, Jinkun; Zhang, Yirong; Ni, Zhongfu; Sun, Qixin

    2013-01-01

    In spite of commercial use of heterosis in agriculture, the molecular basis of heterosis is poorly understood. It was observed that maize hybrid Zong3/87-1 exhibited an earlier onset or heterosis in radicle emergence. To get insights into the underlying mechanism of heterosis in radicle emergence, differential proteomic analysis between hybrid and its parental lines was performed. In total, the number of differentially expressed protein spots between hybrid and its parental lines in dry and 24 h imbibed seed embryos were 134 and 191, respectively, among which 47.01% (63/134) and 34.55% (66/191) protein spots displayed nonadditively expressed pattern. Remarkably, 54.55% of nonadditively accumulated proteins in 24 h imbibed seed embryos displayed above or equal to the level of the higher parent patterns. Moreover, 155 differentially expressed protein spots were identified, which were grouped into eight functional classes, including transcription & translation, energy & metabolism, signal transduction, disease & defense, storage protein, transposable element, cell growth & division and unclassified proteins. In addition, one of the upregulated proteins in F1 hybrids was ZmACT2, a homolog of Arabidopsis thaliana ACT7 (AtACT7). Expressing ZmACT2 driven by the AtACT7 promoter partially complemented the low germination phenotype in the Atact7 mutant. These results indicated that hybridization between two parental lines can cause changes in the expression of a variety of proteins, and it is concluded that the altered pattern of gene expression at translational level in the hybrid may be responsible for the observed heterosis. PMID:23776561

  16. Comparative Biochemical and Proteomic Analyses of Soybean Seed Cultivars Differing in Protein and Oil Content.

    PubMed

    Min, Chul Woo; Gupta, Ravi; Kim, So Wun; Lee, So Eui; Kim, Yong Chul; Bae, Dong Won; Han, Won Young; Lee, Byong Won; Ko, Jong Min; Agrawal, Ganesh Kumar; Rakwal, Randeep; Kim, Sun Tae

    2015-08-19

    This study develops differential protein profiles of soybean (Glycine max) seeds (cv. Saedanbaek and Daewon) varying in protein (47.9 and 39.2%) and oil (16.3 and 19.7%) content using protamine sulfate (PS) precipitation method coupled with a 2D gel electrophoresis (2DGE) approach. Of 71 detected differential spots between Daewon and Saedanbaek, 48 were successfully identified by MALDI-TOF/TOF. Gene ontology analysis revealed that up-regulated proteins in Saedanbaek were largely associated with nutrient reservoir activity (42.6%), which included mainly seed-storage proteins (SSPs; subunits of glycinin and β-conglycinin). Similar results were also obtained in two cultivars of wild soybean (G. soja cv. WS22 and WS15) differing in protein content. Western blots confirmed higher accumulation of SSPs in protein-rich Saedanbaek. Findings presented and discussed in this study highlight a possible involvement of the urea cycle for increased accumulation of SSPs and hence the higher protein content in soybean seeds. PMID:26237057

  17. Studies on the molecular mechanisms of seed germination.

    PubMed

    Han, Chao; Yang, Pingfang

    2015-05-01

    Seed germination that begins with imbibition and ends with radicle emergence is the first step for plant growth. Successful germination is not only crucial for seedling establishment but also important for crop yield. After being dispersed from mother plant, seed undergoes continuous desiccation in ecosystem and selects proper environment to trigger germination. Owing to the contribution of transcriptomic, proteomic, and molecular biological studies, molecular aspect of seed germination is elucidated well in Arabidopsis. Recently, more and more proteomic and genetic studies concerning cereal seed germination were performed on rice (Oryza sativa) and barley (Hordeum vulgare), which possess completely different seed structure and domestication background with Arabidopsis. In this review, both the common features and the distinct mechanisms of seed germination are compared among different plant species including Arabidopsis, rice, and maize. These features include morphological changes, cell and its related structure recovery, metabolic activation, hormone behavior, and transcription and translation activation. This review will provide more comprehensive insights into the molecular mechanisms of seed germination. PMID:25597791

  18. Reboot the system thanks to protein post-translational modifications and proteome diversity: How quiescent seeds restart their metabolism to prepare seedling establishment.

    PubMed

    Arc, Erwann; Galland, Marc; Cueff, Gwendal; Godin, Béatrice; Lounifi, Imen; Job, Dominique; Rajjou, Loïc

    2011-05-01

    Once liberated in their environment, orthodox seeds live in a quiescent dehydrated state not totally exempt of essential molecular events as, for example, the capacity of breaking dormancy during after-ripening. Upon imbibition, if internal regulatory padlocks are released and given adequate external conditions, the quiescent seed is able to "reboot" its system and, thus, germinate. Recent studies unraveled the crucial importance of protein PTMs in seed dormancy, longevity and vigor. As compared to other plant developmental stages, the seed proteome appears quite unique and diverse. Seed proteins encompass several functional classes from primary and secondary metabolism to structural and antimicrobial defense. In the dry state, oxidative damages can occur due to reactive oxygen and nitrogen species produced by non-enzymatic reactions. These reactive species can affect proteins by the oxidation of their amino acids in a post-translational manner. The hormone abscisic acid regulates major aspects of seed life including dormancy and germination. This signaling pathway has been shown to rely on several PTMs such as protein phosphorylation or ubiquitination. PMID:21433284

  19. The associations between Vrs1 alleles and grain quality traits in spring barley Hordeum vulgare L.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley head row type is a major trait affecting end use quality. Six rowed forms emerged due to mutations in the Vrs1 gene in two rowed barleys. Whether barley is two (Vrs1) or six rowed (vrs1) directly affects a wide range of morphological traits related to seed yield and grain quality. Vrs1 has be...

  20. Pseudomonas fluorescens PICF7 displays an endophytic lifestyle in cultivated cereals and enhances yield in barley.

    PubMed

    Mercado-Blanco, Jesús; Alós, Enriqueta; Rey, María Dolores; Prieto, Pilar

    2016-08-01

    Pseudomonas fluorescens PICF7, an indigenous inhabitant of olive roots, displays an endophytic lifestyle in this woody crop and exerts biocontrol against the fungal phytopathogen Verticillium dahliae Here we report microscopy evidence that the strain PICF7 is also able to colonize and persist on or in wheat and barley root tissues. Root colonization of both cereal species followed a similar pattern to that previously reported in olive, including inner colonization of the root hairs. This demonstrates that strain PICF7 can colonize root systems of distant botanical species. Barley plants germinated from PICF7-treated seeds showed enhanced vegetative growth. Moreover, significant increases in the number of grains (up to 19.5%) and grain weight (up to 20.5%) per plant were scored in this species. In contrast, growth and yield were not significantly affected in wheat plants by the presence of PICF7. Proteomics analysis of the root systems revealed that different proteins were exclusively found depending on the presence or absence of PICF7 and only one protein with hydrogen ion transmembrane transporter activity was exclusively found in both PICF7-inoculated barley and wheat plants but not in the controls. PMID:27130938

  1. Hardness locus sequence variation and endosperm texture in spring barley

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Grain texture is an important quality parameter in both wheat and barley. Cultivars with good malting quality tend to be softer than their poor malting counterparts. Harder textured barley seeds with lower dry matter digestibility may potentially be better in feed quality for beef cattle. Most gr...

  2. TRANSFORMATION TO PRODUCE BARLEY RESISTANT TO FUSARIUM GRAMINEARUM

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungal pathogen Fusarium graminearum destroys barley and wheat crops by causing scab disease (Fusarium head blight, FHB). Spores infect seed spike tissues, leading to production of mycotoxins. There are no known barleys with biochemical resistance to Fusarium, although some have various levels ...

  3. Barley stripe mosaic and Barley yellow stripe

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley stripe mosaic was described in Wisconsin as "barley false stripe" in 1910, making it perhaps the first cereal virus disease described in the United States. The disease has been reported from most barley-producing areas of the world, including North and South America, Asia, Africa, Europe, an...

  4. Serine proteinases from barley malt may degrade beta-amylase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley seed proteinases are critically important to seed germination and malting in that they generate amino acids from seed N reserves, supporting embryo growth during germination and yeast fermentation during brewing. However, relatively little is known regarding the endogenous protein substrate ...

  5. Unraveling Regulation of the Small Heat Shock Proteins by the Heat Shock Factor HvHsfB2c in Barley: Its Implications in Drought Stress Response and Seed Development

    PubMed Central

    Reddy, Palakolanu Sudhakar; Kavi Kishor, Polavarapu B.; Seiler, Christiane; Kuhlmann, Markus; Eschen-Lippold, Lennart; Lee, Justin; Reddy, Malireddy K.; Sreenivasulu, Nese

    2014-01-01

    The rapid increase in heat shock proteins upon exposure to damaging stresses and during plant development related to desiccation events reveal their dual importance in plant development and stress tolerance. Genome-wide sequence survey identified 20 non-redundant small heat shock proteins (sHsp) and 22 heat shock factor (Hsf) genes in barley. While all three major classes (A, B, C) of Hsfs are localized in nucleus, the 20 sHsp gene family members are localized in different cell organelles like cytoplasm, mitochondria, plastid and peroxisomes. Hsf and sHsp members are differentially regulated during drought and at different seed developmental stages suggesting the importance of chaperone role under drought as well as seed development. In silico cis-regulatory motif analysis of Hsf promoters showed an enrichment with abscisic acid responsive cis-elements (ABRE), implying regulatory role of ABA in mediating transcriptional response of HvsHsf genes. Gene regulatory network analysis identified HvHsfB2c as potential central regulator of the seed-specific expression of several HvsHsps including 17.5CI sHsp. These results indicate that HvHsfB2c is co-expressed in the central hub of small Hsps and therefore it may be regulating the expression of several HvsHsp subclasses HvHsp16.88-CI, HvHsp17.5-CI and HvHsp17.7-CI. The in vivo relevance of binding specificity of HvHsfB2C transcription factor to HSE-element present in the promoter of HvSHP17.5-CI under heat stress exposure is confirmed by gel shift and LUC-reporter assays. Further, we isolated 477 bp cDNA from barley encoding a 17.5 sHsp polypeptide, which was predominantly upregulated under drought stress treatments and also preferentially expressed in developing seeds. Recombinant HvsHsp17.5-CI protein was expressed in E. coli and purified to homogeneity, which displayed in vitro chaperone activity. The predicted structural model of HvsHsp-17.5-CI protein suggests that the α-crystallin domain is evolutionarily highly

  6. Thionin antifungal peptide synthesis in transgenic barley

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In seeds and vegetative organs of barley and other cereals, thionins are processed into peptides with pronounced anti-microbial properties. In vitro studies demonstrated the toxicity of a- and ß-hordothionins (HTHs) to the fungal pathogen Fusarium graminearum. Increasing the expression of thionin g...

  7. Proteomic comparison between maturation drying and prematurely imposed drying of Zea mays seeds reveals a potential role of maturation drying in preparing proteins for seed germination, seedling vigor, and pathogen resistance.

    PubMed

    Wang, Wei-Qing; Ye, Jian-Qing; Rogowska-Wrzesinska, Adelina; Wojdyla, Katarzyna I; Jensen, Ole Nørregaard; Møller, Ian Max; Song, Song-Quan

    2014-02-01

    We have studied the role(s) of maturation drying in the acquisition of germinability, seedling vigor and pathogen resistance by comparing the proteome changes in maize embryo and endosperm during mature and prematurely imposed drying. Prematurely imposed dried seeds at 40 days after pollination (DAP) germinated almost as well as mature seeds (at 65 DAP), but their seedling growth was slower and they were seriously infected by fungi. A total of 80 and 114 proteins were identified to change at least two-fold (p < 0.05) in abundance during maturation drying in embryo and endosperm, respectively. Fewer proteins (48 and 59 in embryo and endosperm, respectively) changed in abundance during prematurely imposed drying. A number of proteins, 33 and 38 in embryo and endosperm, respectively, changed similarly in abundance during both maturation and prematurely imposed drying. Storage proteins were abundant in this group and may contribute to the acquisition of seed germinability. However, a relatively large number of proteins changed in the embryo (47 spots) and endosperm (76 spots) specifically during maturation drying. Among these proteins, storage proteins in the embryo and defense proteins in the endosperm may be particularly important for seedling vigor and resistance to fungal infection, respectively. PMID:24341390

  8. Metabolomic and Proteomic Profiles Reveal the Dynamics of Primary Metabolism during Seed Development of Lotus (Nelumbo nucifera)

    PubMed Central

    Wang, Lei; Fu, Jinlei; Li, Ming; Fragner, Lena; Weckwerth, Wolfram; Yang, Pingfang

    2016-01-01

    Sacred lotus (Nelumbo nucifera) belongs to the Nelumbonaceae family. Its seeds are widely consumed in Asian countries as snacks or even medicine. Besides the market value, lotus seed also plays a crucial role in the lotus life cycle. Consequently, it is essential to gain a comprehensive understanding of the development of lotus seed. During its development, lotus seed undergoes cell division, expansion, reserve accumulation, desiccation, and maturation phases. We observed morphological and biochemical changes from 10 to 25 days after pollination (DAP) which corresponded to the reserve synthesis and accumulation phase. The volume of the seed expanded until 20 DAP with the color of the seed coat changing from yellow-green to dark green and gradually fading again. Starch and protein rapidly accumulated from 15 to 20 DAP. To further reveal metabolic adaptation, primary metabolites and proteins profiles were obtained using mass spectrometry based platforms. Metabolites and enzymes involved in sugar metabolism, glycolysis, TCA cycle and amino acid metabolism showed sequential dynamics enabling the clear separation of the different metabolic states during lotus seed development. The integration of the data revealed a highly significant metabolic switch at 15 DAP going through a transition of metabolically highly active tissue to the preparation of storage tissue. The results provide a reference data set for the evaluation of primary metabolism during lotus seed development. PMID:27375629

  9. Metabolomic and Proteomic Profiles Reveal the Dynamics of Primary Metabolism during Seed Development of Lotus (Nelumbo nucifera).

    PubMed

    Wang, Lei; Fu, Jinlei; Li, Ming; Fragner, Lena; Weckwerth, Wolfram; Yang, Pingfang

    2016-01-01

    Sacred lotus (Nelumbo nucifera) belongs to the Nelumbonaceae family. Its seeds are widely consumed in Asian countries as snacks or even medicine. Besides the market value, lotus seed also plays a crucial role in the lotus life cycle. Consequently, it is essential to gain a comprehensive understanding of the development of lotus seed. During its development, lotus seed undergoes cell division, expansion, reserve accumulation, desiccation, and maturation phases. We observed morphological and biochemical changes from 10 to 25 days after pollination (DAP) which corresponded to the reserve synthesis and accumulation phase. The volume of the seed expanded until 20 DAP with the color of the seed coat changing from yellow-green to dark green and gradually fading again. Starch and protein rapidly accumulated from 15 to 20 DAP. To further reveal metabolic adaptation, primary metabolites and proteins profiles were obtained using mass spectrometry based platforms. Metabolites and enzymes involved in sugar metabolism, glycolysis, TCA cycle and amino acid metabolism showed sequential dynamics enabling the clear separation of the different metabolic states during lotus seed development. The integration of the data revealed a highly significant metabolic switch at 15 DAP going through a transition of metabolically highly active tissue to the preparation of storage tissue. The results provide a reference data set for the evaluation of primary metabolism during lotus seed development. PMID:27375629

  10. Global Proteome Analyses of Lysine Acetylation and Succinylation Reveal the Widespread Involvement of both Modification in Metabolism in the Embryo of Germinating Rice Seed.

    PubMed

    He, Dongli; Wang, Qiong; Li, Ming; Damaris, Rebecca Njeri; Yi, Xingling; Cheng, Zhongyi; Yang, Pingfang

    2016-03-01

    Regulation of rice seed germination has been shown to mainly occur at post-transcriptional levels, of which the changes on proteome status is a major one. Lysine acetylation and succinylation are two prevalent protein post-translational modifications (PTMs) involved in multiple biological processes, especially for metabolism regulation. To investigate the potential mechanism controlling metabolism regulation in rice seed germination, we performed the lysine acetylation and succinylation analyses simultaneously. Using high-accuracy nano-LC-MS/MS in combination with the enrichment of lysine acetylated or succinylated peptides from digested embryonic proteins of 24 h after imbibition (HAI) rice seed, a total of 699 acetylated sites from 389 proteins and 665 succinylated sites from 261 proteins were identified. Among these modified lysine sites, 133 sites on 78 proteins were commonly modified by two PTMs. The overlapped PTM sites were more likely to be in polar acidic/basic amino acid regions and exposed on the protein surface. Both of the acetylated and succinylated proteins cover nearly all aspects of cellular functions. Ribosome complex and glycolysis/gluconeogenesis-related proteins were significantly enriched in both acetylated and succinylated protein profiles through KEGG enrichment and protein-protein interaction network analyses. The acetyl-CoA and succinyl-CoA metabolism-related enzymes were found to be extensively modified by both modifications, implying the functional interaction between the two PTMs. This study provides a rich resource to examine the modulation of the two PTMs on the metabolism pathway and other biological processes in germinating rice seed. PMID:26767346

  11. New Insight into Quinoa Seed Quality under Salinity: Changes in Proteomic and Amino Acid Profiles, Phenolic Content, and Antioxidant Activity of Protein Extracts.

    PubMed

    Aloisi, Iris; Parrotta, Luigi; Ruiz, Karina B; Landi, Claudia; Bini, Luca; Cai, Giampiero; Biondi, Stefania; Del Duca, Stefano

    2016-01-01

    highest TPC and AA under non-saline conditions. Salinity increased TPC in all three landraces, with the strongest increase occurring in R49, and enhanced radical scavenging capacity in R49 and VR. Overall, results show that salinity deeply altered the seed proteome and amino acid profiles and, in general, increased the concentration of bioactive molecules and AA of protein extracts in a genotype-dependent manner. PMID:27242857

  12. New Insight into Quinoa Seed Quality under Salinity: Changes in Proteomic and Amino Acid Profiles, Phenolic Content, and Antioxidant Activity of Protein Extracts

    PubMed Central

    Aloisi, Iris; Parrotta, Luigi; Ruiz, Karina B.; Landi, Claudia; Bini, Luca; Cai, Giampiero; Biondi, Stefania; Del Duca, Stefano

    2016-01-01

    highest TPC and AA under non-saline conditions. Salinity increased TPC in all three landraces, with the strongest increase occurring in R49, and enhanced radical scavenging capacity in R49 and VR. Overall, results show that salinity deeply altered the seed proteome and amino acid profiles and, in general, increased the concentration of bioactive molecules and AA of protein extracts in a genotype-dependent manner. PMID:27242857

  13. Proteomic approaches to study structure, functions and toxicity of legume seeds lectins. Perspectives for the assessment of food quality and safety.

    PubMed

    Nasi, Antonella; Picariello, Gianluca; Ferranti, Pasquale

    2009-04-13

    Lectins are a structurally diverse class of (glyco)proteins which bind mono- and oligosaccharides with high specificity and in a reversible way. For many years, the unique sugar binding properties of plant lectins have been exploited for the development of biochemical tools for glycoprotein isolation and characterisation, and the use of lectins as a glycoprofiling tool has became much more sophisticated with the advent of lectin microarrays, in which a panel of lectins are immobilized on a single chip for glycomic analysis. Among the numerous lectins studied so far, those from legumes represent the largest family. They can be present at relatively high amounts depending on genetic as well as environmental factors, and are accumulated especially in the seeds. For this reason, some lectins as the phytohemagglutinin from the common bean Phaseolus vulgaris constitute a possible risk, since consumption of raw or incorrectly processed beans has been shown to cause outbreaks of gastroenteritis, nausea and diarrhoea. On the other hand, for these anti-nutritional properties, bean extracts enriched in lectins or in lectin-related amylase inhibitors are also finding a growing use as active ingredients of "weight-blockers" in dietetic preparations for obesity treatment. Current methods to determine the lectin levels in foods are based on immunoenzymatic or toxicity tests, which are largely aspecific. Very recently, the availability of proteomic methodologies has allowed to start development and validation of sensitive and specific assays for detecting trace amounts of harmful lectins in either raw or processed foods. In this review, the main aspects of current and perspective applications of mass spectrometry and proteomic technologies to the structural characterisation of legumes are presented, with focus on issues related to detection, identification, and quantification of phytohemagglutinins relevant for their biochemical, immunological and toxicological aspects. PMID

  14. Effect of aluminum treatment on proteomes of radicles of seeds derived from Al-treated tomato plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aluminum (Al) toxicity is a major constraint to plant growth and crop yield in acid soils. Tomato cultivars are especially susceptible to excessive A1 3+ accumulated in the root zone. In this study, tomato plants were grown in a hydroponic culture system supplemented with 50 uM AlK(SO4)2. Seeds harv...

  15. A Comparison Of Barley Malt Amylolytic Enzyme Thermostabilities As Indicators Of Malt Sugar Concentrations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to test the hypothesis that barley malt amylolytic enzyme thermostabilities would correlate negatively with malt sugar concentrations. Seeds of four two-row and four six-row North American elite barley cultivars were steeped and germinated in a micromalter for 6 days. At 2...

  16. A Comparison of Barley Malt Amylolytic Enzyme Activities and Malt Sugar Concentrations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to test the hypothesis that barley malt alpha-amylase activity would correlate better with malt sugar concentrations than the activities of beta-amylase, or limit dextrinase. Seeds of four two-row and four six-row North American elite barley cultivars were steeped and germi...

  17. A Comparison of Barley Malt Amylolytic Enzyme Activities and Malt Sugar Concentrations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to test the hypothesis that barley malt alpha-amylase activity would correlate better with malt sugar concentrations than the activities of beta-amylase, or limit dextrinase. Seeds of four two-row and four six-row North American elite barley cultivars were steeped and germin...

  18. Proteomic Analysis of Sauvignon Blanc Grape Skin, Pulp and Seed and Relative Quantification of Pathogenesis-Related Proteins

    PubMed Central

    Tian, Bin; Harrison, Roland; Morton, James; Deb-Choudhury, Santanu

    2015-01-01

    Thaumatin-like proteins (TLPs) and chitinases are the main constituents of so-called protein hazes which can form in finished white wine and which is a great concern of winemakers. These soluble pathogenesis-related (PR) proteins are extracted from grape berries. However, their distribution in different grape tissues is not well documented. In this study, proteins were first separately extracted from the skin, pulp and seed of Sauvignon Blanc grapes, followed by trypsin digestion and analysis by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Proteins identified included 75 proteins from Sauvignon Blanc grape skin, 63 from grape pulp and 35 from grape seed, mostly functionally classified as associated with metabolism and energy. Some were present exclusively in specific grape tissues; for example, proteins involved in photosynthesis were only detected in grape skin and proteins found in alcoholic fermentation were only detected in grape pulp. Moreover, proteins identified in grape seed were less diverse than those identified in grape skin and pulp. TLPs and chitinases were identified in both Sauvignon Blanc grape skin and pulp, but not in the seed. To relatively quantify the PR proteins, the protein extracts of grape tissues were seperated by HPLC first and then analysed by SDS-PAGE. The results showed that the protein fractions eluted at 9.3 min and 19.2 min under the chromatographic conditions of this study confirmed that these corresponded to TLPs and chitinases seperately. Thus, the relative quantification of TLPs and chitinases in protein extracts was carried out by comparing the area of corresponding peaks against the area of a thamautin standard. The results presented in this study clearly demonstrated the distribution of haze-forming PR proteins in grape berries, and the relative quantification of TLPs and chitinases could be applied in fast tracking of changes in PR proteins during grape growth and determination of PR

  19. Proteomic analysis of oil body membrane proteins accompanying the onset of desiccation phase during sunflower seed development.

    PubMed

    Thakur, Anita; Bhatla, Satish C

    2015-01-01

    A noteworthy metabolic signature accompanying oil body (OB) biogenesis during oilseed development is associated with the modulation of the oil body membranes proteins. Present work focuses on 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE)-based analysis of the temporal changes in the OB membrane proteins analyzed by LC-MS/MS accompanying the onset of desiccation (20-30 d after anthesis; DAA) in the developing seeds of sunflower (Helianthus annuus L.). Protein spots unique to 20-30 DAA stages were picked up from 2-D gels for identification and the identified proteins were categorized into 7 functional classes. These include proteins involved in energy metabolism, reactive oxygen scavenging, proteolysis and protein turnover, signaling, oleosin and oil body biogenesis-associated proteins, desiccation and cytoskeleton. At 30 DAA stage, exclusive expressions of enzymes belonging to energy metabolism, desiccation and cytoskeleton were evident which indicated an increase in the metabolic and enzymatic activity in the cells at this stage of seed development (seed filling). Increased expression of cruciferina-like protein and dehydrin at 30 DAA stage marks the onset of desiccation. The data has been analyzed and discussed to highlight desiccation stage-associated metabolic events during oilseed development. PMID:26786011

  20. Proteomic analysis of oil body membrane proteins accompanying the onset of desiccation phase during sunflower seed development

    PubMed Central

    Thakur, Anita; Bhatla, Satish C

    2015-01-01

    A noteworthy metabolic signature accompanying oil body (OB) biogenesis during oilseed development is associated with the modulation of the oil body membranes proteins. Present work focuses on 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE)-based analysis of the temporal changes in the OB membrane proteins analyzed by LC-MS/MS accompanying the onset of desiccation (20–30 d after anthesis; DAA) in the developing seeds of sunflower (Helianthus annuus L.). Protein spots unique to 20–30 DAA stages were picked up from 2-D gels for identification and the identified proteins were categorized into 7 functional classes. These include proteins involved in energy metabolism, reactive oxygen scavenging, proteolysis and protein turnover, signaling, oleosin and oil body biogenesis-associated proteins, desiccation and cytoskeleton. At 30 DAA stage, exclusive expressions of enzymes belonging to energy metabolism, desiccation and cytoskeleton were evident which indicated an increase in the metabolic and enzymatic activity in the cells at this stage of seed development (seed filling). Increased expression of cruciferina-like protein and dehydrin at 30 DAA stage marks the onset of desiccation. The data has been analyzed and discussed to highlight desiccation stage-associated metabolic events during oilseed development. PMID:26786011

  1. Association genetics in barley

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Applied and basic barley geneticists have begun to use association genetics as a tool to identify and fine map polymorphisms directly in breeding populations or diversity panels. Barley presents an ideal system because its populations present different extents of LD, from long-range LD in elite cult...

  2. A role for seed storage proteins in Arabidopsis seed longevity

    PubMed Central

    Nguyen, Thu-Phuong; Cueff, Gwendal; Hegedus, Dwayne D; Rajjou, Loïc; Bentsink, Leónie

    2015-01-01

    Proteomics approaches have been a useful tool for determining the biological roles and functions of individual proteins and identifying the molecular mechanisms that govern seed germination, vigour and viability in response to ageing. In this work the dry seed proteome of four Arabidopsis thaliana genotypes, that carry introgression fragments at the position of seed longevity quantitative trait loci and as a result display different levels of seed longevity, was investigated. Seeds at two physiological states, after-ripened seeds that had the full germination ability and aged (stored) seeds of which the germination ability was severely reduced, were compared. Aged dry seed proteomes were markedly different from the after-ripened and reflected the seed longevity level of the four genotypes, despite the fact that dry seeds are metabolically quiescent. Results confirmed the role of antioxidant systems, notably vitamin E, and indicated that protection and maintenance of the translation machinery and energy pathways are essential for seed longevity. Moreover, a new role for seed storage proteins (SSPs) was identified in dry seeds during ageing. Cruciferins (CRUs) are the most abundant SSPs in Arabidopsis and seeds of a triple mutant for three CRU isoforms (crua crub cruc) were more sensitive to artificial ageing and their seed proteins were highly oxidized compared with wild-type seeds. These results confirm that oxidation is involved in seed deterioration and that SSPs buffer the seed from oxidative stress, thus protecting important proteins required for seed germination and seedling formation. PMID:26184996

  3. Combined transcript, proteome, and metabolite analysis of transgenic maize seeds engineered for enhanced carotenoid synthesis reveals pleotropic effects in core metabolism

    PubMed Central

    Decourcelle, Mathilde; Perez-Fons, Laura; Baulande, Sylvain; Steiger, Sabine; Couvelard, Linhdavanh; Hem, Sonia; Zhu, Changfu; Capell, Teresa; Christou, Paul; Fraser, Paul; Sandmann, Gerhard

    2015-01-01

    The aim of this study was to assess whether endosperm-specific carotenoid biosynthesis influenced core metabolic processes in maize embryo and endosperm and how global seed metabolism adapted to this expanded biosynthetic capacity. Although enhancement of carotenoid biosynthesis was targeted to the endosperm of maize kernels, a concurrent up-regulation of sterol and fatty acid biosynthesis in the embryo was measured. Targeted terpenoid analysis, and non-targeted metabolomic, proteomic, and transcriptomic profiling revealed changes especially in carbohydrate metabolism in the transgenic line. In-depth analysis of the data, including changes of metabolite pools and increased enzyme and transcript concentrations, gave a first insight into the metabolic variation precipitated by the higher up-stream metabolite demand by the extended biosynthesis capacities for terpenoids and fatty acids. An integrative model is put forward to explain the metabolic regulation for the increased provision of terpenoid and fatty acid precursors, particularly glyceraldehyde 3-phosphate and pyruvate or acetyl-CoA from imported fructose and glucose. The model was supported by higher activities of fructokinase, glucose 6-phosphate isomerase, and fructose 1,6-bisphosphate aldolase indicating a higher flux through the glycolytic pathway. Although pyruvate and acetyl-CoA utilization was higher in the engineered line, pyruvate kinase activity was lower. A sufficient provision of both metabolites may be supported by a by-pass in a reaction sequence involving phosphoenolpyruvate carboxylase, malate dehydrogenase, and malic enzyme. PMID:25796085

  4. Next-generation protein-rich potato expressing the seed protein gene AmA1 is a result of proteome rebalancing in transgenic tuber

    PubMed Central

    Chakraborty, Subhra; Chakraborty, Niranjan; Agrawal, Lalit; Ghosh, Sudip; Narula, Kanika; Shekhar, Shubhendu; Naik, Prakash S.; Pande, P. C.; Chakrborti, Swarup Kumar; Datta, Asis

    2010-01-01

    Protein deficiency is the most crucial factor that affects physical growth and development and that increases morbidity and mortality especially in developing countries. Efforts have been made to improve protein quality and quantity in crop plants but with limited success. Here, we report the development of transgenic potatoes with enhanced nutritive value by tuber-specific expression of a seed protein, AmA1 (Amaranth Albumin 1), in seven genotypic backgrounds suitable for cultivation in different agro-climatic regions. Analyses of the transgenic tubers revealed up to 60% increase in total protein content. In addition, the concentrations of several essential amino acids were increased significantly in transgenic tubers, which are otherwise limited in potato. Moreover, the transgenics also exhibited enhanced photosynthetic activity with a concomitant increase in total biomass. These results are striking because this genetic manipulation also resulted in a moderate increase in tuber yield. The comparative protein profiling suggests that the proteome rebalancing might cause increased protein content in transgenic tubers. Furthermore, the data on field performance and safety evaluation indicate that the transgenic potatoes are suitable for commercial cultivation. In vitro and in vivo studies on experimental animals demonstrate that the transgenic tubers are also safe for human consumption. Altogether, these results emphasize that the expression of AmA1 is a potential strategy for the nutritional improvement of food crops. PMID:20855595

  5. Proteome profiling of seed storage proteins reveals the nutritional potential of Salicornia brachiata Roxb., an extreme halophyte.

    PubMed

    Jha, Bhavanath; Singh, Nater Pal; Mishra, Avinash

    2012-05-01

    Salicornia brachiata is an extreme halophyte that grows in salty marshes and is considered to be a potential alternative crop for seawater agriculture. Salicornia seeds are rich in protein, and its tender shoots are eaten as salad greens. Seed storage proteins were fractionated by sequential extraction using different solvents, including distilled water for albumins, NaCl (1.0 M) for globulins, NaOH (0.1 N) for glutelins, and ethanol (70% v/v) for prolamins. Globulins accounted for 54.75% of the total seed storage proteins followed by albumins (34.30%) and glutelins (8.70%). The fractionated proteins were characterized using 2D-diagonal SDS-PAGE and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. The globulin fraction, composed of seven intermolecular disulfide-linked polypeptide pairs of molecular mass 63.5, 62.5, 54.7, 53.0, 43.2, 38.5, and 35.1 kDa, encompassed a basic and an acidic subunit. Two-dimensional gels revealed approximately 32 spots, with isoelectric points and molecular masses ranging from 4.93 to 11.6 and from ∼5.2 to ∼109.4 kDa, respectively. Protein spots were identified by MALDI-TOF MS peptide mass fingerprint analysis and further classified. Homology analysis demonstrated that 19% of the proteins were involved in metabolism, 16% were involved in signaling, and 15% were regulatory proteins. Peptide mass fingerprint analysis confirmed the presence of inter- and intramolecular disulfide linkages in the globulin fraction. Sulfur-rich proteins are of high nutritional value, and disulfides make S. brachiata a potential source of dietary supplementation. PMID:22494338

  6. Comparing the Effect of Visceral Fat and Barley Seed Ash (Hordeum Vulgare L) With Silversulfadiazine on Burn Wound Healing in Rats

    PubMed Central

    Azadi, Mohammad; Foruozandeh, Hossein; Karami, Leila; Khodayar, Mohammad Javad; Rashidi Nooshabadi, Mohamadreza; Kalantar, Mojtaba; Gudarzi, Mehdi; Pirouzi, Aliyar

    2015-01-01

    Background: Skin burn is one of the most common complications and remains a major public health issue worldwide. Objectives: This experiment was conducted to study the effects of traditional medicine (Visceral Fat and Barely Seed Ash) compared with silversulfadiazine (SSD) cream on healing burn wounds in rats. Materials and Methods: Sixty adult male Wistar rats were randomly divided into four groups of equal numbers; each group consisted of 15 animals. After sedation, type II of skin burn with 1.5 cm diameter circle was created on the back of rats with a heated metal in boiling water. Group one was not treated and considered as control. The burned areas in the second, third and fourth groups were applied twice a day with normal saline, SSD cream and traditional preparation, respectively. Percentage of the burn wound concentration and histopathological examinations were used as parameters of our study on days 4, 9and 14. Obtained data were compared between the groups and days. Results: SSD cream and traditional preparation had better effects on burnt wound healing compared with control group. Furthermore, on the final day of study, the average percentage of wound concentration in traditional medicine group was significantly greater than other groups (P < 0.05). This finding was supported and confirmed by histological examination as well. Conclusions: Traditional preparation significantly decreased inflammation and accelerated wound healing in treated rats. Furthermore, the findings of this study can be applied clinically in the future. PMID:25866721

  7. Physiological and molecular changes in barley and wheat under salinity.

    PubMed

    Temel, Aslihan; Gozukirmizi, Nermin

    2015-03-01

    In this study, it was aimed to compare salinity-induced changes in barley (Hordeum vulgare L. cv. Bornova-92) and bread wheat (Triticum aestivum L. cv. Gerek-79). Seeds were germinated under saline conditions (0, 50, 100, 250, and 500 mM NaCl) for 2 days and recovered under non-saline conditions for 2 days. At the end of the salt treatment, germination, water content (WC), total soluble protein content, and catalase (CAT, EC 1.11.1.6) activity were affected in both species, while superoxide dismutase (SOD, EC 1.15.1.1) activity was affected in barley. Salinity affected WC, protein content, and CAT activity in both species, while it affected germination in barley and affected fresh weight and SOD activity in wheat after recovery. Physiological responses of both species were correlated. Expression of α-tubulin, Atls1, and Lls1 genes was down-regulated in barley after 250 mM NaCl treatment. HVA1 gene was highly (more than 50-fold) stimulated by salinity in barley. However, α-tubulin and Atls1 genes were down-regulated, and Lls1 gene was up-regulated in wheat after recovery from 250-mM NaCl treatment. Increase in HVA1 expression was not significant in wheat. The expression profiles of barley and wheat under salinity are different, and barley tended to regulate gene expression faster than wheat. PMID:25578157

  8. The Importance of Barley Genetics and Domestication in a Global Perspective

    PubMed Central

    Pourkheirandish, Mohammad; Komatsuda, Takao

    2007-01-01

    Background Archaeological evidence has revealed that barley (Hordeum vulgare) is one of the oldest crops used by ancient farmers. Studies of the time and place of barley domestication may help in understanding ancient human civilization. Scope The studies of domesticated genes in crops have uncovered the mechanisms which converted wild and unpromising wild species to the most important food for humans. In addition to archaeological studies, molecular studies are finding new insights into the process of domestication. Throughout the process of barley domestication human selection on wild species resulted in plants with more harvestable seeds. One of the remarkable changes during barley domestications was the appearance of six-rowed barley. The gene associated with this trait results in three times more seed per spike compared with ancestral wild barley. This increase in number of seed resulted in a major dichotomy in the evolution of barley. The identification of the six-rowed spike gene provided a framework for understanding how this character was evolved. Some important barley domestication genes have been discovered and many are currently being investigated. Conclusions Identification of domestication genes in crops revealed that most of the drastic changes during domestication are the result of functional impairments in transcription factor genes, and creation of new functions is rare. Isolation of the six-rowed spike gene revealed that this trait was domesticated more than once in the domestication history of barley. Six-rowed barley is derived from two-rowed ancestral forms. Isolation of photoperiod-response genes in barley and rice revealed that different genes belonging to similar genetic networks partially control this trait. PMID:17761690

  9. 2012 North Dakota Transgenic Barley FHB Nursery Report

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The 2012 North Dakota transgenic field trials consisted of 23 barley lines, tested in three misted and three non-misted replicates. Plots were sown on May 9, 2012 in hill plots with 10 seed per hill spaced at 30 cm, and all plots were inoculated using the grain spawn method at heading. Lines include...

  10. RECENT ADVANCES IN BARLEY TRANSFORMATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley, an important member of the cereals, has been successfully transformed through various methods such as particle bombardment, Agrobacterium-tumefaciens, DNA uptake, and electroporation. Initially, the transformation in barley concentrated on developing protocols using marker genes such as gus,...

  11. The Barley Phytomer

    PubMed Central

    Forster, Brian P.; Franckowiak, Jerome D.; Lundqvist, Udda; Lyon, Jackie; Pitkethly, Ian; Thomas, William T. B.

    2007-01-01

    Background and Aims Morphological mutants have been useful in elucidating the phytomeric structure of plants. Recently described mutants have shed new light on the ontogeny (development of plant structures) and the phytomeric system of barley (Hordeum vulgare). Since the current model for barley phytomers was not adequate to explain the nature of some mutants, a new model is proposed. Methods New phytomer mutants were detected by visual assessment of mutant families in the Optic barley mutation grid population. This was done at various growth stages using laboratory, glasshouse and field screens. Simple explanations were adopted to account for aberrant phytomer phenotypes and a thesis for a new phytomer model was developed. Key Results and Conclusions A barley phytomer model is presented, in which the origins of vegetative and generative structures can be explained by a single repeating phytomer unit. Organs on the barley plant are divided into two classes, single or paired, depending on their origin. Paired structures are often fused together to create specific organs. The model can be applied to wheat (Triticum aestivum) and related grasses. PMID:17901062

  12. Linkage mapping of putative regulator genes of barley grain development characterized by expression profiling

    PubMed Central

    Pietsch, Christof; Sreenivasulu, Nese; Wobus, Ulrich; Röder, Marion S

    2009-01-01

    Background Barley (Hordeum vulgare L.) seed development is a highly regulated process with fine-tuned interaction of various tissues controlling distinct physiological events during prestorage, storage and dessication phase. As potential regulators involved within this process we studied 172 transcription factors and 204 kinases for their expression behaviour and anchored a subset of them to the barley linkage map to promote marker-assisted studies on barley grains. Results By a hierachical clustering of the expression profiles of 376 potential regulatory genes expressed in 37 different tissues, we found 50 regulators preferentially expressed in one of the three grain tissue fractions pericarp, endosperm and embryo during seed development. In addition, 27 regulators found to be expressed during both seed development and germination and 32 additional regulators are characteristically expressed in multiple tissues undergoing cell differentiation events during barley plant ontogeny. Another 96 regulators were, beside in the developing seed, ubiquitously expressed among all tissues of germinating seedlings as well as in reproductive tissues. SNP-marker development for those regulators resulted in anchoring 61 markers on the genetic linkage map of barley and the chromosomal assignment of another 12 loci by using wheat-barley addition lines. The SNP frequency ranged from 0.5 to 1.0 SNP/kb in the parents of the various mapping populations and was 2.3 SNP/kb over all eight lines tested. Exploration of macrosynteny to rice revealed that the chromosomal orders of the mapped putative regulatory factors were predominantly conserved during evolution. Conclusion We identified expression patterns of major transcription factors and signaling related genes expressed during barley ontogeny and further assigned possible functions based on likely orthologs functionally well characterized in model plant species. The combined linkage map and reference expression map of regulators

  13. The effect of gamma irradiation on the viscosity of two barley cultivars for broiler chicks

    NASA Astrophysics Data System (ADS)

    Al-Kaisey, Mahdi T.; Mohammed, Mahmoud A.; Alwan, Abdul-Kader H.; Mohammed, Manal H.

    2002-03-01

    Seeds of two barley cultivars (Local Black and Shoaa) were gamma irradiated at 0, 10, 50, 100, 150 and 200 kGy doses using Cobalt-60 source to decrease the viscosity. The viscosity was determined in the flour of the seeds using Ostwald U-tube viscometer. The viscosity values were reduced by 25%, 50%, 65%, 72% and 74% in Local Black barley cultivar, while, in Shoaa cultivar the reductions were 15%, 30%, 52%, 69% and 67% at 10, 50, 100, 150 and 200 kGy, respectively. The chemical compositions of the seeds were determined in all treatments.

  14. Loss-of-function of DELLA protein SLN1 activates GA signaling in barley aleurone

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gibberellic acid (GA) is a key phytohormone regulating seed germination and seedling growth. In cereal species, GA is synthesized in embyros and then translocated to aleurone tissues where it induces production of hydrolytic enzymes for mobolization of seed storage reserve. De-embyronated barley a...

  15. A comparison of barley malt amylolytic enzyme thermostabilities and wort sugars produced during mashing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The industrial process that converts seed starch to fermentable sugars, known as mashing, takes place at high temperatures. Barley seed is typically germinated for 4 – 6 days during the malting process during which time the enzymes that convert starch to sugars are either synthetized and/or activat...

  16. Biochemistry, Structure and Function of Non-Wheat Proteins: Case Study of Barley ß-Amylase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The importance of a protein is not always evident and may be due to its multifunctional nature. ß-Amylase in seeds of barley (Hordeum vulgare L.) constitutes approximately 2% of the total protein in mature seeds and is assumed to be important when storage proteins are mobilized to support protein s...

  17. Barley Yellow Dwarf Virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley yellow dwarf (BYD) is the most widespread and economically important virus disease of cereals. The viruses causing BYD were initially grouped based on common biological properties, including persistent and often strain-specific transmission by aphids and induction of yellowing symptoms. The...

  18. REGISTRATION OF 'HERALD' BARLEY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    'Herald' (Reg. No. xxx, P.I. 642403 ) is a low-phytate six-rowed spring feed barley (Hordeum vulgare L.) cultivar developed cooperatively and released in 2006 by the Agricultural Research Service, U.S. Department of Agriculture, and the Idaho Agricultural Experiment Station. It is the first released...

  19. Registration of Endeavor Barley

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ‘Endeavor’ (Reg. No. ______PI 654824); a two-rowed winter malting barley (Hordeum vulgare L.) was developed and submitted for release in 2007 by the Agricultural Research Service-USDA, Aberdeen, ID, in cooperation with the University of Idaho Agricultural Experiment Station. Endeavor is a selection...

  20. Barley Yellow Dwarf

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley yellow dwarf is the most economically important virus disease affecting most cereal crops world wide. This manuscript summarizes the current knowledge of the disease etiology, epidemiology and management. This information is incorporated into the latest revision of the American Phytopathologi...

  1. Microgeographic edaphic differentiation in hordein polymorphisms of wild barley.

    PubMed

    Nevo, E; Beiles, A; Storch, N; Doll, H; Andersen, B

    1983-01-01

    Genetic diversity in the storage protein hordein encoded by two loci, Hor1 and Hor2, was analyzed electrophoretically in seeds from 123 individual plants of wild barley, Hordeum spontaneum, the progenitor of cultivated barley. The test was conducted in two topographically different 100 meter transects in Israel, each equally divided into basalt and terra rossa soil types. Altogether 15 Hor1 and 16 Hor2 phenotypes were detected; 7 phenotypes in Hor1 and 5 in Hor2 were common. Significant differentiation of both Hor1 and Hor2 phenotypes and their associations was found with soil type and topography. Likewise, significant correlations were found between hordein phenotypes and allozyme types detected in a previous study. Our results suggest that at least part of the hordein polymorphisms in wild barley is adaptive and selected by soil and topographic differences over very short distances. PMID:24264871

  2. Proteomic analysis of common bean seed with storage protein deficiency reveals up-regulation of sulfur-rich proteins and starch and raffinose metabolic enzymes, and down-regulation of the secretory pathway.

    PubMed

    Marsolais, Frédéric; Pajak, Agnieszka; Yin, Fuqiang; Taylor, Meghan; Gabriel, Michelle; Merino, Diana M; Ma, Vanessa; Kameka, Alexander; Vijayan, Perumal; Pham, Hai; Huang, Shangzhi; Rivoal, Jean; Bett, Kirstin; Hernández-Sebastià, Cinta; Liu, Qiang; Bertrand, Annick; Chapman, Ralph

    2010-06-16

    A deficiency in major seed storage proteins is associated with a nearly two-fold increase in sulfur amino acid content in genetically related lines of common bean (Phaseolus vulgaris). Their mature seed proteome was compared by an approach combining label-free quantification by spectral counting, 2-DE, and analysis of selective extracts. Lack of phaseolin, phytohemagglutinin and arcelin was mainly compensated by increases in legumin, alpha-amylase inhibitors and mannose lectin FRIL. Along with legumin, albumin-2, defensin and albumin-1 were major contributors to the elevated sulfur amino acid content. Coordinate induction of granule-bound starch synthase I, starch synthase II-2 and starch branching enzyme were associated with minor alteration of starch composition, whereas increased levels of UDP-glucose 4-epimerase were correlated with a 30% increase in raffinose content. Induction of cell division cycle protein 48 and ubiquitin suggested enhanced ER-associated degradation. This was not associated with a classical unfolded protein response as the levels of ER HSC70-cognate binding protein were actually reduced in the mutant. Repression of rab1 GTPase was consistent with decreased traffic through the secretory pathway. Collectively, these results have implications for the nutritional quality of common bean, and provide information on the pleiotropic phenotype associated with storage protein deficiency in a dicotyledonous seed. PMID:20353836

  3. Proteomics analysis in mature seed of four peanut cultivars using two-dimensional gel electrophoresis reveals distinct differential expression of storage, anti-nutritive, and allergenic proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protein profiles of total seed proteins isolated from mature seeds of four peanut cultivars, New Mexico Valencia C (NM Valencia C), Tamspan 90, Georgia Green, and NC-7, were studied using two-dimensional gel electrophoresis coupled with nano electrospray ionization liquid chromatography tandem mass ...

  4. Comparisons of barley malt amylolytic enzyme thermostabilities to wort osmolyte concentrations, malt extract, ASBC measures of malt quality, and initial enzyme activities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study the hypothesis that wort osmolyte concentration (OC) would correlate much better than malt extract (ME) with barley amylolytic enzyme thermostabilities of malts produced over several days of germination was tested. Seeds of 4 two-row and 4 six-row North American elite barley cultivars ...

  5. Integrated analysis of seed proteome and mRNA oxidation reveals distinct post-transcriptional features regulating dormancy in wheat (Triticum aestivum L.).

    PubMed

    Gao, Feng; Rampitsch, Christof; Chitnis, Vijaya R; Humphreys, Gavin D; Jordan, Mark C; Ayele, Belay T

    2013-10-01

    Wheat seeds can be released from a dormant state by after-ripening; however, the underlying molecular mechanisms are still mostly unknown. We previously identified transcriptional programmes involved in the regulation of after-ripening-mediated seed dormancy decay in wheat (Triticum aestivum L.). Here, we show that seed dormancy maintenance and its release by dry after-ripening in wheat is associated with oxidative modification of distinct seed-stored mRNAs that mainly correspond to oxidative phosphorylation, ribosome biogenesis, nutrient reservoir and α-amylase inhibitor activities, suggesting the significance of post-transcriptional repression of these biological processes in regulating seed dormancy. We further show that after-ripening induced seed dormancy release in wheat is mediated by differential expression of specific proteins in both dry and hydrated states, including those involved in proteolysis, cellular signalling, translation and energy metabolism. Among the genes corresponding to these proteins, the expression of those encoding α-amylase/trypsin inhibitor and starch synthase appears to be regulated by mRNA oxidation. Co-expression analysis of the probesets differentially expressed and oxidized during dry after-ripening along with those corresponding to proteins differentially regulated between dormant and after-ripened seeds produced three co-expressed gene clusters containing more candidate genes potentially involved in the regulation of seed dormancy in wheat. Two of the three clusters are enriched with elements that are either abscisic acid (ABA) responsive or recognized by ABA-regulated transcription factors, indicating the association between wheat seed dormancy and ABA sensitivity. PMID:23745731

  6. Analysis of barley by NIRS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Development of a rapid method of analysis of barley for moisture, starch, protein, oil, ash and Beta-glucan was attempted. One hundred forty-three barley grain samples of 3 types (hulled, hulless and malt) over 2 growing seasons and from various locations in the United States were utilized in the s...

  7. Barley yellow dwarf viruses.

    PubMed

    Miller, W A; Rasochová, L

    1997-01-01

    Barley yellow dwarf viruses represent one of the most economically important and ubiquitous groups of plant viruses. This review focuses primarily on four research areas in which progress has been most rapid. These include (a) evidence supporting reclassification of BYDVs into two genera; (b) elucidation of gene function and novel mechanisms controlling gene expression; (c) initial forays into understanding the complex interactions between BYDV virions and their aphid vectors; and (d) replication of a BYDV satellite RNA. Economic losses, symptomatology, and means of control of BYD are also discussed. PMID:15012520

  8. Potential for Production of Dryland Barley for Human Food: Quality and Agronomic Performance of Spring Habit Germplasm in Oregon

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Grain ß-glucan percentage is the most important attribute for barley (Hordeum vulgare L.) varieties destined for the human food market. This trait is important because of the cholesterol-reducing properties of ß-glucan. High levels of grain protein, test weight and seed size may also add value. Seed...

  9. Genetics of barley hooded suppression.

    PubMed Central

    Roig, Cristina; Pozzi, Carlo; Santi, Luca; Müller, Judith; Wang, Yamei; Stile, Maria Rosaria; Rossini, Laura; Stanca, Michele; Salamini, Francesco

    2004-01-01

    The molecular basis of the barley dominant Hooded (K) mutant is a duplication of 305 bp in intron IV of the homeobox gene Bkn3. A chemical mutagenesis screen was carried out to identify genetical factors that participate in Bkn3 intron-mediated gene regulation. Plants from recurrently mutagenized KK seeds were examined for the suppression of the hooded awn phenotype induced by the K allele and, in total, 41 suK (suppressor of K) recessive mutants were identified. Complementation tests established the existence of five suK loci, and alleles suKB-4, suKC-33, suKD-25, suKE-74, and suKF-76 were studied in detail. All K-suppressed mutants showed a short-awn phenotype. The suK loci have been mapped by bulked segregant analysis nested in a standard mapping procedure based on AFLP markers. K suppressor loci suKB, B, E, and F all map in a short interval of chromosome 7H, while the locus suKD is assigned to chromosome 5H. A complementation test between the four suK mutants mapping on chromosome 7H and the short-awn mutant lks2, located nearby, excluded the allelism between suK loci and lks2. The last experiment made clear that the short-awn phenotype of suK mutants is due to a specific dominant function of the K allele, a function that is independent from the control on hood formation. The suK loci are discussed as candidate participants in the regulation of Bkn3 expression. PMID:15166167

  10. Assessment of Genetic Diversity among Barley Cultivars and Breeding Lines Adapted to the US Pacific Northwest, and Its Implications in Breeding Barley for Imidazolinone-Resistance

    PubMed Central

    Mejías, Jaime H.; Gemini, Richa; Brew-Appiah, Rhoda A. T.; Wen, Nuan; Osorio, Claudia; Ankrah, Nii; Murphy, Kevin M.; von Wettstein, Diter

    2014-01-01

    Extensive application of imidazolinone (IMI) herbicides had a significant impact on barley productivity contributing to a continuous decline in its acreage over the last two decades. A possible solution to this problem is to transfer IMI-resistance from a recently characterized mutation in the ‘Bob’ barley AHAS (acetohydroxy acid synthase) gene to other food, feed and malting barley cultivars. We focused our efforts on transferring IMI-resistance to barley varieties adapted to the US Pacific Northwest (PNW), since it comprises ∼23% (335,000 ha) of the US agricultural land under barley production. To effectively breed for IMI-resistance, we studied the genetic diversity among 13 two-rowed spring barley cultivars/breeding-lines from the PNW using 61 microsatellite markers, and selected six barley genotypes that showed medium to high genetic dissimilarity with the ‘Bob’ AHAS mutant. The six selected genotypes were used to make 29–53 crosses with the AHAS mutant and a range of 358–471 F1 seeds were obtained. To make informed selection for the recovery of the recipient parent genome, the genetic location of the AHAS gene was determined and its genetic nature assessed. Large F2 populations ranging in size from 2158–2846 individuals were evaluated for herbicide resistance and seedling vigor. Based on the results, F3 lines from the six most vigorous F2 genotypes per cross combination were evaluated for their genetic background. A range of 20%–90% recovery of the recipient parent genome for the carrier chromosome was observed. An effort was made to determine the critical dose of herbicide to distinguish between heterozygotes and homozygotes for the mutant allele. Results suggested that the mutant can survive up to the 10× field recommended dose of herbicide, and the 8× and 10× herbicide doses can distinguish between the two AHAS mutant genotypes. Finally, implications of this research in sustaining barley productivity in the PNW are discussed. PMID

  11. Albinism in barley androgenesis.

    PubMed

    Makowska, Katarzyna; Oleszczuk, Sylwia

    2014-03-01

    Androgenesis is highly useful for plant breeding, significantly reducing breeding cycle times, as well as in a wide range of biological research. However, for widespread use this process must be efficient. Despite several decades of research on the phenomenon of androgenesis, many processes involved are obscure and there is much to be understood about androgenesis. One of the problems inherent in androgenesis, and reducing its efficiency, is albinism. This article reviews albinism in barley anthers and microspores in vitro cultures. Of special interest is the fate of plastids throughout androgenesis, which is important at several levels, including the genes responsible for driving the green-to-albino ratios. We also summarize the external factors that reduce the incidence of albino plants that are regenerated via androgenesis. PMID:24326697

  12. The TRI101 story: engineering wheat and barley to resist Fusarium head blight

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fusarium Head Blight (FHB), caused primarily by Fusarium graminearum, is a major disease of wheat and barley in the United States and Canada. FHB epidemics have been on the increase since 1993, and have caused severe monetary damage for the growers and the seed industry. Along with reduced yields,...

  13. Near-Infrared Spectroscopy Using a Supercontinuum Laser: Application to Long Wavelength Transmission Spectra of Barley Endosperm and Oil.

    PubMed

    Ringsted, Tine; Dupont, Sune; Ramsay, Jacob; Jespersen, Birthe Møller; Sørensen, Klavs Martin; Keiding, Søren Rud; Engelsen, Søren Balling

    2016-07-01

    The supercontinuum laser is a new type of light source, which combines the collimation and intensity of a laser with the broad spectral region of a lamp. Using such a source therefore makes it possible to focus the light onto small sample areas without losing intensity and thus facilitate either rapid or high-intensity measurements. Single seed transmission analysis in the long wavelength (LW) near-infrared (NIR) region is one area that might benefit from a brighter light source such as the supercontinuum laser. This study is aimed at building an experimental spectrometer consisting of a supercontinuum laser source and a dispersive monochromator in order to investigate its capability to measure the barley endosperm using transmission experiments in the LW NIR region. So far, barley and wheat seeds have only been studied using NIR transmission in the short wavelength region up to 1100 nm. However, the region in the range of 2260-2380 nm has previously shown to be particularly useful in differentiating barley phenotypes using NIR spectroscopy in reflectance mode. In the present study, 350 seeds (consisting of 70 seeds from each of five barley genotypes) in 1 mm slices were measured by NIR transmission in the range of 2235-2381 nm and oils from the same five barley genotypes were measured in a cuvette with a 1 mm path length in the range of 2003-2497 nm. The spectra of the barley seeds could be classified according to genotypes by principal component analysis; and spectral covariances with reference analysis of moisture, β-glucan, starch, protein and lipid were established. The spectral variations of the barley oils were compared to the fatty acid compositions as measured using gas chromotography-mass spectrometry (GC-MS). PMID:27340221

  14. Hulless winter barley for ethanol production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hulless barley is viable feedstock alternative to corn for ethanol production in areas where small grains are produced. The first barley-based ethanol plant in the US is currently under construction by Osage BioEnergy LLC in Hopewell, VA. New hulless winter barley varieties developed by Virginia T...

  15. Development of endosperm transfer cells in barley

    PubMed Central

    Thiel, Johannes

    2014-01-01

    Endosperm transfer cells (ETCs) are positioned at the intersection of maternal and filial tissues in seeds of cereals and represent a bottleneck for apoplasmic transport of assimilates into the endosperm. Endosperm cellularization starts at the maternal-filial boundary and generates the highly specialized ETCs. During differentiation barley ETCs develop characteristic flange-like wall ingrowths to facilitate effective nutrient transfer. A comprehensive morphological analysis depicted distinct developmental time points in establishment of transfer cell (TC) morphology and revealed intracellular changes possibly associated with cell wall metabolism. Embedded inside the grain, ETCs are barely accessible by manual preparation. To get tissue-specific information about ETC specification and differentiation, laser microdissection (LM)-based methods were used for transcript and metabolite profiling. Transcriptome analysis of ETCs at different developmental stages by microarrays indicated activated gene expression programs related to control of cell proliferation and cell shape, cell wall and carbohydrate metabolism reflecting the morphological changes during early ETC development. Transporter genes reveal distinct expression patterns suggesting a switch from active to passive modes of nutrient uptake with the onset of grain filling. Tissue-specific RNA-seq of the differentiating ETC region from the syncytial stage until functionality in nutrient transfer identified a high number of novel transcripts putatively involved in ETC differentiation. An essential role for two-component signaling (TCS) pathways in ETC development of barley emerged from this analysis. Correlative data provide evidence for abscisic acid and ethylene influences on ETC differentiation and hint at a crosstalk between hormone signal transduction and TCS phosphorelays. Collectively, the data expose a comprehensive view on ETC development, associated pathways and identified candidate genes for ETC

  16. Nanoscale Proteomics

    SciTech Connect

    Shen, Yufeng; Tolic, Nikola; Masselon, Christophe D.; Pasa-Tolic, Liljiana; Camp, David G.; Anderson, Gordon A.; Smith, Richard D.; Lipton, Mary S.

    2004-02-01

    This paper describes efforts to develop a liquid chromatography (LC)/mass spectrometry (MS) technology for ultra-sensitive proteomics studies, i.e. nanoscale proteomics. The approach combines high-efficiency nano-scale LC with advanced MS, including high sensitivity and high resolution Fourier transform ion cyclotron resonance (FTICR) MS, to perform both single-stage MS and tandem MS (MS/MS) proteomic analyses. The technology developed enables large-scale protein identification from nanogram size proteomic samples and characterization of more abundant proteins from sub-picogram size complex samples. Protein identification in such studies using MS is feasible from <75 zeptomole of a protein, and the average proteome measurement throughput is >200 proteins/h and ~3 h/sample. Higher throughput (>1000 proteins/h) and more sensitive detection limits can be obtained using a “accurate mass and time” tag approach developed at our laboratory. These capabilities lay the foundation for studies from single or limited numbers of cells.

  17. Exploring the Plant-Microbe Interface by Profiling the Surface-Associated Proteins of Barley Grains.

    PubMed

    Sultan, Abida; Andersen, Birgit; Svensson, Birte; Finnie, Christine

    2016-04-01

    Cereal grains are colonized by a microbial community that actively interacts with the plant via secretion of various enzymes, hormones, and metabolites. Microorganisms decompose plant tissues by a collection of depolymerizing enzymes, including β-1,4-xylanases, that are in turn inhibited by plant xylanase inhibitors. To gain insight into the importance of the microbial consortia and their interaction with barley grains, we used a combined gel-based (2-DE coupled to MALDI-TOF-TOF MS) and gel-free (LC-MS/MS) proteomics approach complemented with enzyme activity assays to profile the surface-associated proteins and xylanolytic activities of two barley cultivars. The surface-associated proteome was dominated by plant proteins with roles in defense and stress-responses, while the relatively less abundant microbial (bacterial and fungal) proteins were involved in cell-wall and polysaccharide degradation and included xylanases. The surface-associated proteomes showed elevated xylanolytic activity and contained several xylanases. Integration of proteomics with enzyme assays is a powerful tool for analysis and characterization of the interaction between microbial consortia and plants in their natural environment. PMID:26928395

  18. Evaluation of the Effect of Two Volatile Organic Compounds on Barley Pathogens.

    PubMed

    Kaddes, Amine; Parisi, Olivier; Berhal, Chadi; Ben Kaab, Sofiene; Fauconnier, Marie-Laure; Nasraoui, Bouzid; Jijakli, M Haissam; Massart, Sébastien; De Clerck, Caroline

    2016-01-01

    This study aimed to determine the effect of Volatile Organic Compounds (VOCs) on some pathogens, these VOCs were emitted during interactions of barley with Fusarium culmorum Schltdl and/or Cochliobolus sativus Shoemaker, two common root rot pathogens. Our work shows that two organic esters: methyl propanoate (MP) and methyl prop-2-enoate (MA) significantly reduced the development of fungi in vitro. Additional tests showed that the esters significantly inhibited spore germination of these pathogens. The activity of these VOCs on a wide range of fungal and bacterial pathogens was also tested in vitro and showed inhibitory action. The effect of the VOCs on infected barley seeds also showed plantlets growing without disease symptoms. MA and MP seem to have potential value as alternative plant protection compounds against barley bioagressors. PMID:27571054

  19. Mutations in Barley Row Type Genes Have Pleiotropic Effects on Shoot Branching

    PubMed Central

    Liller, Corinna Brit; Neuhaus, René; von Korff, Maria; Koornneef, Maarten; van Esse, Wilma

    2015-01-01

    Cereal crop yield is determined by different yield components such as seed weight, seed number per spike and the tiller number and spikes. Negative correlations between these traits are often attributed to resource limitation. However, recent evidence suggests that the same genes or regulatory modules can regulate both inflorescence branching and tillering. It is therefore important to explore the role of genetic correlations between different yield components in small grain cereals. In this work, we studied pleiotropic effects of row type genes on seed size, seed number per spike, thousand grain weight, and tillering in barley to better understand the genetic correlations between individual yield components. Allelic mutants of nine different row type loci (36 mutants), in the original spring barley varieties Barke, Bonus and Foma and introgressed in the spring barley cultivar Bowman, were phenotyped under greenhouse and outdoor conditions. We identified two main mutant groups characterized by their relationships between seed and tillering parameters. The first group comprises all mutants with an increased number of seeds and significant change in tiller number at early development (group 1a) or reduced tillering only at full maturity (group 1b). Mutants in the second group are characterized by a reduction in seeds per spike and tiller number, thus exhibiting positive correlations between seed and tiller number. Reduced tillering at full maturity (group 1b) is likely due to resource limitations. In contrast, altered tillering at early development (groups 1a and 2) suggests that the same genes or regulatory modules affect inflorescence and shoot branching. Understanding the genetic bases of the trade-offs between these traits is important for the genetic manipulation of individual yield components. PMID:26465604

  20. NASA crop calendars: Wheat, barley, oats, rye, sorghum, soybeans, corn

    NASA Technical Reports Server (NTRS)

    Stuckey, M. R.; Anderson, E. N.

    1975-01-01

    Crop calenders used to determine when Earth Resources Technology Satellite ERTS data would provide the most accurate wheat acreage information and to minimize the amount of ground verified information needed are presented. Since barley, oats, and rye are considered 'confusion crops, i.e., hard to differentiate from wheat in ERTS imagery, specific dates are estimated for these crops in the following stages of development: (1) seed-bed operation, (2) planting or seeding, (3) intermediate growth, (4) dormancy, (5) development of crop to full ground cover, (6) heading or tasseling, and flowering, (7) harvesting, and (8) posting-harvest operations. Dormancy dates are included for fall-snow crops. A synopsis is given of each states' growing conditions, special cropping practices, and other characteristics which are helpful in identifying crops from ERTS imagery.

  1. Jekyll encodes a novel protein involved in the sexual reproduction of barley.

    PubMed

    Radchuk, Volodymyr; Borisjuk, Ljudmilla; Radchuk, Ruslana; Steinbiss, Hans-Henning; Rolletschek, Hardy; Broeders, Sylvia; Wobus, Ulrich

    2006-07-01

    Cereal seed development depends on the intimate interaction of filial and maternal tissues, ensuring nourishment of the new generation. The gene jekyll, which was identified in barley (Hordeum vulgare), is preferentially expressed in the nurse tissues. JEKYLL shares partial similarity with the scorpion Cn4 toxin and is toxic when ectopically expressed in Escherichia coli and tobacco (Nicotiana tabacum). In barley, jekyll is upregulated in cells destined for autolysis. The gene generates a gradient of expression in the nucellar projection, which mediates the maternal-filial interaction during seed filling. Downregulation of jekyll by the RNA interference technique in barley decelerates autolysis and cell differentiation within the nurse tissues. Flower development and seed filling are thereby extended, and the nucellar projection no longer functions as the main transport route for assimilates. A slowing down in the proliferation of endosperm nuclei and a severely impaired ability to accumulate starch in the endosperm leads to the formation of irregular and small-sized seeds at maturity. Overall, JEKYLL plays a decisive role in the differentiation of the nucellar projection and drives the programmed cell death necessary for its proper function. We further suggest that cell autolysis during the differentiation of the nucellar projection allows the optimal provision of basic nutrients for biosynthesis in endosperm and embryo. PMID:16766690

  2. Application of Molecular Genetics and Transformation to Barley Improvement

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter of the new barley monograph summarizes current applications of molecular genetics and transformation to barley improvement. The chapter describes recent applications of molecular markers including association genetics, QTL mapping and marker assisted selection in barley programs, and in...

  3. Differential expression of two ß-amylase genes (Bmy1 and Bmy2) in developing and mature barley grain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley (Hordeum vulgare L.) endosperm-specific (Bmy1) and ubiquitous (Bmy2) ß-amylase were studied during the late maturation phase of seed development in four genotypes. Sequencing of Bmy2 from genomic DNA revealed six polymorphisms in the introns and two synonymous SNPs in the coding region. Acc...

  4. Identification of the differentially accumulated proteins associated with low phosphorus tolerance in a Tibetan wild barley accession.

    PubMed

    Nadira, Umme Aktari; Ahmed, Imrul Mosaddek; Zeng, Jianbin; Wu, Feibo; Zhang, Guoping

    2016-07-01

    Low phosphorus (LP) in soil is a widely-occurred limiting factor for crop production in the world. In a previous study we identified a highly LP-tolerant Tibetan wild barley accession (XZ99). Here, a comparatively proteomic analysis was conducted using three barley genotypes differing in LP tolerance to reveal the mechanisms underlying the LP tolerance of XZ99. Totally, 31 differentially accumulated proteins were identified in the roots and leaves of the three genotypes using 2-dimensional gel electrophoresis coupled with mass spectrometry. They were involved in the various biological processes, including carbon and energy metabolism, signal transduction, cell growth and division, secondary metabolism, and stress defense. In comparison with XZ100 (LP sensitive) and ZD9 (LP moderately-tolerant), XZ99 had a more developed root system, which is mainly attributed to enhanced carbohydrate metabolizing proteins under LP conditions. The current results showed that Tibetan wild barley XZ99 and cultivated barley cultivar ZD9 differ in the mechanism of LP tolerance. The changes of the proteins associated with carbohydrate metabolism could account for the difference between the LP-tolerant and LP-sensitive genotypes. In addition, the mRNA expression levels of 9 LP responsive proteins were verified by qRT-PCR. The current results may open a new avenue of understanding the LP tolerance in plants on the proteomic basis. PMID:27111503

  5. Impact of carbon nanotube exposure to seeds of valuable crops.

    PubMed

    Lahiani, Mohamed H; Dervishi, Enkeleda; Chen, Jihua; Nima, Zeid; Gaume, Alain; Biris, Alexandru S; Khodakovskaya, Mariya V

    2013-08-28

    Multiwalled carbon nanotubes (MWCNTs) affected seed germination, growth, and the development of three important crops (barley, soybean, corn). Early seed germination and activation of growth in exposed seedlings was observed when MWCNTs were added to sterile agar medium. Similarly, seed germination was activated for all tested crop species when MWCNTs were deposited on seed surfaces. The ability of MWCNTs to penetrate the seed coats of corn, barley, and soybean was proven by detection of nanotube agglomerates inside MWCNT-exposed seeds using Raman spectroscopy and transmission electron microscopy (TEM). Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that the expression of genes encoding several types of water channel proteins was increased in soybean, corn, and barley seeds coated with MWCNTs compared with uncoated control seeds. Our results indicate that the positive effect of MWCNTs on the germination and growth of seedlings is reproducible between crop species and can be observed for different methods of delivering carbon nanotubes. Such studies could prove the significant potential of carbon nanotubes as regulators of germination and plant growth. PMID:23834323

  6. Control of stripe rust of spring barley with foliar fungicides, 2009

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The study was conducted in a field with Palouse silt loam near Pullman, WA. Urea (46-0-0) was applied at 60 lb/A at the time of cultivation. Susceptible ‘Morex’ spring barley was seeded in rows spaced 14 in. apart at 60 lb/A (99% germination rate) with a drill planter on 30 Apr 09. Harmony Extra 0.3...

  7. Fuel Ethanol Production from Barley Straw

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley straw used in this study contained 34.1±0.6% cellulose, 22.6±0.4% hemicelluloses, and 13.3±0.2% lignin (moisture, 6.5±0.0%). Several pretreatments (dilute acid, lime, and alkaline peroxide) and enzymatic saccharification procedures were evaluated for the conversion of barley straw to monomer...

  8. New leaf diseases of barley in Egypt.

    PubMed

    Mehiar, F F; El-Deen, E; Wasfy, H; El-Samra, I A

    1976-01-01

    Leaf diseases of barley were observed also in Egypt. From leaves of barley were isolated: Helminthosporium teres, H. gramineum, Stemphylium vesicarium, Alternaria triticina, Vlocladium chartarum, Acnemonium kiliense, Stemphylium spec. accompanied with the Pleospora stage. Inoculations on both attached and detached leaves showed that all the tested fungi were pathogenic, except Acremonium kiliense and Ulocladium chartarum. PMID:1037183

  9. Descriptions of Barley Genetic Stocks For 2007

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Barley Genetics Stocks Database provides information on hundreds of morphological markers. We recently characterized and mapped 27 brachytic (brh) semidwarf mutants in barley. The brachytic lines were evaluated for ten phenotypic traits: height, awn, peduncle, rachis internode length, leaf lengt...

  10. Rebirth and Death: Nitric Oxide and Reactive Oxygen Species in Seeds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Most seeds are dormant at maturity, and dormancy must be lost before germination can occur. The emergence of the embryo and its early growth depend on the coordinated activities of the embryo itself and the tissues surrounding it. We have used Arabidopsis seeds and barley grains to study the roles o...

  11. Barley as a green factory for the production of functional Flt3 ligand.

    PubMed

    Erlendsson, Lýdur S; Muench, Marcus O; Hellman, Ulf; Hrafnkelsdóttir, Soffía M; Jonsson, Anders; Balmer, Yves; Mäntylä, Einar; Orvar, Björn L

    2010-02-01

    Biologically active recombinant human Flt3 ligand was expressed and isolated from transgenic barley seeds. Its expression is controlled by a tissue specific promoter that confines accumulation of the recombinant protein to the endosperm tissue of the seed. The recombinant Flt3 ligand variant expressed in the seeds contains an HQ-tag for affinity purification on immobilized metal ion affinity chromatography (IMAC) resin. The tagged protein was purified from seed extracts to near homogeneity using sequential chromatography on IMAC affinity resin and cation exchange resin. We also show that the recombinant Flt3 ligand protein undergoes posttranslational modifications: it is a glycoprotein containing alpha-1,3-fucose and alpha-1,2-xylose. The HQ-tagged Flt3 ligand variant exhibits comparable biological activity to commercial Flt3 ligand. This is the first report showing expression and accumulation of recombinant human growth factor in barley seeds with a yield of active protein similar to a bacterial expression system. The present results demonstrate that plant molecular farming is a viable approach for the bioproduction of human-derived growth factors. PMID:19844912

  12. New Starch Phenotypes Produced by TILLING in Barley

    PubMed Central

    Sparla, Francesca; Falini, Giuseppe; Botticella, Ermelinda; Pirone, Claudia; Talamè, Valentina; Bovina, Riccardo; Salvi, Silvio; Tuberosa, Roberto; Sestili, Francesco; Trost, Paolo

    2014-01-01

    Barley grain starch is formed by amylose and amylopectin in a 1∶3 ratio, and is packed into granules of different dimensions. The distribution of granule dimension is bimodal, with a majority of small spherical B-granules and a smaller amount of large discoidal A-granules containing the majority of the starch. Starch granules are semi-crystalline structures with characteristic X-ray diffraction patterns. Distinct features of starch granules are controlled by different enzymes and are relevant for nutritional value or industrial applications. Here, the Targeting-Induced Local Lesions IN Genomes (TILLING) approach was applied on the barley TILLMore TILLING population to identify 29 new alleles in five genes related to starch metabolism known to be expressed in the endosperm during grain filling: BMY1 (Beta-amylase 1), GBSSI (Granule Bound Starch Synthase I), LDA1 (Limit Dextrinase 1), SSI (Starch Synthase I), SSIIa (Starch Synthase IIa). Reserve starch of nine M3 mutant lines carrying missense or nonsense mutations was analysed for granule size, crystallinity and amylose/amylopectin content. Seven mutant lines presented starches with different features in respect to the wild-type: (i) a mutant line with a missense mutation in GBSSI showed a 4-fold reduced amylose/amylopectin ratio; (ii) a missense mutations in SSI resulted in 2-fold increase in A:B granule ratio; (iii) a nonsense mutation in SSIIa was associated with shrunken seeds with a 2-fold increased amylose/amylopectin ratio and different type of crystal packing in the granule; (iv) the remaining four missense mutations suggested a role of LDA1 in granule initiation, and of SSIIa in determining the size of A-granules. We demonstrate the feasibility of the TILLING approach to identify new alleles in genes related to starch metabolism in barley. Based on their novel physicochemical properties, some of the identified new mutations may have nutritional and/or industrial applications. PMID:25271438

  13. Calcium homeostasis in barley aleurone

    SciTech Connect

    Jones, R.L.

    1990-02-21

    Under the auspices of the Department of Energy we investigated calcium homeostasis in aleurone cells of barley. This investigation was initiated to explore the role played by extracellular Ca{sup 2+} in gibberellic acid (GA)-induced synthesis and secretion of hydrolases in the aleurone layer. We have focused our attention on four topics that relate to the role of Ca{sup 2+} in regulating the synthesis of {alpha}-amylase. First, we determined the stoichiometry of Ca{sup 2+} binding to the two principal classes of barley {alpha}-amylase and examined some of the biochemical and physical properties of the native and Ca{sup 2+}-depleted forms of the enzyme. Second, since {alpha}-amylase is a Ca{sup 2+} containing metalloenzyme that binds one atom of Ca{sup 2+} per molecule, we developed methods to determine the concentration of Ca{sup 2+} in the cytosol of the aleurone cell. We developed a technique for introducing Ca{sup 2+}-sensitive dyes into aleurone protoplasts that allows the measurement of Ca{sup 2+} in both cytosol and endoplasmic reticulum (ER). Third, because the results of our Ca{sup 2+} measurements showed higher levels of Ca{sup 2+} in the ER than in the cytosol, we examined Ca{sup 2+} transport into the ER of control and GA-treated aleurone tissue. And fourth, we applied the technique of patch-clamping to the barley aleurone protoplast to examine ion transport at the plasma membrane. Our results with the patch-clamp technique established the presence of K{sup +} channels in the plasma membrane of the aleurone protoplast, and they showed that this cell is ideally suited for the application of this methodology for studying ion transport. 34 refs.

  14. Lunasin in cereal seeds: What is the origin?

    PubMed

    Mitchell, Rowan A C; Lovegrove, Alison; Shewry, Peter R

    2013-05-01

    Lunasin is a peptide from soybean seeds which has been demonstrated to have anticancer properties. It has also been reported in cereal seeds: wheat, rye, barley and Triticale. However, extensive searches of transcriptome and DNA sequence databases for wheat and other cereals have failed to identify sequences encoding either the lunasin peptide or a precursor protein. This raises the question of the origin of the lunasin reported in cereal grain. PMID:24817784

  15. Proteomic patterns associated with heterosis.

    PubMed

    Xing, Jiewen; Sun, Qixin; Ni, Zhongfu

    2016-08-01

    Heterosis is characterized by higher seed yields, plant biomass or other traits in heterozygotes or hybrids compared with their genetically divergent parents, which are often homozygous. Despite extensive investigation of heterosis and its wide application in crops such as maize, rice, wheat and sorghum, its molecular basis is still enigmatic. In the past century, some pioneers have proposed multigene models referring to the complementation of allelic and gene expression variation, which is likely to be an important contributor to heterosis. In addition, there are potential interactions of epigenetic variation involved in heterosis via novel mechanisms. At the level of gene expression, many recent studies have revealed that the heterosis phenomenon can be deciphered not only at the transcriptional level but also at the proteomic level. This review presents an update on the information supporting the involvement of proteomic patterns in heterosis and a possible future direction of the field. This article is part of a Special Issue entitled: Plant Proteomics - a bridge between fundamental processes and crop production, edited by Dr. Hans-Peter Mock. PMID:26721744

  16. Seed thioredoxin h.

    PubMed

    Hägglund, Per; Finnie, Christine; Yano, Hiroyuki; Shahpiri, Azar; Buchanan, Bob B; Henriksen, Anette; Svensson, Birte

    2016-08-01

    Thioredoxins are nearly ubiquitous disulfide reductases involved in a wide range of biochemical pathways in various biological systems, and also implicated in numerous biotechnological applications. Plants uniquely synthesize an array of thioredoxins targeted to different cell compartments, for example chloroplastic f- and m-type thioredoxins involved in regulation of the Calvin-Benson cycle. The cytosolic h-type thioredoxins act as key regulators of seed germination and are recycled by NADPH-dependent thioredoxin reductase. The present review on thioredoxin h systems in plant seeds focuses on occurrence, reaction mechanisms, specificity, target protein identification, three-dimensional structure and various applications. The aim is to provide a general background as well as an update covering the most recent findings. This article is part of a Special Issue entitled: Plant Proteomics--a bridge between fundamental processes and crop production, edited by Dr. Hans-Peter Mock. PMID:26876537

  17. Comparative transcriptional and proteomic profiling of bread wheat cultivar and its derived transgenic line over-expressing a low molecular weight glutenin subunit gene in the endosperm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have carried out a parallel transcriptional and proteomic comparison of seeds from a transformed bread wheat line that over-expresses a transgenic low molecular weight glutenin subunit gene relative to the corresponding non-transformed genotype. Proteomic analyses showed that, during seed develop...

  18. Seed storage at elevated partial pressure of oxygen, a fast method for analysing seed ageing under dry conditions

    PubMed Central

    Groot, S. P. C.; Surki, A. A.; de Vos, R. C. H.; Kodde, J.

    2012-01-01

    Background and Aims Despite differences in physiology between dry and relative moist seeds, seed ageing tests most often use a temperature and seed moisture level that are higher than during dry storage used in commercial practice and gene banks. This study aimed to test whether seed ageing under dry conditions can be accelerated by storing under high-pressure oxygen. Methods Dry barley (Hordeum vulgare), cabbage (Brassica oleracea), lettuce (Lactuca sativa) and soybean (Glycine max) seeds were stored between 2 and 7 weeks in steel tanks under 18 MPa partial pressure of oxygen. Storage under high-pressure nitrogen gas or under ambient air pressure served as controls. The method was compared with storage at 45 °C after equilibration at 85 % relative humidity and long-term storage at the laboratory bench. Germination behaviour, seedling morphology and tocopherol levels were assessed. Key Results The ageing of the dry seeds was indeed accelerated by storing under high-pressure oxygen. The morphological ageing symptoms of the stored seeds resembled those observed after ageing under long-term dry storage conditions. Barley appeared more tolerant of this storage treatment compared with lettuce and soybean. Less-mature harvested cabbage seeds were more sensitive, as was the case for primed compared with non-primed lettuce seeds. Under high-pressure oxygen storage the tocopherol levels of dry seeds decreased, in a linear way with the decline in seed germination, but remained unchanged in seeds deteriorated during storage at 45 °C after equilibration at 85 % RH. Conclusions Seed storage under high-pressure oxygen offers a novel and relatively fast method to study the physiology and biochemistry of seed ageing at different seed moisture levels and temperatures, including those that are representative of the dry storage conditions as used in gene banks and commercial practice. PMID:22967856

  19. Proteomics, peptidomics, and immunogenic potential of wheat beer (Weissbier).

    PubMed

    Picariello, Gianluca; Mamone, Gianfranco; Cutignano, Adele; Fontana, Angelo; Zurlo, Lucia; Addeo, Francesco; Ferranti, Pasquale

    2015-04-01

    Wheat beer is a traditional light-colored top-fermenting beer brewed with at least 50% malted (e.g., German Weissbier) or unmalted (e.g., Belgian Witbier) wheat (Triticum aestivum) as an adjunct to barley (Hordeum vulgare) malt. For the first time, we explored the proteome of three Weissbier samples, using both 2D electrophoresis (2DE)-based and 2DE-free strategies. Overall, 58 different gene products arising from barley, wheat, and yeast (Saccharomyces spp.) were identified in the protein fraction of a representative Weissbier sample analyzed in detail. Analogous to all-barley-malt beers (BMB), barley and wheat Z-type serpins and nonspecific lipid transfer proteins dominated the proteome of Weissbier. Several α-amylase/trypsin inhibitors also survived the harsh brewing conditions. During brewing, hundreds of peptides are released into beer. By liquid chromatography-electrospray tandem mass spectrometry (LC-ESI MS/MS) analysis, we characterized 167 peptides belonging to 44 proteins, including gliadins, hordeins, and high- and low-molecular-weight glutenin subunits. Because of the interference from the overabundant yeast-derived peptides, we identified only a limited number of epitopes potentially triggering celiac disease. However, Weissbier samples contained 374, 372, and 382 ppm gliadin-equivalent peptides, as determined with the competitive G12 ELISA, which is roughly 10-fold higher than a lager BMB (41 ppm), thereby confirming that Weissbier is unsuited for celiacs. Western blot analysis demonstrated that Weissbier also contained large-sized prolamins immunoresponsive to antigliadin IgA antibodies from the pooled sera of celiac patients (n = 4). PMID:25793656

  20. Supplementing barley or rapeseed meal to dairy cows fed grass-red clover silage: I. Rumen degradability and microbial flow.

    PubMed

    Ahvenjärvi, S; Vanhatalo, A; Huhtanen, P

    2002-08-01

    The present study was conducted to measure the flow of microbial and nonmicrobial N fractions entering the omasal canal of lactating dairy cows fed grass-red clover silage supplemented with barley and rapeseed meal. Four ruminally cannulated Finnish Ayrshire dairy cows were fed, in a 4 x 4 Latin square design, grass-red clover silage alone or supplemented with (on DM basis) 5.1 kg/d of barley, 1.9 kg/d of rape-seed meal or 5.1 kg/d of barley and 1.9 kg/d rapeseed meal. Nonammonia N flow entering the omasal canal was fractionated into microbial and nonmicrobial N using 15N. Microbial N was fractionated into N associated with liquid-associated bacteria, particle-associated bacteria, and protozoa. Supplementation of diets with barley increased microbial N flow entering the omasal canal (P < 0.01) but had no effect on nonmicrobial N flow. Increased microbial N flow was attributed to liquid-associated bacteria and protozoa. Barley had no effect on apparent ruminal N degradability, but increased true ruminal N degradability (P < 0.01). Barley had no effect on urinary N excretion, but increased daily N retention (P = 0.03). Furthermore, barley supplementation decreased ruminal (P = 0.02) and total tract (P < 0.01) NDF digestibility. Supplementation of diets with rapeseed meal increased apparent ruminal N degradability (P < 0.01) and nonmicrobial N flow entering the omasal canal (P < 0.01), but had no effect on true ruminal N degradability. Despite higher N excretion in urine, rapeseed meal improved daily N retention (P < 0.01). Milk yield was increased (P < 0.01) by barley and rapeseed meal supplements, with the responses being additive. Responses attained with barley were primarily due to increased energy supply for ruminal microbes and improvements in energy and protein supply for the animal. However, provision of readily digestible carbohydrates in barley did not improve microbial capture of ruminal ammonia. Benefits associated with rapeseed meal supplementation were

  1. Seed Germination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Initiation of seed germination is a critical decision for plants. It is important for seed populations under natural conditions to spread the timing of germination of individual seeds to maximize the probability of species survival. Therefore, seeds have evolved the multiple layers of mechanisms tha...

  2. Polyploidy and the proteome.

    PubMed

    Soltis, Douglas E; Misra, Biswapriya B; Shan, Shengchen; Chen, Sixue; Soltis, Pamela S

    2016-08-01

    Although major advances have been made during the past 20 years in our understanding of the genetic and genomic consequences of polyploidy, our knowledge of polyploidy and the proteome is in its infancy. One of our goals is to stimulate additional study, particularly broad-scale proteomic analyses of polyploids and their progenitors. Although it may be too early to generalize regarding the extent to which transcriptomic data are predictive of the proteome of polyploids, it is clear that the proteome does not always reflect the transcriptome. Despite limited data, important observations on the proteomes of polyploids are emerging. In some cases, proteomic profiles show qualitatively and/or quantitatively non-additive patterns, and proteomic novelty has been observed. Allopolyploids generally combine the parental contributions, but there is evidence of parental dominance of one contributing genome in some allopolyploids. Autopolyploids are typically qualitatively identical to but quantitatively different from their parents. There is also evidence of parental legacy at the proteomic level. Proteomes clearly provide insights into the consequences of genomic merger and doubling beyond what is obtained from genomic and/or transcriptomic data. Translating proteomic changes in polyploids to differences in morphology and physiology remains the holy grail of polyploidy--this daunting task of linking genotype to proteome to phenotype should emerge as a focus of polyploidy research in the next decade. This article is part of a Special Issue entitled: Plant Proteomics--a bridge between fundamental processes and crop production, edited by Dr. Hans-Peter Mock. PMID:26993527

  3. Differential RNA Expression of Two Barley ß-Amylase Genes (Bmy1 and Bmy2) in Developing Grains and Their Association with ß-Amylase Activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    RNA expression from the barley ß-amylase1 (Bmy1) gene was determined during seed development in four genotypes (Legacy, Harrington, Ashqelon, and PI 296897). The Bmy1 transcript amount in Legacy and Harrington was not significantly different at 17, 19, or 21 days after anthesis (DAA). Ashqelon Bmy...

  4. Water mobility in the endosperm of high beta-glucan barley mutants as studied by nuclear magnetic resonance imaging.

    PubMed

    Fast Seefeldt, Helene; van den Berg, Frans; Köckenberger, Walter; Engelsen, Søren Balling; Wollenweber, Bernd

    2007-04-01

    (1)H NMR imaging (MRI) was used as a noninvasive technique to study water distribution and mobility in hydrated barley (Hordeum vulgare L.) seeds of accessions with varying content of beta glucan (BG), a highly hygroscopic cell wall component. High contents of BG in barley are unfavorable in malting where it leads to clotting of filters and hazing of beer as well as in animal feed where it hinders the rapid uptake of energy. However, a high content of BG has a positive nutritional effect, as it lowers the cholesterol and the glycaemic index. It was studied whether water distribution and mobility were related to content and location of BG. Water mobility was investigated by following the rate and mode of desiccation in hydrated single seeds. In order to determine the different water components, a multispin echo experiment was set up to reveal the T(2) transverse relaxation rates of water within the seeds. A principal component analysis (PCA) discriminated control seeds from the high-BG mutant seeds. MRI proved efficient in tracing the differences in water-holding capacity of contrasting barley seeds. All accessions showed nonuniform distribution of water at full hydration as well as during desiccation. The embryo retained water even after 36 h of drying, whereas the endosperm showed low and heterogeneous mobility of the water after drying. The relaxation time constants indicated that the BG mutants had regions of much higher water mobility around the ventral crease compared to the control. It is concluded that MRI can be applied to investigate temporal and spatial differences in the location of specific chemical compounds in single seeds. PMID:17371735

  5. Drought acclimation in wild and cultivated barley lines. [Hordeum spontaneum; Hordeum vulgare

    SciTech Connect

    Glinka, Z. ); Gunasekera, D.; Mane, S.; Berkowitz, G. )

    1991-05-01

    Wild barley (Hordeum spontaneum) seeds collected from arid and temperate regions in Israel were used, along with cultivated barley (H. vulgare) in a study to evaluate the range of acclimation responses to low leaf water potential ({Psi}w). Stress was imposed on plants by withholding water until {Psi}w was {minus}2 megapascals (MPa). Protoplast volume (PV) was measured at {minus}0.2 and {minus}2 MPa (imposed in vitro) in leaf tissue from well-watered and stressed plants. In well-watered plants, PV declined at {minus}2, as compared to {minus}0.2 MPa in all lines. With tissue from in situ stressed plants, PV reduction at {minus}2 MPa was not as great in some lines. The change in the extent of PV reduction occurring at {minus}2 MPa was used as an index of drought acclimation. The 13 wild barley lines were separated into high, medium, and low acclimation groups. Lines collected from arid regions scored in the high acclimation group. The cultivated barley lines scored in the medium and low groups. Relative water content decline at low leaf {Psi}w in situ was not a good indicator of acclimation; all lines responded similarly. Photosynthesis in situ was measured at high and low leaf {Psi}w in lines from the three groupings. Photosynthetic sensitivity to low {Psi}w was twice as great in low acclimation, as compared to high acclimation lines. It was concluded that PV response to low {Psi}w is a good indicator of drought acclimation in barley, and that wild lines offer a range of acclimation potential which could be used in breeding programs.

  6. The Staphylococcus aureus proteome.

    PubMed

    Otto, Andreas; van Dijl, Jan Maarten; Hecker, Michael; Becher, Dörte

    2014-03-01

    Staphylococcus aureus is a Gram-positive commensal bacterium that is regarded as a major threat for modern health care systems. This relates both to the ability of S. aureus to overcome antibiotic therapy by developing high-level resistance against multiple antibiotics and this bacterium's extensive arsenal of virulence factors. Understanding the mechanisms of resistance and functional studies on stress and starvation responses are the main goals of proteomics in staphylococcal research. This review high-lights recent advances in gel-based and gel-free proteomics analyses of S. aureus and pinpoints the importance of location-specific proteomics studies targeting the cytosol, the membrane, the cell surface and the extracellular milieu in combination with integrated global proteome studies. Emerging hot topics in staphylococcal proteomics are discussed with special focus on in vivo proteomics, membrane vesicles, biofilm formation and the acquisition of absolute proteome data for systems biological modeling approaches. PMID:24439828

  7. 7 CFR 810.204 - Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... barley and Six-rowed Blue Malting barley. 810.204 Section 810.204 Agriculture Regulations of the... Standards for Barley Principles Governing the Application of Standards § 810.204 Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley. Grade Minimum limits of—...

  8. 7 CFR 810.204 - Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... barley and Six-rowed Blue Malting barley. 810.204 Section 810.204 Agriculture Regulations of the... Standards for Barley Principles Governing the Application of Standards § 810.204 Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley. Grade Minimum limits of—...

  9. 7 CFR 810.204 - Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... barley and Six-rowed Blue Malting barley. 810.204 Section 810.204 Agriculture Regulations of the... Standards for Barley Principles Governing the Application of Standards § 810.204 Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley. Grade Minimum limits of—...

  10. 7 CFR 810.204 - Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... barley and Six-rowed Blue Malting barley. 810.204 Section 810.204 Agriculture Regulations of the... Standards for Barley Principles Governing the Application of Standards § 810.204 Grades and grade requirements for Six-rowed Malting barley and Six-rowed Blue Malting barley. Grade Minimum limits of—...