WorldWideScience

Sample records for barley mlo protein

  1. Interaction between powdery mildew and barley with ¤mlo5¤ mildew resistance

    DEFF Research Database (Denmark)

    Lyngkjær, M.F.; Østergård, Hanne

    1998-01-01

    Powdery mildew infection of barley with the mlo5 barley powdery mildew resistance gene was examined, using near-isogenic barley lines, with and without mlo5 resistance, and two near-isogenic powdery mildew isolates, HL3/5 and GE3 with high (virulent) or low (avirulent) penetration efficiency...

  2. The enhanced callose deposition in barley with ml-o powdery mildew resistance genes

    DEFF Research Database (Denmark)

    Skou, Jens-Peder

    1985-01-01

    Carborundum treatment of barley leaves induced a callose deposition which was detected as diffuse blotches in the epidermal cells of susceptible barleys and as deeply stained tracks along the scratches in barleys with the ml-o powdery mildew resistance gene. Subsequent inoculation with powdery...... mildew resulted in appositions that enlarged inversely to their size in the respective varieties when inoculated without carborundum treatment. Aphids sucking the leaves resulted in rows of callose containing spots along the anticlinal cell walls. The spots were larger in the ml-o mutant than...... in the mother variety. Callose was deposited in connection with the pleiotropic necrotic spotting in barleys with the ml-o gene. Modification of the necrotic spotting by crossing the ml-o gene into other gene backgrounds did not result in any change in the size of appositions upon inoculation with powdery...

  3. Tempered mlo broad-spectrum resistance to barley powdery mildew in an Ethiopian landrace.

    Science.gov (United States)

    Ge, Xintian; Deng, Weiwei; Lee, Zheng Zhou; Lopez-Ruiz, Francisco J; Schweizer, Patrick; Ellwood, Simon R

    2016-07-12

    Recessive mutations in the Mlo gene confer broad spectrum resistance in barley (Hordeum vulgare) to powdery mildew (Blumeria graminis f. sp. hordei), a widespread and damaging disease. However, all alleles discovered to date also display deleterious pleiotropic effects, including the naturally occurring mlo-11 mutant which is widely deployed in Europe. Recessive resistance was discovered in Eth295, an Ethiopian landrace, which was developmentally controlled and quantitative without spontaneous cell wall appositions or extensive necrosis and loss of photosynthetic tissue. This resistance is determined by two copies of the mlo-11 repeat units, that occur upstream to the wild-type Mlo gene, compared to 11-12 in commonly grown cultivars and was designated mlo-11 (cnv2). mlo-11 repeat unit copy number-dependent DNA methylation corresponded with cytological and macroscopic phenotypic differences between copy number variants. Sequence data indicated mlo-11 (cnv2) formed via recombination between progenitor mlo-11 repeat units and the 3' end of an adjacent stowaway MITE containing region. mlo-11 (cnv2) is the only example of a moderated mlo variant discovered to date and may have arisen by natural selection against the deleterious effects of the progenitor mlo-11 repeat unit configuration.

  4. A Japanese powdery mildew isolate with exceptionally large infection efficience on Mlo-resistant barley

    DEFF Research Database (Denmark)

    Lyngkjær, M.F.; Jensen, H.P.; Østergård, Hanne

    1995-01-01

    A Japanese field isolate (Race I) of Erysiphe graminis f,sp. hordei was tested on 17 barley lines carrying the mlo powdery mildew resistance gene. Race I produced many successful infections with infection type larger than or equal to 2 on six lines (M66, MC20, SR1, SR7, Atem and Totem). On the re......A Japanese field isolate (Race I) of Erysiphe graminis f,sp. hordei was tested on 17 barley lines carrying the mlo powdery mildew resistance gene. Race I produced many successful infections with infection type larger than or equal to 2 on six lines (M66, MC20, SR1, SR7, Atem and Totem...

  5. Suppression of resistance to Erysiphe graminis f.sp. hordei conferred by the mlo5 barley powdery mildew resistance gene

    DEFF Research Database (Denmark)

    Lyngkjær, M.F.; Carver, T.L.W.; Zeyen, R.J.

    1997-01-01

    . Additional suppression of mlo5 penetration resistance against the avirulent E. graminis isolate was achieved by using DDG, mannose, or glucose in combination with the phenylalanine ammonia lyase inhibitor alpha-aminooxy-beta-phenylpropionic acid (AOPP). A mlo virulent isolate of E. graminis was also tested...... phenolic compound synthesis, but that phenolics are not responsible for the primary mechanism of mlo5 penetration resistance. Sequestration of phosphate ions caused by complexing with DDG or mannose may lower the energy available for penetration resistance in these barley lines, obviating both inherent...

  6. A Compromised Mlo Pathway Affects the Response of Barley to the Necrotrophic Fungus Bipolaris sorokiniana (Teleomorph: Cochliobolus sativus) and Its Toxins.

    Science.gov (United States)

    Kumar, J; Hückelhoven, R; Beckhove, U; Nagarajan, S; Kogel, K H

    2001-02-01

    ABSTRACT In search of new durable disease resistance traits in barley to control leaf spot blotch disease caused by the necrotrophic fungus Bipolaris sorokiniana (teleomorph: Cochliobolus sativus), we developed macroscopic and microscopic scales to judge spot blotch disease development on barley. Infection of barley was associated with cell wall penetration and accumulation of hydrogen peroxide. The latter appeared to take place in cell wall swellings under fungal penetration attempts as well as during cell death provoked by the necrotrophic pathogen. Additionally, we tested the influence of a compromised Mlo pathway that confers broad resistance against powdery mildew fungus (Blumeria graminis f. sp. hordei). Powdery mildew-resistant genotypes with mutations at the Mlo locus (mlo genotypes) showed a higher sensitivity to infiltration of toxic culture filtrate of Bipolaris sorokiniana as compared with wild-type barley. Mutants defective in Ror, a gene required for mlo-specified powdery mildew resistance, were also more sensitive to Bipolaris sorokiniana toxins than wild-type barley but showed less symptoms than mlo5 parents. Fungal culture filtrates induced an H2O2 burst in all mutants, whereas wild-type (Mlo) barley was less sensitive. The results support the hypothesis that the barley Mlo gene product functions as a suppresser of cell death. Therefore, a compromised Mlo pathway is effective for control of biotrophic powdery mildew fungus but not for necrotrophic Bipolaris sorokiniana. We discuss the problem of finding resistance traits that are effective against both biotrophic and necrotrophic pathogens with emphasis on the role of the anti-oxidative system of plant cells.

  7. Comparative analysis of genome-wide Mlo gene family in Cajanus cajan and Phaseolus vulgaris.

    Science.gov (United States)

    Deshmukh, Reena; Singh, V K; Singh, B D

    2016-04-01

    The Mlo gene was discovered in barley because the mutant 'mlo' allele conferred broad-spectrum, non-race-specific resistance to powdery mildew caused by Blumeria graminis f. sp. hordei. The Mlo genes also play important roles in growth and development of plants, and in responses to biotic and abiotic stresses. The Mlo gene family has been characterized in several crop species, but only a single legume species, soybean (Glycine max L.), has been investigated so far. The present report describes in silico identification of 18 CcMlo and 20 PvMlo genes in the important legume crops Cajanus cajan (L.) Millsp. and Phaseolus vulgaris L., respectively. In silico analysis of gene organization, protein properties and conserved domains revealed that the C. cajan and P. vulgaris Mlo gene paralogs are more divergent from each other than from their orthologous pairs. The comparative phylogenetic analysis classified CcMlo and PvMlo genes into three major clades. A comparative analysis of CcMlo and PvMlo proteins with the G. max Mlo proteins indicated close association of one CcMlo, one PvMlo with two GmMlo genes, indicating that there was no further expansion of the Mlo gene family after the separation of these species. Thus, most of the diploid species of eudicots might be expected to contain 15-20 Mlo genes. The genes CcMlo12 and 14, and PvMlo11 and 12 are predicted to participate in powdery mildew resistance. If this prediction were verified, these genes could be targeted by TILLING or CRISPR to isolate powdery mildew resistant mutants.

  8. Calcium transport in protoplasts isolated from ml-o barley isolines resistant and susceptible to powdery mildew. [Hordeum vulgare L

    Energy Technology Data Exchange (ETDEWEB)

    Wrona, A.F.; Spanswick, R.M.; Aist, J.R. (Cornell Univ., Ithaca, NY (USA))

    1988-12-01

    Free cytoplasmic calcium has been postulated to play a role in preventing powdery mildew in a series of homozygous ml-o mutants of barley, Hordeum vulgare L. Protoplasts isolated from 7-day-old plants of the ml-o resistant-susceptible (R-S) barley isolines, Riso 5678/3* {times} Carlsberg II R and S, were used to test for differences in fluxes of Ca{sup 2+} across the plasmalemma. Greater influx or lesser efflux might account for a higher free cytosolic Ca{sup 2+} postulated to exist in ml-o R mutants. Uniform patterns of uptake were maintained for 3 hours from solutions of 0.2 and 2 millimolar Ca{sup 2+}. Washout curves of {sup 45}Ca{sup 2+} from R and S protoplasts revealed three compartments - presumed to represent release from the vacuole, organelles, and the cytoplasm (which included bound as well as free Ca{sup 2+}). Uptake and washout did not differ between isolines. On the basis of recent determinations of submicromolar levels of free cytoplasmic Ca{sup 2+} and their initial rates of {sup 45}ca-labeled Ca{sup 2+} uptake, they show that measurement of the unidirectional influx of Ca{sup 2+} across the plasmalemma is not feasible because the specific activity of the pool of free cytoplasmic calcium increases almost instantaneously to a level that would result in a significant, but unknown, efflux of label. Similarly, measurement of the efflux of Ca{sup 2+} across the plasmalemma is not possible since the activity of the pool of free cytoplasmic calcium is a factor of 350 smaller than the most rapid component of the washout experiment. This pool of cytoplasmic free Ca{sup 2+} will wash out too rapidly and be too small to detect under the conditions of these experiments.

  9. Colonization of Barley by the Broad-Host Hemibiotrophic Pathogen Phytophthora palmivora Uncovers a Leaf Development-Dependent Involvement of Mlo.

    Science.gov (United States)

    Le Fevre, Ruth; O'Boyle, Bridget; Moscou, Matthew J; Schornack, Sebastian

    2016-05-01

    The discovery of barley Mlo demonstrated that filamentous pathogens rely on plant genes to achieve entry and lifecycle completion in barley leaves. While having a dramatic effect on foliar pathogens, it is unclear whether overlapping or distinct mechanisms affect filamentous pathogen infection of roots. To remove the bias connected with using different pathogens to understand colonization mechanisms in different tissues, we have utilized the aggressive hemibiotrophic oomycete pathogen Phytophthora palmivora. P. palmivora colonizes root as well as leaf tissues of barley (Hordeum vulgare). The infection is characterized by a transient biotrophy phase with formation of haustoria. Barley accessions varied in degree of susceptibility, with some accessions fully resistant to leaf infection. Notably, there was no overall correlation between degree of susceptibility in roots compared with leaves, suggesting that variation in different genes influences host susceptibility above and below ground. In addition, a developmental gradient influenced infection, with more extensive colonization observed in mature leaf sectors. The mlo5 mutation attenuates P. palmivora infection but only in young leaf tissues. The barley-P. palmivora interaction represents a simple system to identify and compare genetic components governing quantitative colonization in diverse barley tissue types.

  10. Cloning, Physical Mapping and Expression Analysis of a Wheat Mlo-like Gene

    Institute of Scientific and Technical Information of China (English)

    Ling YU; Ji-Shan NIU; Pei-Du CHEN; Zheng-Qiang MA; Da-Jun LIU

    2005-01-01

    Reverse transcription-polymerase chain reaction (RT-PCR) was performed to isolate a cDNA clone using specific primers designed based on the barley (Hordeum vulgare L.) Mlo gene cDNA sequence.A full-length cDNA encoding an Mlo-like protein was isolated and characterized in a Triticum aestivum L.-Haynaldia villosa L. 6VS/6AL translocation line. The putative protein consists of 534 amino acid residues,which contain a nuclear localization motif (NLS), nine casein kinase Ⅱ motifs (S/T-X-X-D/E) and seven protein kinase C motifs (S/T-X-R/K). It is highly homologous to other plant Mlo proteins. Thus, this clone was designated as Ta-Mlo (GenBank accession No. AF384144). Northern blotting analysis showed that the transcription of Ta-Mlo was enhanced slightly by Blumeria graminis (DC) EO Speer f. sp. tritici. Western blotting analysis showed that the protein expression product of the Ta-Mlo gene in wheat seedling leaves is a membrane-bound protein. The protein could be induced by B. graminis. Southern blotting analysis indicated that there is one copy of the Ta-Mlo gene in each wheat genome. Ta-Mlo was localized on specific chromosomal regions of 2AL, 2BL, and 2DL in wheat.

  11. Inhibition of ABCA1 Protein Expression and Cholesterol Efflux by TNF α in MLO-Y4 Osteocytes.

    Science.gov (United States)

    Wehmeier, Kent R; Kurban, William; Chandrasekharan, Chandrikha; Onstead-Haas, Luisa; Mooradian, Arshag D; Haas, Michael J

    2016-06-01

    Hip fracture and myocardial infarction cause significant morbidity and mortality. In vivo studies raising serum cholesterol levels as well as pro-inflammatory cytokines such as TNF α manifest bone loss and atherosclerotic vascular disease, suggesting that abnormalities of cholesterol transport may contribute to osteoporosis. We used the mouse osteocyte cell line (MLO-Y4) to investigate the effects of TNF α on the expression of cholesterol acceptor proteins such as apolipoprotein A-I (apo A-I) and apolipoprotein E (apo E), as well as on the cholesterol transporters ATP-binding cassette-1 (ABCA1), scavenger receptor class B type 1 (SRB1), and cluster of differentiation 36 (CD36). MLO-Y4 cells do not express apo A-I or apo E; however, they do express all three cholesterol transporters (ABCA1, SRB1, and CD36). Treatment of MLO-Y4 cells with TNF α had no effect on SRB1, CD36, and osteocalcin levels; however, TNF α reduced ABCA1 protein levels in a dose-dependent manner and cholesterol efflux to apo A-I. Interestingly, TNF α treatment increased ABCA1 promoter activity and ABCA1 mRNA levels, and increased liver X receptor α protein expression, but had no effect on retinoid X receptor α and retinoic acid receptor α levels. Pharmacological inhibition of p38 mitogen-activated protein (MAP) kinase, but not c-jun-N-terminal kinase 1 or mitogen-activated protein kinase (MEK), restored ABCA1 protein levels in TNF α-treated cells. These results suggest that pro-inflammatory cytokines regulate cholesterol metabolism in osteocytes in part by suppressing ABCA1 levels post-translationally in a p38 MAP kinase-dependent manner.

  12. Host cell entry of powdery mildew is correlated with endosomal transport of antagonistically acting VvPEN1 and VvMLO to the papilla.

    Science.gov (United States)

    Feechan, A; Jermakow, A M; Ivancevic, A; Godfrey, D; Pak, H; Panstruga, R; Dry, I B

    2013-10-01

    Challenge by a nonadapted powdery mildew fungal pathogen leads to the formation of a local cell-wall apposition (papilla) beneath the point of attempted penetration. Several plasma membrane (PM) proteins with opposing roles in powdery mildew infection, including Arabidopsis thaliana PENETRATION1 (PEN1) and barley (Hordeum vulgare) MILDEW RESISTANCE LOCUS O (MLO), are localized to the site of powdery mildew attack. PEN1 contributes to penetration resistance to nonadapted powdery mildews, whereas MLO is a susceptibility factor required by adapted powdery mildew pathogens for host cell entry. Our previous studies have demonstrated that the vesicle and endosomal trafficking inhibitors, brefeldin A and wortmannin, have opposite effects on the penetration rates of adapted and nonadapted powdery mildews on grapevine. These findings prompted us to study the pathogen-induced intracellular trafficking of grapevine variants of MLO and PEN1. We first identified grapevine (Vitis vinifera) VvPEN1 and VvMLO orthologs that rescue Arabidopsis Atpen1 and Atmlo2 mlo6 mlo12 null mutants, respectively. By using endomembrane trafficking inhibitors in combination with fluorescence microscopy, we demonstrate that VvMLO3/VvMLO4 and VvPEN1 are co-trafficked together from the PM to the site of powdery mildew challenge. This focal accumulation of VvMLO3/VvMLO4 and VvPEN1 to the site of attack seems to be required for their opposing functions during powdery mildew attack, because their subcellular localization is correlated with the outcome of attempted powdery mildew penetration.

  13. Modification of ¤mlo5¤ resistance to ¤Blumeria graminis¤ attack in barley as a consequence of induced accessibility and inaccessibility

    DEFF Research Database (Denmark)

    Lyngkjær, M.F.; Carver, T.L.W.

    1999-01-01

    and formed haustoria within epidermal cells. When either isolate was inoculated onto the same leaves as challenger 48 h later, attacks on cells containing an inducer haustorium were almost invariably successful. Thus, cells containing an inducer haustorium showed almost complete induced accessibility...... to challenge attacks, even by the avirulent isolate. Accessibility was also induced to someextent in adjacent cells, but the effect was localized. By contrast, where inducer attacks failed, and a papilla was formed in the attacked cell, later challenge attacks on the same cells always failed. These......" followed by "challenger") using two fungal isolates. Isolate GE3 is a wild-type, unable to infect Riso-R, and is termed "mlo-avirulent". Isolate HL3/5, selected from GE3, is capable of infecting Riso-R, and is termed "mlo-virulent". When HL3/5 was used as inducer, some attacks penetrated successfully...

  14. Genome-wide identification, characterization, and expression analysis of the MLO gene family in Cucumis sativus.

    Science.gov (United States)

    Zhou, S J; Jing, Z; Shi, J L

    2013-12-11

    Mildew resistance locus o (MLO) is a plant-specific seven-transmembrane (TM) gene family. Several studies have revealed that certain members of the MLO gene family mediate powdery mildew susceptibility in three plant species, namely, Arabidopsis, barley, and tomato. The sequenced cucumber genome provides an opportunity to conduct a comprehensive overview of the MLO gene family. Fourteen genes (designated CsMLO01 through CsMLO14) have been identified within the Cucumis sativus genome by using an in silico cloning method with the MLO amino acid sequences of Arabidopsis thaliana and rice as probes. Sequence alignment revealed that numerous features of the gene family, such as TMs, a calmodulin-binding domain, peptide domains I and II, and 30 important amino acid residues for MLO function, are well conserved. Phylogenetic analysis of the MLO genes from cucumber and other plant species reveals seven different clades (I through VII). Three of these clades comprised MLO genes from A. thaliana, rice, maize, and cucumber, suggesting that these genes may have evolved after the divergence of monocots and dicots. In silico mapping showed that these CsMLOs were located on chromosomes 1, 2, 3, 4, 5, and 6 without any obvious clustering, except CsMLO01. To our knowledge, this paper is the first comprehensive report on MLO genes in C. sativus. These findings will facilitate the functional characterization of the MLOs related to powdery mildew susceptibility and assist in the development of disease resistance in cucumber.

  15. Regulation of gene expression and subcellular protein distribution in MLO-Y4 osteocytic cells by lysophosphatidic acid: Relevance to dendrite outgrowth.

    Energy Technology Data Exchange (ETDEWEB)

    Waters, Katrina M.; Jacobs, Jon M.; Gritsenko, Marina A.; Karin, Norman J.

    2011-02-26

    Osteoblastic and osteocytic cells are highly responsive to the lipid growth factor lysophosphatidic acid (LPA) but the mechanisms by which LPA alters bone cell functions are largely unknown. A major effect of LPA on osteocytic cells is the stimulation of dendrite membrane outgrowth, a process that we predicted to require changes in gene expression and protein distribution. We employed DNA microarrays for global transcriptional profiling of MLO-Y4 osteocytic cells grown for 6 and 24h in the presence or absence of LPA. We identified 932 transcripts that displayed statistically significant changes in abundance of at least 1.25-fold in response to LPA treatment. Gene ontology (GO) analysis revealed that the regulated gene products were linked to diverse cellular processes, including DNA repair, response to unfolded protein, ossification, protein-RNA complex assembly, and amine biosynthesis. Gene products associated with the regulation of actin microfilament dynamics displayed the most robust expression changes, and LPA-induced dendritogenesis in vitro was blocked by the stress fiber inhibitor cytochalasin D. Mass spectrometry-based proteomic analysis of MLO-Y4 cells revealed significant LPA-induced changes in the abundance of 284 proteins at 6h and 844 proteins at 24h. GO analysis of the proteomic data linked the effects of LPA to cell processes that control of protein distribution and membrane outgrowth, including protein localization, protein complex assembly, Golgi vesicle transport, cytoskeleton-dependent transport, and membrane invagination/endocytosis. Dendrites were isolated from LPA-treated MLO-Y4 cells and subjected to proteomic analysis to quantitatively assess the subcellular distribution of proteins. Sets of 129 and 36 proteins were enriched in the dendrite fraction as compared to whole cells after 6h and 24h of LPA exposure, respectively. Protein markers indicated that membranous organelles were largely excluded from the dendrites. Highly represented among

  16. Regulation of gene expression and subcellular protein distribution in MLO-Y4 osteocytic cells by lysophosphatidic acid: Relevance to dendrite outgrowth.

    Science.gov (United States)

    Waters, Katrina M; Jacobs, Jon M; Gritsenko, Marina A; Karin, Norman J

    2011-06-01

    Osteoblastic and osteocytic cells are highly responsive to the lipid growth factor lysophosphatidic acid (LPA) but the mechanisms by which LPA alters bone cell functions are largely unknown. A major effect of LPA on osteocytic cells is the stimulation of dendrite membrane outgrowth, a process that we predicted to require changes in gene expression and protein distribution. We employed DNA microarrays for global transcriptional profiling of MLO-Y4 osteocytic cells grown for 6 and 24h in the presence or absence of LPA. We identified 932 transcripts that displayed statistically significant changes in abundance of at least 1.25-fold in response to LPA treatment. Gene ontology (GO) analysis revealed that the regulated gene products were linked to diverse cellular processes, including DNA repair, response to unfolded protein, ossification, protein-RNA complex assembly, and amine biosynthesis. Gene products associated with the regulation of actin microfilament dynamics displayed the most robust expression changes, and LPA-induced dendritogenesis in vitro was blocked by the stress fiber inhibitor cytochalasin D. Mass spectrometry-based proteomic analysis of MLO-Y4 cells revealed significant LPA-induced changes in the abundance of 284 proteins at 6h and 844 proteins at 24h. GO analysis of the proteomic data linked the effects of LPA to cell processes that control of protein distribution and membrane outgrowth, including protein localization, protein complex assembly, Golgi vesicle transport, cytoskeleton-dependent transport, and membrane invagination/endocytosis. Dendrites were isolated from LPA-treated MLO-Y4 cells and subjected to proteomic analysis to quantitatively assess the subcellular distribution of proteins. Sets of 129 and 36 proteins were enriched in the dendrite fraction as compared to whole cells after 6h and 24h of LPA exposure, respectively. Protein markers indicated that membranous organelles were largely excluded from the dendrites. Highly represented among

  17. Biotin Carboxyl Carrier Protein in Barley Chloroplast Membranes

    DEFF Research Database (Denmark)

    Kannangara, C. G.; Jense, C J

    1975-01-01

    Biotin localized in barley chloroplast lamellae is covalently bound to a single protein with an approximate molecular weight of 21000. It contains one mole of biotin per mole of protein and functions as a carboxyl carrier in the acetyl-CoA carboxylase reaction. The protein was obtained by solubil......Biotin localized in barley chloroplast lamellae is covalently bound to a single protein with an approximate molecular weight of 21000. It contains one mole of biotin per mole of protein and functions as a carboxyl carrier in the acetyl-CoA carboxylase reaction. The protein was obtained...

  18. Functional characterization of the powdery mildew susceptibility gene SmMLO1 in eggplant (Solanum melongena L.).

    Science.gov (United States)

    Bracuto, Valentina; Appiano, Michela; Ricciardi, Luigi; Göl, Deniz; Visser, Richard G F; Bai, Yuling; Pavan, Stefano

    2017-01-09

    Eggplant (Solanum melongena L.) is one of the most important vegetables among the Solanaceae and can be a host to fungal species causing powdery mildew (PM) disease. Specific homologs of the plant Mildew Locus O (MLO) gene family are PM susceptibility factors, as their loss of function results in a recessive form of resistance known as mlo resistance. In a previous work, we isolated the eggplant MLO homolog SmMLO1. SmMLO1 is closely related to MLO susceptibility genes characterized in other plant species. However, it displays a peculiar non-synonymous substitution that leads to a T → M amino acid change at protein position 422, in correspondence of the MLO calmodulin-binding domain. In this study, we performed the functional characterization of SmMLO1. Transgenic overexpression of SmMLO1 in a tomato mlo mutant compromised resistance to the tomato PM pathogen Oidium neolycopersici, thus indicating that SmMLO1 is a PM susceptibility factor in eggplant. PM susceptibility was also restored by the transgenic expression of a synthetic gene, named s-SmMLO1, encoding a protein identical to SmMLO1, except for the presence of T at position 422. This indicates that the T → M polymorphism does not affect the protein role as PM susceptibility factor. Overall, the results of this work are of interest for the functional characterization of MLO proteins and the introduction of PM resistance in eggplant using reverse genetics.

  19. Lysine Rich Proteins in the Salt-Soluble Protein Fraction of Barley

    DEFF Research Database (Denmark)

    Ingversen, J.; Køie, B.

    1973-01-01

    Fractionation of the protein complex from Emir barley showed that the salt-soluble fraction accounts for 44% of the total lysine content but only for 2.......Fractionation of the protein complex from Emir barley showed that the salt-soluble fraction accounts for 44% of the total lysine content but only for 2....

  20. Loss of function in Mlo orthologs reduces susceptibility of pepper and tomato to powdery mildew disease caused by Leveillula taurica.

    Directory of Open Access Journals (Sweden)

    Zheng Zheng

    Full Text Available Powdery mildew disease caused by Leveillula taurica is a serious fungal threat to greenhouse tomato and pepper production. In contrast to most powdery mildew species which are epiphytic, L. taurica is an endophytic fungus colonizing the mesophyll tissues of the leaf. In barley, Arabidopsis, tomato and pea, the correct functioning of specific homologues of the plant Mlo gene family has been found to be required for pathogenesis of epiphytic powdery mildew fungi. The aim of this study was to investigate the involvement of the Mlo genes in susceptibility to the endophytic fungus L. taurica. In tomato (Solanum lycopersicum, a loss-of-function mutation in the SlMlo1 gene results in resistance to powdery mildew disease caused by Oidium neolycopersici. When the tomato Slmlo1 mutant was inoculated with L. taurica in this study, it proved to be less susceptible compared to the control, S. lycopersicum cv. Moneymaker. Further, overexpression of SlMlo1 in the tomato Slmlo1 mutant enhanced susceptibility to L. taurica. In pepper, the CaMlo2 gene was isolated by applying a homology-based cloning approach. Compared to the previously identified CaMlo1 gene, the CaMlo2 gene is more similar to SlMlo1 as shown by phylogenetic analysis, and the expression of CaMlo2 is up-regulated at an earlier time point upon L. taurica infection. However, results of virus-induced gene silencing suggest that both CaMlo1 and CaMlo2 may be involved in the susceptibility of pepper to L. taurica. The fact that overexpression of CaMlo2 restored the susceptibility of the tomato Slmlo1 mutant to O. neolycopersici and increased its susceptibility to L. taurica confirmed the role of CaMlo2 acting as a susceptibility factor to different powdery mildews, though the role of CaMlo1 as a co-factor for susceptibility cannot be excluded.

  1. Golgi localized barley MTP8 proteins facilitate Mn transport.

    Science.gov (United States)

    Pedas, Pai; Schiller Stokholm, Michaela; Hegelund, Josefine Nymark; Ladegård, Anne Hald; Schjoerring, Jan Kofod; Husted, Søren

    2014-01-01

    Many metabolic processes in plants are regulated by manganese (Mn) but limited information is available on the molecular mechanisms controlling cellular Mn homeostasis. In this study, a yeast assay was used to isolate and characterize two genes, MTP8.1 and MTP8.2, which encode membrane-bound proteins belonging to the cation diffusion facilitator (CDF) family in the cereal species barley (Hordeum vulgare). Transient expression in onion epidermal cells showed that MTP8.1 and MTP8.2 proteins fused to the green fluorescent protein (GFP) are localized to Golgi. When heterologously expressed in yeast, MTP8.1 and MTP8.2 were found to be Mn transporters catalysing Mn efflux in a similar manner as the Golgi localized endogenous yeast protein Pmr1p. The level of MTP8.1 transcripts in barley roots increased with external Mn supply ranging from deficiency to toxicity, while MTP8.2 transcripts decreased under the same conditions, indicating non-overlapping functions for the two genes. In barley leaves, the expression of both MTP8 genes declined in response to toxic Mn additions to the roots suggesting a role in ensuring proper delivery of Mn to Golgi. Based on the above we suggest that barley MTP8 proteins are involved in Mn loading to the Golgi apparatus and play a role in Mn homeostasis by delivering Mn to Mn-dependent enzymes and/or by facilitating Mn efflux via secretory vesicles. This study highlights the importance of MTP transporters in Mn homeostasis and is the first report of Golgi localized Mn2+ transport proteins in a monocot plant species.

  2. mlo-based powdery mildew resistance in hexaploid bread wheat generated by a non-transgenic TILLING approach.

    Science.gov (United States)

    Acevedo-Garcia, Johanna; Spencer, David; Thieron, Hannah; Reinstädler, Anja; Hammond-Kosack, Kim; Phillips, Andrew L; Panstruga, Ralph

    2017-03-01

    Wheat is one of the most widely grown cereal crops in the world and is an important food grain source for humans. However, wheat yields can be reduced by many abiotic and biotic stress factors, including powdery mildew disease caused by Blumeria graminis f.sp. tritici (Bgt). Generating resistant varieties is thus a major effort in plant breeding. Here, we took advantage of the non-transgenic Targeting Induced Lesions IN Genomes (TILLING) technology to select partial loss-of-function alleles of TaMlo, the orthologue of the barley Mlo (Mildew resistance locus o) gene. Natural and induced loss-of-function alleles (mlo) of barley Mlo are known to confer durable broad-spectrum powdery mildew resistance, typically at the expense of pleiotropic phenotypes such as premature leaf senescence. We identified 16 missense mutations in the three wheat TaMlo homoeologues, TaMlo-A1, TaMlo-B1 and TaMlo-D1 that each lead to single amino acid exchanges. Using transient gene expression assays in barley single cells, we functionally analysed the different missense mutants and identified the most promising candidates affecting powdery mildew susceptibility. By stacking of selected mutant alleles we generated four independent lines with non-conservative mutations in each of the three TaMlo homoeologues. Homozygous triple mutant lines and surprisingly also some of the homozygous double mutant lines showed enhanced, yet incomplete, Bgt resistance without the occurrence of discernible pleiotropic phenotypes. These lines thus represent an important step towards the production of commercial non-transgenic, powdery mildew-resistant bread wheat varieties.

  3. Origin of worldwide cultivated barley revealed by NAM-1 gene and grain protein content

    Directory of Open Access Journals (Sweden)

    Yonggang eWang

    2015-09-01

    Full Text Available The origin, evolution and distribution of cultivated barley provides powerful insights into the historic origin and early spread of agrarian culture. Here, population-based genetic diversity and phylogenetic analyses were performed to determine the evolution and origin of barley and how domestication and subsequent introgression have affected the genetic diversity and changes in cultivated barley on a worldwide scale. A set of worldwide cultivated and wild barleys from Asia and Tibet of China were analyzed using the sequences for NAM-1 gene and gene-associated traits-GPC (grain protein content. Our results showed Tibetan wild barley distinctly diverged from Near Eastern barley, and confirmed that Tibet is one of the origin and domestication centers for cultivated barley, and in turn supported a polyphyletic origin of domesticated barley. Comparison of haplotype composition among geographic regions revealed gene flow between Eastern and Western barley populations, suggesting that the Silk Road might have played a crucial role in the spread of genes. The GPC in the 118 cultivated and 93 wild barley accessions ranged from 6.73% to 12.35% with a mean of 9.43%. Overall, wild barley had higher averaged GPC (10.44% than cultivated barley. Two unique haplotypes (Hap2 and Hap7 caused by a base mutations (at position 544 in the coding region of the NAM-1 gene might have a significant impact on the GPC. SNPs and haplotypes of NAM-1 associated with GPC in barley could provide a useful method for screening GPC in barley germplasm. The Tibetan wild accessions with lower GPC could be useful for malt barley breeding

  4. Golgi localized barley MTP8 proteins facilitate Mn transport

    DEFF Research Database (Denmark)

    Pedas, Pai Rosager; Schiller, Michaela; Hegelund, Josefine Nymark

    2014-01-01

    8 proteins are involved in Mn loading to the Golgi apparatus and play a role in Mn homeostasis by delivering Mn to Mn-dependent enzymes and/or by facilitating Mn efflux via secretory vesicles. This study highlights the importance of MTP transporters in Mn homeostasis and is the first report of Golgi......-bound proteins belonging to the cation diffusion facilitator (CDF) family in the cereal species barley (Hordeum vulgare). Transient expression in onion epidermal cells showed that MTP8.1 and MTP8.2 proteins fused to the green fluorescent protein (GFP) are localized to Golgi. When heterologously expressed...... in yeast, MTP8.1 and MTP8.2 were found to be Mn transporters catalysing Mn efflux in a similar manner as the Golgi localized endogenous yeast protein Pmr1p. The level of MTP8.1 transcripts in barley roots increased with external Mn supply ranging from deficiency to toxicity, while MTP8.2 transcripts...

  5. Genome-Wide Study of the Tomato SlMLO Gene Family and Its Functional Characterization in Response to the Powdery Mildew Fungus Oidium neolycopersici.

    Science.gov (United States)

    Zheng, Zheng; Appiano, Michela; Pavan, Stefano; Bracuto, Valentina; Ricciardi, Luigi; Visser, Richard G F; Wolters, Anne-Marie A; Bai, Yuling

    2016-01-01

    The MLO (Mildew Locus O) gene family encodes plant-specific proteins containing seven transmembrane domains and likely acting in signal transduction in a calcium and calmodulin dependent manner. Some members of the MLO family are susceptibility factors toward fungi causing the powdery mildew disease. In tomato, for example, the loss-of-function of the MLO gene SlMLO1 leads to a particular form of powdery mildew resistance, called ol-2, which arrests almost completely fungal penetration. This type of penetration resistance is characterized by the apposition of papillae at the sites of plant-pathogen interaction. Other MLO homologs in Arabidopsis regulate root response to mechanical stimuli (AtMLO4 and AtMLO11) and pollen tube reception by the female gametophyte (AtMLO7). However, the role of most MLO genes remains unknown. In this work, we provide a genome-wide study of the tomato SlMLO gene family. Besides SlMLO1, other 15 SlMLO homologs were identified and characterized with respect to their structure, genomic organization, phylogenetic relationship, and expression profile. In addition, by analysis of transgenic plants, we demonstrated that simultaneous silencing of SlMLO1 and two of its closely related homologs, SlMLO5 and SlMLO8, confer higher level of resistance than the one associated with the ol-2 mutation. The outcome of this study provides evidence for functional redundancy among tomato homolog genes involved in powdery mildew susceptibility. Moreover, we developed a series of transgenic lines silenced for individual SlMLO homologs, which lay the foundation for further investigations aimed at assigning new biological functions to the MLO gene family.

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

    Science.gov (United States)

    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

  7. Targeted modification of storage protein content resulting in improved amino acid composition of barley grain

    DEFF Research Database (Denmark)

    Sikdar, Md. Shafiqul Islam; Bowra, S; Schmidt, Daiana;

    2016-01-01

    family members. Analysis of the AA composition of the transgenic lines showed that the level of essential amino acids increased with a concomitant reduction in proline and glutamine. Both the barley C-hordein and wheat ω-gliadin genes proved successful for RNAi-gene mediated suppression of barley C......C-hordein in barley and ω-gliadins in wheat are members of the prolamins protein families. Prolamins are the major component of cereal storage proteins and composed of non-essential amino acids (AA) such as proline and glutamine therefore have low nutritional value. Using double stranded RNAi...... silencing technology directed towards C-hordein we obtained transgenic barley lines with up to 94.7 % reduction in the levels of C-hordein protein relative to the parental line. The composition of the prolamin fraction of the barley parental line cv. Golden Promise was resolved using SDS...

  8. Early changes in protein expression of barley following inoculation with erysiphe graminis f. sp. hordei

    Energy Technology Data Exchange (ETDEWEB)

    Simons, S.P.; Somerville, S.C. (Michigan State Univ., East Lansing (USA))

    1989-04-01

    Erysiphe graminis f. sp. hordei is an obligate pathogen of barley causing the powdery mildew disease. Resistance to this disease is the product of a highly specific interaction between barley lines with specific resistance alleles and pathogen races carrying complementary avirulence alleles. Using congenic barley lines which differ at the M1-a disease reaction locus, we hope to define the early molecular events of this interaction. Accordingly, resistant and susceptible barley seedlings were labelled with {sup 35}S-methionine and examined by two-dimensional electrophoresis at two hour intervals following inoculation. Infection related changes were observed with both isolines during the four to twelve hour time period. Additional differences existed constitutively between the barley lines. These differences have been quantified. Further characterization of these proteins will yield useful markers for events preceding or coinciding with cytological responses any may lead to identification and cloning of the M1-a gene.

  9. Identification of thioredoxin target disulfides in proteins released from barley aleurone layers

    DEFF Research Database (Denmark)

    Hägglund, Per; Bunkenborg, J.; Yang, Fen

    2010-01-01

    Thioredoxins are ubiquitous disulfide reductases involved in a wide range of cellular processes including DNA synthesis, oxidative stress response and apoptosis. In cereal seeds thioredoxins are proposed to facilitate the germination process by reducing disulfide bonds in storage proteins and other...... targets in the starchy endosperm. Here we have applied a thiol-specific labeling approach to identify specific disulfide targets of barley thioredoxin in proteins released from barley aleurone layers incubated in buffer containing gibberellic acid....

  10. Mutation analysis of barley malt protein Z4 and protein Z7 on beer foam stability.

    Science.gov (United States)

    Iimure, Takashi; Kimura, Tatsuji; Araki, Shigeki; Kihara, Makoto; Sato, Masahide; Yamada, Shinji; Shigyou, Tatsuro; Sato, Kazuhiro

    2012-02-15

    Beer foam stability is an important characteristic. It has been suggested that isoforms of protein Z, that is, protein Z4 and protein Z7, contribute to beer foam stability. We investigated the relationship between beer foam stability and protein Z4 and protein Z7 using their deficient mutants. As a protein Z4-deficient mutant, cv. Pirkka was used. Protein Z7 deficiency was screened in 1564 barley accessions in the world collection of Okayama University, Japan. The barley samples from normal, protein Z4-deficient, protein Z7-deficient, and double-deficient were genotyped in F(2) populations and then pooled based on the DNA marker genotypes of protein Z4 and protein Z7. For a brewing trial, F(5) pooled subpopulations were used. After malting and brewing, the foam stability was determined, and the results showed that the levels of foam stability in the four samples were comparable. Two-dimensional gel electrophoresis was used to investigate the proteome in these beer samples. The results showed that low molecular weight proteins, including lipid transfer protein (LTP2), in the deficient mutants were higher than those in the normal sample. Our results suggest that the contribution of protein Z4 and protein Z7 to beer foam stability was not greater than that of other beer proteins.

  11. Structure and Composition of Protein Bodies from Wild-Type and High-Lysine Barley Endosperm

    DEFF Research Database (Denmark)

    Ingversen, J.

    1975-01-01

    Protein bodies were isolated from 13 and 28 day old endosperms of barley mutant 1508 and its wild type, Bomi barley. The fine structure of the isolated protein bodies was determined by electron microscopy, and the proteins present in the preparations characterized by amino-acid analysis and SDS...... with a granular component. Particles with the same structure were present in the protein body preparation from the mutant, where, however, the granular component was the most prominent. Amino-acid composition and SDS-polyacrylamide gel electrophoresis of the proteins from the protein body preparation revealed...

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

    Science.gov (United States)

    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...

  13. Secretomics identifies Fusarium graminearum proteins involved in the interaction with barley and wheat

    DEFF Research Database (Denmark)

    Yang, Fen; Jensen, Jens D.; Svensson, Birte;

    2012-01-01

    of cell walls, starch and proteins. Of these proteins, 35% had not been identified in previous in planta or in vitro studies, 70% were predicted to contain signal peptides and a further 16% may be secreted in a nonclassical manner. Proteins identified in the 72 spots showing differential appearance...... between wheat and barley flour medium were mainly involved in fungal cell wall remodelling and the degradation of plant cell walls, starch and proteins. The in planta expression of corresponding F. graminearum genes was confirmed by quantitative reverse transcriptase‐polymerase chain reaction in barley...... and wheat spikelets harvested at 2−6 days after inoculation. In addition, a clear difference in the accumulation of fungal biomass and the extent of fungal‐induced proteolysis of plant β‐amylase was observed in barley and wheat. The present study considerably expands the current database of F. graminearum...

  14. Do 14-3-3 proteins and plasma membrane H+-AtPases interact in the barley epidermis in response to the barley powdery mildew fungus?

    DEFF Research Database (Denmark)

    Finni, Christine; Andersen, Claus H; Borch, Jonas;

    2002-01-01

    , or treatment with fusicoccin, results in an increase in fusicoccin binding ability of barley leaf membranes. Overlay assays show a fungus-induced increase in binding of digoxygenin-labelled 14-3-3 protein to several proteins including a 100 kDa membrane protein, probably the plasma membrane H...

  15. Do 14-3-3 proteins and plasma membrane H+-ATPases interact in the barley epidermis in response to the barley powdery mildew fungus?

    DEFF Research Database (Denmark)

    Finnie, C.; Andersen, C.H.; Borch, J.;

    2002-01-01

    , or treatment with fusicoccin, results in an increase in fusicoccin binding ability of barley leaf membranes. Overlay assays show a fungus-induced increase in binding of digoxygenin-labelled 14-3-3 protein to several proteins including a 100 kDa membrane protein, probably the plasma membrane H...

  16. A Systematic View of the MLO Family in Rice Suggests Their Novel Roles in Morphological Development, Diurnal Responses, the Light-Signaling Pathway, and Various Stress Responses

    Science.gov (United States)

    Nguyen, Van N. T.; Vo, Kieu T. X.; Park, Hyon; Jeon, Jong-Seong; Jung, Ki-Hong

    2016-01-01

    The Mildew resistance Locus O (MLO) family is unique to plants, containing genes that were initially identified as a susceptibility factor to powdery mildew pathogens. However, little is known about the roles and functional diversity of this family in rice, a model crop plant. The rice genome has 12 potential MLO family members. To achieve systematic functional assignments, we performed a phylogenomic analysis by integrating meta-expression data obtained from public sources of microarray data and real-time expression data into a phylogenic tree. Subsequently, we identified 12 MLO genes with various tissue-preferred patterns, including leaf, root, pollen, and ubiquitous expression. This suggested their functional diversity for morphological agronomic traits. We also used these integrated transcriptome data within a phylogenetic context to estimate the functional redundancy or specificity among OsMLO family members. Here, OsMLO12 showed preferential expression in mature pollen; OsMLO4, in the root tips; OsMLO10, throughout the roots except at the tips; and OsMLO8, expression preferential to the leaves and trinucleate pollen. Of particular interest to us was the diurnal expression of OsMLO1, OsMLO3, and OsMLO8, which indicated that they are potentially significant in responses to environmental changes. In osdxr mutants that show defects in the light response, OsMLO1, OsMLO3, OsMLO8, and four calmodulin genes were down-regulated. This finding provides insight into the novel functions of MLO proteins associated with the light-responsive methylerythritol 4-phosphate pathway. In addition, abiotic stress meta-expression data and real-time expression analysis implied that four and five MLO genes in rice are associated with responses to heat and cold stress, respectively. Upregulation of OsMLO3 by Magnaporthe oryzae infection further suggested that this gene participates in the response to pathogens. Our analysis has produced fundamental information that will enhance future

  17. Synthesis of the major storage protein, hordein, in barley

    DEFF Research Database (Denmark)

    Giese, Nanna Henriette; Andersen, B.; Doll, Hans

    1983-01-01

    A liquid culture system for culturing detached spikes of barley (Hordeum vulgare L.) at different nutritional levels was established. The synthesis of hordein polypeptides was studied by pulse-labeling with [14C]sucrose at different stages of development and nitrogen (N) nutrition. All polypeptid...

  18. Antisense expression of peach mildew resistance locus O (PpMlo1) gene confers cross-species resistance to powdery mildew in Fragaria x ananassa.

    Science.gov (United States)

    Jiwan, Derick; Roalson, Eric H; Main, Dorrie; Dhingra, Amit

    2013-12-01

    Powdery mildew (PM) is one of the major plant pathogens. The conventional method of PM control includes frequent use of sulfur-based fungicides adding to production costs and potential harm to the environment. PM remains a major scourge for Rosaceae crops where breeding approaches mainly resort to gene-for-gene resistance. We have tested an alternate source of PM resistance in Rosaceae. Mildew resistance locus O (MLO) has been well studied in barley due to its role in imparting broad spectrum resistance to PM. We identified PpMlo1 (Prunus persica Mlo) in peach and characterized it further to test if a similar mechanism of resistance is conserved in Rosaceae. Due to its recalcitrance in tissue culture, reverse genetic studies involving PpMloI were not feasible in peach. Therefore, Fragaria x ananassa LF9 line, a taxonomic surrogate, was used for functional analysis of PpMlo1. Agrobacterium-mediated transformation yielded transgenic strawberry plants expressing PpMlo1 in sense and antisense orientation. Antisense expression of PpMlo1 in transgenic strawberry plants conferred resistance to Fragaria-specific powdery mildew, Podosphaera macularis. Phylogenetic analysis of 208 putative Mlo gene copies from 35 plant species suggests a large number of duplications of this gene family prior to the divergence of monocots and eudicots, early in eudicot diversification. Our results indicate that the Mlo-based resistance mechanism is functional in Rosaceae, and that Fragaria can be used as a host to test mechanistic function of genes derived from related tree species. To the best of our knowledge, this work is one of the first attempts at testing the potential of using a Mlo-based resistance strategy to combat powdery mildew in Rosaceae.

  19. Improving the nutritional quality of the barley and wheat grain storage proteins by antisense technology

    DEFF Research Database (Denmark)

    Sikdar, Md. Shafiqul Islam; Lange, Mette; Aaslo, Per

    2011-01-01

    Prolamins are the predominant storage proteins in barley and wheat grains, accounting for 50 to 80% of total seed protein. However, the prolamins are not optimal feed for monogastric animals as they have a low content of certain essential amino acids such as lysine, threonine and tryptophan...... gliadins) are also available from Germany and UK. We have grown them under different N regimes (high, medium and low N) in semi-field conditions. Previously five different antisense C-hordein lines of barley have been characterized in our laboratory. The analyses revealed that the lysine, threonine...

  20. Accelerated rates of protein evolution in barley grain and pistil biased genes might be legacy of domestication.

    Science.gov (United States)

    Shi, Tao; Dimitrov, Ivan; Zhang, Yinling; Tax, Frans E; Yi, Jing; Gou, Xiaoping; Li, Jia

    2015-10-01

    Traits related to grain and reproductive organs in grass crops have been under continuous directional selection during domestication. Barley is one of the oldest domesticated crops in human history. Thus genes associated with the grain and reproductive organs in barley may show evidence of dramatic evolutionary change. To understand how artificial selection contributes to protein evolution of biased genes in different barley organs, we used Digital Gene Expression analysis of six barley organs (grain, pistil, anther, leaf, stem and root) to identify genes with biased expression in specific organs. Pairwise comparisons of orthologs between barley and Brachypodium distachyon, as well as between highland and lowland barley cultivars mutually indicated that grain and pistil biased genes show relatively higher protein evolutionary rates compared with the median of all orthologs and other organ biased genes. Lineage-specific protein evolutionary rates estimation showed similar patterns with elevated protein evolution in barley grain and pistil biased genes, yet protein sequences generally evolve much faster in the lowland barley cultivar. Further functional annotations revealed that some of these grain and pistil biased genes with rapid protein evolution are related to nutrient biosynthesis and cell cycle/division. Our analyses provide insights into how domestication differentially shaped the evolution of genes specific to different organs of a crop species, and implications for future functional studies of domestication genes.

  1. Differential Synthesis in Vitro of Barley Aleurone and Starchy Endosperm Proteins

    DEFF Research Database (Denmark)

    Mundy, John; Hejgaard, Jørn; Hansen, Annette

    1986-01-01

    RNAs from isolated endosperm and aleurone tissues (developing and mature grain) and from cultured (germinating) aleurone layers treated with abscisic acid (ABA) and GA(3). B and C hordein polypeptides and the salt-soluble proteins beta-amylase, protein Z, protein C, the chymotrypsin inhibitors (CI-1 and 2......To widen the selection of proteins for gene expression studies in barley seeds, experiments were performed to identify proteins whose synthesis is differentially regulated in developing and germinating seed tissues. The in vitro synthesis of nine distinct barley proteins was compared using m......, expression of genes encoding ASI, PSI, protein C, and PAPI is tissue and stage-specific during seed development. Only ASI, CI-1, and PAPI were synthesized in significant amounts with mRNA from cultured aleurone layers. The levels of synthesis of PAPI and CI-1 were independent of hormone treatment...

  2. Interaction of barley powdery mildew effector candidate CSEP0055 with the defence protein PR17c

    DEFF Research Database (Denmark)

    Zhang, Wenjing; Pedersen, Carsten; Kwaaitaal, Mark Adrianus Cornelis J;

    2012-01-01

    A large number of effector candidates have been identified recently in powdery mildew fungi. However, their roles and how they perform their functions remain unresolved. In this study, we made use of host-induced gene silencing and confirmed that the secreted barley powdery mildew effector...... with PR17c was confirmed by bimolecular fluorescence complementation analyses. Down-regulation and over-expression of PR17c in epidermal cells of barley confirmed that this protein is important for penetration resistance against the powdery mildew fungus. In line with this, PR17c was found...

  3. Synthesis of Salt Soluble Proteins in Barley. Pulse-Labeling Study of Grain Filling in Liquid-Cultured Detached Spikes

    DEFF Research Database (Denmark)

    Giese, Nanna Henriette; Hejgaard, Jørn

    1984-01-01

    The accumulation of salt-soluble proteins in the endosperm of developing barley (Hordeum vulgare L.) grains was examined. Detached spikes of barley were cultured at different levels of nitrogen nutrition and pulse-labeled with [14C] sucrose at specific times after anthesis. Proteins were extracted...... to increased nitrogen nutrition. Two major components, β-amylase and protein Z in particular, had a synthesis profile almost identical to that of the endosperm storage protein, hordein....

  4. Durable broad-spectrum powdery mildew resistance in pea er1 plants is conferred by natural loss-of-function mutations in PsMLO1

    NARCIS (Netherlands)

    Humphry, M.; Reinstädler, A.; Ivanov, S.; Bisseling, T.; Panstruga, R.

    2011-01-01

    Loss-of-function alleles of plant-specific MLO (Mildew Resistance Locus O) genes confer broad-spectrum powdery mildew resistance in monocot (barley) and dicot (Arabidopsis thaliana, tomato) plants. Recessively inherited powdery mildew resistance in pea (Pisum sativum) er1 plants is, in many aspects,

  5. Grain protein concentration and harvestable protein under future climate conditions. A study of 108 spring barley accessions

    DEFF Research Database (Denmark)

    Ingvordsen, Cathrine Heinz; Gislum, René; Jørgensen, Johannes Ravn;

    2016-01-01

    In the present study a set of 108 spring barley (H. vulgare L.) accessions were cultivated under predicted future levels of temperature and [CO2] as single factors and in combination (IPCC, AR5, RCP8.5). Across all genotypes, elevated [CO2] (700 ppm day/night) slightly decreased protein concentra......In the present study a set of 108 spring barley (H. vulgare L.) accessions were cultivated under predicted future levels of temperature and [CO2] as single factors and in combination (IPCC, AR5, RCP8.5). Across all genotypes, elevated [CO2] (700 ppm day/night) slightly decreased protein...

  6. Effect of pH and Recombinant Barley (Hordeum vulgare L.) Endoprotease B2 on Degradation of Proteins in Soaked Barley

    DEFF Research Database (Denmark)

    Christensen, Jesper Bjerg; Dionisio, Giuseppe; Poulsen, Hanne Damgaard

    2014-01-01

    . Proteolysis evaluated by SDS-PAGE and differential proteomics confirmed a powerful effect of adding rec-HvEP-B2 to the soaked barley, regardless of the genotype. Our study addresses the use of rec-HvEP-B2 as an effective feed enzyme protease. HvEP-B2 has the potential to increase the digestibility of protein...

  7. Surprisingly high stability of barley lipid transfer protein, LTP1, towards denaturant, heat and proteases

    DEFF Research Database (Denmark)

    Lindorff-Larsen, Kresten; Winther, J R

    2001-01-01

    Barley LTP1 belongs to a large family of plant proteins termed non-specific lipid transfer proteins. The in vivo function of these proteins is unknown, but it has been suggested that they are involved in responses towards stresses such as pathogens, drought, heat, cold and salt. Also, the proteins...... have been suggested as transporters of monomers for cutin synthesis. We have analysed the stability of LTP1 towards denaturant, heat and proteases and found it to be a highly stable protein, which apparently does not denature at temperatures up to 100 degrees C. This high stability may be important...

  8. Transcriptome study of storage protein genes of field-grown barley in response to inorganic nitrogen fertilizers

    DEFF Research Database (Denmark)

    Hansen, Michael; Bowra, Steve; Lange, Mette;

    2010-01-01

    The storage proteins of barley, in terms of both amino acid profile and quantity, are traits strongly influenced by the amount of nitrogen applied. Given this, we performed a developmental expression analysis of the genes from barley grains grown under field conditions to further our understanding...... of the molecular and biochemical mechanisms underpinning nitrogen utilization. A barley grain specific micro-array, where a comprehensive set of genes involved in nitrogen mobilization, storage protein synthesis and amino acid metabolism were assembled, was used to obtain a global but focused gene expression...

  9. Quality Control System for Beer Developed with Monoclonal Antibodies Specific to Barley Lipid Transfer Protein

    Directory of Open Access Journals (Sweden)

    Yukie Murakami-Yamaguchi

    2012-10-01

    Full Text Available Non-specific lipid transfer protein (LTP in barley grain reacted with the IgE in sera drawn from food allergy patients. A sandwich-type of enzyme-linked immunosorbent assay (ELISA was developed with mouse monoclonal antibodies raised against LTP purified with barley flour. This ELISA showed a practical working range of 0.3–3 ng/mL and no cross-reactivity with wheat, adlay and rye. Using this ELISA, LTP was determined in several types of barley-foods, including fermented foods such as malt vinegar, barley-malt miso and beer. LTP content in beer of the same kind was approximately constant, even if manufacturing factory and production days were different. Not only as a factor of foam formation and stability but also as an allergen, controlling and monitoring of LTP in beer should be considered. Taken together, our LTP-detecting ELISA can be proposed as an appropriate system for the quality control of beer.

  10. Barley Yellow Mosaic Virus VPg Is the Determinant Protein for Breaking eIF4E-Mediated Recessive Resistance in Barley Plants

    Science.gov (United States)

    Li, Huangai; Kondo, Hideki; Kühne, Thomas; Shirako, Yukio

    2016-01-01

    In this study, we investigated the barley yellow mosaic virus (BaYMV, genus Bymovirus) factor(s) responsible for breaking eIF4E-mediated recessive resistance genes (rym4/5/6) in barley. Genome mapping analysis using chimeric infectious cDNA clones between rym5-breaking (JT10) and rym5-non-breaking (JK05) isolates indicated that genome-linked viral protein (VPg) is the determinant protein for breaking the rym5 resistance. Likewise, VPg is also responsible for overcoming the resistances of rym4 and rym6 alleles. Mutational analysis identified that amino acids Ser-118, Thr-120, and His-142 in JT10 VPg are the most critical residues for overcoming rym5 resistance in protoplasts. Moreover, the rym5-non-breaking JK05 could accumulate in the rym5 protoplasts when eIF4E derived from a susceptible barley cultivar was expressed from the viral genome. Thus, the compatibility between VPg and host eIF4E determines the ability of BaYMV to infect barley plants. PMID:27746794

  11. 巴西橡胶树(Hevea brasiliensis)HbMlo7基因克隆与表达分析%Molecular cloning and characterization ofHbMlo7 gene in rubber tree (Hevea brasiliensis)

    Institute of Scientific and Technical Information of China (English)

    何海霞; 张宇; 王萌; 杨叶; 郑服丛

    2016-01-01

    Mlo基因是一种植物防卫相关基因,负调控植物白粉病抗性。为解析巴西橡胶树(Hevea brasiliensis)中Mlo基因的结构与功能,从橡胶树品种‘热研7-33-97’胶乳中扩增了一个Mlo cDNA序列,将其命名为HbMlo7。该基因编码499个氨基酸,分子量为56.85 kDa。蛋白结构分析发现,该蛋白含有Mlo保守结构域,有一个核定位信号和7个跨膜结构区域,无信号肽。通过qRT-PCR发现HbMlo7基因主要在橡胶树树皮中表达,其表达量在干旱、机械伤害和白粉菌侵染处理下显著上调。脱落酸(ABA)、乙烯利(ETH)、茉莉酸(JA)和过氧化氢(H2O2)均能诱导橡胶树叶片中HbMlo7基因显著上调表达。研究认为HbMlo7是植物Mlo家族成员,参与橡胶树的抗逆反应机制和植物激素信号传导途径。%Mildew resistance locus o(Mlo) is a defence-related gene in plant, which negatively regulates plant resistance to powdery mildew. To analyze the structure and function ofMlo gene inHevea brasiliensis, we cloned a cDNA sequence of Mlo from latex of the rubber tree clone ‘Reyan 7-33-97’. We deifned it asHbMlo7. HbMlo7 gene encoded 499 amino acids with a predicted molecular mass of 56.85 kDa. The protein structure analysis revealed that HbMLO7 has one conserved domain of Mlo, one nuclear localization signal (NLS), seven transmembrane domains, and no signal peptide. Gene expression analysis by quantitative and reverse transcrip-tion PCR (qRT-PCR) indicated thatHbMlo7expressed mainly in the bark of the rubber tree. The expression level ofHbMlo7 was induced by drought, wounding and powdery mildew infection. Abscisic acid (ABA), ethe-phon (ETH), jasmonic acid (JA) and hydrogen peroxide (H2O2) signiifcantly upregulatedHbMlo7 geneexpres-sion in rubber tree leaves. The results suggest thatHbMlo7 is one of the members ofMlo family, and involved in response mechanisms of stress and signal transduction pathway of plant hormones in rubber tree.

  12. Interactions between Barley a-Amylases, Substrates, Inhibitors and Regulatory Proteins

    DEFF Research Database (Denmark)

    Hachem, Maher Abou; Bozonnet, Sophie; Willemoës, Martin

    2006-01-01

    Barley a-amylase binds sugars at two sites on the enzyme surface in addition to the active site. Crystallography and site-directed mutagenesis highlight the importance of aromatic residues at these surface sites as demonstrated by Kd values determined for ß-cyclodextrin by surface plasmon resonance......, a fully hydrated calcium ion at the protein interface mediates contact between inhibitor residues and the enzyme catalytic groups in a manner that depends on calcium and which can be suppressed by site-directed mutagenesis of Glu168 in BASI. Finally certain inhibitors and enzymes are targets...... of the disulphide reductase thioredoxin h that attacks a specific disulphide bond in BASI and, remarkably, reduces two different disulphide bonds in the barley monomeric and dimeric amylase inhibitors that both belong to the CM-proteins and inhibit animal a-amylase....

  13. Non-wheat pasta based on pearl millet flour containing barley and whey protein concentrate.

    Science.gov (United States)

    Yadav, Deep N; Balasubramanian, S; Kaur, Jaspreet; Anand, Tanupriya; Singh, Ashish K

    2014-10-01

    Non-wheat pasta was prepared with pearl millet supplemented with 10-30 % barley flour, 5-15 % whey protein concentrate, 2.5-4 % carboxy methyl cellulose and 27-33 % water using response surface methodology (RSM) following central composite rotatable design (CCRD). Results showed that barley flour and whey protein concentrate (WPC) had significant (p ≤ 0.05) positive effect on lightness and negative effect on stickiness of pasta, thus improved the overall acceptability (OAA). Carboxymethyl cellulose (CMC) improved the textural attributes i.e. increased firmness and decreased stickiness significantly (P ≤ 0.05) and caused a significant (P ≤ 0.05) reduction in solids losses in gruel. Based upon the experiments, the optimized level of ingredients were barley flour 13.80 g 100 g(-1) pearl millet flour (PMF), WPC 12.27 g 100 g(-1) PMF, CMC 3.45 g 100 g(-1) PMF and water 27.6 mL 100 g(-1) ingredients premix with 88 % desirability. The developed pasta had protein 16.47 g, calcium 98.53 mg, iron 5.43 mg, phosphorus 315.5 mg and β-glucan 0.33 g 100 g(-1) pasta (db).

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

    DEFF Research Database (Denmark)

    Sultan, Abida; Andersen, Birgit; Svensson, Birte

    2016-01-01

    -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....

  15. Barley alpha-amylase/subtilisin inhibitor: structure, biophysics and protein engineering

    DEFF Research Database (Denmark)

    Nielsen, P.K.; Bønsager, Birgit Christine; Fukuda, Kenji;

    2004-01-01

    Bifunctional alpha-amylase/subtilisin inhibitors have been implicated in plant defence and regulation of endogenous alpha-amylase action. The barley alpha-amylase/subtilisin inhibitor (BASI) inhibits the barley alpha-amylase 2 (AMY2) and subtilisin-type serine proteases. BASI belongs to the Kunitz...... Ca2+-modulated kinetics of the AMY2/BASl interaction and found that the complex formation involves minimal structural changes. The modulation of the interaction by calcium ions makes it unique among the currently known binding mechanisms of proteinaceous alpha-amylase inhibitors.......-type trypsin inhibitor family of the beta-trefoil fold proteins. Diverse approaches including site-directed mutagenesis, hybrid constructions, and crystallography have been used to characterise the structures and contact residues in the AMY2/BASI complex. The three-dimensional structure of the AMY2/BASI...

  16. Barley germination

    DEFF Research Database (Denmark)

    Daneri-Castro, Sergio N.; Svensson, Birte; Roberts, Thomas H.

    2016-01-01

    conditions continue to be key to discovering the roles of individual protein forms and posttranslational modifications, such as glycosylation. Activity-based proteomics, particularly in combination with new gene editing technologies, has great potential to elucidate the network of enzymes in barley...

  17. Mixed cropping of annual feed legumes with barley improves feed quantity and crude protein content under dry-land conditions

    Directory of Open Access Journals (Sweden)

    Khoshnood Alizadeh

    2013-01-01

    Full Text Available The objective of this research is to determine a suitable mixture of annual feed legumes and barley as a winter crop under dry-land conditions. Seeds of Hungarian vetch (cv. 2670, smooth vetch (cv. Maragheh, and local varieties of grass pea and field pea were mixed with barley (cv. Abidar in a 1:1 ratio and were tested, along with related monoculture. All legumes in the mixture survived winter while legumes alone, except Hungarian vetch, did not survive in the cold areas. The maximum fresh and dry forage yields (56 and 15 ton ha-1 respectively were obtained from a mixture of smooth vetch and barley in provinces with mild winter and more than 400 mm of rainfall. The mixture of barley and smooth vetch resulted in the highest mean crude protein content (17%. Autumn seeding of smooth vetch and barley in a 1:1 ratio produced more than 2 ton ha-1 of dry biomass with good quality in all studied areas and thus could serve as an alternative cropping system after wheat/barley in cold and semi-cold dry land.

  18. Protein and Carbohydrate Accumulation in Normal and High-Lysine Barley in Spike Culture

    DEFF Research Database (Denmark)

    Mather, D.E; Giese, Nanna Henriette

    1984-01-01

    -soluble, hordein, and non-protein N fractions appeared to be normal. Endosperm dry weight and starch were lower than in intact plants and declined at higher N levels. A linear relationship was observed between starch content and the concentration of sucrose in the endosperm water. Uptake of culture medium......Spikes of barley cv. Bomi and high-lysine mutants Riso 1508 and Riso 56 were cultured on liquid media at varying N and sucrose levels. Bomi accumulated N in response to increasing N levels in the medium and a higher level was reached than in spikes of intact plants. The distribution of N in salt...

  19. Preparation of Barley Storage Protein, Hordein, for Analytical Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

    DEFF Research Database (Denmark)

    Doll, Hans; Andersen, Bente

    1981-01-01

    The extraction, reduction, and alkylation of barley hordein for routine electrophoresis in sodium dodecyl sulfate-polyacrylamide gels were studied to set up a simple preparation procedure giving well-resolved bands in the electrophoresis gel. Hordein was extracted from single crushed seeds or flour...... by aqueous 50% propan-2-ol containing a Tris-borate buffer, pH 8.6. The presence of the buffer facilitates the consecutive complete reduction of the extracted protein in the alcohol. Reduction and alkylation in the buffer containing propan-2-ol give sharper bands in the electrophoresis than reduction...

  20. A functional genomics approach to understand the control and regulation of storage protein biosynthesis in barley grain

    DEFF Research Database (Denmark)

    Vincze, É; Hansen, M; Bowra, S;

    2008-01-01

    The aim of the study was to obtain an insight into amino acid and storage protein metabolism in the developing barley grain at the molecular level. Our strategy was to analyse the transcriptome of relevant pathways in developing grains of field grown barley using a grain specific microarray...... reference to the temporal expression levels of a range of genes involved in the photosynthesis process, amino acid and storage protein metabolism. We conclude that the grain specific microarray is a reliable and cost-effective tool for monitoring temporal changes in the transcriptome of the major metabolic...... pathways in the barley grain. The study described here could provide a strong complement to existing knowledge assisting further  understanding of seed development and thereby provide a foundation for plant breeding towards storage protein with improved nutritional quality....

  1. Barley lipid transfer protein, LTP1, contains a new type of lipid-like post-translational modification

    DEFF Research Database (Denmark)

    Lindorff-Larsen, Kresten; Lerche, Mathilde H.; Poulsen, Flemming Martin;

    2001-01-01

    the formation of cutin and involvement in stress and pathogen responses, but there is yet no direct demonstration of an in vivo function. We have found and characterized a novel post-translational modification of the barley nonspecific lipid transfer protein, LTP1. The protein-modification bond is of a new type...

  2. In situ identification and quantification of protein-hydrolyzing ruminal bacteria associated with the digestion of barley and corn grain.

    Science.gov (United States)

    Xia, Yun; Kong, Yunhong; Huang, Heping; Yang, Hee Eun; Forster, Robert; McAllister, Tim A

    2016-12-01

    In this study, BODIPY FL DQ™ casein staining combined with fluorescence in situ hybridization (FISH) was used to detect and identify protein-hydrolyzing bacteria within biofilms that produced active cell-surface-associated serine- and metallo-proteases during the ruminal digestion of barley and corn grain in cows fed barley-based diets at 2 different levels. A doublet coccoid bacterial morphotype associated with barley and corn grain particles fluoresced after BODIPY FL DQ™ casein staining. Bacteria with this morphotype accounted for 3%-10% of the total bacteria attached to surface of cereal grain particles, possibly indicative of an important role in the hydrolysis of the protein matrix within the endosperm. However, the identity of these predominant proteolytic bacteria could not be determined using FISH. Quantitative FISH revealed that known proteolytic species, Prevotella ruminicola, Ruminobacter amylophilus, and Butyrivibrio fibrisolvens, were attached to particles of various cultivars of barley grain and corn, confirming their role in the proteolysis of cereal grains. Differences in chemical composition among different barley cultivars did not affect the composition of proteolytic bacterial populations. However, the concentrate level in the basal diet did have an impact on the relative abundance of proteolytic bacteria and thus possibly their overall contribution to the proteolysis of cereal grains.

  3. Molecular identification based on coat protein sequences of the Barley yellow dwarf virus from Brazil

    Directory of Open Access Journals (Sweden)

    Talita Bernardon Mar

    2013-12-01

    Full Text Available Yellow dwarf disease, one of the most important diseases of cereal crops worldwide, is caused by virus species belonging to the Luteoviridae family. Forty-two virus isolates obtained from oat (Avena sativa L., wheat (Triticum aestivum L., barley (Hordeum vulgare L., corn (Zea mays L., and ryegrass (Lolium multiflorum Lam. collected between 2007 and 2008 from winter cereal crop regions in southern Brazil were screened by polymerase chain reaction (PCR with primers designed on ORF 3 (coat protein - CP for the presence of Barley yellow dwarf virus and Cereal yellow dwarf virus (B/CYDV. PCR products of expected size (~357 bp for subgroup II and (~831 bp for subgroup I were obtained for three and 39 samples, respectively. These products were cloned and sequenced. The subgroup II 3' partial CP amino acid deduced sequences were identified as BYDV-RMV (92 - 93 % of identity with "Illinois" Z14123 isolate. The complete CP amino acid deduced sequences of subgroup I isolates were confirmed as BYDV-PAV (94 - 99 % of identity and established a very homogeneous group (identity higher than 99 %. These results support the prevalence of BYDV-PAV in southern Brazil as previously diagnosed by Enzyme-Linked Immunosorbent Assay (ELISA and suggest that this population is very homogeneous. To our knowledge, this is the first report of BYDV-RMV in Brazil and the first genetic diversity study on B/CYDV in South America.

  4. Multi-method research strategy for understanding changes in storage protein composition in developing barley grain to improve nutritional profile

    DEFF Research Database (Denmark)

    Kaczmarczyk, Agnieszka Ewa

    , the increases in the nitrogen fertiliser are not accompanied by a good quallity of protein composition in barley grain. Hodeins are coded by highly polymorphic genes which are clustered into four main families. B.C.D and γ. The high N-dosage promotes C-hordein synthesis, the class with the lowest content...

  5. Proteome analysis of barley seeds: Identification of major proteins from two-dimensional gels (pl 4-7)

    DEFF Research Database (Denmark)

    Østergaard, O.; Finnie, Christine; Laugesen, S.;

    2004-01-01

    -soluble proteins in extracts of mature barley (Hordeum vulgare) seeds and to follow their fate during germination. About 1200 and 600 spots of p/ 4-7 were detected on 2-D gels by silver staining and colloidal Coomassie Brilliant Blue staining, respectively. About 300 spots were selected for in-gel digestion...

  6. Activity-based protein profiling of hydrolytic enzymes induced by gibberellic acid in isolated aleurone layers of malting barley.

    Science.gov (United States)

    Daneri-Castro, Sergio N; Chandrasekar, Balakumaran; Grosse-Holz, Friederike M; van der Hoorn, Renier A L; Roberts, Thomas H

    2016-09-01

    During barley germination, the aleurone layer secretes most of the enzymes required to degrade the endosperm, many of which are yet to be characterized. We used activity-based protein profiling (ABPP) to detect a range of active enzymes extracted from aleurone layers isolated from grains of a commercial malting barley variety incubated with or without gibberellic acid (GA). Enzymes found to be induced by GA were putative aleurains, cathepsin-B-like proteases and serine hydrolases. By using an inhibitory sugar panel, a specific active retaining β-glycosidase in the barley aleurone was identified as a putative xylanase. Our results show that ABPP can be used rapidly to identify a variety of active enzyme isoforms in cereal aleurone without the need for enzyme purification.

  7. Heat-stable proteins and abscisic acid action in barley aleurone cells

    Energy Technology Data Exchange (ETDEWEB)

    Jacobsen, J.V. (CSIRO, Canberra (Australia)); Shaw, D.C. (Australian National Univ., Canberra (Australia))

    1989-12-01

    ({sup 35}S)Methionine labeling experiments showed that abscisic acid (ABA) induced the synthesis of at least 25 polypeptides in mature barley (Hordeum vulgare) aleurone cells. The polypeptides were not secreted. Whereas most of the proteins extracted from aleurone cells were coagulated by heating to 100{degree}C for 10 minutes, most of the ABA-induced polypeptides remained in solution (heat-stable). ABA had little effect on the spectrum of polypeptides that were synthesized and secreted by aleurone cells, and most of these secreted polypeptides were also heat-stable. Coomassie blue staining of sodium dodecyl sulfate polyacrylamide gels indicated that ABA-induced polypeptides already occurred in high amounts in mature aleurone layers having accumulated during grain development. About 60% of the total protein extracted from mature aleurone was heat stable. Amino acid analyses of total preparations of heat-stable and heat-labile proteins showed that, compared to heat-labile proteins, heat-stable intracellular proteins were characterized by higher glutamic acid/glutamine (Glx) and glycine levels and lower levels of neutral amino acids. Secreted heat-stable proteins were rich in Glx and proline. The possibilities that the accumulation of the heat-stable polypeptides during grain development is controlled by ABA and that the function of these polypeptides is related to their abundance and extraordinary heat stability are considered.

  8. Isolation, molecular characterisation and mapping of four rose MLO orthologues

    Directory of Open Access Journals (Sweden)

    Helgard eKaufmann

    2012-11-01

    Full Text Available Powdery mildew is a major disease of economic importance in cut and pot roses. As an alternative to conventional resistance breeding strategies utilising single-dominant genes or QTLs, mildew resistance locus o (MLO-based resistance might offer some advantages. In dicots such as Arabidopsis, pea and tomato, loss-of-function mutations in MLO genes confer high levels of broad-spectrum resistance. Here, we report the isolation and characterisation of four MLO homologues from a large rose EST collection isolated from leaves. These genes are phylogenetically closely related to other dicot MLO genes that are involved in plant powdery mildew interactions. Therefore, they are candidates for MLO genes involved in rose powdery mildew interactions. Two of the four isolated genes contain all of the sequence signatures considered to be diagnostic for MLO genes. We mapped all four genes to three linkage groups and conducted the first analysis of alternative alleles. This information is discussed in regards to a reverse genetics approach aimed at the selection of rose plants that are homozygous for loss of function in one or more MLO genes.

  9. Isolation, Molecular Characterization, and Mapping of Four Rose MLO Orthologs.

    Science.gov (United States)

    Kaufmann, Helgard; Qiu, Xianqin; Wehmeyer, Juliane; Debener, Thomas

    2012-01-01

    Powdery mildew is a major disease of economic importance in cut and pot roses. As an alternative to conventional resistance breeding strategies utilizing single-dominant genes or QTLs, mildew resistance locus o (MLO)-based resistance might offer some advantages. In dicots such as Arabidopsis, pea, and tomato, loss-of-function mutations in MLO genes confer high levels of broad-spectrum resistance. Here, we report the isolation and characterization of four MLO homologs from a large rose EST collection isolated from leaves. These genes are phylogenetically closely related to other dicot MLO genes that are involved in plant powdery mildew interactions. Therefore, they are candidates for MLO genes involved in rose powdery mildew interactions. Two of the four isolated genes contain all of the sequence signatures considered to be diagnostic for MLO genes. We mapped all four genes to three linkage groups and conducted the first analysis of alternative alleles. This information is discussed in regards to a reverse genetics approach aimed at the selection of rose plants that are homozygous for loss-of-function in one or more MLO genes.

  10. Factorial combinations of protein interactions generate a multiplicity of florigen activation complexes in wheat and barley.

    Science.gov (United States)

    Li, Chengxia; Lin, Huiqiong; Dubcovsky, Jorge

    2015-10-01

    The FLOWERING LOCUS T (FT) protein is a central component of a mobile flowering signal (florigen) that is transported from leaves to the shoot apical meristem (SAM). Two FT monomers and two DNA-binding bZIP transcription factors interact with a dimeric 14-3-3 protein bridge to form a hexameric protein complex. This complex, designated as the 'florigen activation complex' (FAC), plays a critical role in flowering. The wheat homologue of FT, designated FT1 (= VRN3), activates expression of VRN1 in the leaves and the SAM, promoting flowering under inductive long days. In this study, we show that FT1, other FT-like proteins, and different FD-like proteins, can interact with multiple wheat and barley 14-3-3 proteins. We also identify the critical amino acid residues in FT1 and FD-like proteins required for their interactions, and demonstrate that 14-3-3 proteins are necessary bridges to mediate the FT1-TaFDL2 interaction. Using in vivo bimolecular fluorescent complementation (BiFC) assays, we demonstrate that the interaction between FT1 and 14-3-3 occurs in the cytoplasm, and that this complex is then translocated to the nucleus, where it interacts with TaFDL2 to form a FAC. We also demonstrate that a FAC including FT1, TaFDL2 and Ta14-3-3C can bind to the VRN1 promoter in vitro. Finally, we show that relative transcript levels of FD-like and 14-3-3 genes vary among tissues and developmental stages. Since FD-like proteins determine the DNA specificity of the FACs, variation in FD-like gene expression can result in spatial and temporal modulation of the effects of mobile FT-like signals.

  11. The influence of barley malt protein modification on beer foam stability and their relationship to the barley dimeric alpha-amylase inhibitor-I (BDAI-I) as a possible foam-promoting protein.

    Science.gov (United States)

    Okada, Yoshihiro; Iimure, Takashi; Takoi, Kiyoshi; Kaneko, Takafumi; Kihara, Makoto; Hayashi, Katsuhiro; Ito, Kazutoshi; Sato, Kazuhiro; Takeda, Kazuyoshi

    2008-02-27

    The foam stability of beer is one of the important key factors in evaluating the quality of beer. The purpose of this study was to investigate the relationship between the level of malt modification (degradation of protein, starch, and so on) and the beer foam stability. This was achieved by examining foam-promoting proteins using two-dimensional gel electrophoresis (2DE). We found that the foam stability of beer samples brewed from the barley malts of cultivars B and C decreased as the level of malt modification increased; however, the foam stability of cultivar A did not change. To identify the property providing the increased foam stability of cultivar A, we analyzed beer proteins using 2DE. We analyzed three fractions that could contain beer foam-promoting proteins, namely, beer whole proteins, salt-precipitated proteins, and the proteins concentrated from beer foam. As a result, we found that in cultivar A, some protein spots did not change in any of these three protein fractions even when the level of malt modification increased, although the corresponding protein spots in cultivars B and C decreased. We analyzed these protein spots by peptide mass finger printing using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. As a result, all of these spots were identified as barley dimeric alpha-amylase inhibitor-I (BDAI-I). These results suggest that BDAI-I is an important contributor to beer foam stability.

  12. Proteomes of the barley aleurone layer: A model system for plant signalling and protein secretion.

    Science.gov (United States)

    Finnie, Christine; Andersen, Birgit; Shahpiri, Azar; Svensson, Birte

    2011-05-01

    The cereal aleurone layer is of major importance due to its nutritional properties as well as its central role in seed germination and industrial malting. Cereal seed germination involves mobilisation of storage reserves in the starchy endosperm to support seedling growth. In response to gibberellic acid produced by the embryo, the aleurone layer synthesises hydrolases that are secreted to the endosperm for the degradation of storage products. The barley aleurone layer can be separated from the other seed tissues and maintained in culture, allowing the study of the effect of added signalling molecules in an isolated system. These properties have led to its use as a model system for the study of plant signalling and germination. More recently, proteome analysis of the aleurone layer has provided new insight into this unique tissue including identification of plasma membrane proteins and targeted analysis of germination-related changes and the thioredoxin system. Here, analysis of intracellular and secreted proteomes reveals features of the aleurone layer system that makes it promising for investigations of plant protein secretion mechanisms.

  13. Cy5 maleimide labelling for sensitive detection of free thiols in native protein extracts: identification of seed proteins targeted by barley thioredoxin h isoforms.

    Science.gov (United States)

    Maeda, Kenji; Finnie, Christine; Svensson, Birte

    2004-03-01

    Barley thioredoxin h isoforms HvTrxh1 and HvTrxh2 differ in temporal and spatial distribution and in kinetic properties. Target proteins of HvTrxh1 and HvTrxh2 were identified in mature seeds and in seeds after 72 h of germination. Improvement of the established method for identification of thioredoxin-targeted proteins based on two-dimensional electrophoresis and fluorescence labelling of thiol groups was achieved by application of a highly sensitive Cy5 maleimide dye and large-format two-dimensional gels, resulting in a 10-fold increase in the observed number of labelled protein spots. The technique also provided information about accessible thiol groups in the proteins identified in the barley seed proteome. In total, 16 different putative target proteins were identified from 26 spots using tryptic in-gel digestion, matrix-assisted laser-desorption ionization-time-of-flight MS and database search. HvTrxh1 and HvTrxh2 were shown to have similar target specificity. Barley alpha-amylase/subtilisin inhibitor, previously demonstrated to be reduced by both HvTrxh1 and HvTrxh2, was among the identified target proteins, confirming the suitability of the method. Several alpha-amylase/trypsin inhibitors, some of which are already known as target proteins of thioredoxin h, and cyclophilin known as a target protein of m-type thioredoxin were also identified. Lipid transfer protein, embryospecific protein, three chitinase isoenzymes, a single-domain glyoxalase-like protein and superoxide dismutase were novel identifications of putative target proteins, suggesting new physiological roles of thioredoxin h in barley seeds.

  14. Proteomes of the barley aleurone layer: A model system for plant signalling and protein secretion

    DEFF Research Database (Denmark)

    Finnie, Christine; Andersen, Birgit; Shahpiri, Azar

    2011-01-01

    The cereal aleurone layer is of major importance due to its nutritional properties as well as its central role in seed germination and industrial malting. Cereal seed germination involves mobilisation of storage reserves in the starchy endosperm to support seedling growth. In response to gibberel......The cereal aleurone layer is of major importance due to its nutritional properties as well as its central role in seed germination and industrial malting. Cereal seed germination involves mobilisation of storage reserves in the starchy endosperm to support seedling growth. In response...... to gibberellic acid produced by the embryo, the aleurone layer synthesises hydrolases that are secreted to the endosperm for the degradation of storage products. The barley aleurone layer can be separated from the other seed tissues and maintained in culture, allowing the study of the effect of added signalling...... molecules in an isolated system. These properties have led to its use as a model system for the study of plant signalling and germination. More recently, proteome analysis of the aleurone layer has provided new insight into this unique tissue including identification of plasma membrane proteins and targeted...

  15. Cy5 maleimide labelling for sensitive detection of free thiols in native protein extracts: identification of seed proteins targeted by barley thioredoxin h isoforms

    DEFF Research Database (Denmark)

    Maeda, K.; Finnie, Christine; Svensson, Birte

    2004-01-01

    search. HvTrxh1 and HvTrxh2 were shown to have similar target specificity. Barley alpha-amylase/subtilisin inhibitor, previously demonstrated to be reduced by both HvTrxh1 and HvTrxh2, was among the identified target proteins, confirming the suitability of the method. Several alpha-amylase/trypsin...... inhibitors, some of which are already known as target proteins of thioredoxin h, and cyclophilin known as a target protein of m-type thioredoxin were also identified. Lipid transfer protein, embryo-specific protein, three chitinase isoenzymes, a single-domain glyoxalase-like protein and superoxide dismutase...

  16. Case Study of Somaclonal Variation in Resistance Genes Mlo and Pme3 in Flaxseed (Linum usitatissimum L.) Induced by Nanoparticles

    Science.gov (United States)

    Kokina, Inese; Jermaļonoka, Marija; Bankovska, Linda; Gerbreders, Vjačeslavs; Ogurcovs, Andrejs; Jahundoviča, Inese

    2017-01-01

    Nanoparticles influence on genome is investigated worldwide. The appearance of somaclonal variation is a cause of great concern for any micropropagation system. Somaclonal variation describes the tissue-culture-induced phenotypic and genotypic variations. This paper shows the results of somaclonal variation in two resistance genes pectin methylesterase and Mlo-like protein in all tissue culture development stages, as donor plant, calluses, and regenerants of Linum usitatissimum induced by gold and silver nanoparticles. In this paper, it was essential to obtain DNA material from all tissue culture development stages from one donor plant to record changes in each nucleotide sequence. Gene region specific primers were developed for resistance genes such as Mlo and Pme3 to define the genetic variability in tissue culture of L. usitatissimum. In recent years, utilization of gold and silver nanoparticles in tissue culture is increased and the mechanisms of changes in genome induced by nanoparticles still remain unclear. Obtained data show the somaclonal variation increase in calluses obtained from one donor plant and grown on medium supplemented by gold nanoparticles (Mlo 14.68 ± 0.98; Pme3 2.07 ± 0.87) or silver nanoparticles (Mlo 12.01 ± 0.43; Pme3 10.04 ± 0.46) and decrease in regenerants. Morphological parameters of calluses showed a number of differences between each investigated culture group. PMID:28326314

  17. Genomic identification, phylogeny, and expression analysis of MLO genes involved in susceptibility to powdery mildew in Fragaria vesca.

    Science.gov (United States)

    Miao, L X; Jiang, M; Zhang, Y C; Yang, X F; Zhang, H Q; Zhang, Z F; Wang, Y Z; Jiang, G H

    2016-08-05

    The MLO (powdery mildew locus O) gene family is important in resistance to powdery mildew (PM). In this study, all of the members of the MLO family were identified and analyzed in the strawberry (Fragaria vesca) genome. The strawberry contains at least 20 members of the MLO family, and the protein sequence contained between 171 and 1485 amino acids, with 0-34 introns. Chromosomal localization showed that the MLOs were unevenly distributed on each of the chromosomes, except for chromosome 4. The greatest number of MLOs (seven) was found on chromosome 3. A phylogenetic tree showed that the MLOs were divided into seven groups (I-VII), four of which consisted of MLOs from strawberry, Arabidopsis thaliana, rice, and maize, suggesting that these genes may have evolved after the divergence of monocots and dicots. Multiple sequence alignment showed that strawberry MLO candidates related to powdery mildew resistance possessed seven highly conserved transmembrane domains, a calmodulin-binding domain, and two conserved regions, all of which are important domains for powdery mildew resistance genes. Expressed sequence tag analysis revealed that the MLOs were induced by multiple abiotic stressors, including low and high temperature, drought, and high salinity. These findings will contribute to the functional characterization of MLOs related to PM susceptibility, and will assist in the development of disease resistance in strawberries.

  18. Mechanism for the increased defecation and jejunum mucosal protein content in rats by feeding germinated barley foodstuff.

    Science.gov (United States)

    Kanauchi, O; Agata, K; Fushiki, T

    1997-03-01

    We investigated the effects of germinated barley foodstuff (GBF) on the fecal excretion and jejunum mucosal protein content in male Sprague-Dawley rats fed on various diets with the same protein and dietary fiber levels. Under these experimental conditions, GBF was confirmed to induce greater fecal output compared with commercial water-soluble or -insoluble dietary fibers. While the dietary fiber extracted from GBF increased the fecal output and mucosal protein content, the protein fraction of GBF degraded to the peptide form did not increase the fecal output or mucosal protein content. Increased mucosal protein and fecal output were thus found to require the presence of the dietary fiber fraction or possibly the protein fraction bound tightly to the dietary fiber of GBF. GBF feeding increased the volatile fatty acids concentration in the cecum, indicating that GBF may be efficiently fermented in the intestinal tract.

  19. Microwave irradiation induced changes in protein molecular structures of barley grains: relationship to changes in protein chemical profile, protein subfractions, and digestion in dairy cows.

    Science.gov (United States)

    Yan, Xiaogang; Khan, Nazir A; Zhang, Fangyu; Yang, Ling; Yu, Peiqiang

    2014-07-16

    The objectives of this study were to evaluate microwave irradiation (MIR) induced changes in crude protein (CP) subfraction profiles, ruminal CP degradation characteristics and intestinal digestibility of rumen undegraded protein (RUP), and protein molecular structures in barley (Hordeum vulgare) grains. Samples from hulled (n = 1) and hulless cultivars (n = 2) of barley, harvested from four replicate plots in two consecutive years, were evaluated. The samples were either kept as raw or irradiated in a microwave for 3 min (MIR3) or 5 min (MIR5). Compared to raw grains, MIR5 decreased the contents of rapidly degradable CP subfraction (from 45.22 to 6.36% CP) and the ruminal degradation rate (from 8.16 to 3.53%/h) of potentially degradable subfraction. As a consequence, the effective ruminal degradability of CP decreased (from 55.70 to 34.08% CP) and RUP supply (from 43.31 to 65.92% CP) to the postruminal tract increased. The MIR decreased the spectral intensities of amide 1, amide II, α-helix, and β-sheet and increased their ratios. The changes in protein spectral intensities were strongly correlated with the changes in CP subfractions and digestive kinetics. These results show that MIR for a short period (5 min) with a lower energy input can improve the nutritive value and utilization of CP in barely grains.

  20. UNcleProt (Universal Nuclear Protein database of barley): The first nuclear protein database that distinguishes proteins from different phases of the cell cycle.

    Science.gov (United States)

    Blavet, Nicolas; Uřinovská, Jana; Jeřábková, Hana; Chamrád, Ivo; Vrána, Jan; Lenobel, René; Beinhauer, Jana; Šebela, Marek; Doležel, Jaroslav; Petrovská, Beáta

    2017-01-02

    Proteins are the most abundant component of the cell nucleus, where they perform a plethora of functions, including the assembly of long DNA molecules into condensed chromatin, DNA replication and repair, regulation of gene expression, synthesis of RNA molecules and their modification. Proteins are important components of nuclear bodies and are involved in the maintenance of the nuclear architecture, transport across the nuclear envelope and cell division. Given their importance, the current poor knowledge of plant nuclear proteins and their dynamics during the cell's life and division is striking. Several factors hamper the analysis of the plant nuclear proteome, but the most critical seems to be the contamination of nuclei by cytosolic material during their isolation. With the availability of an efficient protocol for the purification of plant nuclei, based on flow cytometric sorting, contamination by cytoplasmic remnants can be minimized. Moreover, flow cytometry allows the separation of nuclei in different stages of the cell cycle (G1, S, and G2). This strategy has led to the identification of large number of nuclear proteins from barley (Hordeum vulgare), thus triggering the creation of a dedicated database called UNcleProt, http://barley.gambrinus.ueb.cas.cz/ .

  1. Starch granule protein (SGP) polymorphism in cultivated naked barley from Qinghai-Tibet Plateau in China and relationship between SGPs and starch/amylose content

    Institute of Scientific and Technical Information of China (English)

    Zhifen PAN; Yixing ZOU; Tao ZHAO; Guangbing DENG; Xuguang ZHAI; Fang WU; Maoqun YU

    2008-01-01

    Starch granule proteins (SGPs) are minor components bound with starch granule, whose variation could impact starch properties. This study investigated, for the first time, the variation of SGPs in the cultivated naked barley from Qinghai-Tibet Plateau in China. The relation-ship between SGPs and starch content was preliminarily dealt with. Ten major SGPs and 16 types of patterns were present in the 66 cultivated naked varieties, indicating that the SGPs in cultivated naked barley from Qinghai-Tibet Plateau in China are polymorphic. The SGPs of naked barley in Tibet and Sichuan were greatly different and the SGP patterns were specific to sampling regions. Significance test analysis demonstrated that the SGPs described in this study, except for SGP1, could be related with the variation of starch content in the different naked barleys.

  2. Identification of a protein which differs in lines of barley resistant or susceptible to Erysiphe graminis f. sp. hordei

    Energy Technology Data Exchange (ETDEWEB)

    Simons, S.P.; Somerville, S.C. (Michigan State Univ., East Lansing (USA))

    1990-05-01

    As yet no resistance genes or gene products have been isolated from a plant. We wish to isolate the Ml-a gene which encodes resistance by barley to Erysiphe graminis f. sp. hordei, the causal agent of the powdery mildew disease. We have utilized near isogenic barley lines, resistant and susceptible to E. g. hordei, race CR3 to provide a homogenous background for examination of protein differences related to resistance. To enrich for epidermal tissue, the site of infection, proteins from {sup 35}S labelled coleoptiles were analyzed by 2D-PAGE. Two major protein differences were observed, one of which is a polymorphism identified by a higher molecular weight form in the susceptible line, with the lower molecular weight form appearing in the resistant line. A partially susceptible mutant line derived from the resistant line shows a 63% reduction in the level of the resistant form of the polypeptide. A likely conclusion from these results is that we have identified the Ml-a gene product. Further characterization of the polymorphic protein and its gene will lead to an understanding of gene-for-gene related resistance processes.

  3. Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials

    Science.gov (United States)

    Schalk, Kathrin; Lexhaller, Barbara; Koehler, Peter

    2017-01-01

    Gluten proteins from wheat, rye, barley and, in rare cases, oats, are responsible for triggering hypersensitivity reactions such as celiac disease, non-celiac gluten sensitivity and wheat allergy. Well-defined reference materials (RM) are essential for clinical studies, diagnostics, elucidation of disease mechanisms and food analyses to ensure the safety of gluten-free foods. Various RM are currently used, but a thorough characterization of the gluten source, content and composition is often missing. However, this characterization is essential due to the complexity and heterogeneity of gluten to avoid ambiguous results caused by differences in the RM used. A comprehensive strategy to isolate gluten protein fractions and gluten protein types (GPT) from wheat, rye, barley and oat flours was developed to obtain well-defined RM for clinical assays and gluten-free compliance testing. All isolated GPT (ω5-gliadins, ω1,2-gliadins, α-gliadins, γ-gliadins and high- and low-molecular-weight glutenin subunits from wheat, ω-secalins, γ-75k-secalins, γ-40k-secalins and high-molecular-weight secalins from rye, C-hordeins, γ-hordeins, B-hordeins and D-hordeins from barley and avenins from oats) were fully characterized using analytical reversed-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), N-terminal sequencing, electrospray-ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS) and untargeted LC-MS/MS of chymotryptic hydrolyzates of the single GPT. Taken together, the analytical methods confirmed that all GPT were reproducibly isolated in high purity from the flours and were suitable to be used as RM, e.g., for calibration of LC-MS/MS methods or enzyme-linked immunosorbent assays (ELISAs). PMID:28234993

  4. Two seven-transmembrane domain MILDEW RESISTANCE LOCUS O proteins cofunction in Arabidopsis root thigmomorphogenesis.

    Science.gov (United States)

    Chen, Zhongying; Noir, Sandra; Kwaaitaal, Mark; Hartmann, H Andreas; Wu, Ming-Jing; Mudgil, Yashwanti; Sukumar, Poornima; Muday, Gloria; Panstruga, Ralph; Jones, Alan M

    2009-07-01

    Directional root expansion is governed by nutrient gradients, positive gravitropism and hydrotropism, negative phototropism and thigmotropism, as well as endogenous oscillations in the growth trajectory (circumnutation). Null mutations in phylogenetically related Arabidopsis thaliana genes MILDEW RESISTANCE LOCUS O 4 (MLO4) and MLO11, encoding heptahelical, plasma membrane-localized proteins predominantly expressed in the root tip, result in aberrant root thigmomorphogenesis. mlo4 and mlo11 mutant plants show anisotropic, chiral root expansion manifesting as tightly curled root patterns upon contact with solid surfaces. The defect in mlo4 and mlo11 mutants is nonadditive and dependent on light and nutrients. Genetic epistasis experiments demonstrate that the mutant phenotype is independently modulated by the Gbeta subunit of the heterotrimeric G-protein complex. Analysis of expressed chimeric MLO4/MLO2 proteins revealed that the C-terminal domain of MLO4 is necessary but not sufficient for MLO4 action in root thigmomorphogenesis. The expression of the auxin efflux carrier fusion, PIN1-green fluorescent protein, the pattern of auxin-induced gene expression, and acropetal as well as basipetal auxin transport are altered at the root tip of mlo4 mutant seedlings. Moreover, addition of auxin transport inhibitors or the loss of EIR1/AGR1/PIN2 function abolishes root curling of mlo4, mlo11, and wild-type seedlings. These results demonstrate that the exaggerated root curling phenotypes of the mlo4 and mlo11 mutants depend on auxin gradients and suggest that MLO4 and MLO11 cofunction as modulators of touch-induced root tropism.

  5. Protein mobilization and malting-specific proteinase expression during barley germination

    Science.gov (United States)

    Robust barley was germinated under three different conditions: 1) a production run in a commercial malting facility, 2) a micromalting protocol developed to generate malt with malting quality metrics similar to that from the commercial malting, and 3) a simple laboratory setting using a humidified ...

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

    Science.gov (United States)

    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...

  7. Two-dimensional gel electrophoresis pattern (pH 6-11) and identification of water-soluble barley seed and malt proteins by mass spectrometry

    DEFF Research Database (Denmark)

    Bak-Jensen, K.S.; Laugesen, S.; Roepstorff, P.

    2004-01-01

    A protocol was established for two-dimensional gel electrophoresis (2-DE) of barley seed and malt proteins in the pH range of 6-11. Proteins extracted from flour in a low-salt buffer were focused after cup-loading onto IPG strips. Successful separation in the second dimension was achieved using...

  8. Extraction, separation, and purification of wheat gluten proteins and related proteins of barley, rye, and oats.

    Science.gov (United States)

    Tatham, A S; Gilbert, S M; Fido, R J; Shewry, P R

    2000-01-01

    The wheat proteins, which are active in celiac disease and other glutenrelated conditions, are defined as prolamins, in that they are soluble as individual subunits in alcohol-water mixtures, such as 50% (v/v) aqueous propan-1-ol or 60-70% (v/v) aqueous ethanol. However, in wheat grain and flour, about half of these subunits are present in polymers that are not soluble in alcohol-water mixtures unless disulfide bonds between the component subunits are reduced using an agent such as 2-mercaptoethanol (2-ME) or dithiothreitol (DTT). These alcohol-insoluble polymers are traditionally called glutenins and the related alcohol-soluble monomers are called gliadins; the two groups of proteins together form the major part of the gluten fraction. Gluten can be readily prepared from wheat by washing dough to remove the bulk of the starch, cell-wall material, and soluble components. It is a cohesive viscoelastic mass that contains, in addition to the gluten proteins, small amounts of other proteins, residual starch (about 25% dry wt), and lipid.

  9. Low-resolution structure of the full-length barley (Hordeum vulgare SGT1 protein in solution, obtained using small-angle X-ray scattering.

    Directory of Open Access Journals (Sweden)

    Michał Taube

    Full Text Available SGT1 is an evolutionarily conserved eukaryotic protein involved in many important cellular processes. In plants, SGT1 is involved in resistance to disease. In a low ionic strength environment, the SGT1 protein tends to form dimers. The protein consists of three structurally independent domains (the tetratricopeptide repeats domain (TPR, the CHORD- and SGT1-containing domain (CS, and the SGT1-specific domain (SGS, and two less conserved variable regions (VR1 and VR2. In the present study, we provide the low-resolution structure of the barley (Hordeum vulgare SGT1 protein in solution and its dimer/monomer equilibrium using small-angle scattering of synchrotron radiation, ab-initio modeling and circular dichroism spectroscopy. The multivariate curve resolution least-square method (MCR-ALS was applied to separate the scattering data of the monomeric and dimeric species from a complex mixture. The models of the barley SGT1 dimer and monomer were formulated using rigid body modeling with ab-initio structure prediction. Both oligomeric forms of barley SGT1 have elongated shapes with unfolded inter-domain regions. Circular dichroism spectroscopy confirmed that the barley SGT1 protein had a modular architecture, with an α-helical TPR domain, a β-sheet sandwich CS domain, and a disordered SGS domain separated by VR1 and VR2 regions. Using molecular docking and ab-initio protein structure prediction, a model of dimerization of the TPR domains was proposed.

  10. Increased Abundance of Proteins Involved in Phytosiderophore Production in Boron-Tolerant Barley1[C][W

    Science.gov (United States)

    Patterson, John; Ford, Kris; Cassin, Andrew; Natera, Siria; Bacic, Antony

    2007-01-01

    Boron (B) phytotoxicity affects cereal-growing regions worldwide. Although B-tolerant barley (Hordeum vulgare) germplasm is available, molecules responsible for this tolerance mechanism have not been defined. We describe and use a new comparative proteomic technique, iTRAQ peptide tagging (iTRAQ), to compare the abundances of proteins from B-tolerant and -intolerant barley plants from a ‘Clipper’ × ‘Sahara’ doubled-haploid population selected on the basis of a presence or absence of two B-tolerance quantitative trait loci. iTRAQ was used to identify three enzymes involved in siderophore production (Iron Deficiency Sensitive2 [IDS2], IDS3, and a methylthio-ribose kinase) as being elevated in abundance in the B-tolerant plants. Following from this result, we report a potential link between iron, B, and the siderophore hydroxymugineic acid. We believe that this study highlights the potency of the iTRAQ approach to better understand mechanisms of abiotic stress tolerance in cereals, particularly when applied in conjunction with bulked segregant analysis. PMID:17478636

  11. Differential disease resistance response in the barley necrotic mutant nec1

    Directory of Open Access Journals (Sweden)

    Kunga Laura

    2011-04-01

    Full Text Available Abstract Background Although ion fluxes are considered to be an integral part of signal transduction during responses to pathogens, only a few ion channels are known to participate in the plant response to infection. CNGC4 is a disease resistance-related cyclic nucleotide-gated ion channel. Arabidopsis thaliana CNGC4 mutants hlm1 and dnd2 display an impaired hypersensitive response (HR, retarded growth, a constitutively active salicylic acid (SA-mediated pathogenesis-related response and elevated resistance against bacterial pathogens. Barley CNGC4 shares 67% aa identity with AtCNGC4. The barley mutant nec1 comprising of a frame-shift mutation of CNGC4 displays a necrotic phenotype and constitutively over-expresses PR-1, yet it is not known what effect the nec1 mutation has on barley resistance against different types of pathogens. Results nec1 mutant accumulated high amount of SA and hydrogen peroxide compared to parental cv. Parkland. Experiments investigating nec1 disease resistance demonstrated positive effect of nec1 mutation on non-host resistance against Pseudomonas syringae pv. tomato (Pst at high inoculum density, whereas at normal Pst inoculum concentration nec1 resistance did not differ from wt. In contrast to augmented P. syringae resistance, penetration resistance against biotrophic fungus Blumeria graminis f. sp. hordei (Bgh, the causal agent of powdery mildew, was not altered in nec1. The nec1 mutant significantly over-expressed race non-specific Bgh resistance-related genes BI-1 and MLO. Induction of BI-1 and MLO suggested putative involvement of nec1 in race non-specific Bgh resistance, therefore the effect of nec1on mlo-5-mediated Bgh resistance was assessed. The nec1/mlo-5 double mutant was as resistant to Bgh as Nec1/mlo-5 plants, suggesting that nec1 did not impair mlo-5 race non-specific Bgh resistance. Conclusions Together, the results suggest that nec1 mutation alters activation of systemic acquired resistance

  12. Barley peroxidase isozymes

    Science.gov (United States)

    Laugesen, Sabrina; Bak-Jensen, Kristian Sass; Hägglund, Per; Henriksen, Anette; Finnie, Christine; Svensson, Birte; Roepstorff, Peter

    2007-12-01

    Thirteen peroxidase spots on two-dimensional gels were identified by comprehensive proteome analysis of the barley seed. Mass spectrometry tracked multiple forms of three different peroxidase isozymes: barley seed peroxidase 1, barley seed-specific peroxidase BP1 and a not previously identified putative barley peroxidase. The presence of multiple spots for each of the isozymes reflected variations in post-translational glycosylation and protein truncation. Complete sequence coverage was achieved by using a series of proteases and chromatographic resins for sample preparation prior to mass spectrometric analysis. Distinct peroxidase spot patterns divided the 16 cultivars tested into two groups. The distribution of the three isozymes in different seed tissues (endosperm, embryo, and aleurone layer) suggested the peroxidases to play individual albeit partially overlapping roles during germination. In summary, a subset of three peroxidase isozymes was found to occur in the seed, whereas products of four other barley peroxidase genes were not detected. The present analysis documents the selective expression profiles and post-translational modifications of isozymes from a large plant gene family.

  13. Protein and hordein fraction content in barley seeds as affected by sowing date and their relations to malting quality

    Institute of Scientific and Technical Information of China (English)

    QI Jun-cong; CHEN Jin-xin; WANG Jun-mei; WU Fei-bo; CAO Lian-pu; ZHANG Guo-ping

    2005-01-01

    The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relationships among these traits were examined. The results showed that grain protein content and B hordein content increased as the sowing date postponed and were significantly affected by sowing date, while C and D hordein contents were less influenced by sowing date. There were significant differences in grain protein and hordein fraction content among the ten cultivars. The coefficient of variation of D hordein content was much larger than that of B and C hordein contents, suggesting its greater variation caused by different sowing dates. Beta-amylase activity and diastatic power were also significantly affected by sowing date, with malt extract being less affected. Significant differences in measured malt quality were found among the ten cultivars. Grain protein was significantly correlated with B hordein and malt extract positively and negatively, respectively. There was no significant correlation between beta-amylase activity or diastatic power and grain protein content. B hordein was negatively and significantly correlated with malt extract, but no significant correlations between C hordein, D hordein and malting quality traits.

  14. PTK1, a mitogen-activated-protein kinase gene, is required for conidiation, appressorium formation, and pathogenicity of Pyrenophora teres on barley.

    Science.gov (United States)

    Ruiz-Roldán, M C; Maier, F J; Schäfer, W

    2001-02-01

    Mitogen-activated protein kinases (MAPKs) are a group of protein kinases that execute a wide variety of roles in cellular signal transduction pathways such as osmoregulation, cell wall biosynthesis, growth, and differentiation. A polymerase chain reaction (PCR) with degenerate primers based on conserved regions of known MAPKs was used to clone the MAPK gene PTK1 from the leaf pathogen Pyrenophora teres (anamorph Drechslera teres), the causal agent of net blotch of barley (Hordeum vulgare L.). The predicted amino acid sequence shows high homology with MAPKs from other phytopathogenic fungi. The gene is present in the genome as a single copy. PTK1 is expressed during in vitro growth on complete medium, under conidiation-inducing conditions and during infection of barley leaves, as shown by reverse transcription-PCR studies. In order to assess the role of PTK1 in the life cycle of P. teres, targeted gene disruption was conducted. Mutants carrying an interrupted copy of the gene were deficient in conidiation, did not form appressoria on glass surfaces or on barley leaves, lost their ability to infect barley leaves, and could not colonize host tissues following artificial wounding.

  15. Cloning and Expression Analysis of Mlo Gene from Pericallis hybrida B. Nord

    Institute of Scientific and Technical Information of China (English)

    Wang Jin-gang; Li Wei; Ren Hong-wei; Lv Yuan-da; Bai Ding; Xu Jing

    2014-01-01

    The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction (RT-PCR) and RACE. The result of sequence analysis indicated that Mlo gene from Pericallis hybrida B. Nord. contained about 1 296 bp open reading frame and encoded 431 amino acids. According to the comparison of the exogenous gene sequences by BLAST analysis and phylogenetic analysis, Mlo gene shared over 85%nucleotide homology and 98%amino acid homology. Finally, through semi-quantitative-PCR and fluorescence quantitative analysis, we found that Mlo gene showed the highest expression levels in leaves and the lowest in roots after inoculated with powdery mildew pathogen for different days.

  16. Molecular cloning of the barley seed protein CMd: a variant member of the alpha-amylase/trypsin inhibitor family of cereals.

    Science.gov (United States)

    Halford, N G; Morris, N A; Urwin, P; Williamson, M S; Kasarda, D D; Lew, E J; Kreis, M; Shewry, P R

    1988-09-07

    The nucleotide and deduced amino-acid sequences of a cDNA clone encoding the barley seed protein CMd are described. The sequence is homologous with those of a family of inhibitors of alpha-amylase and trypsin, except for two short insertions. The longest of these (14 residues) is at the junction between the three proposed ancestral regions that comprise this family of proteins, and has limited identity with alpha-amylases of bacterial origin.

  17. Remodeling of the major light-harvesting antenna protein of PSII protects the young leaves of barley (Hordeum vulgare L.) from photoinhibition under prolonged iron deficiency.

    Science.gov (United States)

    Saito, Akihiro; Iino, Tomohisa; Sonoike, Kintake; Miwa, Eitaro; Higuchi, Kyoko

    2010-12-01

    Because of the high demand for iron of the photosynthetic apparatus in thylakoid membranes, iron deficiency primarily affects the electron transfer between the two photosystems (PSI and PSII), resulting in photooxidative damage in plants. However, in barley, PSII is protected against photoinhibition, and the plant survives even with a low iron content in its chlorotic leaves. In this study, we report an adaptation mechanism of the photosynthetic apparatus in barley to iron deficiency, which is concomitant with the remodeling of a PSII antenna system. Transcriptome analysis revealed that long-term iron deficiency induced the expression of two genes of the major light-harvesting Chl a/b-binding protein of PSII (LHCII), namely HvLhcb1.11 and HvLhcb1.12. Chl fluorescence analysis of the wild type and Lhcb1-less chlorina mutants clearly showed that non-photochemical quenching (NPQ) of the wild type was increased by approximately 200% by iron deficiency, whereas NPQ of chlorina mutants did not change significantly under iron deficiency. The mutant showed severe photodamage in young leaves under prolonged iron deficiency, suggesting that the HvLhcb1 protein is essential for both thermal dissipation and photoprotection in iron-deficient barley. Analysis of thylakoid protein complexes revealed that the proportion of the monomeric form of Lhcb1 significantly increased in barley grown under iron-deficient conditions. We hypothesize that alteration of the HvLhcb1 subpopulations modifies the organization of LHCII in the thylakoid membranes, which is a key step for thermal dissipation to compensate for excess excitation energy and thereby protect the photosystems from serious damage in iron-deficient barley leaves.

  18. Identification of thioredoxin h-reducible disulphides in proteomes by differential labelling of cysteines: insight into recognition and regulation of proteins in barley seeds by thioredoxin h.

    Science.gov (United States)

    Maeda, Kenji; Finnie, Christine; Svensson, Birte

    2005-04-01

    Using thiol-specific fluorescence labelling, over 30 putative target proteins of thioredoxin h with diverse structures and functions have been identified in seeds of barley and other plants. To gain insight at the structural level into the specificity of target protein reduction by thioredoxin h, thioredoxin h-reducible disulphide bonds in individual target proteins are identified using a novel strategy based on differential alkylation of cysteine thiol groups by iodoacetamide and 4-vinylpyridine. This method enables the accessible cysteine side chains in the thiol form (carbamidomethylated) to be distinguished from those inaccessible or disulphide bound form (pyridylethylated) according to the mass difference in the peptide mass maps obtained by matrix-assistend laser desorption/ionisation-time of flight mass spectrometry. Using this approach, in vitro reduction of disulphides in recombinant barley alpha-amylase/subtilisin inhibitor (BASI) by barley thioredoxin h isoform 1 was analysed. Furthermore, the method was coupled with two-dimensional electrophoresis for convenient thioredoxin h-reducible disulphide identification in barley seed extracts without the need for protein purification or production of recombinant proteins. Mass shifts of 15 peptides, induced by treatment with thioredoxin h and differential alkylation, identified specific reduction of nine disulphides in BASI, four alpha-amylase/trypsin inhibitors and a protein of unknown function. Two specific disulphides, located structurally close to the alpha-amylase binding surfaces of BASI and alpha-amylase inhibitor BMAI-1 were demonstrated to be reduced to a particularly high extent. For the first time, specificity of thioredoxin h for particular disulphide bonds is demonstrated, providing a basis to study structural aspects of the recognition mechanism and regulation of target proteins.

  19. Colonization pattern of the biocontrol strain Pseudomonas chlororaphis MA 342 on barley seeds visualized by using green fluorescent protein.

    Science.gov (United States)

    Tombolini, R; van der Gaag, D J; Gerhardson, B; Jansson, J K

    1999-08-01

    Pseudomonas chlororaphis MA 342 is a potent biocontrol agent that can be used against several seed-borne diseases of cereal crops, including net blotch of barley caused by the fungus Drechslera teres. In this study, strain MA 342 was tagged with the gfp gene (encoding the green fluorescent protein) in order to study the fate of cells after seed inoculation. The gfp-tagged strain, MA 342G2, had the same biocontrol efficacy as the wild type when it was applied at high cell concentrations to seeds but was less effective at lower cell concentrations. By comparing cell counts determined by microscopy to the number of CFU, we found that the number of culturable cells was significantly lower than the total number of bacteria on seeds which were inoculated and dried for 20 h. Confocal microscopy and epifluorescence stereomicroscopy were used to determine the pattern of MA 342G2 colonization and cell aggregation on barley seeds. Immediately after inoculation of seeds, bacteria were found mainly under the seed glume, and there was no particular aggregation pattern. However, after the seeds were sown, irregularly distributed areas of bacterial aggregation were found, which reflected epiphytic colonization of glume cells. There was a trend towards bacterial aggregation near the embryo but never within the embryo. Bacterial aggregates were regularly found in the groove of each seed formed by the base of the coleoptile and the scutellum. Based on these results, we suggest that MA 342 colocalizes with the pathogen D. teres, which facilitates the action of the fungistatic compound(s) produced by this strain.

  20. Identification of thioredoxin h-reducible disulphides in proteornes by differential labelling of cysteines: Insight into recognition and regulation of proteins in barley seeds by thioredoxin h

    DEFF Research Database (Denmark)

    Maeda, Kenji; Finnie, Christine; Svensson, Birte

    2005-01-01

    or production of recombinant proteins. Mass shifts of 15 peptides, induced by treatment with thioredoxin h and differential alkylation, identified specific reduction of nine disulphides in BASI, four alpha-amylase/trypsin inhibitors and a protein of unknown function. Two specific disulphides, located...... alpha-amylase/subtilisin inhibitor (BASI) by barley thioredoxin h isoform 1 was analysed. Furthermore, the method was coupled with two-dimensional electrophoresis for convenient thioredoxin h-reducible disulphide identification in barley seed extracts without the need for protein purification...... structurally close to the alpha-amylase binding surfaces of BASI and alpha-amylase inhibitor BMAI-1 were demonstrated to be reduced to a particularly high extent. For the first time, specificity of thioredoxin h for particular disulphide bonds is demonstrated, providing a basis to study structural aspects...

  1. Live Cell Imaging During Germination Reveals Dynamic Tubular Structures Derived from Protein Storage Vacuoles of Barley Aleurone Cells

    Directory of Open Access Journals (Sweden)

    Verena Ibl

    2014-09-01

    Full Text Available The germination of cereal seeds is a rapid developmental process in which the endomembrane system undergoes a series of dynamic morphological changes to mobilize storage compounds. The changing ultrastructure of protein storage vacuoles (PSVs in the cells of the aleurone layer has been investigated in the past, but generally this involved inferences drawn from static pictures representing different developmental stages. We used live cell imaging in transgenic barley plants expressing a TIP3-GFP fusion protein as a fluorescent PSV marker to follow in real time the spatially and temporally regulated remodeling and reshaping of PSVs during germination. During late-stage germination, we observed thin, tubular structures extending from PSVs in an actin-dependent manner. No extensions were detected following the disruption of actin microfilaments, while microtubules did not appear to be involved in the process. The previously-undetected tubular PSV structures were characterized by complex movements, fusion events and a dynamic morphology. Their function during germination remains unknown, but might be related to the transport of solutes and metabolites.

  2. Effects of Blends of Low-Protein Winter Wheat Flour and Barley Byproducts on Quality Changes in Noodles

    Science.gov (United States)

    Lee, Na-Young

    2016-01-01

    The physicochemical characteristics of fresh noodles made with blends of low-protein wheat flour and barley byproduct (BBP, 250 μm) were investigated. The crude protein contents (PC) of flour from Goso and Backjoong cultivars were 7.91% and 7.67%, respectively. PC and β-glucan contents from the BBP were 14.10% and 3.11%, respectively, which were higher than those in wheat flour. The water-holding capacity (WHC) of various blends was increased as a function of BBP but not gluten contents. Goso flour had the highest starch content (78.68%), with peak and final viscosities of 3,099 and 3,563 cp, respectively. Peak and final viscosities, trough, breakdown, and setback of the blends were decreased with the addition of BBP. Noodles made with Backjoong had the highest thickness score, while the hardness of noodles made with blends of Goso or Backjoong and 20% BBP were similar to those made from wheat flour only. The WHC of the samples was strongly correlated with PC, crude fiber, and β-glucan. The PC was not correlated with final viscosity, setback, thickness, hardness, gumminess, or chewiness. PMID:28078259

  3. Live Cell Imaging During Germination Reveals Dynamic Tubular Structures Derived from Protein Storage Vacuoles of Barley Aleurone Cells.

    Science.gov (United States)

    Ibl, Verena; Stoger, Eva

    2014-01-01

    The germination of cereal seeds is a rapid developmental process in which the endomembrane system undergoes a series of dynamic morphological changes to mobilize storage compounds. The changing ultrastructure of protein storage vacuoles (PSVs) in the cells of the aleurone layer has been investigated in the past, but generally this involved inferences drawn from static pictures representing different developmental stages. We used live cell imaging in transgenic barley plants expressing a TIP3-GFP fusion protein as a fluorescent PSV marker to follow in real time the spatially and temporally regulated remodeling and reshaping of PSVs during germination. During late-stage germination, we observed thin, tubular structures extending from PSVs in an actin-dependent manner. No extensions were detected following the disruption of actin microfilaments, while microtubules did not appear to be involved in the process. The previously-undetected tubular PSV structures were characterized by complex movements, fusion events and a dynamic morphology. Their function during germination remains unknown, but might be related to the transport of solutes and metabolites.

  4. Allelic diversity of a beer haze active protein gene in cultivated and Tibetan wild barley and development of allelic specific markers.

    Science.gov (United States)

    Ye, Lingzhen; Dai, Fei; Qiu, Long; Sun, Dongfa; Zhang, Guoping

    2011-07-13

    The formation of haze is a serious quality problem in beer production. It has been shown that the use of silica elute (SE)-ve malt (absence of molecular weight (MW) ∼14000 Da) for brewing can improve haze stability in the resultant beer, and the protein was identified as a barley trypsin inhibitor of the chloroform/methanol type (BTI-CMe). The objectives of this study were to determine (1) the allelic diversity of the gene controlling BTI-CMe in cultivated and Tibetan wild barley and (2) allele-specific (AS) markers for screening SE protein type. A survey of 172 Tibetan annual wild barley accessions and 71 cultivated barley genotypes was conducted, and 104 wild accessions and 35 cultivated genotypes were identified as SE+ve and 68 wild accessions and 36 cultivated genotypes as SE-ve. The allelic diversity of the gene controlling BTI-CMe was investigated by cloning, alignment, and association analysis. It was found that there were significant differences between the SE+ve and SE-ve types in single-nucleotide polymorphisms at 234 (SNP(234)), SNP(313), and SNP(385.) Furthermore, two sets of AS markers were developed to screen SE protein type based on SNP(313). AS-PCR had results very similar to those obtained by immunoblot method. Mapping analysis showed that the gene controlling the MW∼14 kDa band was located on the short arm of chromosome 3H, at the position of marker BPB-0527 (33.302 cM) in the Franklin/Yerong DH population.

  5. Allelic variations of a light harvesting chlorophyll a/b-binding protein gene (Lhcb1 associated with agronomic traits in barley.

    Directory of Open Access Journals (Sweden)

    Yanshi Xia

    Full Text Available Light-harvesting chlorophyll a/b-binding protein (LHCP is one of the most abundant chloroplast proteins in plants. Its main function is to collect and transfer light energy to photosynthetic reaction centers. However, the roles of different LHCPs in light-harvesting antenna systems remain obscure. Exploration of nucleotide variation in the genes encoding LHCP can facilitate a better understanding of the functions of LHCP. In this study, nucleotide variations in Lhcb1, a LHCP gene in barley, were investigated across 292 barley accessions collected from 35 different countries using EcoTILLING technology, a variation of the Targeting Induced Local Lesions In Genomes (TILLING. A total of 23 nucleotide variations were detected including three insert/deletions (indels and 20 single nucleotide polymorphisms (SNPs. Among them, 17 SNPs were in the coding region with nine missense changes. Two SNPs with missense changes are predicted to be deleterious to protein function. Seventeen SNP formed 31 distinguishable haplotypes in the barley collection. The levels of nucleotide diversity in the Lhcb1 locus differed markedly with geographic origins and species of accessions. The accessions from Middle East Asia exhibited the highest nucleotide and haplotype diversity. H. spontaneum showed greater nucleotide diversity than H. vulgare. Five SNPs in Lhcb1 were significantly associated with at least one of the six agronomic traits evaluated, namely plant height, spike length, number of grains per spike, thousand grain weight, flag leaf area and leaf color, and these SNPs may be used as potential markers for improvement of these barley traits.

  6. Purification of barley dimeric α-amylase inhibitor-1 (BDAI-1) and avenin-like protein-a (ALP) from beer and their impact on beer foam stability.

    Science.gov (United States)

    Iimure, Takashi; Kihara, Makoto; Sato, Kazuhiro; Ogushi, Kensuke

    2015-04-01

    Foam stability is a key factor of beer quality for consumers and brewers. Recent beer proteome analyses have suggested that barley dimeric α-amylase inhibitor-1 (BDAI-1) and avenin-like protein-a (ALP) derived from barley are important for beer foam stability. In this study, BDAI-1 and ALP were purified from a Japanese commercial beer sample using salt precipitation and column chromatography. The purification level was verified using two-dimensional gel electrophoresis, mass spectrometry, and database searches. Purified BDAI-1 and ALP were added to a beer sample to compare the foam stability to that of a control beer sample. As a result, beer foam stability was significantly improved by BDAI-1 but not by ALP, thereby suggesting that BDAI-1 affects beer foam stability whereas ALP does not.

  7. A role for arabinogalactan proteins in gibberellin-induced alpha-amylase production in barley aleurone cells.

    Science.gov (United States)

    Suzuki, Yoshihito; Kitagawa, Mamiko; Knox, J Paul; Yamaguchi, Isomaro

    2002-03-01

    Arabinogalactan proteins (AGPs) are plant proteoglycans that have been implicated in plant growth and development. The possible involvement of AGPs in the action of gibberellin (GA), a class of plant hormones, was examined by applying beta-glucosyl Yariv reagent (beta-Glc)3Y, a synthetic phenyl glycoside that interacts selectively with AGPs, to barley aleurone protoplasts. Gibberellin induces transcription and secretion of alpha-amylases in the protoplasts. Induction of alpha-amylase was clearly inhibited by (beta-Glc)3Y but not by (alpha-Gal)3Y, a negative control of the Yariv reagent that does not interact with AGPs. Transfection analysis, using an alpha-amylase promoter-GUS fusion gene in the protoplasts, indicated that the transcriptional activation of the alpha-amylase promoter was inhibited specifically by (beta-Glc)3Y. These observations are the first indication of an involvement of AGPs in a plant hormone function. The inhibitory effect of (beta-Glc)3Y was not observed when aleurone layers or half-seed grains were used. This result, together with the fact that protoplasts do not have cell walls, suggests that the AGPs that function in alpha-amylase induction reside at the plasma membrane. An aleurone-specific AGP was detected by reversed-phase HPLC, and supported the idea that an AGP may play an important role in aleurone-specific events. The possible mechanism of AGP function in gibberellin-induced alpha-amylase production is discussed.

  8. Salinity Stress in Roots of Contrasting BarleyGenotypes Reveals Time-Distinct and Genotype-Specific Patterns for Defined Proteins

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    Soil salinity is one of the most severe abiotic stress factors threatening agriculture worldwide. Hence,particular interest exists in unraveling mechanisms leading to salt tolerance and improved crop plant performance onsaline soils. Barley is considered to be one of the most salinity-tolerant crops, but varying levels of tolerance are wellcharacterized. A proteomic analysis of the roots of two contrasting cultivars (cv. Steptoe and cv. Morex) is presented.Young plants were exposed to a period of 1, 4, 7, or 10 d at 0, 100, or 150mM NaCI. The root proteome was analyzedbased on two-dimensional gel electrophoresis. A number of cultivar-specific and salinity stress-responsive proteins wereidentified. Mass spectrometry-based identification was successful for 74 proteins, and a hierarchical clustering analysisgrouped these into five clusters based on similarity of expression profile. The rank product method was applied to sta-tistically access the early and late responses, and this delivered a number of new candidate proteins underlying salinitytolerance in barley. Among these were some germin-like proteins, some pathogenesis-related proteins, and numerousas-yet uncharacterized proteins. Notably, proteins involved in detoxification pathways and terpenoid biosynthesis weredetected as early responsive to salinity and may function as a means of modulating growth-regulating mechanisms andmembrane stability via fine tuning of phytohormone and secondary metabolism in the root.

  9. Production of extruded barley, cassava, corn and quinoa enriched with whey proteins and cashew pulp

    Science.gov (United States)

    Well-formulated snacks can play a positive role in enhancing health by providing essential nutrients, such as increased protein and fiber, that mitigate metabolic syndrome associated with obesity. Adding whey protein concentrate (WPC80) and cashew pulp (CP) to corn meal, a major ingredient in extru...

  10. Dipeptidyl Peptidase IV Inhibitory Peptides Derived from Oat (Avena sativa L.), Buckwheat (Fagopyrum esculentum), and Highland Barley (Hordeum vulgare trifurcatum (L.) Trofim) Proteins.

    Science.gov (United States)

    Wang, Feng; Yu, Guoyong; Zhang, Yanyan; Zhang, Bolin; Fan, Junfeng

    2015-11-01

    Peptides released from oat, buckwheat, and highland barley proteins were examined for their in vitro inhibitory effects on dipeptidyl peptidase IV (DPP4), an enzyme that deactivates incretin hormones involved in insulin secretion. All of the hydrolysates exhibited DPP4 inhibitory activities, with IC50 values ranging from 0.13 mg/mL (oat glutelin alcalase digestion) to 8.15 mg/mL (highland barley albumin tryptic digestion). The lowest IC50 values in gastrointestinal, alcalase, and tryptic digestions were 0.99 mg/mL (oat flour), 0.13 mg/mL (oat glutelin), and 1.83 mg/mL (highland barley glutelin). In all, 35 peptides of more than seven residues were identified in the tryptic hydrolysates of oat globulin using liquid chromatography-mass spectroscopy. Peptides LQAFEPLR and EFLLAGNNK were synthesized and their DPP4 inhibitory activities determined. LQAFEPLR showed high in vitro DPP4 inhibitory activity with an IC50 value of 103.5 μM.

  11. The promoter of the barley aleurone-specific gene encoding a putative 7 kDa lipid transfer protein confers aleurone cell-specific expression in transgenic rice.

    Science.gov (United States)

    Kalla, R; Shimamoto, K; Potter, R; Nielsen, P S; Linnestad, C; Olsen, O A

    1994-12-01

    This paper describes the aleurone-specific gene Ltp2 from barley, which encodes a putative 7 kDa non-specific lipid transfer protein. As shown by Northern and in situ hybridization analyses, the Ltp2 transcript is present in barley aleurone cells shortly after the initiation of aleurone cell differentiation. The expression of Ltp2 increases until grain mid-maturity, but the mRNA is absent from mature grains. The Ltp2 transcript is undetectable in the embryo and vegetative tissues, confirming the aleurone specificity of the Ltp2 gene. The ability of the isolated 801 bp Ltp2 promoter to direct aleurone-specific expression in immature barley grains is demonstrated by particle bombardment experiments. In these experiments, the activity of the Ltp2 promoter is 5% of the activity of the strong constitutive Actin1 promoter from rice, as quantified by GUS activity measurements. In stably transformed rice plants containing the Ltp2 promoter-Gus construct, the specificity of the Ltp2 promoter is confirmed in vivo by the presence of GUS activity exclusively in the aleurone layer. This study demonstrates the conserved nature of the regulatory signals involved in aleurone-specific gene transcription in cereal grains.

  12. Enrichment and identification of integral membrane proteins from barley aleurone layers by reversed-phase chromatography, SDS-PAGE and LC-MS/MS

    DEFF Research Database (Denmark)

    Hynek, Radovan; Svensson, Birte; Nørregaard Jensen, Ole

    2006-01-01

    The plasma membrane of the cereal aleurone layer is the site of perception of germination signals and release of enzymes to the starchy endosperm. Analysis of membrane proteins is challenging due to their hydrophobicity and low abundance, thus little is known about the membrane proteins involved...... in seed germination. A membrane fraction highly enriched for the plasma membrane H+-ATPase was prepared from barley aleurone layers by aqueous two-phase partitioning. Since detergent and salt washes did not efficiently remove soluble proteins from the membrane preparations, an alternative procedure......+-ATPase, two proteins possibly involved in ion-channel regulation and two proteins of unknown function. This represents the first analysis of membrane proteins involved in seed germination, using a proteomics approach....

  13. Enrichment and identification of integral membrane proteins from barley aleurone layers by reversed-phase chromatography, SDS-PAGE, and LC-MS/MS

    DEFF Research Database (Denmark)

    Hynek, Radovan; Svensson, Birte; Jensen, O.N.

    2006-01-01

    The plasma membrane of the cereal aleurone layer is the site of perception of germination signals and release of enzymes to the starchy endosperm. Analysis of membrane proteins is challenging due to their hydrophobicity and low abundance; thus, little is known about the membrane proteins involved...... in seed germination. A membrane fraction highly enriched for the plasma membrane H+-ATPase was prepared from barley aleurone layers by aqueous two-phase partitioning. Because detergent and salt washes did not efficiently remove soluble proteins from the membrane preparations, an alternative procedure......+-ATPase, two proteins possibly involved in ion-channel regulation, and two proteins of unknown function. This represents the first analysis of membrane proteins involved in seed germination using a proteomics approach....

  14. Application of proteomics to investigate barley-Fusarium graminearum interaction

    DEFF Research Database (Denmark)

    Yang, Fen

    , the molecular mechanisms of barley defense to Fusarium graminearum at the early infection stage were studied. Antibodies against barley β-amylases were shown to be the markers for infection at proteome level and for selection of the time for proteome analysis before extensive degradation caused by the fungus...... proteases which could be responsible for proteolysis of β-amylases in the infected barley. In Chapter 4, the in vitro secretome of F. graminearum on the 2-D gels in the presence of substrates of barley or wheat grain was studied. Totally 69 unique fungal proteins identified were mainly cell....... In addition, sharper increase in fungal biomass was observed in barley than in wheat and fungal induced proteolytic fragments of - amylases were only observed in barley not in wheat. Furthermore, a barley PR17 protein and a fungal hypothetical protein were expressed in E. coli and purified in Chapter 5...

  15. HEALTH BENEFITS OF BARLEY

    Directory of Open Access Journals (Sweden)

    Akula Annapurna

    2013-09-01

    Full Text Available Prevalence of lifestyle diseases is increasing day by day. Mostly the younger generation do not have much awareness about healthy nutritional supplements. One such important cereal grain not used mostly by youngsters is barley It is a good old grain with so many health benefits like weight reduction, decreasing blood pressure, blood cholesterol, blood glucose in Type 2 diabetes and preventing colon cancer. It is easily available and cheap grain. It contains both soluble and insoluble fiber, protein, vitamins B and E, minerals selenium, magnesium and iron, copper, flavonoids and anthocynins. Barley contains soluble fiber, beta glucan binds to bile acids in the intestines and thereby decreasing plasma cholesterol levels. Absorbed soluble fiber decreases cholesterol synthesis by liver and cleansing blood vessels. Insoluble fiber provides bulkiness in the intestines, thereby satiety. decreased appetite. It promotes intestinal movements relieving constipation, cleansing colonic harmful bacteria and reduced incidence of colonic cancer. It is a good source of niacin ,reducing LDL levels and increasing HDL levels. Selenium and vitamin E providing beneficial antioxidant effects. Magnesium, a cofactor for many carbohydrate metabolism enzymes and high fiber content contributes for its blood glucose reducing effect in Type 2 diabetes. It is having good diuretic activity and is useful in urinary tract infections. Barley contains gluten, contraindicated in celiac disease.

  16. Brewing with fractionated barley

    OpenAIRE

    Donkelaar, van, CC René

    2016-01-01

    Brewing with fractionated barley Beer is a globally consumed beverage, which is produced from malted barley, water, hops and yeast. In recent years, the use of unmalted barley and exogenous enzymes have become more popular because they enable simpler processing and reduced environmental impact. Raw barley, however, contains less endogenous enzymes and more undesired components for the use of beer brewing, compared to malted barley.  The overall aim of this thesis is to investigate how ba...

  17. Differential labelling of cysteines for simultaneous identification of thioredoxin h-reducible disulphides in native protein extracts: insight into recognition and regulation of proteins in barley seeds by thioredoxin h

    DEFF Research Database (Denmark)

    Maeda, Kenji; Finnie, Christine; Svensson, Birte

    2005-01-01

    . Mass shifts of 15 peptides, induced by treatment with thioredoxin h and differential alkylation, identified specific reduction of nine disulphides in BASI, four a-amylase/trypsin inhibitors and a protein of unknown function. Two specific disulphides, located structurally close to the alpha......-amylase/subtilisin inhibitor (BASI) by barley thioredoxin h isoform 1 was analysed. Furthermore, the method was coupled with twodimensional electrophoresis for convenient thioredoxin h-reducible disulphide identification in barley seed extracts without the need for protein purification or production of recombinant proteins......-amylase binding surfaces of BASI and alpha-amylase inhibitor BMAI-1 were demonstrated to be reduced to a particularly high extent. For the first time, specificity of thioredoxin h for particular disulphide bonds is demonstrated, providing a basis to study structural aspects of the recognition mechanism...

  18. Invitro Synthesis of Barley Endosperm Proteins on Wild Type and Mutant Templates

    DEFF Research Database (Denmark)

    Brandt, A.; Ingversen, J.

    1976-01-01

    electrophoresis showed that proteins with molecular weights ranging from 200,000 to 10,000 daltons were synthesized. A substantial part of the polypeptides coded for by the template associated with the membrane bound polysomes was identified as hordeins by their solubility in 55% isopropanol and by their co...

  19. A Nearly Non-Functional Mutant Allele of the Storage Protein Locus Hor2 in Barley

    DEFF Research Database (Denmark)

    Doll, Hans

    1980-01-01

    The low content of the storage protein fraction hordein-2 in the high-lysine mutant Risø 56 is due to a mutation at or near the locus Hor2 coding for hordein-2 polypeptides. The mutant gene is recessive in its qualitative effect on the electrophoretic banding pattern of hordein-2, but co...

  20. Thioredoxin reductase from barley: Structure, recognition of thioredoxin, protein engineering and catalytic mechanism

    DEFF Research Database (Denmark)

    Kirkensgaard, Kristine Groth

    mutational study of some of the residues and loops, which were indicated to be important for the binding between the two proteins. Enzyme kinetics analyses of these mutants suggest that the FAD‐loop is critical for binding of HvTrxh2. Especially, Trp42HvNTR2 and Met43HvNTR2 appears to be important...

  1. Heterologous expression of surface-active proteins from barley and filamentous fungi in Pichia pastoris and characterization of their contribution to beer gushing.

    Science.gov (United States)

    Lutterschmid, Georg; Muranyi, Monika; Stübner, Matthias; Vogel, Rudi F; Niessen, Ludwig

    2011-05-14

    The spontaneous over-foaming of beer upon opening, i.e. beer gushing, is an unwanted phenomenon for the brewing industry. Currently, surface-active proteins from filamentous fungi and non-specific lipid transfer proteins (nsLTP1) from barley are discussed as gushing inducers. In our study the class I hydrophobin FcHyd3p from Fusarium culmorum, the class II hydrophobin Hfb2 from Trichoderma reesei, the alkaline foam protein A (AfpA) from F. graminearum and nsLTP1 from Hordeum vulgare cv. Marnie (barley) were heterologously expressed in Pichia pastoris and used in gushing tests. The class I hydrophobin FcHyd3p was unable to induce gushing in beer. The class II hydrophobin Hfb2 was able to induce gushing in beer, but proved to be inhibited by heat treatment as well as by the presence of enriched hop compounds. Both resulted in a reduced gushing potential. AfpA and nsLTP1 exhibited no gushing-inducing potential at the amounts added to beer. Addition of these proteins to beer or carbonated water previously treated with class II hydrophobins revealed a gushing reducing character.

  2. The knock-down of the expression of MdMLO19 reduces susceptibility to powdery mildew (Podosphaera leucotricha) in apple (Malus domestica).

    Science.gov (United States)

    Pessina, Stefano; Angeli, Dario; Martens, Stefan; Visser, Richard G F; Bai, Yuling; Salamini, Francesco; Velasco, Riccardo; Schouten, Henk J; Malnoy, Mickael

    2016-10-01

    Varieties resistant to powdery mildew (PM; caused by Podosphaera leucotricha) are a major component of sustainable apple production. Resistance can be achieved by knocking-out susceptibility S-genes to be singled out among members of the MLO (Mildew Locus O) gene family. Candidates are MLO S-genes of phylogenetic clade V up-regulated upon PM inoculation, such as MdMLO11 and 19 (clade V) and MdMLO18 (clade VII). We report the knock-down through RNA interference of MdMLO11 and 19, as well as the complementation of resistance with MdMLO18 in the Arabidopsis thaliana triple mlo mutant Atmlo2/6/12. The knock-down of MdMLO19 reduced PM disease severity by 75%, whereas the knock-down of MdMLO11, alone or in combination with MdMLO19, did not result in any reduction or additional reduction of susceptibility compared with MdMLO19 alone. The test in A. thaliana excluded a role for MdMLO18 in PM susceptibility. Cell wall appositions (papillae) were present in both PM-resistant and PM-susceptible plants, but were larger in resistant lines. No obvious negative phenotype was observed in plants with mlo genes knocked down. Apparently, MdMLO19 plays the pivotal role in apple PM susceptibility and its knock-down induces a very significant level of resistance.

  3. Functional Analysis of Barley Powdery Mildew Effector Candidates and Identification of their Barley Targets

    DEFF Research Database (Denmark)

    Ahmed, Ali Abdurehim

    The genome of barley powdery mildew fungus (Blumeria graminis f. sp. hordei, Bgh) encodes around 500 Candidate Secreted Effector Proteins (CSEPs), which are believed to be delivered to the barley cells either to interfere with plant defence and/or promote nutrient uptake. So far, little is known...

  4. Abscisic acid prevents the coalescence of protein storage vacuoles by upregulating expression of a tonoplast intrinsic protein gene in barley aleurone.

    Science.gov (United States)

    Lee, Sung-eun; Yim, Hui-kyung; Lim, Mi-na; Yoon, In sun; Kim, Jeong hoe; Hwang, Yong-sic

    2015-03-01

    Tonoplast intrinsic proteins (TIPs) are integral membrane proteins that are known to function in plants as aquaporins. Here, we propose another role for TIPs during the fusion of protein storage vacuoles (PSVs) in aleurone cells, a process that is promoted by gibberellic acid (GA) and prevented by abscisic acid (ABA). Studies of the expression of barley (Hordeum vulgare) TIP genes (HvTIP) showed that GA specifically decreased the abundance of HvTIP1;2 and HvTIP3;1 transcripts, while ABA strongly increased expression of HvTIP3;1. Increased or decreased expression of HvTIP3;1 interfered with the hormonal effects on vacuolation in aleurone protoplasts. HvTIP3;1 gain-of-function experiments delayed GA-induced vacuolation, whereas HvTIP3;1 loss-of-function experiments promoted vacuolation in ABA-treated aleurone cells. These results indicate that TIP plays a key role in preventing the coalescence of small PSVs in aleurone cells. Hormonal regulation of the HvTIP3;1 promoter is similar to the regulation of the endogenous gene, indicating that induction of the transcription of HvTIP3;1 by ABA is a critical factor in the prevention of PSV coalescence in response to ABA. Promoter analysis using deletions and site-directed mutagenesis of sequences identified three cis-acting elements that are responsible for ABA responsiveness in the HvTIP3;1 promoter. Promoter analysis also showed that ABA responsiveness of the HvTIP3;1 promoter is likely to occur via a unique regulatory system distinct from that involving the ABA-response promoter complexes.

  5. Differences in phytase activity and phytic acid content between cultivated and Tibetan annual wild barleys.

    Science.gov (United States)

    Dai, Fei; Qiu, Long; Xu, Yang; Cai, Shengguan; Qiu, Boyin; Zhang, Guoping

    2010-11-24

    The Qinghai-Tibetan Plateau in China is considered to be one of the original centers of cultivated barley. At present, little is known about the phytase activity (Phy) or phytic acid content (PA) in grains of Tibetan annual wild barley. Phy and PA were determined in grains of 135 wild and 72 cultivated barleys. Phy ranged from 171.3 to 1299.2 U kg(-1) and from 219.9 to 998.2 U kg(-1) for wild and cultivated barleys, respectively. PA and protein contents were much higher in wild barley than in cultivated barley. Tibetan annual wild barley showed a larger genetic diversity in phytase activity and phytic acid and protein contents and is of value for barley breeding. There is no significant correlation between phytase activity and phytic acid or protein content in barley grains, indicating that endogenous phytase activity had little effect on the accumulation of phytic acid.

  6. DNA binding sites recognised in vitro by a knotted class 1 homeodomain protein encoded by the hooded gene, k, in barley (Hordeum vulgare)

    DEFF Research Database (Denmark)

    Krusell, L; Rasmussen, I; Gausing, K

    1997-01-01

    The homeodomain of the knotted classes of transcription factors from plants differs from the well characterized Antp/En type homeodomains from Drosophila at key amino acid residues contributing to the DNA binding. A cDNA, Hvh21, derived from the hooded gene and encoding a full length homolog...... of knotted1 from maize was isolated from barley seedlings and expressed as a maltose binding protein fusion in E. coli. The purified HvH21-fusion protein selected DNA fragments with 1-3 copies of the sequence TGAC. Gel shift experiments showed that the TGAC element was required for binding and the results...... further indicate that the HvH21-fusion protein binds DNA as a monomer. Udgivelsesdato: 1997-May-12...

  7. Novel prediction method of beer foam stability using protein Z, barley dimeric alpha-amylase inhibitor-1 (BDAI-1) and yeast thioredoxin.

    Science.gov (United States)

    Iimure, Takashi; Takoi, Kiyoshi; Kaneko, Takafumi; Kihara, Makoto; Hayashi, Katsuhiro; Ito, Kazutoshi; Sato, Kazuhiro; Takeda, Kazuyoshi

    2008-09-24

    Foam stability is an important quality trait of beer. Our previous results of two-dimensional gel electrophoresis (2DE) analyses of beer proteins implied a relationship between barley dimeric alpha-amylase inhibitor-1 (BDAI-1) and beer foam stability as judged by the NIBEM-T analyzer. To develop a novel prediction method of beer foam stability under different conditions of barley cultivar and malt modification, multiple linear regression analysis was applied. The spot intensities of major beer proteins on 2DE gel were quantified and used as explanatory variables. The foam stabilities of 25 beer samples each brewed from malt with different malt modification in one of the three cultivars (cultivars A, B, and C) were explained by the spot intensities of BDAI-1 at the 5% significance level ( r = 0.421). Furthermore, two other major protein spots (b0 and b5) were observed on the 2DE gels of Japanese commercial beer samples with different foam stability. Then, multiple regression for foam stability was calculated using these three spot intensities as explanatory variables. As a result, 72.1% of the beer foam stability in 25 beer samples was explained by a novel multiple regression equation calculated using spot b0 and BDAI-1 as positive explanatory variables and spot b5 as a negative variable. To verify the validity of the multiple regression equation and the explanatory variables, the beer foam stability in practical beer samples was analyzed. As a result, 81.5% of the beer foam stability in 10 Japanese commercial beer samples was also explained by using spot b0 and BDAI-1 as positive explanatory variables and spot b5 as a negative variable. Mass spectrometry analyses followed by database searches revealed that protein spots b0 and b5 were identified as protein Z originated from barley and thioredoxin originated from yeast, respectively. These results confirm that BDAI-1 and protein Z are foam-positive factors and identify yeast thioredoxin as a possible novel foam

  8. Brewing with fractionated barley

    NARCIS (Netherlands)

    Donkelaar, van L.H.G.

    2016-01-01

    Brewing with fractionated barley Beer is a globally consumed beverage, which is produced from malted barley, water, hops and yeast. In recent years, the use of unmalted barley and exogenous enzymes have become more popular because they enable simpler processing and reduced environmental impact. Raw

  9. Barley seed proteomics from spots to structures

    DEFF Research Database (Denmark)

    Finnie, Christine; Svensson, Birte

    2009-01-01

    with information from rice and other cereals facilitate identification of barley proteins. Several hundred barley seed proteins are identified and lower abundance proteins including membrane proteins are now being analysed. In the present review we focus on variation in protein profiles of seed tissues during...... forms on 2D-gels. Specific protein families, including peroxidases and alpha-amylases have been subjected to in-depth analysis resulting in characterisation of different isozymes, post-translational. modifications and processing. A functional proteomics study focusing on the seed thioredoxin system has...

  10. Transgenic barley: a prospective tool for biotechnology and agriculture.

    Science.gov (United States)

    Mrízová, Katarína; Holasková, Edita; Öz, M Tufan; Jiskrová, Eva; Frébort, Ivo; Galuszka, Petr

    2014-01-01

    Barley (Hordeum vulgare L.) is one of the founder crops of agriculture, and today it is the fourth most important cereal grain worldwide. Barley is used as malt in brewing and distilling industry, as an additive for animal feed, and as a component of various food and bread for human consumption. Progress in stable genetic transformation of barley ensures a potential for improvement of its agronomic performance or use of barley in various biotechnological and industrial applications. Recently, barley grain has been successfully used in molecular farming as a promising bioreactor adapted for production of human therapeutic proteins or animal vaccines. In addition to development of reliable transformation technologies, an extensive amount of various barley genetic resources and tools such as sequence data, microarrays, genetic maps, and databases has been generated. Current status on barley transformation technologies including gene transfer techniques, targets, and progeny stabilization, recent trials for improvement of agricultural traits and performance of barley, especially in relation to increased biotic and abiotic stress tolerance, and potential use of barley grain as a protein production platform have been reviewed in this study. Overall, barley represents a promising tool for both agricultural and biotechnological transgenic approaches, and is considered an ancient but rediscovered crop as a model industrial platform for molecular farming.

  11. Nucleotide sequence of coat protein gene for GPV isolate of barley yellow dwarf virus and construction of expression plasmid for plant

    Institute of Scientific and Technical Information of China (English)

    成卓敏; 何小源; 吴茂森; 周广和; Paul Keese; P.M.Waterhouse

    1996-01-01

    GPV is a Chinese serotype isolate of barley yellow dwarf virus (BYDV) that has no reactionwith antiserum of MAV, PAV, SGV, RPV and RMV. The sequence of the coat protein (CP) of GPV isolate of BYDV was identified and its amino acid sequence was deduced. The coding region for the putative GPV CP is 603 bases nucleotides and encodes a Mr 22218 (22 ku) protein. The same as MAV, PAV and RPV, GPV contained a second ORF within the coat protein coding region. This protein of 17024 Mr (17 ku) is thought to correspond to the Virion protein genome linked (Vpg). Sequence comparisons of the CP coding region between the GPV isolate of BYDV and other isolates of BYDV have been done. The nucleotide and ammo acid sequence homology of GPV has a greater identity to the sequence of RPV than those of PAV and MAV. The GPV CP sequence shared 83.7% of nucleotide similarity and 77.5% of deduced amino add similarity, whereas that of the PAV and MAV shared 56.9%. 53.2% and 44.1%. 43.8% respectively. According to BYDV-GPV CP seque

  12. Optimised purification and characterisation of lipid transfer protein 1 (LTP1) and its lipid-bound isoform LTP1b from barley malt.

    Science.gov (United States)

    Nieuwoudt, Melanie; Lombard, Nicolaas; Rautenbach, Marina

    2014-08-15

    In beer brewing, brewers worldwide strive to obtain product consistency in terms of flavour, colour and foam. Important proteins contributing to beer foam are lipid transfer proteins (LTPs), in particular LTP1 and its lipid-bound isoform LTP1b, which are known to transport lipids in vivo and prevent lipids from destabilising the beer foam. LTP1 and LTP1b were successfully purified using only five purification steps with a high purified protein yield (160 mg LTP1 and LTP1b from 200 g barley). Circular dichroism of LTP1 and LTP1b confirmed that both proteins are highly tolerant to high temperatures (>90 °C) and are pH stable, particularly at a neutral to a more basic pH. Only LTP1 exhibited antiyeast and thermo-stable lytic activity, while LTP1b was inactive, indicating that the fatty acid moiety compromised the antimicrobial activity of LTP1. This lack in antiyeast activity and the positive foam properties of LTP1b would benefit beer fermentation and quality.

  13. Molecular dynamics simulation of the cooperative adsorption of barley lipid transfer protein and cis-isocohumulone at the vacuum-water interface.

    Science.gov (United States)

    Euston, S R; Hughes, P; Naser, Md A; Westacott, R E

    2008-11-01

    Molecular dynamic simulations have been carried out on systems containing a mixture of barley lipid transfer protein (LTP) and cis-isocohumulone (a hop derived iso-alpha-acid) in one of its enol forms, in bulk water and at the vacuum-water interface. In solution, the cis-isocohumulone molecules bind to the surface of the LTP molecule. The mechanism of binding appears to be purely hydrophobic in nature via desolvation of the protein surface. Binding of hop acids to the LTP leads to a small change in the 3-D conformation of the protein, but no change in the proportion of secondary structure present in helices, even though there is a significant degree of hop acid binding to the helical regions. At the vacuum-water interface, cis-isocohumulone shows a high surface activity and adsorbs rapidly at the interface. LTP then shows a preference to bind to the preadsorbed hop acid layer at the interface rather than to the bare water-vacuum interface. The free energy of adsorption of LTP at the hop-vacuum-water interface is more favorable than for adsorption at the vacuum-water interface. Our results support the view that hop iso-alpha-acids promote beer foam stability by forming bridges between separate adsorbed protein molecules, thus strengthening the adsorbed protein layer and reducing foam breakdown by lamellar phase drainage. The results also suggest a second mechanism may also occur, whereby the concentration of protein at the interface is increased via enhanced protein adsorption to adsorbed hop acid layers. This too would increase foam stability through its effect on the stabilizing protein layer around the foam bubbles.

  14. 青稞蛋白质凝胶特性的研究%Study on gelation properties of highland barley protein

    Institute of Scientific and Technical Information of China (English)

    武菁菁; 李鑫磊; 张艺; 王辉; 胡国洲; 阚建全

    2013-01-01

    Highland barley protein with weak gelation properties constrained its application.To explore the formation condition of highland barley protein gelation,highland barley protein was extracted by isoelectric precipitation method,and the effect of protein concentration,pH,heating temperature,heating time,urea,neutral salt and propanediol concentration on gelation properties were studied by texture analyzer and rheometer.Results indicated that gel started to form when the concentration was beyond 14% at alkaline condition (pH ≥8).The hardness of gel reached maximum when the heating temperature was 95℃ and heating time was 40min.With the increase of NaCl and propanediol concentrations,the hardness and storage modulus of the protein gel firstly increased and then decreased.However,the hardness and storage modulus significantly increased with the increase of urea concentration and the gel hardness was 90.29g when urea concentration was 7mol/L.Results showed that denaturation by urea destroyed hydrogen bonds between intramolecular and intermolecular chains and hydrophobic interaction with the result of protein unfolding and active groups exposing,which was in favor of gel formation.%青稞蛋白质弱的凝胶特性限制了其的应用.为了探索青稞蛋白质凝胶形成的条件,采用碱溶酸沉法提取青稞中的蛋白质,并研究了青稞蛋白质浓度、加热温度、加热时间、pH、尿素浓度、中性盐浓度以及丙二醇浓度对其凝胶特性的影响.研究结果表明,青稞蛋白质在碱性和质量浓度大于14%的条件下能形成凝胶;凝胶硬度在加热温度95℃和加热时间40min时达到最大;随着NaC1浓度和丙二醇浓度的升高,凝胶硬度和储能模量呈现先升高后降低的趋势;随着尿素浓度的增加,青稞蛋白质凝胶硬度和储能模量增加,当尿素浓度为7mol/L时,凝胶硬度为90.29g,说明尿素的变性作用可以破坏分子内和分子间的氢键以及疏水相互作用力,

  15. Barley grain for ruminants: A global treasure or tragedy

    Directory of Open Access Journals (Sweden)

    Nikkhah Akbar

    2012-07-01

    Full Text Available Abstract Barley grain (Hordeum vulgare L. is characterized by a thick fibrous coat, a high level of ß-glucans and simply-arranged starch granules. World production of barley is about 30 % of that of corn. In comparison with corn, barley has more protein, methionine, lysine, cysteine and tryptophan. For ruminants, barley is the third most readily degradable cereal behind oats and wheat. Due to its more rapid starch fermentation rate compared with corn, barley also provides a more synchronous release of energy and nitrogen, thereby improving microbial nutrient assimilation. As a result, feeding barley can reduce the need for feeding protected protein sources. However, this benefit is only realized if rumen acidity is maintained within an optimal range (e.g., > 5.8 to 6.0; below this range, microbial maintenance requirements and wastage increase. With a low pH, microbial endotoxines cause pro-inflammatory responses that can weaken immunity and shorten animal longevity. Thus, mismanagement in barley processing and feeding may make a tragedy from this treasure or pearl of cereal grains. Steam-rolling of barley may improve feed efficiency and post-rumen starch digestion. However, it is doubtful if such processing can improve milk production and feed intake. Due to the need to process barley less extensively than other cereals (as long as the pericarp is broken, consistent and global standards for feeding and processing barley could be feasibly established. In high-starch diets, barley feeding reduces the need for capacious small intestinal starch assimilation, subsequently reducing hindgut starch use and fecal nutrient loss. With its nutritional exclusivities underlined, barley use will be a factual art that can either matchlessly profit or harm rumen microbes, cattle production, farm economics and the environment.

  16. Barley metallothioneins

    DEFF Research Database (Denmark)

    Hegelund, Josefine Nymark; Schiller, Michaela; Kichey, Thomas

    2012-01-01

    and protein abundance during grain development. MT3 was present in tissues of both maternal and filial origin throughout grain filling. In contrast, MT4 was confined to the embryo and aleurone layer, where it appeared during tissue specialization and remained until maturity. Using state-of-the-art speciation...... storage in developing and mature grains. The localization of MT4 and its discrimination against Cd make it an ideal candidate for future biofortification strategies directed toward increasing food and feed Zn concentrations....

  17. Structure, function and protein engineering in starch debranching enzyme systems. Barley limit dextrinase and its endogenous inhibitor

    DEFF Research Database (Denmark)

    Møller, Marie Sofie

    Starch is the most abundant storage carbohydrate in cereal grains. It is composed primarily of amylopectin, a polymer of glucose in which α-1,4-linked glucan chains are branched with α-1,6-bonds. Enzymatic degradation of starch in germinating barley seeds involves an initial solubilisation, mainly...... by α-amylase, followed by hydrolysis of the resulting dextrins to oligosaccharides and glucose by the concerted action of α- and β-amylase, limit dextrinase (LD), and α-glucosidase. Only LD is able to hydrolyse α-1,6-linkages in limit dextrins. Since LD is the sole debranching enzyme in the germinating...... on these comparisons. In the second part of the thesis I present the crystal structure of the complex between LD and LDI determined to 2.7 Å. The structure revealed a novel mode of inhibition distinctly different from that of the interaction between LDI-like inhibitors and α-1,4-acting enzymes (α-amylases) from...

  18. Glutamate synthase activities and protein changes in relation to nitrogen nutrition in barley: the dependence on different plastidic glucose-6P dehydrogenase isoforms.

    Science.gov (United States)

    Esposito, Sergio; Guerriero, Gea; Vona, Vincenza; Di Martino Rigano, Vittoria; Carfagna, Simona; Rigano, Carmelo

    2005-01-01

    In barley (Hordeum vulgare L. var. Nure), glutamate synthesis and the production of reducing power by the oxidative pentose phosphate pathway (OPPP) are strictly correlated biochemical processes. NADH-GOGAT was the major root isoform, whose activity increased on a medium supplied with NH4+ or NO3-; by contrast, no noticeable variations could be observed in the leaves of plants supplied with nitrogen. In the leaves, the major isoform is Fd-GOGAT, whose activity increased under nitrogen feeding. G6PDH activity increased in the roots supplied with nitrogen; no variations were observed in the leaves. Moreover, an increase of the P2 isoform in the roots was measured, giving 13.6% G6PDH activity localized in the plastids under ammonium, and 25.2% under nitrate feeding conditions. Western blots confirmed that P2-G6PDH protein was induced in the roots by nitrogen. P1-G6PDH protein was absent in the roots and increased in the leaves by nitrogen supply to the plants. The changes measured in cytosolic G6PDH seem correlated to more general cell growth processes, and do not appear to be directly involved in glutamate synthesis. The effects of light on Fd-GOGAT is discussed, together with the possibility for P2-G6PDH to sustain nitrogen assimilation upon illumination.

  19. Implementation of biochemical screening to improve baking quality of barley

    DEFF Research Database (Denmark)

    Vincze, Éva; Dionisio, Giuseppe; Aaslo, Per;

    2011-01-01

    Barley (Hordeum vulgare) has the potential to offer considerable human nutritional benefits, especially as supplement to wheat-based breads. Under current commercial baking conditions it is not possible to introduce more that 20% barley flour to the wheat bread without negative impact...... proteins. Changing the storage protein composition can lessen this problem. Our working hypothesis was that exploiting the substantial genetic variation within the gene pool for storage proteins could enable improving the baking qualities of barley flour. We characterised forty-nine barley cultivars...... for variations in storage protein and AA composition. These cultivars were selected based on their higher protein contents (11.8–17.6%). The results obtained indicated that substantial variation not only in the distribution of the hordein polypeptides but also in the relative proportions of the storage proteins...

  20. Transformation of blackgram (Vigna mungo (L.) Hepper) by barley chitinase and ribosome-inactivating protein genes towards improving resistance to Corynespora leaf spot fungal disease.

    Science.gov (United States)

    Chopra, Rajan; Saini, Raman

    2014-12-01

    Blackgram (Vigna mungo (L.) Hepper), an important grain legume crop, is sensitive to many fungal pathogens including Corynespora cassiicola, the causal agent of corynespora leaf spot disease. In the present study, plasmid pGJ42 harboring neomycin phosphotransferase (nptII) a selectable marker gene, the barley antifungal genes chitinase (AAA56786) and ribosome-inactivating protein (RIP; AAA32951) were used for the transformation, to develop fungal resistance for the first time in blackgram. The presence and integration of transgene into the blackgram genome was confirmed by PCR and Southern analysis with an overall transformation frequency of 10.2 %. Kanamycin selection and PCR analysis of T0 progeny revealed the inheritance of transgene in Mendelian fashion (3:1). Transgenic plants (T1), evaluated for fungal resistance by in vitro antifungal assay, arrested the growth of C. cassiicola up to 25-40 % over the wild-type plants. In fungal bio-assay screening, the transgenic plants (T1) sprayed with C. cassiicola spores showed a delay in onset of disease along with their lesser extent in terms of average number of diseased leaves and reduced number and size of lesions. The percent disease protection among different transformed lines varies in the range of 27-47 % compare to control (untransformed) plants. These results demonstrate potentiality of chitinase and RIP from a heterologous source in developing fungal disease protection in blackgram and can be helpful in increasing the production of blackgram.

  1. A potential new selection criterion for breeding winter barley optimal protein and amino acid profiles for liquid pig feed

    DEFF Research Database (Denmark)

    Christensen, Jesper Bjerg; Blaabjerg, Karoline; Poulsen, Hanne Damgaard

    The hypothesis is that cereal proteases in liquid feed degrade and convert water insoluble storage protein into water soluble protein, which may improve the digestibility of protein in pigs compared with dry feeding. Protein utilization is increased by matching the amino acid (AAs) content...... concentration was analysed in the supernatant after centrifugation. After 15 min., app. 16% of the total protein was soluble and until 8 hours an increase of 5% units was observed. However, from 8 to 48 hours it increased with 10% units for some cultivars. Based on these analyses, cultivars were selected...

  2. MS Based Imaging of Barley Seed Development

    Institute of Scientific and Technical Information of China (English)

    Manuela Peukert; Andrea Matros; Hans-Peter Mock

    2012-01-01

    Spatially resolved analysis of metabolites and proteins is essential to model compartmentalized cellular processes in plants.Within recent years,tremendous progress has been made in MS based imaging (MSI) techniques,mostly MALDI MSI.The technology has been pioneered and is now widely applied in medicinal and pharmacological studies,and in recent years found its way into plant science (Kaspar et al.,2011; Peukert etal.,2012).We are interested in the elucidation of spatially resolved metabolic networks related to barley grain development.An understanding of developmentally and ecologically regulated processes affecting agronomical traits such as final grain weight,seed quality and stress tolerance is of outmost importance,as barley provides one of the staple foods.Barley also serves as a model plant for other cereals such as wheat.The presentation will introduce an untargeted MALDI MSI approach to the analysis of me-tabolite patterns during barley grain development.We analyzed longitudinal and cross sections from developing barley grains (3,7,10 and 14 days after pollination).In the presentation we will address spatial resolution,sensitivity and identification of unknown compounds will also be discussed.A major task is to connect the metabolite patterns to distinct cellular and physiological events.As an example,particular metabolite distributions indicative for nutrient transport into the developing endosperm will be shown.

  3. Evaluation of Barley as Human Food

    Directory of Open Access Journals (Sweden)

    Mehmet Köten

    2013-12-01

    Full Text Available Barley, as animal feed, raw material for malting and human food, constitute an important part among cereal sources in the world. Majority of barley that produced both in Turkey and other countries of the world, is being used as animal feed. Poor baking quality, taste and appearance of barley restricted its use in human nutrition. However, recently high protein, fiber, especially β-glucan and high starch content appeal to food industry. Many scientific researches established that β-glucan, a soluble fiber, has an effect in healing coronary-hearth diseases, lowering blood cholesterol level, balancing blood sugar level, preventing obesity. Being a healthy cereal that can be used in various purposes, and an additive in many food products, barley is considered a very promising cereal, and research to increase possibilities of its use in human nutrition is being increased. In the literature, there has been researches on making noodles, bulgur, kavut (roasted cereal, breakfast cereals. In this study the researches relating to evaluation of barley, importance of which is increased every day, as human food was reviewed.

  4. The barley Jip23b gene

    DEFF Research Database (Denmark)

    Müller-Uri, Frieder; Cameron-Mills, Verena; Mundy, John

    2002-01-01

    The barley gene (Jip23) encoding a 23,000-Da protein of unknown function was isolated and shown to be induced by jasmonate methyl ester (MeJA) in leaves. 5'upstream Jip23 sequence was isolated and fused to the beta-glucuronidase gene (GUS), and this reporter was introduced by particle bombardment...

  5. Regulation of basal resistance by a powdery mildew-induced cysteine-rich receptor-like protein kinase in barley

    DEFF Research Database (Denmark)

    Rayapuram, Cbgowda; Jensen, Michael K; Maiser, Fabian

    2012-01-01

    , followed by a rather short 17-amino-acid transmembrane domain, which includes an AAA motif, two features characteristic of endoplasmic reticulum (ER)-targeted proteins and, finally, a characteristic putative protein kinase domain in the C-terminus. The HvCRK1 transcript was isolated from leaves inoculated...

  6. Differential appearance of isoforms and cultivar variation in protein temporal profiles revealed in the maturing barley grain proteome

    DEFF Research Database (Denmark)

    Finnie, Christine; Bak-Jensen, K.S.; Laugesen, Sabrina

    2006-01-01

    -peroxiredoxin isoform was identified in three spots, one present throughout grain filling, one appearing during desiccation and one observed only in mature seeds. This suggested post-translational modification of the protein to different degrees during seed maturation. Distinct isoforms of several proteins were...

  7. Endoproteolytic activity assay in malting barley

    Directory of Open Access Journals (Sweden)

    Blanca Gómez Guerrero

    2013-12-01

    Full Text Available Hydrolysis of barley proteins into peptides and amino acids is one of the most important processes during barley germination.The degradation of the endosperm stored proteins facilitates water and enzyme movements, enhances modification, liberates starch granules and increases soluble amino nitrogen. Protease activity is the result of the activities of a mixture of exo- and endo-proteases. The barley proteins are initially solubilized by endo-proteases and the further by exo-proteases. Four classes of endo-proteases have been described: serine-proteases, cysteine-proteases, aspartic-proteases and metallo-proteases. The objective of this work was to develop a rapid and colorimetric enzymatic assay to determine the endo-proteolytic activity of the four endo-protease classes using two different substrates: azo-gelatin and azo-casein. Optimum conditions for the assays such as: pH,reaction time and temperature and absorbance scale were determined. Azo-gelatin presented several difficulties in standardizing an “in solution” assay. On the other hand, azo-casein allowed standardization of the assay for the four enzyme classes to produce consistent results. The endo-proteoteolytic method developed was applied to determine the endo-protease activity in barley, malt and wort.

  8. Nucleotide sequence of a cDNA coding for the barley seed protein CMa: an inhibitor of insect α-amylase

    DEFF Research Database (Denmark)

    Rasmussen, Søren Kjærsgård; Johansson, A.

    1992-01-01

    The primary structure of the insect alpha-amylase inhibitor CMa of barley seeds was deduced from a full-length cDNA clone pc43F6. Analysis of RNA from barley endosperm shows high levels 15 and 20 days after flowering. The cDNA predicts an amino acid sequence of 119 residues preceded by a signal...... peptide of 25 amino acids. Ala and Leu account for 55% of the signal peptide. CMa is 60-85% identical with alpha-amylase inhibitors of wheat, but shows less than 50% identity to trypsin inhibitors of barley and wheat. The 10 Cys residues are located in identical positions compared to the cereal inhibitor...

  9. The levels of boron-uptake proteins in roots are correlated with tolerance to boron stress in barley

    Science.gov (United States)

    Boron (B) is an essential micronutrient required for plant growth and development. Recently, two major B-uptake proteins, BOR1 and NIP5;1 have been identified and partially characterized. BOR1 is a high-affinity B transporter involved in xylem loading in roots, and NIP5;1 acts is a major boric-acid ...

  10. Differential control of xanthophylls and light-induced stress proteins, as opposed to light-harvesting chlorophyll a/b proteins, during photosynthetic acclimation of barley leaves to light irradiance

    Science.gov (United States)

    Montane; Tardy; Kloppstech; Havaux

    1998-09-01

    Barley (Hordeum vulgare L.) plants were grown at different photon flux densities ranging from 100 to 1800 &mgr;mol m-2 s-1 in air and/or in atmospheres with reduced levels of O2 and CO2. Low O2 and CO2 partial pressures allowed plants to grow under high photosystem II (PSII) excitation pressure, estimated in vivo by chlorophyll fluorescence measurements, at moderate photon flux densities. The xanthophyll-cycle pigments, the early light-inducible proteins, and their mRNA accumulated with increasing PSII excitation pressure irrespective of the way high excitation pressure was obtained (high-light irradiance or decreased CO2 and O2 availability). These findings indicate that the reduction state of electron transport chain components could be involved in light sensing for the regulation of nuclear-encoded chloroplast gene expression. In contrast, no correlation was found between the reduction state of PSII and various indicators of the PSII light-harvesting system, such as the chlorophyll a-to-b ratio, the abundance of the major pigment-protein complex of PSII (LHCII), the mRNA level of LHCII, the light-saturation curve of O2 evolution, and the induced chlorophyll-fluorescence rise. We conclude that the chlorophyll antenna size of PSII is not governed by the redox state of PSII in higher plants and, consequently, regulation of early light-inducible protein synthesis is different from that of LHCII.

  11. The CELLULOSE-SYNTHASE LIKE C (CSLC) Family of Barley Includes Members that Are Integral Membrane Proteins Targeted to the Plasma Membrane

    Institute of Scientific and Technical Information of China (English)

    Fenny M. Dwivany; Dina Yuli; Rachel A. Burton; Neil J. Shirley; Sarah M. Wilson; Geoffrey B. Fincher; Antony Bacic; Ed Newbigin; Monika S. Doblin

    2009-01-01

    The CELLULOSESYNTHASE-LIKE C(CSLC) family is an ancient lineage within the CELLULOSE SYNTHASE/CEL-LULOSE SYNTHASE-LIKE (CESA/CSL) polysaccharide synthase superfamily that is thought to have arisen before the diver-gence of mosses and vascular plants. As studies in the flowering plant Arabidopsis have suggested synthesis of the (1,4)-β-glucan backbone of xyloglucan (XyG), a wall polysaccharide that tethers adjacent cellulose microfibrils to each other, as a probable function for the CSLCs, CSLC function was investigated in barley (Hordeum vulgare L.), a species with low amounts of XyG in its walls. Four barley CSLC genes were identified (designated HvCSLC1-4). Phylogenetic analysis reveals three well supported clades of CSLCs in flowering plants, with barley having representatives in two of these clades. The four barley CSLCs were expressed in various tissues, with in situ PCR detecting transcripts in all cell types of the coleoptile and root, including cells with primary and secondary cell walls. Co-expression analysis showed that HvCSLC3 was coor-dinately expressed with putative XyG xylosyltransferase genes. Both immuno-EM and membrane fractionation showed that HvCSLC2 was located in the plasma membrane of barley suspension-cultured cells and was not in internal membranes such as endoplasmic reticulum or Golgi apparatus. Based on our current knowledge of the sub-cellular locations of poly-saccharide synthesis, we conclude that the CSLC family probably contains more than one type of polysaccharide synthase.

  12. Study on Functional Properties and Hulless Barley Protein by Ultrasonic-Assisted Extraction%青稞蛋白的超声波辅助提取工艺及其功能特性研究

    Institute of Scientific and Technical Information of China (English)

    杨希娟; 党斌; 吴昆仑; 迟德钊; 刘新红

    2013-01-01

    In order to extend the applications of hulless barley protein prepared by different methods and environmental factors in food industry,effects of Extraction of by ultrasonic wave heating hulless barley protein and the functional properties were studied.The optimal extraction technology condition of hulless barley protein was determined by orthogonal experiment.Effect of protein concentration,temperature,pH,salt,sugar on solubility,water holding capacity,foaming capacity and stability,emulsifying capacity and stability of protein were evaluated.The results showed that the optimal extraction conditions were as follows:solid-liquid 1∶22,pH 10.5,ultrasonic power 550 W,ultrasonic extraction time 20 min at room temperature,the yield and The purity of protein could reach up to 93.15% and 78.67% under such conditions.Solubility,water holding capacity,foaming capacity and stability,emulsifying capacity and stability were the lowest around isoelectric point.Its solubility and water holding capacity were increased by suitable heating.Added salt could damage to the foam stability,emulsifying properties and emulsion stability of hulless barley protein,and sugar enhanced these functional properties.The properties of Hulless barley protein obtained in this way are satisfying,and it is suitable for use in foods such as cakes and hams.%研究超声波辅助提取青稞蛋白的工艺及其功能特性,为青稞蛋白的制备和应用提供理论依据.通过实验设计确定超声波辅助提取青稞蛋白的工艺条件,系统研究蛋白质浓度、pH、温度、离子强度对其功能特性的影响.结果表明:超声波辅助提取青稞蛋白的最佳工艺条件:室温,pH 10.5,料液比1∶22,超声功率550W,时间20min.在此条件下青稞蛋白提取率为93.15%,纯度78.67%.在等电点附近青稞蛋白的溶解度、持水力、泡沫特性和乳化特性均最差;适当的热处理可以增强其溶解度和持水力;盐的添加,损害了青稞蛋

  13. Effects of elevated atmospheric CO2 on protein quality/quantity and Zn content of barley grain

    DEFF Research Database (Denmark)

    Uddin, Mohammad Nasir; Lange, Mette; Schjørring, Jan Kofod;

    2011-01-01

    it is estimated that nearly half of the world’s population, primarily children and women in the developing world, suffer from zinc deficiency as they live on a diet that mostly is based on cereals and other starch-rich staples that have a quite low Zn content and with poor zinc bioavailability. The rise...... the increases of yield is primarily in the form of starch there will be a “dilution” of other nutrients like minerals and proteins in the grain with severe consequences for the nutrition of human and livestock Although the nutritional short-comings could be met by using higher amounts of nitrogen and mineral...... fertilizer it would increase nitrogen and mineral load (in particular for zinc in areas with intensive pig production) so this strategy may not be warranted. Alternative approaches to increase grain Zn content are to create transgenic lines that accumulate more Zn or to exploit natural variation in zinc...

  14. Malting barley BRS Borema

    Directory of Open Access Journals (Sweden)

    Euclydes Minella

    2006-01-01

    Full Text Available BRS Borema is an early maturing, two-rowed spring barley registered in 2003 for commercial production inSouthern Brazil, bred by Embrapa Trigo. It combines good yield potential with superior malting quality and a reasonable levelof disease (net blotch, powdery mildew, leaf rust resistance. It is well-adapted to all major production regions of maltingbarley in Brazil.

  15. Correlation of metabolic changes of infected paulownia tissue culture with PWB-MLO pathogenic mechanism

    Institute of Scientific and Technical Information of China (English)

    田国忠; 黄钦才; 袁巧平; 张锡津

    1995-01-01

    Paulownia sp.tissue culture plantlets infected with paulownia witches’ broom mycoplasma-likeorganism (PWB-MLO) expressed a typical witches’ broom symptom.The results determined by transmissionelectron microscopy and DAPI staining fluorescence microscopy showed that the severity of the symptom wasdirectly related with MLO concentration in the sieve tubes of the phloem.The results of the analysis ofendogenous hormones and phenolic compounds by HPLC revealed that in all cases,the free IAA content ofthe diseased plant apparently lower than that of the healthy,and the free IAA concentration had strikinglypositive correlation with catechol concentration changes (r=0.90,n= 11).The changes of cytokinin (Z +ZR,iPA),GA3 and ABA were shown less relation with MLO infection.The differences of peroxidase andisoenzyme,IAA oxidase and phenylanine ammonia lyase (PAL) between the extract of infected plants andthat of uninfected as well as different effects of NAA and catechol additions on MS media could further re-flect the nature of the relationship and the interaction among MLOs,catechol and the free IAA concentration.

  16. 17β-estradiol rapidly activates calcium release from intracellular stores via the GPR30 pathway and MAPK phosphorylation in osteocyte-like MLO-Y4 cells

    KAUST Repository

    Ren, Jian

    2012-03-06

    Estrogen regulates critical cellular functions, and its deficiency initiates bone turnover and the development of bone mass loss in menopausal females. Recent studies have demonstrated that 17β-estradiol (E 2) induces rapid non-genomic responses that activate downstream signaling molecules, thus providing a new perspective to understand the relationship between estrogen and bone metabolism. In this study, we investigated rapid estrogen responses, including calcium release and MAPK phosphorylation, in osteocyte-like MLO-Y4 cells. E 2 elevated [Ca 2+] i and increased Ca 2+ oscillation frequency in a dose-dependent manner. Immunolabeling confirmed the expression of three estrogen receptors (ERα, ERβ, and G protein-coupled receptor 30 [GPR30]) in MLO-Y4 cells and localized GPR30 predominantly to the plasma membrane. E 2 mobilized calcium from intracellular stores, and the use of selective agonist(s) for each ER showed that this was mediated mainly through the GPR30 pathway. MAPK phosphorylation increased in a biphasic manner, with peaks occurring after 7 and 60 min. GPR30 and classical ERs showed different temporal effects on MAPK phosphorylation and contributed to MAPK phosphorylation sequentially. ICI182,780 inhibited E 2 activation of MAPK at 7 min, while the GPR30 agonist G-1 and antagonist G-15 failed to affect MAPK phosphorylation levels. G-1-mediated MAPK phosphorylation at 60 min was prevented by prior depletion of calcium stores. Our data suggest that E 2 induces the non-genomic responses Ca 2+ release and MAPK phosphorylation to regulate osteocyte function and indicate that multiple receptors mediate rapid E 2 responses. © 2012 Springer Science+Business Media, LLC.

  17. Crystal structures of barley thioredoxin h isoforms HvTrxh1 and HvTrxh2 reveal features involved in protein recognition and possibly in discriminating the isoform specificity

    DEFF Research Database (Denmark)

    Maeda, Kenji; Hägglund, Per; Finnie, Christine;

    2008-01-01

    H-type thioredoxins (Trxs) constitute a particularly large Trx sub-group in higher plants. Here, the crystal structures are determined for the two barley Trx h isoforms, HvTrxh1 and HvTrxh2, in the partially radiation-reduced state to resolutions of 1.7 angstrom, and for HvTrxh2 in the oxidized...... state to 2.0 angstrom. The two Trxs have a sequence identity of 51% and highly similar fold and active-site architecture. Interestingly, the four independent molecules in the crystals of HvTrxh1 form two relatively large and essentially identical protein-protein interfaces. In each interface, a loop......) of the bound loop segment in the proximity of the Cys40 thiol. The interaction involves three characteristic backbone-backbone hydrogen bonds in an antiparallel beta-sheet-like arrangement, similar to the arrangement observed in the structure of an engineered, covalently bound complex between Trx...

  18. In vitro inhibition of platelet aggregation by peptides derived from oat (Avena sativa L.), highland barley (Hordeum vulgare Linn. var. nudum Hook. f.), and buckwheat (Fagopyrum esculentum Moench) proteins.

    Science.gov (United States)

    Yu, Guoyong; Wang, Feng; Zhang, Bolin; Fan, Junfeng

    2016-03-01

    Bioactive compounds present in foods could have beneficial effects on human health. In this study, we report the capacity of peptides released from oat, highland barley, and buckwheat proteins after enzymatic digestion to inhibit platelet aggregation in vitro. All hydrolysates showed high antiplatelet activity, with IC50 values of 0.282mg/ml (oat flour gastrointestinal hydrolysate, 6h) to 2.496mg/ml (highland barley glutelin tryptic hydrolysate, 14h) in a dose-dependent manner. Thirty-eight peptides with more than seven residues were identified in the tryptic hydrolysates of oat globulin. Results of computational modeling revealed that nine peptides, including ALPIDVLANAYR, EFLLAGNNKR, GEEFGAFTPK, QLAQIPR, LQAFEPLR, ALPVDVLANAYR, GEEFDAFTPK, QKEFLLAGNNK, and TNPNSMVSHIAGK bound the cyclooxygenase-1 active centers with low binding energy (-6.5 to -7.5kcal/mol). This is the first report to identify antiplatelet peptides from grain hydrolysates and the binding modes at the molecular level, leading to their possible use as functional food ingredients to prevent thrombosis.

  19. Investigation of the effect of nitrogen on severity of Fusarium Head Blight in barley

    DEFF Research Database (Denmark)

    Yang, Fen; Jensen, J.D.; Spliid, N.H.;

    2010-01-01

    The effect of nitrogen on Fusarium Head Blight (FHB) in a susceptible barley cultivar was investigated using gel-based proteomics. Barley grown with either 15 or 100 kg ha(-1)N fertilizer was inoculated with Fusarium graminearum (Fg). The storage protein fraction did not change significantly...

  20. Localization to Chromosomes of Structural Genes for the Major Protease Inhibitors of Barley Grains

    DEFF Research Database (Denmark)

    Hejgaard, Jørn; Bjørn, S.E.; Nielsen, Gunnar Gissel

    1984-01-01

    Wheat-barley chromosome addition lines were compared by isoelectric focusing of protein extracts to identify chromosomes carrying loci for the major immunochemically distinct protease inhibitors of barley grains. Structural genes for the following inhibitors were localized: an inhibitor of both...

  1. FERTILIZING BREWING BARLEY (Hordeum vulgare L.

    Directory of Open Access Journals (Sweden)

    I. Kádár

    2000-12-01

    Full Text Available Four levels of N, P and K nutrition (poor, moderate, satisfactory and high and all their possible combinations with 64 treatments in two replications (128 plots were studied in a long term field trial on barley yield and malting quality. A standard East-European spring barley "Opal" (bred in Czechoslovakia was grown in 1986, 13th year of the agricultural experiment, involving various crops in previous years, on a calcareous loamy chernozem soil. The optimum fertility levels for yield enhancement resulted in the poorest malting quality: low modification and extract but long saccharification time and high protein. To solve this problem the brewing industry will have to apply the well-known technological methods available since growers are not likely to give up their fertilizers. Applying soil and plant analysis data, having knowledge about both soil and plant optimum values, the danger of the excessive use of fertilizers can be realized and decreased.

  2. Lysine metabolism in antisense C-hordein barley grains

    DEFF Research Database (Denmark)

    Schmidt, Daiana; Rizzi, Vanessa; Gaziola, Salete A;

    2015-01-01

    ) and five antisense C-hordein transgenic barley lines. Considering the amounts of soluble and protein-bound aspartate-derived amino acids together with the analysis of key enzymes of aspartate metabolic pathway, we suggest that the C-hordein suppression did not only alter the metabolism of at least one......The grain proteins of barley are deficient in lysine and threonine due to their low concentrations in the major storage protein class, the hordeins, especially in the C-hordein subgroup. Previously produced antisense C-hordein transgenic barley lines have an improved amino acid composition......, with increased lysine, methionine and threonine contents. The objective of the study was to investigate the possible changes in the regulation of key enzymes of the aspartate metabolic pathway and the contents of aspartate-derived amino acids in the nontransgenic line (Hordeum vulgare L. cv. Golden Promise...

  3. Allele mining in barley genetic resources reveals genes of race-nonspecific powdery mildew resistance

    Directory of Open Access Journals (Sweden)

    Annika eSpies

    2012-01-01

    Full Text Available Race-nonspecific, or quantitative, pathogen resistance is of high importance to plant breeders due to its expected durability. However, it is usually controlled by multiple quantitative trait loci (QTL and therefore difficult to handle in practice. Knowing the genes that underlie race-nonspecific resistance would allow its exploitation in a more targeted manner. Here, we performed an association-genetic study in a customized worlwide collection of spring barley accessions for candidate genes of race-nonspecific resistance to the powdery mildew fungus Blumeria graminis f.sp. hordei (Bgh and combined data with results from QTL-mapping- as well as functional-genomics approaches. This led to the idenfication of 11 associated genes with converging evidence for an important role in race-nonspecific resistance in the presence of the Mlo-gene for basal susceptibility. Outstanding in this respect was the gene encoding the transcription factor WRKY2. The results suggest that unlocking plant genetic resources and integrating functional-genomic with genetic approaches accelerates the discovery of genes underlying race-nonspecific resistance in barley and other crop plants.

  4. EFFECTS OF NITROGEN ON PHENOLIC ACID AND PROTEIN CONTENT IN BARLEY (Hordeum vulgare L. ) GRAIN%氮素对大麦籽粒中酚酸和蛋白质含量的影响

    Institute of Scientific and Technical Information of China (English)

    王祥军; 齐军仓; 贾力群; 王倩; 王琴; 马建峰; 王仙; 曹连莆

    2011-01-01

    Nitrogen is one of the necessary nutrient elements for crop growth. The primary and secondary metabolisms of plant are regulated by nitrogen nutrient levels. As important secondary metabolites, phenolic acids play significant roles in plant resistances, and potentially contribute to barley qualities. However, little is known concerning effects of nitrogen on barley (Hordeum vulgare L. ) phenolic acids. In the present study, two barley cultivars were used, and seven nitrogen concentration treatments (0,412.5, 825.0, 1237.5, 1650, 3300, and 4950mg/L, respectively) under in vitro spike culture were conducted to investigate the effects of nitrogen on phenolic acid and protein contents in barley grain. Results showed that the development of barley grain was inhibited under high nitrogen levels (3300 ~ 4950mg/L), and grain weight decreasing significantly was as the response. Total hydroxybenzoic acid content (THBA), total hydroxycinnamic acid content (THCA), total phenolic acid content (TPA) and THBA/THCA ratio were all significantly affected by nitrogen concentration, cultivar and the interaction between nitrogen concentration and cultivar. Four indexes above were all significantly positively correlated with nitrogen concentration. Except for hordein content ( HC ), the protein indexes of crude protein content (CPC), salt-soluble protein content (SSPC), and glutelin content (GC) were all significantly affected by nitrogen concentration, moreover, SSPC and GC were significantly affected by cultivar and the interaction between nitrogen concentration and cultivar. Excluding HC and GC in the four indexes, CPC and SSPC were significantly positive-correlated with nitrogen concentration. The results also indicated that phenolic acid contents (including THBA, THCA, and TPA) in barley grain increased cooperated with CPC and SSPC when the nitrogen nutrient level went up.%采用穗培养技术在7个氮素(NH4NO3为氮源)浓度水平(N1:0mg/L;N2:412.5 mg/L;N3:825.0mg/L;N4

  5. Barley Transformation Using Biolistic Techniques

    Science.gov (United States)

    Harwood, Wendy A.; Smedley, Mark A.

    Microprojectile bombardment or biolistic techniques have been widely used for cereal transformation. These methods rely on the acceleration of gold particles, coated with plasmid DNA, into plant cells as a method of directly introducing the DNA. The first report of the generation of fertile, transgenic barley plants used biolistic techniques. However, more recently Agrobacterium-mediated transformation has been adopted as the method of choice for most cereals including barley. Biolistic procedures are still important for some barley transformation applications and also provide transient test systems for the rapid checking of constructs. This chapter describes methods for the transformation of barley using biolistic procedures and also highlights the use of the technology in transient assays.

  6. Barley (Hordeum vulgare L.) cysteine proteases: heterologous expression, purification and characterization

    DEFF Research Database (Denmark)

    Rosenkilde, Anne Lind; Dionisio, Giuseppe; Holm, Preben Bach

    2011-01-01

    During germination of barley seeds, mobilization of protein is essential and cysteine proteases accounts for more than 90 % of the total proteolytic activity in the degradation of barley seed storage proteins. Cysteine proteases exist as pro-enzyme and is activated through reduction of the active...... site cysteines and by removal of the pro-domain. The complement of cysteine proteases is comprehensive and for detailed studies of the individual components of this complement, a fast and efficient eukaryotic expression platform is highly desirable. A cDNA clone of the barley key cysteine endoprotease...

  7. Heterologous expression and purification of barley (Hordeum vulgare L.) cysteine protease in yeast

    DEFF Research Database (Denmark)

    Rosenkilde, Anne Lind; Dionisio, Giuseppe; Holm, Preben Bach

    2011-01-01

    The mobilization of protein during germination of barley seeds is essential and Cysteine Proteases accounts for more than 90 % of the total proteolytic activity in the degradation of barley seed storage proteins [1]. Cysteine proteases exist as pro-enzyme until activated through reduction...... of the active site cysteines and via removal of the pro-domain. The complement of cysteine proteases is comprehensive and for detailed studies of the individual components of this complement, a fast and efficient eukaryotic expression platform is highly desirable. The barley key cysteine protease, endoprotease...

  8. Identification of a phytase gene in barley (Hordeum vulgare L..

    Directory of Open Access Journals (Sweden)

    Fei Dai

    Full Text Available BACKGROUND: Endogenous phytase plays a crucial role in phytate degradation and is thus closely related to nutrient efficiency in barley products. The understanding of genetic information of phytase in barley can provide a useful tool for breeding new barley varieties with high phytase activity. METHODOLOGY/PRINCIPAL FINDINGS: Quantitative trait loci (QTL analysis for phytase activity was conducted using a doubled haploid population. Phytase protein was purified and identified by the LC-ESI MS/MS Shotgun method. Purple acid phosphatase (PAP gene was sequenced and the position was compared with the QTL controlling phytase activity. A major QTL for phytase activity was mapped to chromosome 5 H in barley. The gene controlling phytase activity in the region was named as mqPhy. The gene HvPAP a was mapped to the same position as mqPhy, supporting the colinearity between HvPAP a and mqPhy. CONCLUSIONS/SIGNIFICANCE: It is the first report on QTLs for phytase activity and the results showed that HvPAP a, which shares a same position with the QTL, is a major phytase gene in barley grains.

  9. Barley coleoptile peroxidases. Purification, molecular cloning, and induction by pathogens

    DEFF Research Database (Denmark)

    Kristensen, B.K.; Bloch, H.; Rasmussen, Søren Kjærsgård

    1999-01-01

    A cDNA clone encoding the Prx7 peroxidase from barley (Hordeum vulgare L.) predicted a 341-amino acid protein with a molecular weight of 36,515. N- and C-terminal putative signal peptides were present, suggesting a vacuolar location of the peroxidase. Immunoblotting and reverse-transcriptase poly...

  10. Aspects of the barley seed proteome during development and germination

    DEFF Research Database (Denmark)

    Finnie, Christine; Maeda, K.; Østergaard, O.;

    2004-01-01

    Analysis of the water-soluble barley seed proteome has led to the identification of proteins by MS in the major spots on two-dimensional gels covering the pi ranges 4-7 and 6-11. This provides the basis for in-depth studies of proteome changes during seed development and germination, tissue...

  11. Replication of DNA during barley endosperm development

    DEFF Research Database (Denmark)

    Giese, H.

    1992-01-01

    The incorporation of [6-H-3]-thymidine into DNA of developing barley end sperm was examined by autoradiography of cross sections of seeds and DNA analysis. The majority of nuclear divisions took place in the very young endosperm, but as late as 25 days after anthesis there was evidence for DNA...... replication. The DNA content of the endosperm increases during development and in response to nitrogen application in parallel to the storage protein synthesis profile. The hordein genes were hypersensitive to DNase I treatment throughout development....

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

    DEFF Research Database (Denmark)

    Petersen, Jørgen; Rogowska-Wrzesinska, Adelina; Jensen, Ole Nørregaard

    2013-01-01

    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....

  13. 17β estradiol regulation of connexin 43-based gap junction and mechanosensitivity through classical estrogen receptor pathway in osteocyte-like MLO-Y4 cells.

    KAUST Repository

    Ren, Jian

    2013-04-01

    Connexin 43 (Cx43) plays an essential role in osteocyte mechanotransduction. Although estrogen involves in the adaptive responses of bone cells to mechanical loadings, its effects on osteocytic Cx43-based gap junction intercellular communication (GJIC) remain obscure. We found that 17β estradiol (E2) up-regulated Cx43, and enhanced GJIC in osteocyte-like MLO-Y4 cells in fluorescence recovery after photobleaching (FRAP) assay. Combination of E2 pre-treatment and oscillating fluid flow (OFF) further enhanced Cx43 expression and mitogen-activated protein kinase (MAPK) phosphorylation, comparing to E2 or OFF treatment alone. Both blocking of classical estrogen receptors (ERα/β) by fulvestrant and ERα knockdown by small interfering RNA inhibited E2-mediated Cx43 increase, while a GPR30-specific agonist G-1 failed to promote Cx43 expression. Our results suggest that the presence of E2 enhanced Cx43-based GJIC mainly via ERα/β pathway, and sensitized osteocytes to mechanical loading. © 2012 Elsevier Inc. All rights reserved.

  14. Evaluation of qualitative and quantitative immunoassays to detect barley contamination in gluten-free beer with confirmation using LC/MS/MS.

    Science.gov (United States)

    Allred, Laura K; Sealey Voyksner, Jennifer A; Voyksner, Robert D

    2014-01-01

    To meet the need for the detection and quantitation of barley gluten in beer, qualitative screening and quantitative immunoassays based on the monoclonal antigluten antibody 401/21 (Skerritt) were validated in a single laboratory. Sample replicates were tested at each stage of beer production using multiple yeast strains and methods of end-stage protein removal. Quantitation was performed using barley-specific standards based on barley flour extracts. Immunoassay results were confirmed using LC/MS/MS for barley-specific peptides. The LOD for the qualitative screening test was 5 mg/L barley gluten. Recovery for the barley-spiked worts ranged from 81 to 128% in the quantitative ELISA assay; the LOD was methods were found to be fit for the purposes of detection of low levels of barley gluten in beer.

  15. The untranslated leader sequence of the barley lipoxygenase 1 (Lox1) gene confers embryo-specific expression

    NARCIS (Netherlands)

    Rouster, J.; Mechelen J. van; Cameron-Mills, V.

    1998-01-01

    The barley lipoxygenase 1 (Lox1) gene encodes a protein expressed in embryos during grain development and germination and in leaves after methyl-jasmonate (MeJA) treatment. Transient gene expression assays in germinating barley embryos were used to identify cis-regulatory elements involved in the em

  16. Effect of pulsed electric field on the germination of barley seeds

    DEFF Research Database (Denmark)

    Dymek, Katarzyna; Dejmek, Petr; Panarese, Valentina;

    2012-01-01

    This study explores metabolic responses of germinating barley seeds upon the application of pulsed electric fields (PEF). Malting barley seeds were steeped in aerated water for 24 h and PEF-treated at varying voltages (0 (control), 110, 160, 240, 320, 400 and 480 V). The seeds were then allowed...... to finish germination in saturated air. It is shown that exposure of germinating barley to PEF affects radicle emergence without significantly affecting the seeds’ gross metabolic activity, as quantified by isothermal calorimetry. An exploration of protein 2-DE profiles of both the embryo and the starchy...

  17. Involvement of Alternative Splicing in Barley Seed Germination.

    Science.gov (United States)

    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.

  18. The Concentration and Yield of Hordein and some Lysine-Rich Proteins as Influenced by the lys gene of Hiproly Barley

    DEFF Research Database (Denmark)

    Balasaraswathi, R.; Køie, B; Doll, Hans

    1984-01-01

    the corresponding normal gene. The lys lines had 16% lower grain yield and 13% lower single seed weight than the corresponding normal lines. The concentration of hordein in the protein and the yield of this storage protein were strongly reduced in the lys lines. On the contrary, the concentration of protein Z, β...

  19. Alanine aminotransferase controls seed dormancy in barley

    Science.gov (United States)

    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

  20. Environmental and transgene expression effects on the barley seed proteome

    DEFF Research Database (Denmark)

    Finnie, Christine; Steenholdt, T.; Noguera, O.R.;

    2004-01-01

    The barley (Hordeum vulgare) cultivar Golden Promise is no longer widely used for malting, but is amenable to transformation and is therefore a valuable experimental cultivar. Its characteristics include high salt tolerance, however it is also susceptible to several fungal pathogens. Proteome....... Eleven of these were identified by mass spectrometric peptide mass mapping, including an abundant chitinase implicated in defence against fungal pathogens and a small heat-shock protein. To enable a comparison with transgenic seed protein patterns, differences in spot patterns between field...... with extra nitrogen. Finally, the fate of transgene products in barley seeds was followed. Spots containing two green fluorescent protein constructs and the herbicide resistance marker phosphinothricin acetyltransferase were observed in 2D-gel patterns of transgenic seeds and identified by mass spectrometry...

  1. Transcriptomics analysis of hulless barley during grain development with a focus on starch biosynthesis.

    Science.gov (United States)

    Tang, Yawei; Zeng, Xingquan; Wang, Yulin; Bai, Lijun; Xu, Qijun; Wei, Zexiu; Yuan, Hongjun; Nyima, Tashi

    2017-01-01

    Hulless barley, with its unique nutritional value and potential health benefits, has increasingly attracted attentions in recent years. However, the transcription dynamics during hulless barley grain development is not well understood. In the present study, we investigated the transcriptome changes during barley grain development using Illumina paired-end RNA-sequencing. Two datasets of the developing grain transcriptomes from two barley landraces with the differential seed starch synthesis traits were generated, and comparative transcriptome approach in both genotypes was performed. The results showed that 38 differentially expressed genes (DEGs) were found co-modulated in both genotypes during the barley grain development. Of those, the proteins encoded by most of those DGEs were found, such as alpha-amylase-related proteins, lipid-transfer protein, homeodomain leucine zipper (HD-Zip), NUCLEAR FACTOR-Y, subunit B (NF-YBs), as well as MYB transcription factors. More interestingly, two genes Hvulgare_GLEAN_10012370 and Hvulgare_GLEAN_10021199 encoding SuSy, AGPase (Hvulgare_GLEAN_10033640 and Hvulgare_GLEAN_10056301), as well as SBE2b (Hvulgare_GLEAN_10018352) were found to significantly contribute to the regulatory mechanism during grain development in both genotypes. Moreover, six co-expression modules associated with specific biological processes or pathways (M1 to M6) were identified by consensus co-expression network. Significantly enriched pathways of those module genes showed difference in both genotypes. These results will expand our understanding of the complex molecular mechanism of starch synthesis during barley grain development.

  2. Characterization of Gibberellin Receptor Mutants of Barley (Hordeum vulgare L.)

    Institute of Scientific and Technical Information of China (English)

    Peter M.Chandler; Carol A.Harding; Anthony R.Ashton; Mark D.Mulcair; Nicholas E.Dixon; Lewis N.Mander

    2008-01-01

    The sequence of Gidl (a gene for a gibberellin (GA) receptor from rice) was used to identify a putative orthoIogue from barley.This was expressed in E.coil,and produced a protein that was able to bind GA in vitro with both structural specificity and saturability.Its potential role in GA responses was investigated using barley mutants with reduced GA sensitivity (gsel mutants).Sixteen different gsel mutants each carried a unique nucleotide substitution in this sequence.In all but one case,these changes resulted in single amino acid substitutions,and,for the remaining mutant,a substitution in the 5' untranslated region of the mRNA is proposed to interfere with translation initiation.There was perfect linkage in segregating populations between new mutant alleles and the gsel phenotype,leading to the conclusion that the putative GID1 GA receptor sequence in barley corresponds to the Gsel locus.Determination of endogenous GA contents in one of the mutants revealed enhanced accumulation of bioactive GA1,and a deficit of C20 GA precursors.All of the gsel mutants had reduced sensitivity to exogenous GA3,and to AC94377 (a GA analogue) at concentrations that are normally 'saturating',but,at much higher concentrations,there was often a considerable response.The comparison between barley and rice mutants reveals interesting differences between these two cereal species in GA hormonal physiology.

  3. Real-time visualization of conformational changes within single MloK1 cyclic nucleotide-modulated channels.

    Science.gov (United States)

    Rangl, Martina; Miyagi, Atsushi; Kowal, Julia; Stahlberg, Henning; Nimigean, Crina M; Scheuring, Simon

    2016-09-20

    Eukaryotic cyclic nucleotide-modulated (CNM) ion channels perform various physiological roles by opening in response to cyclic nucleotides binding to a specialized cyclic nucleotide-binding domain. Despite progress in structure-function analysis, the conformational rearrangements underlying the gating of these channels are still unknown. Here, we image ligand-induced conformational changes in single CNM channels from Mesorhizobium loti (MloK1) in real-time, using high-speed atomic force microscopy. In the presence of cAMP, most channels are in a stable conformation, but a few molecules dynamically switch back and forth (blink) between at least two conformations with different heights. Upon cAMP depletion, more channels start blinking, with blinking heights increasing over time, suggestive of slow, progressive loss of ligands from the tetramer. We propose that during gating, MloK1 transitions from a set of mobile conformations in the absence to a stable conformation in the presence of ligand and that these conformations are central for gating the pore.

  4. Resistance to Barley Leaf Stripe

    DEFF Research Database (Denmark)

    Nørgaard Knudsen, J. C.

    1986-01-01

    Ten barley [Hordeum vulgare] genotypes were inoculated with twelve isolates of Pyrenophora graminea of diverse European and North African origin. Race specific resistance occurred. Four, possibly five, genetically different sources of race-specific resistance were found, three of them occurring i...

  5. Effects of Pleurotus sapidus (Schulzer Sacc. treatment on nutrient composition and ruminal fermentability of barley straw, barley rootless, and a mixture of the two

    Directory of Open Access Journals (Sweden)

    Alfonso Soto-Sánchez

    2015-09-01

    Full Text Available Barley (Hordeum vulgare L., and its derivatives, ranks fourth in cereal production worldwide, and the Pleurotus species are among the most efficient types of lignocellulolytic white-rot fungi. The objective of this research study was to evaluate the degradation of barley straw and barley rootless with an inoculum of Pleurotus to improve their nutritional availability as a food source for ruminants. Two experiments were conducted; the first was to determine the effects of inoculation of Pleurotus sapidus (Schulzer Sacc. (PS in barley straw (BS, barley rootless (BR, and a 75% BS and 25% BR mixture (M. The second experiment was to evaluate the same substrates in vitro ruminal fermentation. Barley rootless had better organic matter (OM degradability than BS after 24 h incubation with PS. The protein content in BR was higher than in BS (P < 0.01. Enzyme activities had the highest concentration from the start of fermentation, and in vitro dry matter (DM degradability in BS and BR increased after 8 and 24 d fermentation, respectively (P < 0.05. Propionic acid concentration was enhanced after 16 d fermentation in BR (P < 0.5. The use of BS combined with BR exhibited better fermentation; this result provides relevant information for integrating BR with other substrates and improving the use of straw, which can be more nutritionally available for feeding ruminants.

  6. Optimization of extraction method of the water-soluble proteins of barley and malt and construction of two-dimensional electrophoretic map%大麦和麦芽水溶性蛋白提取方案的优化及双向电泳图谱的建立

    Institute of Scientific and Technical Information of China (English)

    夏先锋; 朴永哲; 沈海萍; 翟明昌; 成建国; 赵长新

    2012-01-01

    To develop an efficient and high resolution 2-DE procedure for barley and malt research, in this study, the method of extraction and sample preparation for two dimensional gel electrophoresis (2-DE) were optimized using high quality barley cultivars Gairdner and by changing the type and amount of extraction reagents as well as extraction time. By using the optimized method, protein extraction and sample preparation methods could be achieved. The protein sample of barley was obtained after grinding, buffer-phenol extraction, ethanol washing, and was freeze-dried. Protein spots were separated by 2-DE technique, stained with silver nitrate, recorded and detected by PDQuest software. The results showed that over 700 spots of barley and about 500 spots of malt were separated by 2-DE. The established of those methods may provide powerful tools for developmental proteomics and quality improvement of barley in the future.%为了建立一套适用于大麦和麦芽的双向电泳分析方法,得到更加清晰、全面的双向电泳图谱。以高产优质大麦品种Gairdner为材料,对提取方法以及提取过程中所用的试剂的种类、用量和处理时间等进行比较分析和优化,运用蛋白定量法测定提取样品的蛋白质含量,摸索出一套完整的用于双向电泳分析的大麦蛋白提取和样品制备方法。大麦或麦芽经研磨,缓冲液-苯酚提取,醇洗,冻干后得到的蛋白质样品,双向电泳分离,硝酸银染色,PDQuest分析,图谱结果显示大麦检测到700个蛋白质点,麦芽检测到500个蛋白质点。这一方法的构建可作为大麦蛋白质组研究的重要工具,为今后大麦蛋白质组学以及对大麦品质改良的研究奠定了一定的基础。

  7. The plasma membrane proteome of germinating barley embryos

    DEFF Research Database (Denmark)

    Hynek, Radovan; Svensson, Birte; Jensen, O.N.;

    2009-01-01

    with amphiphilicity and low abundance of membrane proteins. A fraction enriched in plasma membranes was prepared from embryos dissected from 18 h germinated barley seeds using aqueous two-phase partitioning. Reversed-phase chromatography on C-4 resin performed in micro-spin columns with stepwise elution by 2-propanol...... was used to reduce soluble protein contamination and enrich for hydrophobic proteins. Sixty-one proteins in 14 SDS-PAGE bands were identified by LC-MS/MS and database searches. The identifications provide new insight into the plasma membrane functions in seed germination....

  8. Identification of a protein associated with circulative transmission of Barley yellow dwarf virus from cereal aphids, Schizaphis graminum and Sitobion avenae

    Institute of Scientific and Technical Information of China (English)

    WANG Xifeng; ZHOU Guanghe

    2003-01-01

    Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae,two aphid species transmitting BYDV-GAV. P50 in the extracts of S. graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation. After feeding the antiserum through membrane, the transmission efficiencies of S. graminun and S. avenae for BYDV-GAV decreased significantly. It was suggested that P50 should be related with transmission process. Location of P50 was found at the plasma membrane surrounding the accessory salivary gland (ASG) in the head tissues of S. graminum by immunogold-labelling experiment.The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.

  9. Zinc Blotting Assay for Detection of Zinc-Binding Prolamin in Barley (Hordeum vulgare) Grain

    DEFF Research Database (Denmark)

    Uddin, Mohammad Nasir; Langkilde, Ane; Vincze, Éva

    2014-01-01

    In plants, zinc is commonly found bound to proteins. In barley (Hordeum vulgare), major storage proteins are alcohol-soluble prolamins known as hordeins, and some of them have the potential to bind or store zinc. 65Zn overlay and blotting techniques have been widely used for detecting zinc......-binding protein. However, to our knowledge so far this zinc blotting assay has never been applied to detect a prolamin fraction in barley grains. A radioactive zinc (65ZnCl2) blotting technique was optimized to detect zinc-binding prolamins, followed by development of an easy-to-follow nonradioactive colorimetric...... zinc blotting method with a zinc-sensing dye, dithizone. Hordeins were extracted from mature barley grain, separated by SDS-PAGE, blotted on a membrane, renatured, overlaid, and probed with zinc; subsequently, zinc-binding specificity of certain proteins was detected either by autoradiography or color...

  10. Antisense-mediated suppression of C-hordein biosynthesis in the barley grain results in correlated changes in the transcriptome, protein profile, and amino acid composition

    DEFF Research Database (Denmark)

    Hansen, Mette; Lange, Marianne; Friis, Carsten;

    2007-01-01

    Antisense- or RNAi-mediated suppression of the biosynthesis of nutritionally inferior storage proteins is a promising strategy for improving the amino acid profile of seeds. However, the potential pleiotropic effects of this on interconnected pathways and the agronomic quality traits need...

  11. The germinlike protein GLP4 exhibits superoxide dismutase activity and is an important component of quantitative resistance in wheat and barley

    DEFF Research Database (Denmark)

    Christensen, Anders Bentsen; Thordal-Christensen, Hans; Zimmermann, Grit

    2004-01-01

    , was examined in an epidermal transient expression system and in transgenic Arabidopsis thaliana plants overexpressing His-tagged HvGLP4. Leaf extracts of transgenic Arabidopsis overexpressing HvGLP4 contained a novel His-tagged protein with superoxide dismutase activity and HvGLP4 epitopes. Transient...

  12. Cisgenic barley for animal feed

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2011-01-01

    Service and Sejet Plant Breeding based on the Cisgenesis concept. Cisgenesis implies that the plants are transformed only with its own or very closely related genetic material and that the final Cisgenic plants have to be free of any foreign genes. The Cisgenesis concept allows for the introduction...... of extra gene copies of a particular gene whereby a particular trait can be accentuated. Transgenic crops generated by the Cisgenesis concept are accordingly very similar to those generated by conventional breeding. In our part of the project we are focusing on barley phytases as candidate genes...... using a genomic barley lambda library. The clone has been inserted into a Cisgenic Agrobacterium vector where both the gene of interest and the selection gene are flanked by their own T-DNA borders in order to promote integration of the two genes at unlinked places in the plant genome. T0-plants show...

  13. Development of DNA markers associated with beer foam stability for barley breeding.

    Science.gov (United States)

    Iimure, Takashi; Kihara, Makoto; Ichikawa, Seiichiro; Ito, Kazutoshi; Takeda, Kazuyoshi; Sato, Kazuhiro

    2011-01-01

    Traits conferring brewing quality are important objectives in malting barley breeding. Beer foam stability is one of the more difficult traits to evaluate due to the requirement for a relatively large amount of grain to be malted and then the experimental costs for subsequent brewing trials. Consequently, foam stability tends to be evaluated with only advanced lines in the final stages of the breeding process. To simplify the evaluation and selection for this trait, efficient DNA makers were developed in this study. Previous studies have suggested that the level of both of the foam-associated proteins Z4 and Z7 were possible factors that influenced beer foam stability. To confirm the relationship between levels of these proteins in beer and foam stability, 24 beer samples prepared from malt made from 10 barley cultivars, were examined. Regression analyses suggested that beer proteins Z4 and Z7 could be positive and negative markers for beer foam stability, respectively. To develop DNA markers associated with contents of proteins Z4 and Z7 in barley grain, nucleotide sequence polymorphisms in barley cultivars in the upstream region of the translation initiation codon, where the promoter region might be located were compared. As a result, 5 and 23 nucleotide sequence polymorphisms were detected in protein Z4 and protein Z7, respectively. By using these polymorphisms, cleaved amplified polymorphic sequence (CAPS) markers were developed. The CAPS markers for proteins Z4 and Z7 were applied to classify the barley grain content of 23 barley cultivars into two protein Z4 (pZ4-H and pZ4-L) and three protein Z7 (the pZ7-H, pZ7-L and pZ7-L2) haplotypes, respectively. Barley cultivars with pZ4-H showed significantly higher levels of protein Z4 in grain, and those with pZ7-L and pZ7-L2 showed significantly lower levels of protein Z7 in grain. Beer foam stability in the cultivars with pZ4-H and pZ7-L was significantly higher than that with pZ4-L and pZ7-H, respectively. Our

  14. Identification and Expression Analysis of the Barley (Hordeum vulgare L. Aquaporin Gene Family.

    Directory of Open Access Journals (Sweden)

    Runyararo M Hove

    Full Text Available Aquaporins (AQPs are major intrinsic proteins (MIPs that mediate bidirectional flux of water and other substrates across cell membranes, and play critical roles in plant-water relations, dehydration stress responses and crop productivity. However, limited data are available as yet on the contributions of these proteins to the physiology of the major crop barley (Hordeum vulgare. The present work reports the identification and expression analysis of the barley MIP family. A comprehensive search of publicly available leaf mRNA-seq data, draft barley genome data, GenBank transcripts and sixteen new annotations together revealed that the barley MIP family is comprised of at least forty AQPs. Alternative splicing events were likely in two plasma membrane intrinsic protein (PIP AQPs. Analyses of the AQP signature sequences and specificity determining positions indicated a potential of several putative AQP isoforms to transport non-aqua substrates including physiological important substrates, and respond to abiotic stresses. Analysis of our publicly available leaf mRNA-seq data identified notable differential expression of HvPIP1;2 and HvTIP4;1 under salt stress. Analyses of other gene expression resources also confirmed isoform-specific responses in different tissues and/or in response to salinity, as well as some potentially inter-cultivar differences. The work reports systematic and comprehensive analysis of most, if not all, barley AQP genes, their sequences, expression patterns in different tissues, potential transport and stress response functions, and a strong framework for selection and/or development of stress tolerant barley varieties. In addition, the barley data would be highly valuable for genetic studies of the evolutionarily closely related wheat (Triticum aestivum L..

  15. Inactivation of barley limit dextrinase inhibitor by thioredoxin-catalysed disulfide reduction

    DEFF Research Database (Denmark)

    Jensen, Johanne Mørch; Hägglund, Per; Christensen, Hans Erik Mølager;

    2012-01-01

    Barley limit dextrinase (LD) that catalyses hydrolysis of α-1,6 glucosidic linkages in starch-derived dextrins is inhibited by limit dextrinase inhibitor (LDI) found in mature seeds. LDI belongs to the chloroform/methanol soluble protein family (CM-protein family) and has four disulfide bridges a...

  16. Molecular characterization of two lipoxygenases from barley

    NARCIS (Netherlands)

    Mechelen, J.R. van; Schuurink, R.C.; Smits, M.; Graner, A.; Douma, A.C.; Sedee, N.J.A.; Schmitt, N.F.; Valk, B.E.

    1999-01-01

    Two full-length lipoxygenase cDNA sequences (LoxB and LoxC) from barley (Hordeum distichum cv. L. Triumph) are described. The cDNAs share high homology with the barley LoxA cDNA. Southern blotting experiments indicate single copy numbers of the three lipoxygenase genes. RFLP mapping revealed the pre

  17. Issues surrounding health claims for barley.

    Science.gov (United States)

    Ames, Nancy P; Rhymer, Camille R

    2008-06-01

    Government-approved health claims support dietary intervention as a safe and practical approach to improving consumer health and provide industry with regulatory guidelines for food product labels. Claims already allowed in the United States, United Kingdom, Sweden, and The Netherlands for reducing cholesterol through consumption of oat or barley soluble fiber provide a basis for review, but each country may have different criteria for assessing clinical evidence for a physiological effect. For example, the FDA-approved barley health claim was based on a petition that included 39 animal model studies and 11 human clinical trials. Since then, more studies have been published, but with few exceptions, clinical data continue to demonstrate that the consumption of barley products is effective for lowering total and LDL cholesterol. More research is needed to fully understand the mechanism of cholesterol reduction and the role of beta-glucan molecular weight, viscosity, and solubility. In an assessment of the physiological efficacy of a dietary intervention, consideration should also be given to the potential impact of physical and thermal food-processing treatments and genotypic variation in the barley source. New barley cultivars have been generated specifically for food use, possessing increased beta-glucan, desirable starch composition profiles, and improved milling/processing traits. These advances in barley production, coupled with the establishment of a government-regulated health claim for barley beta-glucan, will stimulate new processing opportunities for barley foods and provide consumers with reliable, healthy food choices.

  18. Effects of split nitrogen fertilization on post-anthesis photoassimilates, nitrogen use efficiency and grain yield in malting barley

    DEFF Research Database (Denmark)

    Cai, Jian; Jiang, Dong; Liu, Fulai

    2011-01-01

    photosynthesis after anthesis, dry matter accumulation and assimilates remobilization, nitrogen use efficiency and grain yield to fraction of topdressed nitrogen treatments were investigated in malting barley. Net photosynthetic rate of the penultimate leaf, leaf area index and light extinction coefficient...... and agronomic nitrogen use efficiencies. The enhanced nitrogen use efficiency was corresponding to the improved photosynthetic nitrogen-use efficiency in the leaves at fraction of topdressed nitrogen of 30%. In conclusion, appropriate fraction of topdressed nitrogen application on malting barley improved...... assimilation rate and nitrogen use efficiency resulting in higher grain yields and proper grain protein content in malting barley....

  19. Grain Composition and Functional Ingredients of Barley Varieties Created in Latvia

    Directory of Open Access Journals (Sweden)

    Šterna Vita

    2015-09-01

    Full Text Available Cereals, including barley, have been recognised as functional foods that provide beneficial effect on the health of the consumer and decrease the risk of various diseases. The aim of investigation was to determine the grain composition of barley varieties and perspective breeding lines bred in Latvia and to evaluate its functional ingredients. The results of analysis showed that protein content among varieties ranged from 106.6-146.8 g·kg-1, total dietary fibre 187.4-208.2 g·kg-1, β-glucans 42.8 g-49.4 g·kg-1, and amount of α-tocopherol 6.03-8.93 mg·kg-1. The sum of essential amino acids in barley grain samples was from 32.90 g·kg-1 to 38.71 g·kg-1. All varieties of hulled and hulless barley grain were found to be sources of protein with high biological value. Comparison of barley varieties bred in Latvia suggests that variety ‘Kornelija’ outperforms others in protein, dietary fibre and micronutrient content.

  20. A major baker's asthma allergen from rye flour is considerably more active than its barley counterpart.

    Science.gov (United States)

    García-Casado, G; Armentia, A; Sánchez-Monge, R; Sánchez, L M; Lopez-Otín, C; Salcedo, G

    1995-05-01

    A rye flour protein of about 13.5 kDa, as well as its barley homologue, have been isolated. The rye component was recognized in vitro by IgE of allergic patients and provoked positive responses in 15 out of 21 baker's asthma patients (71%) when skin prick tests were performed. Its barley homologue showed no detectable in vitro reactivity and caused positive responses in only one-third of patients. Although no inhibitory activity against different alpha-amylases or trypsin was found for these two proteins, their N-terminal sequencing revealed considerable similarity to several members of the cereal alpha-amylase/trypsin inhibitor family.

  1. Calcium homeostasis in barley aleurone

    Energy Technology Data Exchange (ETDEWEB)

    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.

  2. Effects of Nitrogen Rate and Planting Density on Yield and Its Components and Protein Content of Barley Variety Supi 6%施氮量和播种密度对苏啤6号产量、产量构成因素及其蛋白质含量的影响

    Institute of Scientific and Technical Information of China (English)

    臧慧; 陈和; 陈健; 沈会权; 陶红; 陈晓静; 乔海龙

    2011-01-01

    以苏啤6号为材料,研究了不同施氮量和播种密度对其产量、产量构成因素及籽粒蛋白质含量的影响.结果表明:施氮量和播种密度对苏啤6号产量、产量构成因素、籽粒蛋白质含量有显著影响.本试验条件下,苏啤6号的适宜施氮量为210 kg/hm2,播种基本苗为300万/hm2.%With the barley variety Supi 6 as material, the effects of nitrogen rate and planting density on yield and yield components and protein content of barley were studied. The results showed that nitrogen appli cation rate and planting density could significantly influent the yield and its components and the protein content in grain of Supi 6. Under the experiment conditions and the optimal nitrogen rate was 210 kg/hm2 and the bas ic seedling number was 3 000 thousand per hectare.

  3. Establishment of action levels for quality control of IMRT flat panel: experience with the algorithm iGRiMLO; Establecimiento de niveles de accion para el control de calidad de IMRT con panel plano: experiencia con el algoritmo iGRiMLO

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez, V.; Dolores, VV. de los; Pastor, V.; Martinez, J.; Gimeno, J.; Guardino, C.; Crispin, V.

    2011-07-01

    Algorithm has been used at our institution iGRiMLO scheduled for individual verification of treatment plans for intensity modulated radiotherapy (IMRT) step and shoot through portal dosimetry pretreatment of non-transmission, triggering the plan directly to a portal imaging device (EPID) of an amorphous silicon flat panel.

  4. Long-term influence of feeding barley treated with lactic acid and heat on performance and energy balance in dairy cows.

    Science.gov (United States)

    Gruber, Leonhard; Khol-Parisini, Annabella; Humer, Elke; Abdel-Raheem, Sherief M; Zebeli, Qendrim

    2017-02-01

    The study evaluated the long-term influence of feeding ground barley treated with lactic acid (LA) alone or with LA and heat on performance, energy and protein balance in dairy cows. Thirty cows were fed three diets differing in the treatment of barley grain, either unprocessed ground barley (Control), ground barley steeped in 1% LA at room temperature (LA-treated barley) or ground barley steeped in 1% LA with an additional heating at 55°C (LAH-treated barley). Cows were studied from week 3 to 17 post-partum. Dry matter intake (DMI), milk yield and composition and body weight (BW) were measured daily. Estimated energy and protein balances were calculated and blood samples were collected three times during the experiment and analysed for common metabolites of energy and lipid metabolism. Digestibility of different treated barley and other dietary ingredients was investigated in vivo using four wethers. The treatment of barley with LA and LAH increased the digestibility of organic matter (OM) by approximately 5% and the content of metabolisable energy by 0.5-0.6 MJ/kg DM. Data showed no effect of feeding diets containing LA- or LAH-treated barley at 39% of DM on overall DMI, BW, BW change, milk production and composition and on the blood variables studied. Diet influenced the estimated balances of net energy of lactation (p balances. In conclusion, feeding diets containing LA- or LAH-treated barley had no influence on performance, milk composition and blood metabolites, but LA treatment without heat seems to improve the energy balance of cows.

  5. The Metabolic Signature of Biomass Formation in Barley.

    Science.gov (United States)

    Ghaffari, Mohammad R; Shahinnia, Fahimeh; Usadel, Björn; Junker, Björn; Schreiber, Falk; Sreenivasulu, Nese; Hajirezaei, Mohammad R

    2016-09-01

    The network analysis of genome-wide transcriptome responses, metabolic signatures and enzymes' relationship to biomass formation has been studied in a diverse panel of 12 barley accessions during vegetative and reproductive stages. The primary metabolites and enzymes involved in central metabolism that determine the accumulation of shoot biomass at the vegetative stage of barley development are primarily being linked to sucrose accumulation and sucrose synthase activity. Interestingly, the metabolic and enzyme links which are strongly associated with biomass accumulation during reproductive stages are related to starch accumulation and tricarboxylic acid (TCA) cycle intermediates citrate, malate, trans-aconitate and isocitrate. Additional significant associations were also found for UDP glucose, ATP and the amino acids isoleucine, valine, glutamate and histidine during the reproductive stage. A network analysis resulted in a combined identification of metabolite and enzyme signatures indicative for grain weight accumulation that was correlated with the activity of ADP-glucose pyrophosphorylase (AGPase), a rate-limiting enzyme involved in starch biosynthesis, and with that of alanine amino transferase involved in the synthesis of storage proteins. We propose that the mechanism related to vegetative and reproductive biomass formation vs. seed biomass formation is being linked to distinct fluxes regulating sucrose, starch, sugars and amino acids as central resources. These distinct biomarkers can be used to engineer biomass production and grain weight in barley.

  6. Dissecting molecular interactions involved in recognition of target disulfides by the barley thioredoxin system

    DEFF Research Database (Denmark)

    Björnberg, Olof; Maeda, Kenji; Svensson, Birte

    2012-01-01

    Thioredoxin reduces disulfide bonds, thus regulating activities of target proteins in various biological systems, e.g., inactivation of inhibitors of starch hydrolases and proteases in germinating plant seeds. In the three-dimensional structure of a complex with barley α-amylase/subtilisin inhibi......Thioredoxin reduces disulfide bonds, thus regulating activities of target proteins in various biological systems, e.g., inactivation of inhibitors of starch hydrolases and proteases in germinating plant seeds. In the three-dimensional structure of a complex with barley α......-amylase/subtilisin inhibitor (BASI), two loops in barley thioredoxin h2 (HvTrxh2), containing an invariant cis-proline (86EAMP89) and a conserved glycine (104VGA106), surround the active site cysteines ( 45WCGPC49) and contribute to binding of BASI through backbone-backbone hydrogen bonds [Maeda, K., Hägglund, P., Finnie, C...

  7. Nutritional assessment of barley, talbina and their germinated products

    Directory of Open Access Journals (Sweden)

    Mohamed kamal El-Sayed Youssef

    2013-02-01

    Full Text Available Talbina is a food product with high potential applications as a functional food. Talbina was prepared from two barley varieties namely: Giza126 and Giza130 by adding whole barley flour to water (1:10 w/v and (1:5 w/v for germinated barley then heating at  80° C for 5 minutes with continuous stirring until reaching a porridge like texture. The present investigation was carried out in an attempt to clearly the nutritional assessment of talbina as a functional food. The study included the determination of gross chemical composition, caloric value, mineral composition, vitamins composition and the amino acids composition. Meanwhile, computation of the chemical scores (CS and A/E ratios were carried out for raw, germinated barley, talbina, germinated talbina and commercial talbina. The data revealed that protein content of the all raw studied and processing treatments ranged from 8.75-18.34g/100g on dry weight basis. Besides, the all treatments recorded rather slight decrease in crude fat content. Likewise, ash and carbohydrates ranged between 2.29-2.86 and 73.40-82.66%, respectively. Whereas crude fiber had an increase after treatments and it ranged from 3.83-4.37%. On the other hand by making talbina iron, manganese, copper and zinc increased especially zinc, which recorded higher value than that recommended daily. Furthermore, germinated talbina130 recorded the highest amounts of vitamins B2, Nicotinic acid, B6 and folic acid. Moreover, the present study indicated that phenylalanine was the highest essential amino acid, followed by leucine.

  8. A laser ablation ICP-MS based method for multiplexed immunoblot analysis: applications to manganese-dependent protein dynamics of photosystem II in barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    de Bang, Thomas Christian; Petersen, Jørgen; Pedas, Pai Rosager

    2015-01-01

    developed a multiplexed antibody-based assay and analysed selected PSII subunits in barley (Hordeum vulgare L.). A selection of antibodies were labelled with specific lanthanides and immunoreacted with thylakoids exposed to Mn deficiency after western blotting. Subsequently, western blot membranes were...... analysed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), which allowed selective and relative quantitative analysis via the different lanthanides. The method was evaluated against established liquid chromatography electrospray ionization tandem mass spectrometry (LC...

  9. Induction of beta-1,3-glucanase in barley in response to infection by fungal pathogens.

    Science.gov (United States)

    Jutidamrongphan, W; Andersen, J B; Mackinnon, G; Manners, J M; Simpson, R S; Scott, K J

    1991-05-01

    The sequence of a partial cDNA clone corresponding to an mRNA induced in leaves of barley (Hordeum vulgare) by infection with fungal pathogens matched almost perfectly with that of a cDNA clone coding for beta-1,-3-glucanase isolated from the scutellum of barley. Western blot analysis of intercellular proteins from near-isogenic barley lines inoculated with the powdery mildew fungus (Erysiphe graminis f. sp. hordei) showed a strong induction of glucanase in all inoculated lines but was most pronounced in two resistant lines. These data were confirmed by beta-1,3-glucanase assays. The barley cDNA was used as a hybridization probe to detect mRNAs in barley, wheat (Triticum aestivum), rice (oryza sativus), and sorghum (Sorghum bicolor), which are induced by infection with the necrotrophic pathogen Bipolaris sorokiniana. These results demonstrate that activation of beta-1,3-glucanase genes may be a general response of cereals to infection by fungal pathogens.

  10. Hordein Variation in Wild (Hordeum Spontaneum) and Cultivated (H. Vulgare) Barley

    DEFF Research Database (Denmark)

    Doll, Hans; Brown, A. H. D.

    1979-01-01

    The storage protein hordein contains two major groups of polypeptides which are highly polymorphic in barley, and in its evolutionary progenitor Hordeum spontaneum Koch. Crosses between the two species showed that the complex electrophoretic phenotypes within the two groups of polypeptides are go...

  11. Primer Dependent and Independent Forms of Soluble Starch Synthetase from Developing Barley Endosperms

    DEFF Research Database (Denmark)

    Kreis, M.

    1980-01-01

    The activity of soluble starch synthetase (ADP-glucose: agr-1,4-glucan agr-4-glucosyltransferase) in the non-purified extract from 16 day-old Bomi barley endosperms (Hordeum vulgare L.) was low and the reaction was non-linear when plotted against protein concentration. Starch synthetase was purif...

  12. Degradability Characteristics of Treated and Untreated Barley Grain Using In situ Technique

    Directory of Open Access Journals (Sweden)

    Akbar Taghizadeh

    2008-01-01

    Full Text Available This study was carried out to determine of degradability characteristics of untreated barely grain (UBG and treated barley grain with autoclaving at 120°C, 5´ (TBG1 and 20' (TBG2, treated barley grain at 100°C, 5´ (TBG3 and 20' (TBG4, using in situ technique in Ghizel sheep’s. The sheep fed diet content 40% alfalfa: 60% concentrate containing 2.9 Mcal kg-1 DM and 14% CP. The incubation times were 0, 4, 8, 16, 24, 36 and 48 h and rumen degradation of cp and DM was measured. The equation of p = a+b (1-e-ct was used for fitting of dry matter and crude protein disappearance data. The dry matter disappearance of TBG1 and TBG2 at 24 and 48 h were lower than the other treatments (p<0.05. The crude protein disappearance of 24 and 48 h of UBG was more than the other treatment (p<0.05. Treating of barley grain of 120°C (5' and 20' can be decreased ruminal crude protein degradability of barley grain resulting high escaped crude protein into lower digestive tract.

  13. Secretory expression of functional barley limit dextrinase by Pichia pastoris using high cell-density fermentation

    DEFF Research Database (Denmark)

    Vester-Christensen, Malene Bech; Abou Hachem, Maher; Næsted, Henrik

    2010-01-01

    Heterologous production of large multidomain proteins from higher plants is often cumbersome. Barley limit dextrinase (LD), a 98 kDa multidomain starch and alpha-limit dextrin debranching enzyme, plays a major role in starch mobilization during seed germination and is possibly involved in starch...

  14. Leaf senescence and nutrient remobilisation in barley and wheat

    DEFF Research Database (Denmark)

    Gregersen, P L; Holm, P B; Krupinska, K

    2008-01-01

    Extensive studies have been undertaken on senescence processes in barley and wheat and their importance for the nitrogen use efficiency of these crop plants. During the senescence processes, proteins are degraded and nutrients are re-mobilised from senescing leaves to other organs, especially...... in degradative, metabolic and regulatory processes that could be used in future strategies aimed at modifying the senescence process. The breeding of crops for characters related to senescence processes, e.g. higher yields and better nutrient use efficiency, is complex. Such breeding has to cope with the dilemma...

  15. Important Quality Attributes of Malt Barley (Hordeum vulgare L. ): Effect of Genetics, Environments and Agronomic Factors

    Institute of Scientific and Technical Information of China (English)

    Virender Sardana; ZHANG Guo-ping

    2003-01-01

    Barley has been in use as food and fodder for thousands of years, and also used currently as themost important raw material in malting and brewing. The quality of malt barley is not much satisfied with therequirement by malting and brewing industry worldwide, although the great effort has been devoted to its im-provement through breeding and agronomy. The quality of malt barley is involved in many physical and chemi-cal traits, including hydrolytic enzymes. Of them, kernel protein content, beta-amylase activity, beta-glucancontent and kernel plumpness have dramatic influence on malting or brewing quality. The expression of thesequality parameters varies greatly in genotypes and is easily changed by diverse environments and agronomicpractices. This paper reviewed the important role of various parameters influencing the quality of malt barleyand the effect of different factors on them.

  16. Chromosomal Location and Expression of Green Fluorescent Protein (gfp) Gene in Microspore Derived Transgenic Barley (Hordeum vulgare L.)%转基因大麦中gfp基因的染色体位置及其表达

    Institute of Scientific and Technical Information of China (English)

    陈建民; Carlson A R; 万建民; Kasha K J

    2003-01-01

    通过对大麦小孢子进行基因枪轰击获得4株转绿色荧光蛋白基因(gfp)的植株(A、C、D、E),以gfp基因为探针进行荧光原位杂交(FISH)研究转化植株中转基因插入位置和基因表达.4个株系在染色体7L(5HL)的不同位置都有一个插入点,而E株系在染色体5S(7HS)还有第2个插入点.所有的转基因T0代植株都是半合子并在T1、T2代发生分离.D株系GFP未表达,但FISH和PCR分析表明gfp基因已成功插入其染色体.各株系在根尖和花粉中的GFP表达水平不同:C株系在花粉表达强而在根尖表达中等;A株系在花粉中等表达而在根尖表达较淡;E株系则在根尖高表达,花粉中等表达.A和C株系在根尖和花粉的GFP分离都表现单位点特性,而E株系的根尖分离表现重叠作用(15∶1)特征,但在花粉中表达GFP的频率低.PCR结果和3个分离株系的根尖表达结果一致.D和E株系的GFP表达不正常可能和gfp基因插入位置或基因的结构有关.%Four doubled haploid barley lines (A,C,D,E) derived from gfp (green fluorescent protein) transformation and selection following particle bombardment of microspores were studied for gene expression pattern and the location of genome inserts.The integration sites were detected by fluorescence in situ hybridization (FISH) using the gfp plasmid DNA as a probe.Plants from events A,C,D and E all have a single insert site on chromosome 7L(5HL) at different locations while line E has a second insert site on chromosome 5S(7HS).All original transgenic plants were hemizygous for the transgenes and segregated in the T1 and T2 generations.Although line D had no GFP expression,FISH and PCR could detect gfp gene on its chromosome in transformed plants.Expression levels of GFP varied with lines and tissues examined.Plants from line C showed good expression in pollen and an intermediate level in root tips.Plants from A have intermediate expression of GFP in the pollen and light expression in the

  17. 2015 nationwide survey revealed Barley stripe mosaic virus in Korean barley fields

    Science.gov (United States)

    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...

  18. Optimization of thermal stability of barley protein-glucose maillard reaction by response surface methodology%响应面分析法在大麦蛋白-葡萄糖Maillard反应物热稳定性研究中的应用

    Institute of Scientific and Technical Information of China (English)

    董学艳; 刘继超; 王晓丹; 聂妤; 赵长新

    2011-01-01

    酿造大麦是啤酒生产过程中最重要的原料,大麦中的蛋白质对麦芽与啤酒的质量起着重要的作用,其中蛋白的热稳定性对啤酒泡沫的形成和稳定均具有重要的作用.国内外的研究表明,通过美拉德反应能够有效地改善蛋白质的功能特性.本论文主要研究美拉德反应对于大麦蛋白热稳定性的影响,通过二次回归通用旋转正交试验设计,应用响应面分析法对大麦蛋白-葡萄糖(Pr-G)Maillard反应过程中的各反应条件进行分析,建立数学模型,优化反应条件参数,以求得到热稳定性佳的Pr-G Maillard反应共聚物.结果表明:在90℃、pH9.08、反应时间为39.00min、大麦蛋白-葡萄糖的配比为1:1.04可以得到高热稳定性的反应共聚物.在此最佳反应条件下对反应共聚物的热稳定性进行了研究,证实了预测值和实测值是一致的.%Brewing barley is the most important raw material in beer production. The protein in the barley plays an important part in the quality of malt and beer, and the heat -stable protein plays important roles on formation and stability of the beer foam. It was reported that the functional properties of protein could be modified by Maillard reaction. The effect of the thermal stability of barley protein by Maillard reaction was studied. The factors such as pH, the ratio of reactants and reaction time affecting the Maillard reaction kinetics were discussed. Response surface methodology was used to optimize Maillard reaction factors in order to get Maillard reaction with the highest thermal stability property.The results indicated that reaction temperature 90 ℃, pH 9.08, reaction time 40 min and weight ratio of barley protein to glucose 1: 1.04 were the optimum reaction conditions. Under the above conditions, the thermal stability of conjugation was studied, and it was confirmed that the predicted and actual value of the thermal stability were identical.

  19. Effect of Barley and Enzyme on Performance, Carcass, Enzyme Activity and Digestion Parameters of Broilers

    Directory of Open Access Journals (Sweden)

    majid kalantar

    2016-04-01

    Full Text Available Introduction Corn has been recently used for producing ethanol fuel in the major corn-producing countries such as the US and Brazil. Recent diversion of corn for biofuel production along with the increased world's demand for this feedstuff has resulted in unprecedented rise in feed cost for poultry worldwide. Alternative grains such as wheat and barley can be successfully replaced for corn in poultry diets. These cereal grains can locally grow in many parts of the world as they have remarkably lower water requirement than corn. Wheat and barley are generally used as major sources of energy in poultry diets. Though the major components of these grains are starch and proteins, they have considerable content of non-starch polysaccharides (NSPs, derived from the cell walls (Olukosi et al. 2007; Mirzaie et al. 2012. NSPs are generally considered as anti-nutritional factors (Jamroz et al. 2002. The content and structure of NSP polymers vary between different grains, which consequently affect their nutritive value (Olukosi et al. 2007.Wheat and barley are generally used as major sources of energy in poultry diets. The major components of these grains are starch and proteins, they have considerable content of non-starch polysaccharides (NSPs, derived from the cell walls. NSPs are generally considered as anti-nutritional factors. The content and structure of NSP polymers vary between different grains, which consequently affect their nutritive value. The major part of NSPs in barley comprises polymers of (1→3 (1→4-β- glucans which could impede growth factors and consequently carcass quality through lowering the rate and amount of available nutrients in the mucosal surface of the intestinal. Materials and Methods This experiment was conducted to evaluate the effect of corn and barley based diets supplemented with multi-enzyme on growth, carcass, pancreas enzyme activity and physiological characteristics of broilers. A total number of 375 one day old

  20. Transcriptional Responses to Gibberellin and Abscisic Acid in Barley Aleurone

    Institute of Scientific and Technical Information of China (English)

    Kegui Chen; Yong-Qiang Charles An

    2006-01-01

    Cereal aleurone has been established as a model system to investigate giberrellin (GA) and abscisic acid (ABA) responses. Using Barley 1 GeneChip, we examined the mRNA accumulation of over 22 000 genes in de-embryonated barley aleurone treated with GA and ABA. We observed that 1328 genes had more than a threefold change in response to GA treatment, whereas 206 genes had a more than threefold change in response to ABA treatment. Interestingly, approximately 2.5-fold more genes were up-regulated than downregulated by ABA. Eighty-three genes were differentially regulated by both GA and ABA. Most of the genes were subject to antagonistic regulation by ABA and GA, particularly for genes related to seed maturation and germination, such as genes encoding late embryogenesis abundant proteins and storage mobilization enzymes. This supports the antagonistic roles of GA and ABA in seed maturation and seed germination.Interestingly, we observed that a significant percentage of the genes were coordinately regulated by both GA and ABA. Some GA-responsive genes encoded proteins involved in ethylene, jasmonate, brassinosteroid and auxin metabolic and signaling transduction pathways, suggesting their potential interaction with the GA response. We also identified a group of transcription factor genes, such as MYB and Homeobox genes, that were differentially regulated by GA. In addition, a number of GA- and/or ABA-responsive genes encoded components potentially involved in GA and ABA signal transduction pathway. Overall, the present study provides a comprehensive and global view of transcript expression accompanying the GA and ABA response in barley aleurone and identifies a group of genes with potential regulatory functions in GA- and ABA-signaling pathways for future functional validation.

  1. 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.).

    Science.gov (United States)

    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.

  2. Fermentation of barley by using Saccharomyces cerevisiae: examination of barley as a feedstock for bioethanol production and value-added products.

    Science.gov (United States)

    Gibreel, Amera; Sandercock, James R; Lan, Jingui; Goonewardene, Laksiri A; Zijlstra, Ruurd T; Curtis, Jonathan M; Bressler, David C

    2009-03-01

    The objective of this study was to examine the ethanol yield potential of three barley varieties (Xena, Bold, and Fibar) in comparison to two benchmarks, corn and wheat. Very high gravity (VHG; 30% solids) fermentations using both conventional and Stargen 001 enzymes for starch hydrolysis were carried out as simultaneous saccharification and fermentation. The grains and their corresponding dried distiller's grain with solubles (DDGS) were also analyzed for nutritional and value-added characteristics. A VHG traditional fermentation approach utilizing jet-cooking fermentation revealed that both dehulled Bold and Xena barley produced ethanol concentrations higher than that produced by wheat (12.3, 12.2, and 11.9%, respectively) but lower than that produced by corn (13.8%). VHG-modified Stargen-based fermentation of dehulled Bold barley demonstrated comparable performance (14.3% ethanol) relative to that of corn (14.5%) and wheat (13.3%). Several important components were found to survive fermentation and were concentrated in DDGS. The highest yield of phenolics was detected in the DDGS (modified Stargen 001, 20% solids) of Xena (14.6 mg of gallic acid/g) and Bold (15.0 mg of gallic acid/g) when the hull was not removed before fermentation. The highest concentration of sterols in DDGS from barley was found in Xena (3.9 mg/g) when the hull was included. The DDGS recovered from corn had the highest concentration of fatty acids (72.6 and 77.5 mg/g). The DDGS recovered from VHG jet-cooking fermentations of Fibar, dehulled Bold, and corn demonstrated similar levels of tocopherols and tocotrienols. Corn DDGS was highest in crude fat but was lowest in crude protein and in vitro energy digestibility. Wheat DDGS was highest in crude protein content, similar to previous studies. The barley DDGS was the highest in in vitro energy digestibility.

  3. Response of barley aleurone layers to abscisic acid

    Energy Technology Data Exchange (ETDEWEB)

    Ho, D.T.H.; Varner, J.E.

    1976-02-01

    Cordycepin, an inhibitor of RNA synthesis in barley (Hordeum vulgare L.) aleurone cells, does not inhibit the gibberellic acid-enhanced ..cap alpha..-amylase (EC 3.2.1.1.) synthesis in barley aleurone layers if it is added 12 hours or more after the addition of the hormone. However, the accumulation of ..cap alpha..-amylase activity after 12 hours of gibberellic acid can be decreased by abscisic acid. The accumulation of ..cap alpha..-amylase activity is sustained or quickly restored when cordycepin is added simultaneously or some time after abscisic acid, indicating that the response of aleurone layers to abscisic acid depends on the continuous synthesis of a short lived RNA. By analysis of the newly synthesized proteins by gel electrophoresis with sodium dodecylsulfate, it was observed that the synthesis of ..cap alpha..-amylase is decreased in the presence of abscisic acid while the synthesis of most of the other proteins remains unchanged. From the rate of resumption of ..cap alpha..-amylase production in the presence of cordycepin and abscisic acid, it appears that abscisic acid does not have a measurable effect on the stability of ..cap alpha..-amylase mRNA.

  4. Triple Hybridization with Cultivated Barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Bothmer, R. von; Claesson, L.; Flink, J.;

    1989-01-01

    represented species closely or distantly related to H. jubatum and H. lechleri. In trispecific crosses with diploid barley, the seed set was 5.7%. Crosses with tetraploid barley were highly unsuccessful (0.2% seed set). Three lines of diploid barley were used in the crosses, i.e. 'Gull', 'Golden Promise...

  5. phiC31 integrase-mediated site-specific recombination in barley.

    Directory of Open Access Journals (Sweden)

    Eszter Kapusi

    Full Text Available The Streptomyces phage phiC31 integrase was tested for its feasibility in excising transgenes from the barley genome through site-specific recombination. We produced transgenic barley plants expressing an active phiC31 integrase and crossed them with transgenic barley plants carrying a target locus for recombination. The target sequence involves a reporter gene encoding green fluorescent protein (GFP, which is flanked by the attB and attP recognition sites for the phiC31 integrase. This sequence disruptively separates a gusA coding sequence from an upstream rice actin promoter. We succeeded in producing site-specific recombination events in the hybrid progeny of 11 independent barley plants carrying the above target sequence after crossing with plants carrying a phiC31 expression cassette. Some of the hybrids displayed fully executed recombination. Excision of the GFP gene fostered activation of the gusA gene, as visualized in tissue of hybrid plants by histochemical staining. The recombinant loci were detected in progeny of selfed F(1, even in individuals lacking the phiC31 transgene, which provides evidence of stability and generative transmission of the recombination events. In several plants that displayed incomplete recombination, extrachromosomal excision circles were identified. Besides the technical advance achieved in this study, the generated phiC31 integrase-expressing barley plants provide foundational stock material for use in future approaches to barley genetic improvement, such as the production of marker-free transgenic plants or switching transgene activity.

  6. Identification of a novel gene (Hsdr4) involved in water-stress tolerance in wild barley.

    Science.gov (United States)

    Suprunova, Tatiana; Krugman, Tamar; Distelfeld, Assaf; Fahima, Tzion; Nevo, Eviatar; Korol, Abraham

    2007-05-01

    Drought is one of the most severe stresses limiting plant growth and yield. Genes involved in water stress tolerance of wild barley (Hordeum spontaneoum), the progenitor of cultivated barley, were investigated using genotypes contrasting in their response to water stress. Gene expression profiles of water-stress tolerant vs. water-stress sensitive wild barley genotypes, under severe dehydration stress applied at the seedling stage, were compared using cDNA-AFLP analysis. Of the 1100 transcript-derived fragments (TDFs) amplified about 70 displayed differential expression between control and stress conditions. Eleven of them showed clear difference (up- or down-regulation) between tolerant and susceptible genotypes. These TDFs were isolated, sequenced and tested by RT-PCR. The differential expression of seven TDFs was confirmed by RT-PCR, and TDF-4 was selected as a promising candidate gene for water-stress tolerance. The corresponding gene, designated Hsdr4 (Hordeum spontaneum dehydration-responsive), was sequenced and the transcribed and flanking regions were determined. The deduced amino acid sequence has similarity to the rice Rho-GTPase-activating protein-like with a Sec14 p-like lipid-binding domain. Analysis of Hsdr4 promoter region that was isolated by screening a barley BAC library, revealed a new putative miniature inverted-repeat transposable element (MITE), and several potential stress-related binding sites for transcription factors (MYC, MYB, LTRE, and GT-1), suggesting a role of the Hsdr4 gene in plant tolerance to dehydration stress. Furthermore, the Hsdr4 gene was mapped using wild barley mapping population to the long arm of chromosome 3H between markers EBmac541 and EBmag705, within a region that previously was shown to affect osmotic adaptation in barley.

  7. Silorane resin supports proliferation, differentiation, and mineralization of MLO-A5 bone cells in vitro and bone formation in vivo

    OpenAIRE

    Eick, J. David; Barragan-Adjemian, Cielo; Rosser, Jennifer; Melander, Jennifer R.; Dusevich, Vladimir; Weiler, Rachel A.; Miller, Bradley D.; Kilway, Kathleen V.; Dallas, Mark R.; Bi, Lianxing; Nalvarte, Elisabet L.; Bonewald, Lynda F.

    2012-01-01

    Methyl methacrylate used in bone cements has drawbacks of toxicity, high exotherm, and considerable shrinkage. A new resin, based on silorane/oxirane chemistry, has been shown to have little toxicity, low exotherm, and low shrinkage. We hypothesized that silorane-based resins may also be useful as components of bone cements as well as other bone applications and began testing on bone cell function in vitro and in vivo. MLO-A5, late osteoblast cells, were exposed to polymerized silorane (SilMi...

  8. A proteomics-based study of endogenous and microbial xylanases and xylanase inhibitors associated with barley grains used for liquid feed

    DEFF Research Database (Denmark)

    Sultan, Abida

    The mature barley grain contains a complement of enzymes that are synthesized during seed development for degradation of seed storage reserves during germination. These enzyme activities (first wave enzymes) are considered important for maximizing nutrient digestibility in food and feed. Several...... of these enzyme activities between barley cultivars, as well as the distribution and composition of the residing commensal fungal community and the grain surface associated proteins. To obtain new insight into the interplay between barley grains and the colonizing fungi, we set out to study the activities...

  9. Nitrogen deficiency in barley (Hordeum vulgare) seedlings induces molecular and metabolic adjustments that trigger aphid resistance.

    Science.gov (United States)

    Comadira, Gloria; Rasool, Brwa; Karpinska, Barbara; Morris, Jenny; Verrall, Susan R; Hedley, Peter E; Foyer, Christine H; Hancock, Robert D

    2015-06-01

    Agricultural nitrous oxide (N2O) pollution resulting from the use of synthetic fertilizers represents a significant contribution to anthropogenic greenhouse gas emissions, providing a rationale for reduced use of nitrogen (N) fertilizers. Nitrogen limitation results in extensive systems rebalancing that remodels metabolism and defence processes. To analyse the regulation underpinning these responses, barley (Horedeum vulgare) seedlings were grown for 7 d under N-deficient conditions until net photosynthesis was 50% lower than in N-replete controls. Although shoot growth was decreased there was no evidence for the induction of oxidative stress despite lower total concentrations of N-containing antioxidants. Nitrogen-deficient barley leaves were rich in amino acids, sugars and tricarboxylic acid cycle intermediates. In contrast to N-replete leaves one-day-old nymphs of the green peach aphid (Myzus persicae) failed to reach adulthood when transferred to N-deficient barley leaves. Transcripts encoding cell, sugar and nutrient signalling, protein degradation and secondary metabolism were over-represented in N-deficient leaves while those associated with hormone metabolism were similar under both nutrient regimes with the exception of mRNAs encoding proteins involved in auxin metabolism and responses. Significant similarities were observed between the N-limited barley leaf transcriptome and that of aphid-infested Arabidopsis leaves. These findings not only highlight significant similarities between biotic and abiotic stress signalling cascades but also identify potential targets for increasing aphid resistance with implications for the development of sustainable agriculture.

  10. Immigration of the barley mildew pathogen into field plots of barley

    DEFF Research Database (Denmark)

    O'Hara, R.B.; Brown, J.K.M.

    1996-01-01

    Immigration of the barley powdery mildew pathogen (Erysiphe graminis f.sp. hordei) into field plots of the spring barley variety Tyra (carrying the resistance allele Mla1) was investigated. Spores were trapped from the top of the plot canopies, as well as from control plots of wheat with no barley...... nearby. Comparison of the frequencies of virulent and avirulent single-colony isolates showed that the amount of immigration, relative to the amount of inoculum being produced within the plot, reduced very rapidly, until it could not be detected in the middle of the growing season (mid-June)....

  11. Response of barley seedlings to oxidative stress generated by treatments with growth hormones

    OpenAIRE

    Zenovia Olteanu; Elena Truta; Lacramiora Oprica; Maria Magdalena Zamfirache

    2009-01-01

    The effects induced by growth hormone regulators on soluble protein level and some oxidoreductases in Hordeum vulgare cv. Madalin seedlings were investigated. The study of superoxide dismutase, catalyse and peroxidase behaviour and of protein synthesis was realized in dynamics to evaluate the response of barley seedlings to oxidative stress generated by exposure to hormone factors. During experiments, peroxidase registered smaller limits of variability than superoxide dismutase an...

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

    Science.gov (United States)

    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 ), (/r/ in ), respectively, as detected by both methods.

  13. Mildew-resistant mutants induced in North American two- and six-rowed malting barley cultivars

    DEFF Research Database (Denmark)

    Molina-Cano, J.L.; Simiand, J.P.; Sopena, A.;

    2003-01-01

    and were shown to have two new alleles at the mlo locus; these were designated, respectively, mlo31 and mlo32. Mutant URS2, showing partial resistance, was inherited as a dominant gene, but was not an allele at the Mla locus. The mean yield of each mutant was higher than that of its parental line......, but both URS1 and URS2 showed lower malt extract than Ursula. This lower extract might be due to the smaller grain size of the mutants that could, in turn, result from necrotic lesions in the leaves, as implied by the effects on grain yield....

  14. Proteome analysis of grain filling and seed maturation in barley.

    Science.gov (United States)

    Finnie, Christine; Melchior, Sabrina; Roepstorff, Peter; Svensson, Birte

    2002-07-01

    In monocotyledonous plants, the process of seed development involves the deposition of reserves in the starchy endosperm and development of the embryo and aleurone layer. The final stages of seed development are accompanied by an increase in desiccation tolerance and drying out of the mature seed. We have used two-dimensional gel electrophoresis for a time-resolved study of the changes in proteins that occur during seed development in barley (Hordeum vulgare). About 1,000 low-salt extractable protein spots could be resolved on the two-dimensional gels. Protein spots were divided into six categories according to the timing of appearance or disappearance during the 5-week period of comparison. Nineteen different proteins or protein fragments in 36 selected spots were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry (MS) or nano-electrospray tandem MS/MS. Some proteins were present throughout development (for example, cytosolic malate dehydrogenase), whereas others were associated with the early grain filling (ascorbate peroxidase) or desiccation (Cor14b) stages. Most noticeably, the development process is characterized by an accumulation of low-M(r) alpha-amylase/trypsin inhibitors, serine protease inhibitors, and enzymes involved in protection against oxidative stress. We present examples of proteins not previously experimentally observed, differential extractability of thiol-bound proteins, and possible allele-specific spot variation. Our results both confirm and expand on knowledge gained from previous analyses of individual proteins involved in grain filling and maturation.

  15. Chemical characterization of Brazilian hulless barley varieties, flour fractionation, and protein concentration Caracterização química de variedades brasileiras de cevada nua, fracionamento da farinha e concentração de proteína

    Directory of Open Access Journals (Sweden)

    Cristiane Vieira Helm

    2004-12-01

    Full Text Available Even though hulless barley is widely known due to its nutritional potential, in Brazil it is only grown at a few agricultural experimental stations. There is no published data about the chemical composition of Brazilian hulless barley varieties; however, research laboratories have studied their agronomical characteristics. The objectives of this study were to present the chemical characterization and effect of flour fractionation on protein concentration of six Brazilian hulless barley varieties, namely IAC IBON 214/82, IAC 8612/421, IAC 8501/31, IAC 8501/12, IAPAR 39-Acumaí, and IAC 8501/22. The analyses included: ash, ether extract, total protein, starch, total insoluble and soluble dietary fiber, and beta-glucans. Flour fractionation was carried out by sieving. The flour fractions were evaluated for crude protein, protein, and protein and non-protein nitrogen. Chemical composition varied (P Apesar de a cevada nua ser amplamente conhecida por seu potencial nutricional, no Brasil é apenas cultivada em poucas estações experimentais agronômicas. Em relação as variedades brasileiras de cevada nua, não se têm dados sobre a composição química, entretanto instituições de pesquisa têm estudado suas características agronômicas. Os objetivos deste estudo foram apresentar a caracterização química e o efeito no fracionamento da farinha obtida visando à concentração de proteína de seis variedades brasileiras de cevada nua: IAC IBON 214/82, IAC 8612/421, IAC 8501/31, IAC 8501/12, IAPAR 39-Acumaí, e IAC 8501/22. Foram realizadas análises de: cinzas, extrato etéreo, proteína total, amido, fibra alimentar total, solúvel e insolúvel e beta-glucanas. As frações das farinhas foram obtidas por peneiramento e avaliadas quanto aos teores de proteína bruta, proteína, nitrogênio protéico e não protéico. Houve variação entre as variedades testadas (P < 0,05. As variedades IAC 8501/22, IAC 8501/31 e IAC 8501/12 apresentaram o

  16. Effects of high glucose and advanced glycation end products on the expressions of sclerostin and RANKL as well as apoptosis in osteocyte-like MLO-Y4-A2 cells

    Energy Technology Data Exchange (ETDEWEB)

    Tanaka, Ken-ichiro, E-mail: ken1nai@med.shimane-u.ac.jp; Yamaguchi, Toru, E-mail: yamaguch@med.shimane-u.ac.jp; Kanazawa, Ippei, E-mail: ippei.k@med.shimane-u.ac.jp; Sugimoto, Toshitsugu, E-mail: sugimoto@med.shimane-u.ac.jp

    2015-05-29

    In diabetes mellitus (DM), high glucose (HG) and advanced glycation end products (AGEs) are involved in bone quality deterioration. Osteocytes produce sclerostin and receptor activator of nuclear factor-kB ligand (RANKL) and regulate osteoblast and osteoclast function. However, whether HG or AGEs directly affect osteocytes and regulate sclerostin and RANKL production is unknown. Here, we examined the effects of HG, AGE2, and AGE3 on the expression of sclerostin and RANKL and on apoptosis in osteocyte-like MLO-Y4-A2 cells. Treatment of the cells with 22 mM glucose, 100 μg/mL either AGE2 or AGE3 significantly increased the expression of sclerostin protein and mRNA; however, both AGEs, but not glucose, significantly decreased the expression of RANKL protein and mRNA. Moreover, treatment of the cells with HG, AGE2, or AGE3 for 72 h induced significant apoptosis. These detrimental effects of HG, AGE2, and AGE3 on sclerostin and RANKL expressions and on apoptosis were antagonized by pretreatment of the cells with 10{sup −8} M human parathyroid hormone (PTH)-(1–34). Thus, HG and AGEs likely suppress bone formation by increasing sclerostin expression in osteocytes, whereas AGEs suppress bone resorption by decreasing RANKL expression. Together, these processes may cause low bone turnover in DM. In addition, HG and AGEs may cause cortical bone deterioration by inducing osteocyte apoptosis. PTH may effectively treat these pathological processes and improve osteocyte function. - Highlights: • AGEs are involved in bone quality deterioration in diabetes mellitus (DM). • AGEs increased sclerostin as well as apoptosis, and decreased RANKL in osteocytes. • The effects of AGEs on osteocyte function were antagonized by human PTH-(1–34). • AGEs may cause low bone turnover and cortical porosity in DM. • PTH may be effective in bone quality deterioration by improving osteocyte function.

  17. Abscisic acid effects on activity and expression of barley (Hordeum vulgare) plastidial glucose-6-phosphate dehydrogenase.

    Science.gov (United States)

    Cardi, Manuela; Chibani, Kamel; Cafasso, Donata; Rouhier, Nicolas; Jacquot, Jean-Pierre; Esposito, Sergio

    2011-07-01

    Total glucose-6-phosphate dehydrogenase (G6PDH) activity, protein abundance, and transcript levels of G6PDH isoforms were measured in response to exogenous abscisic acid (ABA) supply to barley (Hordeum vulgare cv Nure) hydroponic culture. Total G6PDH activity increased by 50% in roots treated for 12 h with exogenous 0.1 mM ABA. In roots, a considerable increase (35%) in plastidial P2-G6PDH transcript levels was observed during the first 3 h of ABA treatment. Similar protein variations were observed in immunoblotting analyses. In leaves, a 2-fold increase in total G6PDH activity was observed after ABA treatment, probably related to an increase in the mRNA level (increased by 50%) and amount of protein (increased by 85%) of P2-G6PDH. Together these results suggest that the plastidial P2-isoform plays an important role in ABA-treated barley plants.

  18. Barley β-glucans extraction and partial characterization.

    Science.gov (United States)

    Limberger-Bayer, Valéria M; de Francisco, Alicia; Chan, Aline; Oro, Tatiana; Ogliari, Paulo J; Barreto, Pedro L M

    2014-07-01

    Barley is rarely used in the food industry, even though it is a main source of β-glucans, which have important health benefits and a technological role in food. This work evaluated the humid extraction of barley β-glucans and partially characterized them. The extraction was studied using surface response methodology with both temperature and pH as variables. The extracted β-glucans were characterized by chemical and rheological analysis, infrared spectroscopy and scanning electron microscopy. The effect on extraction of linear and quadratic terms of pH and temperature corresponding to the regression model was significant, and we obtained a maximum concentration of 53.4% at pH 7.56 and temperature 45.5°C, with protein and mainly starch contamination. The extracted β-glucans presented a higher apparent viscosity than the commercial ones, the behavior of the commercial and extracted samples can be described as Newtonian and pseudoplastic, respectively. The results of infrared spectroscopy and scanning electron microscopy were characteristic of commercial β-glucans, indicating that this method is efficient for extracting β-glucans.

  19. Cell layer-specific distribution of transiently expressed barley ESCRT-III component HvVPS60 in developing barley endosperm.

    Science.gov (United States)

    Hilscher, Julia; Kapusi, Eszter; Stoger, Eva; Ibl, Verena

    2016-01-01

    The significance of the endosomal sorting complexes required for transport (ESCRT)-III in cereal endosperm has been shown by the identification of the recessive mutant supernumerary aleurone layer1 (SAL1) in maize. ESCRT-III is indispensable in the final membrane fission step during biogenesis of multivesicular bodies (MVBs), responsible for protein sorting to vacuoles and to the cell surface. Here, we annotated barley ESCRT-III members in the (model) crop Hordeum vulgare and show that all identified members are expressed in developing barley endosperm. We used fluorescently tagged core ESCRT-III members HvSNF7a/CHMP4 and HvVPS24/CHMP3 and the associated ESCRT-III component HvVPS60a/CHMP5 for transient localization studies in barley endosperm. In vivo confocal microscopic analyses show that the localization of recombinantly expressed HvSNF7a, HvVPS24 and HvVPS60a differs within barley endosperm. Whereas HvSNF7a induces large agglomerations, HvVPS24 shows mainly cytosolic localization in aleurone and subaleurone. In contrast, HvVPS60a localizes strongly at the plasma membrane in aleurone. In subaleurone, HvVPS60a was found to a lesser extent at the plasma membrane and at vacuolar membranes. These results indicate that the steady-state association of ESCRT-III may be influenced by cell layer-specific protein deposition or trafficking and remodelling of the endomembrane system in endosperm. We show that sorting of an artificially mono-ubiquitinated Arabidopsis plasma membrane protein is inhibited by HvVPS60a in aleurone. The involvement of HvVPS60a in different cell layer-specific trafficking pathways, reflected by localization of HvVPS60a at the plasma membrane in aleurone and at the PSV membrane in subaleurone, is discussed.

  20. Chromosomal loci associated with endosperm hardness in a malting barley cross.

    Science.gov (United States)

    Walker, Cassandra K; Panozzo, J F; Ford, R; Eckermann, P; Moody, D; Lehmensiek, A; Appels, R

    2011-01-01

    A breeding objective for the malting barley industry is to produce lines with softer, plumper grain containing moderate protein content (9-12%) as they are more likely to imbibe water readily and contain more starch per grain, which in turn produces higher levels of malt extract. In a malting barley mapping population, 'Arapiles' × 'Franklin', the most significant and robust quantitative trait locus (QTL) for endosperm hardness was observed on the short arm of chromosome 1H, across three environments over two growing seasons. This accounted for 22.6% (Horsham 2000), 26.8% (Esperance 2001), and 12.0% (Tarranyurk 2001) of the genetic variance and significantly increased endosperm hardness by 2.06-3.03 SKCS hardness units. Interestingly, Arapiles and Franklin do not vary in Ha locus alleles. Therefore, this region, near the centromere on chromosome 1H, may be of great importance when aiming to manipulate endosperm hardness and malting quality. Interestingly, this region, close to the centromere on chromosome 1H, in our study, aligns with the region of the genome that includes the HvCslF9 and the HvGlb1 genes. Potentially, one or both of these genes could be considered to be candidate genes that influence endosperm hardness in the barley grain. Additional QTLs for endosperm hardness were detected on chromosomes 2H, 3H, 6H and 7H, confirming that the hardness trait in barley is complex and multigenic, similar to many malting quality traits of interest.

  1. Structure and function of the bacterial root microbiota in wild and domesticated barley.

    Science.gov (United States)

    Bulgarelli, Davide; Garrido-Oter, Ruben; Münch, Philipp C; Weiman, Aaron; Dröge, Johannes; Pan, Yao; McHardy, Alice C; Schulze-Lefert, Paul

    2015-03-11

    The microbial communities inhabiting the root interior of healthy plants, as well as the rhizosphere, which consists of soil particles firmly attached to roots, engage in symbiotic associations with their host. To investigate the structural and functional diversification among these communities, we employed a combination of 16S rRNA gene profiling and shotgun metagenome analysis of the microbiota associated with wild and domesticated accessions of barley (Hordeum vulgare). Bacterial families Comamonadaceae, Flavobacteriaceae, and Rhizobiaceae dominate the barley root-enriched microbiota. Host genotype has a small, but significant, effect on the diversity of root-associated bacterial communities, possibly representing a footprint of barley domestication. Traits related to pathogenesis, secretion, phage interactions, and nutrient mobilization are enriched in the barley root-associated microbiota. Strikingly, protein families assigned to these same traits showed evidence of positive selection. Our results indicate that the combined action of microbe-microbe and host-microbe interactions drives microbiota differentiation at the root-soil interface.

  2. SNP Analysis and Haplotype Identification in Chymotrypsin Inhibitor-2 (CI-2) Gene of Barley

    Institute of Scientific and Technical Information of China (English)

    ZENG Xiang-hui; WEI Yu-ming; JIANG Qian-tao; QI Peng-fei; ZHENG You-liang

    2009-01-01

    Barley chymotrypsin inhibitor-2 (CI-2) was considered to be a promising candidate for enhancing the nutritional value of other cereals by increasing its concentration as it is rich in lysine than any other storage protein. Also, it was proposed that CI-2 might play an important role in the inhibition of proteolytic enzymes from pests or pathogens as CI-2 can strongly inhibit chymotrypsin and subtilisin. In this study, a total of 93 CI-2 gene sequences were isolated from wild and cultivated barley. 48 SNPs and 4 indels were detected across the entire sequences. The frequency of SNPs in the non-coding region (1 out of 9 bases) was slightly higher than that in the coding region (1 out of 10.7 bases). In all, 33.3% of the candidate cSNPs resulted in amino acid changes. As a total, the 24 cSNPs resulted in 15 amino acid changes. Ten distinguishable haplotypes were detected, among which 3 haplotypes were shared in the most barley accessions, whereas the rest of the haplotypes appeared at a lower frequency. In addition, three haplotypes (haplotype 4, 8, and 9) were unique for single accessions. These results suggested that low diversity at the CI-2 locus was detected among the cultivated and wild barley.

  3. Effects of Microbial Additives on Chemical Composition and Fermentation Characteristics of Barley Silage

    Science.gov (United States)

    Amanullah, S. M.; Kim, D. H.; Lee, H. J.; Joo, Y. H.; Kim, S. B.; Kim, S. C.

    2014-01-01

    This study examined the effects of bacterial inoculants on chemical composition and fermentation indices of barley silage. Barley forage (Youngyang) was harvested at 24% dry matter (DM) and wilted to 47.9% DM. The wilted barley forage was chopped to 3–5 cm length and applied with no inoculant (CON), L. plantarum (1×1010 cfu/g, LP) or Effective Microorganisms (0.5×109 cfu/g, EM). Then the forages were ensiled in four replications for each treatment in 20 L mini silos and stored for 100 days. The contents of crude protein and ether extract were higher in CON silage ensiled for 100-d, while the contents of DM and crude ash were higher in EM silage (p0.05). The pH, ammonia-N concentration and lactate to acetate ratio were higher (p<0.05) in CON silage, while lactate concentrations were higher (p<0.05) in CON and LP silage. Acetate concentration and lactic acid bacteria was increased (p<0.05) by both inoculants (LP and EM), but propionate concentration and yeast was increased (p<0.05) by EM and LP, respectively. These results indicated that the fermentation quality of barley silage was improved by the application of bacterial inoculants. PMID:25049981

  4. Adaptation of barley to harsh Mediterranean environments.

    NARCIS (Netherlands)

    Oosterom, van E.

    1993-01-01

    Research ObjectivesBarley is in Syria the dominant crop in areas receiving less than 300 mm annual precipitation. Grain yield is often below 1 ton ha -1, and is reduced by low temperatures in winter and terminal drought stress in spring. Variation i

  5. THE STIMULATING EFFECT OF LASER RED LIGHT, FAR RED LIGHT AND SODIUM CARBONATE AT THE INITIAL STAGES OF BARLEY ONTOGENESIS

    Directory of Open Access Journals (Sweden)

    G. P. Dudin

    2014-01-01

    Full Text Available Summary. Modern ecological state of the environment and human unhealthy diet cause many diseases. A healthy diet is the one that contains adequate amounts of proteins, fats, carbohydrates, vitamins, macronutrients and micronutrients. Photosynthesis i. e. the process by which plants produce organic compounds from carbon dioxide and water, is the source of life, the source of evolution and proliferation of life forms on the Earth. Thus, the juice made from sprouted barley provides physiologically active chlorophyll, macronutrients and micronutrients, vitamins А, В2 , В3 , В5 , В6 , В8 , Е and К. It is well known that light from a red laser with a wavelength of 638.2 nm has a stimulating action on the germination energy, germination ability and productivity of seeds, and on the crop yields. Therefore, this research is of primary importance today. The research result produced a sharp decline in plant vigor and germinating capacity of barley when soaking in 1n sodium carbonate solution, as well as changes in the ratio of potassium-sodium balance in plants. Thus at lower concentrations of sodium carbonate and 0.1 n sodium increasing of pigment content in barley is observed on the seventh day. The red laser light has a similar stimulating action: the chlorophyll content of barley plants increased after the red laser treatment of barley seeds. However, the chlorophyll contents were depressed when the seeds were exposed to far red light with wavelengths of 754±10 nm. Using these factors, one can manage the content of chlorophyll and sodium-potassium balance in the initial stages of barley ontogenesis in the technology of barley juice or the powder for a healthy and proper human diet.

  6. Immunological characterization of the gluten fractions and their hydrolysates from wheat, rye and barley.

    Science.gov (United States)

    Rallabhandi, Prasad; Sharma, Girdhari M; Pereira, Marion; Williams, Kristina M

    2015-02-18

    Gluten proteins in wheat, rye and barley cause celiac disease, an autoimmune disorder of the small intestine, which affects approximately 1% of the world population. Gluten is comprised of prolamin and glutelin. Since avoidance of dietary gluten is the only option for celiac patients, a sensitive gluten detection and quantitation method is warranted. Most regulatory agencies have set a threshold of 20 ppm gluten in foods labeled gluten-free, based on the currently available ELISA methods. However, these methods may exhibit differences in gluten quantitation from different gluten-containing grains. In this study, prolamin and glutelin fractions were isolated from wheat, rye, barley, oats and corn. Intact and pepsin-trypsin (PT)-digested prolamin and glutelin fractions were used to assess their immunoreactivity and gluten recovery by three sandwich and two competitive ELISA kits. The Western blots revealed varied affinity of ELISA antibodies to gluten-containing grain proteins and no reactivity to oat and corn proteins. ELISA results showed considerable variation in gluten recoveries from both intact and PT-digested gluten fractions among different kits. Prolamin fractions showed higher gluten recovery compared to their respective glutelin fractions. Among prolamins, barley exhibited higher recovery compared to wheat and rye with most of the ELISA kits used. Hydrolysis resulted in reduced gluten recovery of most gluten fractions. These results suggest that the suitability of ELISA for accurate gluten quantitation is dependent upon various factors, such as grain source, antibody specificity, gluten proteins and the level of their hydrolysis in foods.

  7. [SGP polymorphism in cultivated naked barley from Qinghai-Tibet plateau in China and the relationship between SGPs and starch content].

    Science.gov (United States)

    Pan, Zhi-Fen; Zhou, Yi-Xing; Zhao, Tao; Deng, Guang-Bing; Zhai, Xu-Guang; Wu, Fang; Yu, Mao-Qun

    2007-05-01

    Starch granule proteins (SGPs) are minor components bound with starch granule, which mutation may be related to starch properties. This study investigated the variation of SGPs in cultivated naked barley from Qinghai-Tibet Plateau in China for the first time, and the relationship between SGPs and starch content was preliminarily done. Ten major SGPs and 16 types of patterns were present in 66 cultivated naked varieties, indicating SGPs in cultivated naked barley from Qinghai-Tibet Plateau in China are polymorphic. SGPs in Tibet and Sichuan naked barley were greatly different and SGPs were specific to origin of site. Significance test analysis demonstrates SGPs described in this study except for SGP1 may be related with the variation of starch content in different naked barley.

  8. Mapping of the Hor-3 locus encoding D hordein in Barley.

    Science.gov (United States)

    Blake, T K; Ullrich, S E; Nilan, R A

    1982-12-01

    The hordein storage proteins of barley (Hordeum vulgare L.) are of intense interest due to their genetic diversity and prominence and impact on the industrial and agricultural uses of the seed. Two major hordein loci have been previously mapped on chromosome 5 (Hor-1 and Hor-2 encoding the C and B hordeins, respectively). A third major locus, Hor-3, which codes for D hordein, has been located in the centromeric region of chromosome 5, probably on the long arm. Two allelic variants with apparent molecular weights of 83,000 and 91,000 and similar isoelectric points of 8.0 comprise the products of this locus in the barley varieties 'Advance' and 'Triple Awned Lemma'. The D hordein examined is similar in molecular weight and isoelectric point to the high molecular weight (HMW) glutenin proteins encoded by the 1B chromosome of wheat (Triticum aestivum L.).

  9. Single nucleotide polymorphisms in HSP17.8 and their association with agronomic traits in barley.

    Directory of Open Access Journals (Sweden)

    Yanshi Xia

    Full Text Available Small heat shock protein 17.8 (HSP17.8 is produced abundantly in plant cells under heat and other stress conditions and may play an important role in plant tolerance to stress environments. However, HSP17.8 may be differentially expressed in different accessions of a crop species exposed to identical stress conditions. The ability of different genotypes to adapt to various stress conditions resides in their genetic diversity. Allelic variations are the most common forms of genetic variation in natural populations. In this study, single nucleotide polymorphisms (SNPs of the HSP17.8 gene were investigated across 210 barley accessions collected from 30 countries using EcoTILLING technology. Eleven SNPs including 10 from the coding region of HSP17.8 were detected, which form nine distinguishable haplotypes in the barley collection. Among the 10 SNPs in the coding region, six are missense mutations and four are synonymous nucleotide changes. Five of the six missense changes are predicted to be deleterious to HSP17.8 function. The accessions from Middle East Asia showed the higher nucleotide diversity of HSP17.8 than those from other regions and wild barley (H. spontaneum accessions exhibited greater diversity than the cultivated barley (H. vulgare accessions. Four SNPs in HSP17.8 were found associated with at least one of the agronomic traits evaluated except for spike length, namely number of grains per spike, thousand kernel weight, plant height, flag leaf area and leaf color. The association between SNP and these agronomic traits may provide new insight for study of the gene's potential contribution to drought tolerance of barley.

  10. Single nucleotide polymorphisms in HSP17.8 and their association with agronomic traits in barley.

    Science.gov (United States)

    Xia, Yanshi; Li, Ronghua; Ning, Zhengxiang; Bai, Guihua; Siddique, Kadambot H M; Yan, Guijun; Baum, Michael; Varshney, Rajeev K; Guo, Peiguo

    2013-01-01

    Small heat shock protein 17.8 (HSP17.8) is produced abundantly in plant cells under heat and other stress conditions and may play an important role in plant tolerance to stress environments. However, HSP17.8 may be differentially expressed in different accessions of a crop species exposed to identical stress conditions. The ability of different genotypes to adapt to various stress conditions resides in their genetic diversity. Allelic variations are the most common forms of genetic variation in natural populations. In this study, single nucleotide polymorphisms (SNPs) of the HSP17.8 gene were investigated across 210 barley accessions collected from 30 countries using EcoTILLING technology. Eleven SNPs including 10 from the coding region of HSP17.8 were detected, which form nine distinguishable haplotypes in the barley collection. Among the 10 SNPs in the coding region, six are missense mutations and four are synonymous nucleotide changes. Five of the six missense changes are predicted to be deleterious to HSP17.8 function. The accessions from Middle East Asia showed the higher nucleotide diversity of HSP17.8 than those from other regions and wild barley (H. spontaneum) accessions exhibited greater diversity than the cultivated barley (H. vulgare) accessions. Four SNPs in HSP17.8 were found associated with at least one of the agronomic traits evaluated except for spike length, namely number of grains per spike, thousand kernel weight, plant height, flag leaf area and leaf color. The association between SNP and these agronomic traits may provide new insight for study of the gene's potential contribution to drought tolerance of barley.

  11. Digestibility of whole-crop barley and oat silages in dairy heifers.

    Science.gov (United States)

    Wallsten, J; Bertilsson, J; Nadeau, E; Martinsson, K

    2010-03-01

    This study evaluated the digestibility of whole-crop cereal silage (WCCS) made from oats and six-rowed barley harvested at the heading, early milk and early dough stages, and two-rowed barley harvested at the early milk and early dough stages of maturity. The eight WCCSs were fed to 32 Swedish Red heifers in a changeover design over three periods of 28 days each. The heifers were first fed ad libitum for 17 days and then at 0.95 of ad libitum for 11 days of each period. During the last 5 days all faeces and orts were collected to determine the digestibility of the silages. Only the maturity stage effect was significant for the WCCS organic matter (OM) digestibility and the average OM digestibility was higher at the heading stage (698 g/kg) than at early milk (647 g/kg) and early dough (652 g/kg) stages of maturity. For neutral detergent fibre (NDF) digestibility the crop × maturity stage effect was significant. The NDF digestibility decreased from the heading to the early milk stage for both six-rowed barley (746 to 607 g/kg) and oats (698 to 596 g/kg). There was no further significant decrease in NDF digestibility for six-rowed barley at the early dough stage (577 g/kg), but for two-rowed barley it decreased from the early milk (682 g/kg) to the early dough (573 g/kg) stage, and also for oats the NDF digestibility was lowest at the early dough stage (507 g/kg). The decrease in NDF digestibility during maturation was to a large extent compensated by an increase in starch concentration in the crops. The starch digestibility was lower for six-rowed barley at early dough stage (948 g/kg) than at early milk stage (977 g/kg), and was also lower compared with oats (979 g/kg) at early dough stage. The average crude protein (CP) digestibility was higher at the heading (646 g/kg) and the early milk (642 g/kg) stages than at the early dough stage (599 g/kg), and oats had higher average CP digestibility (650 g/kg) than six-rowed (613 g/kg) and two-rowed (624 g/kg) barley

  12. Efficient secretory expression of functional barley limit dextrinase inhibitor by high cell-density fermentation of Pichia pastoris

    DEFF Research Database (Denmark)

    Jensen, Johanne Mørch; Vester-Christensen, Malene Bech; Møller, Marie Sofie;

    2011-01-01

    The limit dextrinase inhibitor (LDI) from barley seeds acts specifically on limit dextrinase (LD), an endogenous starch debranching enzyme. LDI is a 14kDa hydrophobic protein containing four disulfide bonds and one unpaired thiol group previously found to be either glutathionylated or cysteinylat...

  13. High-level expression of the native barley alpha-amylase/subtilisin inhibitor in Pichia pastoris

    DEFF Research Database (Denmark)

    Micheelsen, Pernille Ollendorff; Ostergaard, Peter Rahbek; Lange, Lene;

    2008-01-01

    An expression system for high-level expression of the native Hordeum vulgare alpha-amylase/subtilisin inhibitor (BASI) has been developed in Pichia pastoris, using the methanol inducible alcohol oxidase 1 (AOX1) promoter. To optimize expression, two codon-optimized coding regions have been designed...... and characterized by Edman degradation, liquid chromatography mass spectrometry and insoluble blue starch assay, and was shown to possess the same characteristics as wild-type protein purified from barley grains....

  14. Effects of Different Storage Conditions on the Nutritional Qualities of Barley and Maize

    OpenAIRE

    POLAT, Havva Eylem

    2015-01-01

    The effects of different types of storage on nutritional quality values of barley and maize are examined. For this purpose, 3 different storage types, including horizontal-reinforced concrete storage, vertical-reinforced concrete silo and verticalsteel silo were taken into consideration. Nutritional quality values (dry matter, crude protein, crude fat, crude fiber, crude ash, and metabolizable energy) of samples were analyzed in the laboratory in four replicates. Total of 438 samples were col...

  15. Inhibition of coagulation factors by recombinant barley serpin BSZx

    DEFF Research Database (Denmark)

    Dahl, Søren Weis; Rasmussen, S.K.; Petersen, L..C.;

    1996-01-01

    Barley serpin BSZx is a potent inhibitor of trypsin and chymotrypsin at overlapping reactive sites (Dahl, S.W., Rasmussen, S.K. and Hejgaard, J. (1996) J. Biol, Chem., in press), We have now investigated the interactions of BSZx with a range of serine proteinases from human plasma, pancreas...... as substrate, Complexes of these proteinase with BSZx resisted boiling in SDS, and amino acid sequencing showed that cleavage in the reactive center loop only occurred after P-1 Arg. Activated protein C and leukocyte elastase were slowly inhibited by BSZx (k(ass) = 1-2 x 10(2) M(-1) s(-1)) whereas factor XIIa......, urokinase and tissue type plasminogen activator, plasmin and pancreas kallikrein and elastase were not or only weakly affected, The inhibition pattern with mammalian proteinases reveal a specificity of BSZx similar to that of antithrombin III. Trypsin from Fusarium was not inhibited while interaction...

  16. Barley polyamine oxidase: Characterisation and analysis of the cofactor and the N-terminal amino acid sequence

    DEFF Research Database (Denmark)

    Radova, A.; Sebela, M.; Galuszka, P.

    2001-01-01

    This paper reports the first purification method developed for the isolation of an homogeneous polyamine oxidase (PAO) from etiolated barley seedlings. The crude enzyme preparation was obtained after initial precipitation of the extract with protamine sulphate and ammonium sulphate. The enzyme...... was further purified to a final homogeneity (by the criteria of isoelectric focusing and SDS-PAGE) using techniques of low pressure chromatography followed by two FPLC steps. The purified yellow enzyme showed visible absorption maxima of a flavoprotein at 380 and 450 nm: the presence of FAD as the cofactor...... was further confirmed by measuring the fluorescence spectra, Barley PAO is an acidic protein (pI 5.4) containing 3% of neutral sugars: its molecular mass determined by SDS-PAGE was 56 kDa, whilst gel permeation chromatography revealed the higher value of 76 kDa. The N-terminal amino acid sequence of barley...

  17. THE DIFFERENCES BETWEEN THE OLD AND NEW BARLEY VARIETIES ON THE BASIS OF HORDEIN POLYMORPHISM WITH RESPECT TO QUALITATIVE PARAMETERS

    Directory of Open Access Journals (Sweden)

    Marián Tomka

    2015-02-01

    Full Text Available The current registered barley varieties are classified according to their biological and economic characters meeting the growing conditions. Varieties are characterized using molecular markers on the level of protein and DNA. Using starch gel electrophoresis we studied the frequency of hordein alleles, which are potential markers of economically important traits of 30 barley varieties. Among the 26 alleles detected in two main loci (Hrd A, Hrd B, we identified 7 potential markers (A3, A12, A23, B8, B17, B19, B47 with the frequency ranged from 1.18% to 12.14%. Number of these alleles differs over the years and together with number of lines per variety has decreasing tendency. On the basis of the detected alleles we are able to predict malting quality parameters and resistance to most common barley pathogens.

  18. In vivo allergenic activities of eleven purified members of a major allergen family from wheat and barley flour.

    Science.gov (United States)

    Armentia, A; Sanchez-Monge, R; Gomez, L; Barber, D; Salcedo, G

    1993-05-01

    Eleven purified members of the alpha-amylase/trypsin inhibitor family from wheat and barley that showed very different IgE-binding capacities when previously assayed in vitro, were used in double blind in vivo diagnostic tests to further evaluate their allergenic activity. These tests were carried out in 31 patients who showed allergic sensitization to wheat flour as verified by skin test, RAST and challenge test. The three members of the protein family with highest IgE binding in vitro (the glycosylated subunits of tetrameric alpha-amylase inhibitors CM16* from wheat and CMb* from barley, and the barley monomeric inhibitor BMAI-1) were found to be the strongest allergens as indicated by skin sensitivity in prick tests.

  19. Agrobacterium-mediated transformation of barley (Hordeum vulgare L.).

    Science.gov (United States)

    Ismagul, Ainur; Mazonka, Iryna; Callegari, Corinne; Eliby, Serik

    2014-01-01

    Barley biotechnology requires efficient genetic engineering tools for producing transgenic plants necessary for conducting reverse genetics analyses in breeding and functional genomics research. Agrobacterium-mediated genetic transformation is an important technique for producing barley transgenics with simple low-copy number transgenes. This chapter reports a refined protocol for the systematic high-throughput transformation of the advanced Australian spring barley breeding line WI4330.

  20. Barley Genes as Tools to Confer Abiotic Stress Tolerance in Crops.

    Science.gov (United States)

    Gürel, Filiz; Öztürk, Zahide N; Uçarlı, Cüneyt; Rosellini, Daniele

    2016-01-01

    Barley is one of the oldest cultivated crops in the world with a high adaptive capacity. The natural tolerance of barley to stress has led to increasing interest in identification of stress responsive genes through small/large-scale omics studies, comparative genomics, and overexpression of some of these genes by genetic transformation. Two major categories of proteins involved in stress tolerance are transcription factors (TFs) responsible from the re-programming of the metabolism in stress environment, and genes encoding Late Embryogenesis Abundant (LEA) proteins, antioxidant enzymes, osmolytes, and transporters. Constitutive overexpression of several barley TFs, such as C-repeat binding factors (HvCBF4), dehydration-responsive element-binding factors (HvDREB1), and WRKYs (HvWRKY38), in transgenic plants resulted in higher tolerance to drought and salinity, possibly by effectively altering the expression levels of stress tolerance genes due to their higher DNA binding affinity. Na(+)/H(+) antiporters, channel proteins, and lipid transporters can also be the strong candidates for engineering plants for tolerance to salinity and low temperatures.

  1. Barley genes as a tool to confer abiotic stress tolerance in crops

    Directory of Open Access Journals (Sweden)

    Filiz Gürel

    2016-08-01

    Full Text Available Barley is one of the oldest cultivated crops in the world with a high adaptive capacity. The natural tolerance of barley to stress has led to increasing interest in identification of stress responsive genes through small/large-scale omics studies, comparative genomics, and overexpression of some of these genes by genetic transformation. Two major categories of proteins involved in stress tolerance are transcription factors (TFs responsible from the re-programming of the metabolism in stress environment, and genes encoding Late Embryogenesis Abundant (LEA proteins, antioxidant enzymes, osmolytes and transporters. Constitutive overexpression of several barley TFs, such as C-repeat binding factors (HvCBF4, dehydration-responsive element-binding factors (HvDREB1 and WRKYs (HvWRKY38, in transgenic plants resulted in higher tolerance to drought and salinity, possibly by effectively altering the expression levels of stress tolerance genes due to their higher DNA binding affinity. Na+/H+ antiporters, channel proteins, and lipid transporters can also be the strong candidates for engineering plants for tolerance to salinity and low temperatures.

  2. A semidwarf phenotype of barley uzu results from a nucleotide substitution in the gene encoding a putative brassinosteroid receptor.

    Science.gov (United States)

    Chono, Makiko; Honda, Ichiro; Zeniya, Haruko; Yoneyama, Koichi; Saisho, Daisuke; Takeda, Kazuyoshi; Takatsuto, Suguru; Hoshino, Tsuguhiro; Watanabe, Yoshiaki

    2003-11-01

    Brassinosteroids (BRs) play important roles throughout plant growth and development. Despite the importance of clarifying the mechanism of BR-related growth regulation in cereal crops, BR-related cereal mutants have been identified only in rice (Oryza sativa). We previously found that semidwarf barley (Hordeum vulgare) accessions carrying the "uzu" gene, called "uzu" barley in Japan, are non-responding for brassinolide (BL). We then performed chemical and molecular analyses to clarify the mechanisms of uzu dwarfism using isogenic line pairs of uzu gene. The response of the uzu line to BL was significantly lower than that of its corresponding normal line. Measurement of BRs showed that the uzu line accumulates BRs, similar to known BR-insensitive mutants. The marker synteny of rice and barley chromosomes suggests that the uzu gene may be homologous to rice D61, a rice homolog of Arabidopsis BR-insensitive 1 (BRI1), encoding a BR-receptor protein. A barley homolog of BRI1, HvBRI1, was isolated by using degenerate primers. A comparison of HvBRI1 sequences in uzu and normal barley varieties showed that the uzu phenotype is correlated with a single nucleotide substitution. This substitution results in an amino acid change at a highly conserved residue in the kinase domain of the BR-receptor protein. These results may indicate that uzu dwarfism is caused by the missense mutation in HvBRI1. The uzu gene is being introduced into all hull-less barley cultivars in Japan as an effective dwarf gene for practical use, and this is the first report about an agronomically important mutation related to BRs.

  3. Analysis of Pregerminated Barley Using Hyperspectral Image Analysis

    DEFF Research Database (Denmark)

    Arngren, Morten; Hansen, Per Waaben; Eriksen, Birger

    2011-01-01

    Pregermination is one of many serious degradations to barley when used for malting. A pregerminated barley kernel can under certain conditions not regerminate and is reduced to animal feed of lower quality. Identifying pregermination at an early stage is therefore essential in order to segregate...... the barley kernels into low or high quality. Current standard methods to quantify pregerminated barley include visual approaches, e.g. to identify the root sprout, or using an embryo staining method, which use a time-consuming procedure. We present an approach using a near-infrared (NIR) hyperspectral...

  4. Changes of Limiting Dextrinase in Germinating Process of Malting Barley

    Institute of Scientific and Technical Information of China (English)

    LIANG Xiu-mei; LI Fen; WANG Hong-zhen; WANG Xing-zhi

    2002-01-01

    Based on five different species of barley, the foot layer analytic method was used to examine the activity and heat-resistance of the limiting dextrinase. The study was conducted on the dynamic changes of several types of the dextrinase in barley germinating process, the effect of temperature on the dextrinase and the divergence of dextrinase in different barley variety. The probability of the dextrinase that as reference index is used for screening and evaluating beer barley was discussed. The importance of dextrinase in brewing and its significant function was also discussed.

  5. Cisgenic Barley with Improved Phytase Activity

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2010-01-01

    based on the cisgenesis concept. In contrast to transgenesis, cisgenesis implies that the plant is transformed only with its own or very closely related genetic material. Furthermore, all “helper” genes and gene sequences of foreign nature are removed from the transformed plant lines. Cisgenic crops...... are accordingly very similar to those generated by conventional breeding. The cisgenesis concept allows for the introduction of extra gene copies of a particular gene to accentuate the trait. We are using a barley purple acid phosphatase expressed during grain filling as candidate gene for cisgenesis. A genomic...... barley lambda library has been used to isolate the genomic clone of this phytase including 2.3 kb of the promoter region and 600 bp of the terminator region. The clone has been inserted into a cisgenic Agrobacterium vector where both the gene of interest and the selection gene are flanked by their own T...

  6. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2012-01-01

    The cisgenesis concept implies that plants are transformed only with their own genetic materials or genetic materials from closely related species capable of sexual hybridization. Furthermore, foreign sequences such as selection genes and vector-backbone sequences should be absent. We used a barley...... phytase gene (HvPAPhy_a) expressed during grain filling to evaluate the cisgenesis concept in barley. The marker gene elimination method was used to obtain marker-free plant lines. Here, the gene of interest and the selection gene are flanked by their own T-DNA borders to allow unlinked integration...... of the two genes. We analysed the transformants for co-transformation efficiency, increased phytase activities in the grain, integration of the kanamycin resistance gene of the vector-backbone and segregation between the HvPAPhy_a insert and the hygromycin resistance gene. The frequencies of the four...

  7. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2010-01-01

    based on the cisgenesis concept. In contrast to transgenesis, cisgenesis implies that the plant is transformed only with its own or very closely related genetic material. Furthermore, all “helper” genes and gene sequences of foreign nature are removed from the transformed plant lines. Cisgenic crops...... are accordingly very similar to those generated by conventional breeding. The cisgenesis concept allows for the introduction of extra gene copies of a particular gene to accentuate the trait. We are using a barley purple acid phosphatase expressed during grain filling as candidate gene for cisgenesis. A genomic...... barley lambda library has been used to isolate the genomic clone of this phytase including 2.3 kb of the promoter region and 600 bp of the terminator region. The clone has been inserted into a cisgenic Agrobacterium vector where both the gene of interest and the selection gene are flanked by their own T...

  8. Silorane resin supports proliferation, differentiation, and mineralization of MLO-A5 bone cells in vitro and bone formation in vivo.

    Science.gov (United States)

    Eick, J David; Barragan-Adjemian, Cielo; Rosser, Jennifer; Melander, Jennifer R; Dusevich, Vladimir; Weiler, Rachel A; Miller, Bradley D; Kilway, Kathleen V; Dallas, Mark R; Bi, Lianxing; Nalvarte, Elisabet L; Bonewald, Lynda F

    2012-04-01

    Methyl methacrylate used in bone cements has drawbacks of toxicity, high exotherm, and considerable shrinkage. A new resin, based on silorane/oxirane chemistry, has been shown to have little toxicity, low exotherm, and low shrinkage. We hypothesized that silorane-based resins may also be useful as components of bone cements as well as other bone applications and began testing on bone cell function in vitro and in vivo. MLO-A5, late osteoblast cells, were exposed to polymerized silorane (SilMix) resin (and a standard polymerized bisGMA/TEGDMA methacrylate (BT) resin and compared to culture wells without resins as control. A significant cytotoxic effect was observed with the BT resin resulting in no cell growth, whereas in contrast, SilMix resin had no toxic effects on MLO-A5 cell proliferation, differentiation, nor mineralization. The cells cultured with SilMix produced increasing amounts of alkaline phosphatase (1.8-fold) compared to control cultures. Compared to control cultures, an actual enhancement of mineralization was observed in the silorane resin-containing cultures at days 10 and 11 as determined by von Kossa (1.8-2.0 fold increase) and Alizarin red staining (1.8-fold increase). A normal bone calcium/phosphate atomic ratio was observed by elemental analysis along with normal collagen formation. When used in vivo to stabilize osteotomies, no inflammatory response was observed, and the bone continued to heal. In conclusion, the silorane resin, SilMix, was shown to not only be non cytototoxic, but actually supported bone cell function. Therefore, this resin has significant potential for the development of a nontoxic bone cement or bone stabilizer.

  9. Silorane resin supports proliferation, differentiation, and mineralization of MLO-A5 bone cells in vitro and bone formation in vivo

    Science.gov (United States)

    Eick, J. David; Barragan-Adjemian, Cielo; Rosser, Jennifer; Melander, Jennifer R.; Dusevich, Vladimir; Weiler, Rachel A.; Miller, Bradley D.; Kilway, Kathleen V.; Dallas, Mark R.; Bi, Lianxing; Nalvarte, Elisabet L.; Bonewald, Lynda F.

    2015-01-01

    Methyl methacrylate used in bone cements has drawbacks of toxicity, high exotherm, and considerable shrinkage. A new resin, based on silorane/oxirane chemistry, has been shown to have little toxicity, low exotherm, and low shrinkage. We hypothesized that silorane-based resins may also be useful as components of bone cements as well as other bone applications and began testing on bone cell function in vitro and in vivo. MLO-A5, late osteoblast cells, were exposed to polymerized silorane (SilMix) resin (and a standard polymerized bisGMA/TEGDMA methacrylate (BT) resin and compared to culture wells without resins as control. A significant cytotoxic effect was observed with the BT resin resulting in no cell growth, whereas in contrast, SilMix resin had no toxic effects on MLO-A5 cell proliferation, differentiation, nor mineralization. The cells cultured with SilMix produced increasing amounts of alkaline phosphatase (1.8-fold) compared to control cultures. Compared to control cultures, an actual enhancement of mineralization was observed in the silorane resin-containing cultures at days 10 and 11 as determined by von Kossa (1.8–2.0 fold increase) and Alizarin red staining (1.8-fold increase). A normal bone calcium/phosphate atomic ratio was observed by elemental analysis along with normal collagen formation. When used in vivo to stabilize osteotomies, no inflammatory response was observed, and the bone continued to heal. In conclusion, the silorane resin, SilMix, was shown to not only be non cytototoxic, but actually supported bone cell function. Therefore, this resin has significant potential for the development of a nontoxic bone cement or bone stabilizer. PMID:22278990

  10. Search for endophytic diazotrophs in barley seeds

    Directory of Open Access Journals (Sweden)

    Myriam S. Zawoznik

    2014-06-01

    Full Text Available 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.

  11. Search for endophytic diazotrophs in barley seeds.

    Science.gov (United States)

    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.

  12. The NAC transcription factors of barley

    DEFF Research Database (Denmark)

    Wagner, Michael; Holm, Preben Bach; Gregersen, Per L.

    2011-01-01

    ). From these data we have identified not only putative regulators of leaf senescence (HvNAC005, HvNAC027 and HvNAC029), but also possible regulators of secondary wall synthesis (HvNAC033, HvNAC034 and HvNAC039), lateral root formation (HvNAC022) and seed development (HvNAC017, HvNAC018, HvNAC019 and Hv...... genes characterized so far have regulatory functions in a broad range of plant developmental processes and tolerances to both biotic and abiotic stresses. This makes the NAC family highly interesting target genes for plant researchers and breeders. As part of a larger project on the identification...... of Hordeum vulgare (barley) leaf senescence regulators, we have attempted to characterize for the first time all presently available barley NAC genes (HvNACs). By searching the NCBI barley EST database using the tBLASTn function, with all known NAC genes from Brachypodium and rice as input, in combination...

  13. Expression of a barley cystatin gene in maize enhances resistance against phytophagous mites by altering their cysteine-proteases.

    Science.gov (United States)

    Carrillo, Laura; Martinez, Manuel; Ramessar, Koreen; Cambra, Inés; Castañera, Pedro; Ortego, Felix; Díaz, Isabel

    2011-01-01

    Phytocystatins are inhibitors of cysteine-proteases from plants putatively involved in plant defence based on their capability of inhibit heterologous enzymes. We have previously characterised the whole cystatin gene family members from barley (HvCPI-1 to HvCPI-13). The aim of this study was to assess the effects of barley cystatins on two phytophagous spider mites, Tetranychus urticae and Brevipalpus chilensis. The determination of proteolytic activity profile in both mite species showed the presence of the cysteine-proteases, putative targets of cystatins, among other enzymatic activities. All barley cystatins, except HvCPI-1 and HvCPI-7, inhibited in vitro mite cathepsin L- and/or cathepsin B-like activities, HvCPI-6 being the strongest inhibitor for both mite species. Transgenic maize plants expressing HvCPI-6 protein were generated and the functional integrity of the cystatin transgene was confirmed by in vitro inhibitory effect observed against T. urticae and B. chilensis protein extracts. Feeding experiments impaired on transgenic lines performed with T. urticae impaired mite development and reproductive performance. Besides, a significant reduction of cathepsin L-like and/or cathepsin B-like activities was observed when the spider mite fed on maize plants expressing HvCPI-6 cystatin. These findings reveal the potential of barley cystatins as acaricide proteins to protect plants against two important mite pests.

  14. 两种麦类土传花叶病毒外壳蛋白叶绿体离体跨膜运输%Analysis on the Import of Coat Proteins of Chinese Wheat Mosaic Virus and Barley Yellow Mosaic Virus into Intact Chloroplasts in Vitro

    Institute of Scientific and Technical Information of China (English)

    任春梅; 程兆榜; 魏利辉; 范永坚; 周益军

    2012-01-01

    根据Banerjees等(1992)的方法,建立了中国小麦花叶病毒(CWMV)和大麦黄花叶病毒(BaYMV)外壳蛋白(CP)叶绿体离体跨膜运输体系,研究了孵育时间、CP浓度对跨膜运输效率的影响。结果表明,CWMV-CP和BaYMV-CP可分别快速进入离体的小麦与大麦叶绿体中,其跨膜运输所需的孵育时间均最低为5min,孵育时间超过15min后进入叶绿体中CP的量不受影响;加入跨膜体系中CP的浓度与跨膜后进入叶绿体的CP浓度呈正相关,能够实现跨膜的最低CWMV-CP和BaYMV-CP浓度分别为4.2和37.8μg.mL^-1。%Referred to Banerjee's method, the systems of importing the coat proteins of Chinese wheat mosaic virus and Barley yellow mosaic virus (CWMV CP and BaYMV-CP) into intact chloroplasts in vitro were established, the effects of incubation duration, the concentration of CWMV-CP and BaYMV-CP on import efficiency were studied. The results showed that both CWMV-CP and BaYMV- CP was able to rapidly import into isolated chloroplasts of wheat and barley. In the test, the lowest incubation duration of CWMV-CP and BaYMV-CP importing into the chloroplasts were both 5 min, the amounts of CWMV-CP and BaYMV-CP imported into the chloroplasts were not increased with the extension of incubation time after 15 min. Besides, the relation between the amounts of CWMV-CP and BaYMV-CP imported into the chloroplasts and the amounts of CWMV-CP and BaYMV-CP added in the import system was significantly positive, the lowest concentrations of CWMV CP and BaYMV- CP in the import system was 4.2 and 37.8 μg. mL^-1 , respectively. The establishment of CWMV-CP and BaYMV-CP importing into intact chloroplasts in vitro could supply foundation for studying the pathogenesis of CWMV and BaYMV.

  15. Survival of the biocontrol agents Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 on the spikes of barley in the field

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xin; ZHANG Bing-xin; ZHANG Zhen; SHEN Wei-feng; YANG Ching-hong; YU Jing-quan; ZHAO Yu-hua

    2005-01-01

    Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents, Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of the Brevibacillus brevis ZJY- 1 and Bacillus subtilis ZJY- 116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treatment of barley spikes with biocontrol agents before inoculation with F. graminearum.

  16. Milk production is unaffected by replacing barley or sodium hydroxide wheat with maize cob silage in rations for dairy cows.

    Science.gov (United States)

    Hymøller, L; Hellwing, A L F; Lund, P; Weisbjerg, M R

    2014-05-01

    Starch is an important energy-providing nutrient for dairy cows that is most commonly provided from cereal grains. However, ruminal fermentation of large amounts of easily degradable starch leads to excessive production and accumulation of volatile fatty acids (VFA). VFA not only play a vital role in the energy metabolism of dairy cows but are also the main cause of ruminal acidosis and depressed feed intake. The aim of the present study was to compare maize cob silage (MCS) as an energy supplement in rations for dairy cows with highly rumen-digestible rolled barley and with sodium hydroxide wheat (SHW), which has a higher proportion of by-pass starch than barley. Two studies were carried out: (1) a production study on 45 Danish Holstein cows and (2) an intensive study to determine digestibilities, rumen fermentation patterns and methane emission using three rumen-cannulated Danish Holstein cows. Both studies were organised as a 3×3 Latin square with three experimental periods and three different mixed rations. The rations consisted of grass-clover silage and maize silage (~60% of dry matter (DM)), rapeseed cake, soybean meal, sugar beet pulp and one of three different cereals as a major energy supplement: MCS, SHW or rolled barley (~25% of DM). When MCS replaced barley or SHW as an energy supplement in the mixed rations, it resulted in a lower dry matter intake; however, the apparent total tract digestibilities of DM, organic matter, NDF, starch and protein were not different between treatments. The energy-corrected milk yield was unaffected by treatment. The fat content of the milk on the MCS ration was not different from the SHW ration, whereas it was higher on the barley ration. The protein content of the milk decreased when MCS was used in the ration compared with barley and SHW. From ruminal VFA patterns and pH measures, it appeared that MCS possessed roughage qualities with respect to rumen environment, while at the same time being sufficiently energy rich

  17. Transgenic Production of an Anti HIV Antibody in the Barley Endosperm.

    Directory of Open Access Journals (Sweden)

    Goetz Hensel

    Full Text Available Barley is an attractive vehicle for producing recombinant protein, since it is a readily transformable diploid crop species in which doubled haploids can be routinely generated. High amounts of protein are naturally accumulated in the grain, but optimal endosperm-specific promoters have yet to be perfected. Here, the oat GLOBULIN1 promoter was combined with the legumin B4 (LeB4 signal peptide and the endoplasmic reticulum (ER retention signal (SEKDEL. Transgenic barley grain accumulated up to 1.2 g/kg dry weight of recombinant protein (GFP, deposited in small roundish compartments assumed to be ER-derived protein bodies. The molecular farming potential of the system was tested by generating doubled haploid transgenic lines engineered to synthesize the anti-HIV-1 monoclonal antibody 2G12 with up to 160 μg recombinant protein per g grain. The recombinant protein was deposited at the periphery of protein bodies in the form of a mixture of various N-glycans (notably those lacking terminal N-acetylglucosamine residues, consistent with their vacuolar localization. Inspection of protein-A purified antibodies using surface plasmon resonance spectroscopy showed that their equilibrium and kinetic rate constants were comparable to those associated with recombinant 2G12 synthesized in Chinese hamster ovary cells.

  18. Inhibitory properties of cysteine protease pro-peptides from barley confer resistance to spider mite feeding.

    Directory of Open Access Journals (Sweden)

    M Estrella Santamaria

    Full Text Available C1A plant cysteine proteases are synthesized as pre-pro-enzymes that need to be processed to become active by the pro-peptide claves off from its cognate enzyme. These pro-sequences play multifunctional roles including the capacity to specifically inhibit their own as well as other C1A protease activities from diverse origin. In this study, it is analysed the potential role of C1A pro-regions from barley as regulators of cysteine proteases in target phytophagous arthropods (coleopteran and acari. The in vitro inhibitory action of these pro-sequences, purified as recombinant proteins, is demonstrated. Moreover, transgenic Arabidopsis plants expressing different fragments of HvPap-1 barley gene containing the pro-peptide sequence were generated and the acaricide function was confirmed by bioassays conducted with the two-spotted spider mite Tetranychus urticae. Feeding trials resulted in a significant reduction of leaf damage in the transgenic lines expressing the pro-peptide in comparison to non-transformed control and strongly correlated with an increase in mite mortality. Additionally, the analysis of the expression levels of a selection of potential mite targets (proteases and protease inhibitors revealed a mite strategy to counteract the inhibitory activity produced by the C1A barley pro-prodomain. These findings demonstrate that pro-peptides can control mite pests and could be applied as defence proteins in biotechnological systems.

  19. Inhibitory properties of cysteine protease pro-peptides from barley confer resistance to spider mite feeding.

    Science.gov (United States)

    Santamaria, M Estrella; Arnaiz, Ana; Diaz-Mendoza, Mercedes; Martinez, Manuel; Diaz, Isabel

    2015-01-01

    C1A plant cysteine proteases are synthesized as pre-pro-enzymes that need to be processed to become active by the pro-peptide claves off from its cognate enzyme. These pro-sequences play multifunctional roles including the capacity to specifically inhibit their own as well as other C1A protease activities from diverse origin. In this study, it is analysed the potential role of C1A pro-regions from barley as regulators of cysteine proteases in target phytophagous arthropods (coleopteran and acari). The in vitro inhibitory action of these pro-sequences, purified as recombinant proteins, is demonstrated. Moreover, transgenic Arabidopsis plants expressing different fragments of HvPap-1 barley gene containing the pro-peptide sequence were generated and the acaricide function was confirmed by bioassays conducted with the two-spotted spider mite Tetranychus urticae. Feeding trials resulted in a significant reduction of leaf damage in the transgenic lines expressing the pro-peptide in comparison to non-transformed control and strongly correlated with an increase in mite mortality. Additionally, the analysis of the expression levels of a selection of potential mite targets (proteases and protease inhibitors) revealed a mite strategy to counteract the inhibitory activity produced by the C1A barley pro-prodomain. These findings demonstrate that pro-peptides can control mite pests and could be applied as defence proteins in biotechnological systems.

  20. CDNA cloning, characterization and expression of an endosperm-specific barley peroxidase

    DEFF Research Database (Denmark)

    Rasmussen, Søren Kjærsgård; Welinder, K.G.; Hejgaard, J.

    1991-01-01

    A barley peroxidase (BP 1) of pI ca. 8.5 and M(r) 37000 has been purified from mature barley grains. Using antibodies towards peroxidase BP 1, a cDNA clone (pcR7) was isolated from cDNA expression library. The nucleotide sequence of pcR7 gave a derived amino acid sequence identical to the 158 C......-terminal amino acid residues of mature BP 1. The clone pcR7 encodes an additional C-terminal sequence of 22 residues, which apparently are removed during processing. BP 1 is less than 50% identical to other sequenced plant peroxidases. Analyses of RNA and protein from aleurone, endosperm and embryo tissue showed...

  1. The barley Frost resistance-H2 locus.

    Science.gov (United States)

    Pasquariello, Marianna; Barabaschi, Delfina; Himmelbach, Axel; Steuernagel, Burkhard; Ariyadasa, Ruvini; Stein, Nils; Gandolfi, Francesco; Tenedini, Elena; Bernardis, Isabella; Tagliafico, Enrico; Pecchioni, Nicola; Francia, Enrico

    2014-03-01

    Frost resistance-H2 (Fr-H2) is a major QTL affecting freezing tolerance in barley, yet its molecular basis is still not clearly understood. To gain a better insight into the structural characterization of the locus, a high-resolution linkage map developed from the Nure × Tremois cross was initially implemented to map 13 loci which divided the 0.602 cM total genetic distance into ten recombination segments. A PCR-based screening was then applied to identify positive bacterial artificial chromosome (BAC) clones from two genomic libraries of the reference genotype Morex. Twenty-six overlapping BACs from the integrated physical-genetic map were 454 sequenced. Reads assembled in contigs were subsequently ordered, aligned and manually curated in 42 scaffolds. In a total of 1.47 Mbp, 58 protein-coding sequences were identified, 33 of which classified according to similarity with sequences in public databases. As three complete barley C-repeat Binding Factors (HvCBF) genes were newly identified, the locus contained13 full-length HvCBFs, four Related to AP2 Triticeae (RAPT) genes, and at least five CBF pseudogenes. The final overall assembly of Fr-H2 includes more than 90 % of target region: all genes were identified along the locus, and a general survey of Repetitive Elements obtained. We believe that this gold-standard sequence for the Morex Fr-H2 will be a useful genomic tool for structural and evolutionary comparisons with Fr-H2 in winter-hardy cultivars along with Fr-2 of other Triticeae crops.

  2. Combining unmalted barley and pearling gives good quality brewing

    NARCIS (Netherlands)

    Donkelaar, van Laura H.G.; Hageman, Jos A.; Oguz, Serhat; Noordman, Tom R.; Boom, Remko M.; Goot, van der Atze Jan

    2016-01-01

    Brewing with unmalted barley can reduce the use of raw materials, thereby increasing the efficiency of the brewing process. However, unmalted barley contains several undesired components for brewing and has a low enzymatic activity. Pearling, an abrasive milling method, has been proposed as a pre

  3. Low Phytic Acid Barley Responses to Phosphorus Rates

    Science.gov (United States)

    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...

  4. Progressive hull removal from barley using the Fitzpatrick comminuting mill

    Science.gov (United States)

    The objective of the study was to explore an alternative use of the Fitzpatrick Comminuting Machine: to use it to remove the hull from hulled barley while keeping the barley kernel intact. Traditionally, this mill is used to grind material, but we have recently discovered that it also has the abili...

  5. Some quality attributes of low fat ice cream substituted with hulless barley flour and barley ß-glucan.

    Science.gov (United States)

    Abdel-Haleem, Amal M H; Awad, R A

    2015-10-01

    The purpose of this paper is to investigate some quality attributes of low fat ice cream (LFIC) substituted with hulless barley flour (HBF) and barley ß-glucan (BBG). The methodology included in this paper is based on adding HBF (1, 2, 3 and 4 %) as a partial substitution of skim milk powder (SMP) and BBG (0.40 %) as a complete substitution of carboxy methyl cellulose (CMC). All mixes and resultant ice cream samples were evaluated for their physicochemical properties as well as the sensory quality attributes.The results indicated that substitution of SMP with HBF significantly increased total solids (TS), fat and crude fiber, while crude protein and ash significantly decreased in ice cream mixes. BBG exhibited the same manner of control. Specific gravity was gradually increased with adding HBFand BBG in the mixes and therefore the overrun percent was significantly changed in the resultant ice cream. Adding HBF in ice cream formula led to significant decrease in acidity with higher freezing point and the product showed higher ability to meltdown. BBG treatment showed the same trend of control. Values of flow time and viscosity significantly increased with increasing HBF in the ice cream mixes, but these values significantly decreased in BBG mix. The time required to freeze ice cream mixes was decreased with increasing the ratio of HBF but, increased in BBG treatment. The substitution of SMP with 1 and 2 % HBF significantly (P ≤ 0.05) enhanced sensory attributes of ice cream samples. While, BBG treatment achieved mild score and acceptability.

  6. Secretion, purification, and characterisation of barley alpha-amylase produced by heterologous gene expression in Aspergillus niger.

    Science.gov (United States)

    Juge, N; Svensson, B; Williamson, G

    1998-04-01

    Efficient production of recombinant barley alpha-amylase has been achieved in Aspergillus niger. The cDNA encoding alpha-amylase isozyme 1 (AMY1) and its signal peptide was placed under the control of the Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and the A. nidulans trpC gene terminator. Secretion yields up to 60 mg/l were obtained in media optimised for alpha-amylase activity and low protease activity. The recombinant AMY1 (reAMY1) was purified to homogeneity and found to be identical to native barley AMY1 with respect to size, pI, and immunoreactivity. N-terminal sequence analysis of the recombinant protein indicated that the endogenous plant signal peptide is correctly processed in A. niger. Electrospray ionisation/mass spectrometry gave a molecular mass for the dominant form of 44,960 Da, in accordance with the loss of the LQRS C-terminal residues; glycosylation apparently did not occur. The activities of recombinant and native barley alpha-amylases are very similar towards insoluble and soluble starch as well as 2-chloro-4-nitrophenol beta-D-maltoheptaoside and amylose (degree of polymerisation = 17). Barley alpha-amylase is the first plant protein efficiently secreted and correctly processed by A. niger using its own signal sequence.

  7. Barley Stripe Mosaic Virus and the Frequency of Triploids and Aneuploids in Barley

    DEFF Research Database (Denmark)

    Sandfær, J.

    1973-01-01

    BSMV infection caused a pronounced increase in the frequency of triploid and aneuploid seeds in eleven barley varieties, but with considerable variation in frequency among varieties. In some of the varieties triploids exceeded three per cent. In virus-free material a few triploids were found in m...

  8. Stability of Barley stripe mosaic virus induced gene silencing in barley

    DEFF Research Database (Denmark)

    Bruun-Rasmussen, Marianne; Madsen, Christian Toft; Jessing, Stine;

    2007-01-01

    Virus-induced gene silencing (VIGS) can be used as a powerful tool for functional genomics studies in plants. With this approach, it is possible to target most genes and downregulate the messenger (m)RNA in a sequence-specific manner. Barley stripe mosaic virus (BSMV) is an established VIGS vecto...... inoculation, although large parts of the insert had been lost from the virus vector. The instability of the insert, observed consistently throughout our experiments, offers an explanation for the transient nature of silencing when using BSMV as a VIGS vector.......Virus-induced gene silencing (VIGS) can be used as a powerful tool for functional genomics studies in plants. With this approach, it is possible to target most genes and downregulate the messenger (m)RNA in a sequence-specific manner. Barley stripe mosaic virus (BSMV) is an established VIGS vector...... for barley and wheat; however, silencing using this vector is generally transient, with efficient silencing often being confined to the first two or three systemically infected leaves. To investigate this further, part of the barley Phytoene desaturase (PDS) gene was inserted into BSMV and the resulting...

  9. Giemsa C-banding of Barley Chromosomes. IV. Chromosomal Constitution of Autotetraploid Barley

    DEFF Research Database (Denmark)

    Linde-Laursen, Ib

    1984-01-01

    The progeny of an autotetraploid barley plant (C1) consisted of 45 tetraploids and 33 aneuploids. Giemsa C-banding was used to identify each of the chromosomes in 20 euploid and 31 aneuploid C2--seedlings, and in 11 C3--offspring of aneuploid C2--plants. The euploid C2--seedlings all had four hom...

  10. A barley flour inhibitor of insect alpha-amylase is a major allergen associated with baker's asthma disease.

    Science.gov (United States)

    Barber, D; Sánchez-Monge, R; Gómez, L; Carpizo, J; Armentia, A; López-Otín, C; Juan, F; Salcedo, G

    1989-05-08

    A barley salt-soluble protein of 14.5 kDa, which inhibits the alpha-amylase from the insect Tenebrio molitor, has been identified as a major IgE-binding component of sera from baker's asthma patients. The N-terminal amino acid sequence of this protein indicates that it is a member of a previously described family of alpha-amylase/trypsin inhibitors.

  11. Structural lability of Barley stripe mosaic virus virions.

    Directory of Open Access Journals (Sweden)

    Valentin V Makarov

    Full Text Available Virions of Barley stripe mosaic virus (BSMV were neglected for more than thirty years after their basic properties were determined. In this paper, the physicochemical characteristics of BSMV virions and virion-derived viral capsid protein (CP were analyzed, namely, the absorption and intrinsic fluorescence spectra, circular dichroism spectra, differential scanning calorimetry curves, and size distributions by dynamic laser light scattering. The structural properties of BSMV virions proved to be intermediate between those of Tobacco mosaic virus (TMV, a well-characterized virus with rigid rod-shaped virions, and flexuous filamentous plant viruses. The BSMV virions were found to be considerably more labile than expected from their rod-like morphology and a distant sequence relation of the BSMV and TMV CPs. The circular dichroism spectra of BSMV CP subunits incorporated into the virions, but not subunits of free CP, demonstrated a significant proportion of beta-structure elements, which were proposed to be localized mostly in the protein regions exposed on the virion outer surface. These beta-structure elements likely formed during virion assembly can comprise the N- and C-terminal protein regions unstructured in the non-virion CP and can mediate inter-subunit interactions. Based on computer-assisted structure modeling, a model for BSMV CP subunit structural fold compliant with the available experimental data was proposed.

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

    Science.gov (United States)

    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, (13)C 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.

  13. A novel major gene on chromosome 6H for resistance of barley against the barley yellow dwarf virus

    NARCIS (Netherlands)

    Niks, R.E.; Habekuss, A.; Bekele, B.; Ordon, F.

    2004-01-01

    In a mapping population derived from the Ethiopian barley line L94 x Vada, natural infection by barley yellow dwarf virus (BYDV) occurred. While line L94 hardly showed symptoms, Vada was severely affected. The 103 recombinant inbred lines segregated bimodally. The major gene responsible for this res

  14. Differentially expressed genes between drought-tolerant and drought-sensitive barley genotypes in response to drought stress during the reproductive stage

    Science.gov (United States)

    Guo, Peiguo; Baum, Michael; Grando, Stefania; Ceccarelli, Salvatore; Bai, Guihua; Li, Ronghua; von Korff, Maria; Varshney, Rajeev K.; Graner, Andreas; Valkoun, Jan

    2009-01-01

    Drought tolerance is a key trait for increasing and stabilizing barley productivity in dry areas worldwide. Identification of the genes responsible for drought tolerance in barley (Hordeum vulgare L.) will facilitate understanding of the molecular mechanisms of drought tolerance, and also facilitate the genetic improvement of barley through marker-assisted selection or gene transformation. To monitor the changes in gene expression at the transcriptional level in barley leaves during the reproductive stage under drought conditions, the 22K Affymetrix Barley 1 microarray was used to screen two drought-tolerant barley genotypes, Martin and Hordeum spontaneum 41-1 (HS41-1), and one drought-sensitive genotype Moroc9-75. Seventeen genes were expressed exclusively in the two drought-tolerant genotypes under drought stress, and their encoded proteins may play significant roles in enhancing drought tolerance through controlling stomatal closure via carbon metabolism (NADP malic enzyme, NADP-ME, and pyruvate dehydrogenase, PDH), synthesizing the osmoprotectant glycine-betaine (C-4 sterol methyl oxidase, CSMO), generating protectants against reactive-oxygen-species scavenging (aldehyde dehydrogenase,ALDH, ascorbate-dependent oxidoreductase, ADOR), and stabilizing membranes and proteins (heat-shock protein 17.8, HSP17.8, and dehydrin 3, DHN3). Moreover, 17 genes were abundantly expressed in Martin and HS41-1 compared with Moroc9-75 under both drought and control conditions. These genes were possibly constitutively expressed in drought-tolerant genotypes. Among them, seven known annotated genes might enhance drought tolerance through signalling [such as calcium-dependent protein kinase (CDPK) and membrane steroid binding protein (MSBP)], anti-senescence (G2 pea dark accumulated protein, GDA2), and detoxification (glutathione S-transferase, GST) pathways. In addition, 18 genes, including those encoding Δl-pyrroline-5-carboxylate synthetase (P5CS), protein phosphatase 2C

  15. Transglycosylation by barley α-amylase 1

    DEFF Research Database (Denmark)

    Mótyán, János A.; Fazekas, Erika; Mori, Haruhide;

    2011-01-01

    The transglycosylation activity of barley α-amylase 1 (AMY1) and active site AMY1 subsite mutant enzymes was investigated. We report here the transferase ability of the V47A, V47F, V47D and S48Y single mutants and V47K/S48G and V47G/S48D double mutant AMY1 enzymes in which the replaced amino acids...... DP 2, DP 3 and DP 5 were successfully applied to detect activity of Bacillus stearothermophilus maltogenic α-amylase, human salivary α-amylase and Bacillus licheniformis α-amylase, respectively in a fast and simple fluorometric assay....

  16. Dynamic Allocation of Sugars in Barley

    Science.gov (United States)

    Cumberbatch, L. C.; Crowell, A. S.; Fallin, B. A.; Howell, C. R.; Reid, C. D.; Weisenberger, A. G.; Lee, S. J.; McKisson, J. E.

    2014-03-01

    Allocation of carbon and nitrogen is a key factor for plant productivity. Measurements are carried out by tracing 11C-tagged sugars using positron emission tomography and coincidence counting. We study the mechanisms of carbon allocation and transport from carbohydrate sources (leaves) to sinks (stem, shoot, roots) under various environmental conditions such as soil nutrient levels and atmospheric CO2 concentration. The data are analyzed using a transfer function analysis technique to model transport and allocation in barley plants. The experimental technique will be described and preliminary results presented. This work was supported in part by USDOE Grant No. DE-FG02-97-ER41033 and DE-SC0005057.

  17. Liquid digestate from anaerobic treatment of source-separated household waste as fertilizer to barley.

    Science.gov (United States)

    Haraldsen, Trond Knapp; Andersen, Uno; Krogstad, Tore; Sørheim, Roald

    2011-12-01

    This study examined the efficiency of different organic waste materials as NPK fertilizer, in addition to the risk for leaching losses related to shower precipitation in the first part of the growing season. The experiment was tested in a pot trial on a sandy soil in a greenhouse. Six organic fertilizers were evaluated: liquid anaerobic digestate (LAD) sourced from separated household waste, nitrified liquid anaerobic digestate (NLAD) of the same origin as LAD, meat and bone meal (MBM), hydrolysed salmon protein (HSP), reactor-composted catering waste (CW) and cattle manure (CM). An unfertilized control, calcium nitrate (CN) and Fullgjødsel® 21-4-10 were used as reference fertilizers. At equal amounts of mineral nitrogen both LAD and Fullgjødsel® gave equal yield of barley in addition to equal uptake of N, P, and K in barley grain. NLAD gave significantly lower barley yield than the original LAD due to leaching of nitrate-N after a simulated surplus of precipitation (28 mm) at Zadoks 14. There was significantly increased leaching of nitrate N from the treatments receiving 160 kg N ha(-1) of CN and NLAD in comparison with all the other organic fertilizers. In this study LAD performed to the same degree as Fullgjødsel® NPK fertilizer and it was concluded that LAD can be recommended as fertilizer for cereals. Nitrification of the ammonium N in the digestate caused significantly increased nitrate leaching, and cannot be recommended.

  18. The white barley mutant albostrians shows enhanced resistance to the biotroph Blumeria graminis f. sp. hordei.

    Science.gov (United States)

    Jain, Sanjay Kumar; Langen, Gregor; Hess, Wolfgang; Börner, Thomas; Hückelhoven, Ralph; Kogel, Karl-Heinz

    2004-04-01

    We performed cytological and molecular analyses of the interaction between the biotrophic barley powdery mildew fungus Blumeria graminis f. sp. hordei and white and green leaves of the barley albostrians mutant. The leaves have the same nuclear genotype but differ from each other in respect to plastid differentiation. White leaves showed enhanced penetration resistance to B. graminis f. sp. hordei, associated with higher epidermal H2O2 accumulation beneath the appressorial germ tubes and protein cross-linking in papillae. Very low basal salicylic acid content was found in white leaves, which further confirmed that H2O2 accumulation and penetration resistance in barley are independent of salicylic acid. Expression analysis of stress and defense-related genes, including such being involved in reactive oxygen species production and cell death regulation, revealed stronger constitutive or pathogen-induced transcript accumulation in white leaves. We discuss the data on the basis of the finding that white albostrians leaves exhibit a supersusceptible interaction phenotype with the hemibiotrophic fungus Bipolaris sorokiniana.

  19. Purification and characterization of three chitinases and one beta-1,3-glucanase accumulating in the medium of cell suspension cultures of barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Kragh, K.M.; Jacobsen, S.; Dalgaard Mikkelsen, J.;

    1991-01-01

    chromatography. Two of the chitinases were identified as the previously described endochitinases T and C from barley grain. The third and novel chitinase, designated K, was the major basic chitinase in the medium constituting 4% of the soluble protein. Chitinase K was found to be a 33-kDa endochitinase with p...... chitinases from barley aleurone and barley, bean and potato leaves. The purified beta-1,3-glucanase with a molecular weight (MW) of 32 kDa and pI greater-than-or-equal-to 9.8 constituted 1% of the soluble protein in the medium. Based on similar MW, pI and amino acid composition as well as identical N...

  20. Rye inhibitors of animal alpha-amylases show different specificities, aggregative properties and IgE-binding capacities than their homologues from wheat and barley.

    Science.gov (United States)

    García-Casado, G; Sánchez-Monge, R; López-Otín, C; Salcedo, G

    1994-09-01

    Three new members of the alpha-amylase/trypsin-inhibitor family of cereal endosperm have been isolated from rye. N-terminal amino acid sequence comparison revealed that two of the purified proteins were the rye homologues of the barley monomeric inhibitor (BMAI-1) previously described, while the other rye protein corresponded to one of the subunits of the barley and wheat heterotetrameric inhibitors. However, the inhibitory specificities (active against human salivary alpha-amylase), aggregative behaviours (mainly as dimeric forms) and IgE-binding capacities (not recognized by sera from allergic patients) of the rye inhibitors were clearly different from those of their wheat and barley counterparts. These results indicate that homologous inhibitors may have distinctive properties in different cereal species.

  1. HvZIP7 mediates zinc accumulation in barley (Hordeum vulgare) at moderately high zinc supply

    DEFF Research Database (Denmark)

    Tiong, Jingwen; Mcdonald, Glenn K.; Genc, Yusuf

    2014-01-01

    , barley (Hordeum vulgare) HvZIP7 was investigated for its functions in Zn transport. The functions of HvZIP7 in planta were studied using in situ hybridization and transient analysis of subcellular localization with a green fluorescent protein (GFP) reporter. Transgenic barley lines overexpressing HvZIP7......Summary: High expression of zinc (Zn)-regulated, iron-regulated transporter-like protein (ZIP) genes increases root Zn uptake in dicots, leading to high accumulation of Zn in shoots. However, none of the ZIP genes tested previously in monocots could enhance shoot Zn accumulation. In this report...... were also generated to further understand the functions of HvZIP7 in metal transport. HvZIP7 is strongly induced by Zn deficiency, primarily in vascular tissues of roots and leaves, and its protein was localized in the plasma membrane. These properties are similar to its closely related homologs...

  2. Improving drought and salinity tolerance in barley by application of salicylic acid and potassium nitrate

    Directory of Open Access Journals (Sweden)

    Khalaf Ali Fayez

    2014-01-01

    Full Text Available Growth and physiological activities of barley (Hordeum vulgare L. cv. Gustoe grown in soil cultures were evaluated to recognize the ameliorative role of salicylic acid (SA and KNO3 against the negative effects of salt and water deficit stresses. Barley plants were subjected to three levels of NaCl (50, 100 and 150 mM, three levels of water stress (80%, 70% and 50% of the soil water content (SWC and the combination of 150 mM NaCl + 50 μM SA, 150 mM NaCl + 10 mM KNO3, 50% SWC + 50 μM SA and 50% SWC + 10 mM KNO3 for two weeks. Salt and water deficit stresses reduced the shoot growth, leaf photosynthetic pigments, K+ contents and provoked oxidative stress in leaves confirmed by considerable changes in soluble carbohydrate, proline, malondialdehyde (MDA, total phenolic compounds, antioxidant activity and Na+ contents. Leaf soluble protein of salt and water deficit treated plants was unaffected. The Na+/K+ ratio increased with increasing salt and water deficit treated plants. Application of 50 μM SA or 10 mM KNO3 to150 mM NaCl and/or 50% SWC treated plants improved these attributes under salt and water stresses. Soluble carbohydrates in stressed plants may have a significant role in osmotic adjustment. It can be concluded that the addition of SA or KNO3 can ameliorate the oxidative stress in barley stressed plants. This ameliorative effect might be maintained through low MDA contents and decreased Na+/K+ ratio in leaves. This study also provided evidence for the ability of barley cultivation in salt and water deficit soils due to its capacity for osmotic adjustment.

  3. Influence of Temperature on the Extractibility of Polysaccharides in Barley

    Directory of Open Access Journals (Sweden)

    Rodica Căpriţă

    2011-10-01

    Full Text Available Barley contains substantial amounts of both soluble and insoluble non-starch polysaccharides (NSP. The main watersoluble NSP in barley are highly viscous β-glucans. Monogastric animals, including humans and birds, cannotsynthesize β-glucanase, and the amount of β-glucanase derived from barley grain and bacteria in the gastrointestinaltract is insufficient to completely hydrolyze β-glucans. In the present investigation, we have studied the influence oftemperature and heating time on the extractibility of soluble polysaccharides in barley. Heating the barley samples at60°C and 80°C before extraction has the effect of lowering the soluble fraction of the polysaccharides. The dynamicviscosity values of water extracts from barley decreased up to 21.68% when heating at 60ºC for 15 minutes, and upto 25.30% when heating at 80ºC for 15 minutes, when the determinations were made immediately after extractseparation. Heating the barley samples for 15 minutes at 80°C deactivates the endogenous hydrolytic enzymes.

  4. The barley anion channel, HvALMT1, has multiple roles in guard cell physiology and grain metabolism.

    Science.gov (United States)

    Xu, Muyun; Gruber, Benjamin D; Delhaize, Emmanuel; White, Rosemary G; James, Richard A; You, Jiangfeng; Yang, Zhenming; Ryan, Peter R

    2015-01-01

    The barley (Hordeum vulgare) gene HvALMT1 encodes an anion channel in guard cells and in certain root tissues indicating that it may perform multiple roles. The protein localizes to the plasma membrane and facilitates malate efflux from cells when constitutively expressed in barley plants and Xenopus oocytes. This study investigated the function of HvALMT1 further by identifying its tissue-specific expression and by generating and characterizing RNAi lines with reduced HvALMT1 expression. We show that transgenic plants with 18-30% of wild-type HvALMT1 expression had impaired guard cell function. They maintained higher stomatal conductance in low light intensity and lost water more rapidly from excised leaves than the null segregant control plants. Tissue-specific expression of HvALMT1 was investigated in developing grain and during germination using transgenic barley lines expressing the green fluorescent protein (GFP) with the HvALMT1 promoter. We found that HvALMT1 is expressed in the nucellar projection, the aleurone layer and the scutellum of developing barley grain. Malate release measured from isolated aleurone layers prepared from imbibed grain was significantly lower in the RNAi barley plants compared with control plants. These data provide molecular and physiological evidence that HvALMT1 functions in guard cells, in grain development and during germination. We propose that HvALMT1 releases malate and perhaps other anions from guard cells to promote stomatal closure. The likely roles of HvALMT1 during seed development and grain germination are also discussed.

  5. Molecular analysis of Korean isolate of barley yellow mosaic virus.

    Science.gov (United States)

    Lee, Kui Jae; Choi, Min Kyung; Lee, Wang Hyu; Rajkumar, Mani

    2006-04-01

    The complete sequences of both RNAs of an isolate of barley yellow mosaic virus (BaYMV) from Haenam, Korea, were determined. RNA1 is 7639 nucleotides long [excluding the 3'-poly(A)], and codes for a 270 kDa polyprotein of 2411 amino acids which contains the capsid protein (CP) at the C terminus and seven putative non-structural proteins. RNA2 is 3582 nucleotides long and codes for a polyprotein of 890 amino acids, which contains a 28 kDa putative proteinase (P1) and a 73 kDa polypeptide (P2). The whole sequences of Korean isolate (BaYMV-K) closely resembled those of an isolate from Japan (BaYMV-J) (99.6 identical nucleotides for RNA1; 99.4 for RNA2) and china (BaYMV-C) (96.7 and 96.2%, respectively) than from Germany (BaYMV-G) (93.6 and 90.4%, respectively). The greatest differences between the BaYMV-K and BaYMV-J isolates were in the 3'-NCRs of RNA1 and 5' NCRs of RNA2 and there were also some other regions of difference in Nib Pro (RNA1) and P1 (RNA2). Further, the phylogenetic analysis of CP region showed that Asian and European isolates formed distinct clusters. However, molecular variations between isolates could not be linked to earlier results showing differences in cultivar response.

  6. Methane emissions from feedlot cattle fed barley or corn diets.

    Science.gov (United States)

    Beauchemin, K A; McGinn, S M

    2005-03-01

    Methane emitted from the livestock sector contributes to greenhouse gas emissions worldwide. Understanding the variability in enteric methane production related to diet is essential to decreasing uncertainty in greenhouse gas emission inventories and to identifying viable greenhouse gas reduction strategies. Our study focused on measuring methane in growing beef cattle fed corn- or barley-based diets typical of those fed to cattle in North American feedlots. The experiment was designed as a randomized complete block (group) design with two treatments, barley and corn. Angus heifer calves (initial BW = 328 kg) were allocated to two groups (eight per group), with four cattle in each group fed a corn or barley diet. The experiment was conducted over a 42-d backgrounding phase, a 35-d transition phase and a 32-d finishing phase. Backgrounding diets consisted of 70% barley silage or corn silage and 30% concentrate containing steam-rolled barley or dry-rolled corn (DM basis). Finishing diets consisted of 9% barley silage and 91% concentrate containing barley or corn (DM basis). All diets contained monensin (33 mg/kg of DM). Cattle were placed into four large environmental chambers (two heifers per chamber) during each phase to measure enteric methane production for 3 d. During the backgrounding phase, DMI was greater by cattle fed corn than for those fed barley (10.2 vs. 7.6 kg/d, P cattle were in the chambers; thus, methane emissions (g/d) reported may underestimate those of the feedlot industry. Methane emissions per kilogram of DMI and as a percentage of GE intake were not affected by grain source during the backgrounding phase (24.6 g/kg of DMI; 7.42% of GE), but were less (P methane emissions of cattle fed high-forage backgrounding diets and barley-based finishing diets. Mitigating methane losses from cattle will have long-term environmental benefits by decreasing agriculture's contribution to greenhouse gas emissions.

  7. Enhancing the aluminium tolerance of barley by expressing the citrate transporter genes SbMATE and FRD3.

    Science.gov (United States)

    Zhou, Gaofeng; Pereira, Jorge F; Delhaize, Emmanuel; Zhou, Meixue; Magalhaes, Jurandir V; Ryan, Peter R

    2014-06-01

    Malate and citrate efflux from root apices is a mechanism of Al(3+) tolerance in many plant species. Citrate efflux is facilitated by members of the MATE (multidrug and toxic compound exudation) family localized to the plasma membrane of root cells. Barley (Hordeum vulgare) is among the most Al(3+)-sensitive cereal species but the small genotypic variation in tolerance that is present is correlated with citrate efflux via a MATE transporter named HvAACT1. This study used a biotechnological approach to increase the Al(3+) tolerance of barley by transforming it with two MATE genes that encode citrate transporters: SbMATE is the major Al(3+)-tolerance gene from sorghum whereas FRD3 is involved with Fe nutrition in Arabidopsis. Independent transgenic and null T3 lines were generated for both transgenes. Lines expressing SbMATE showed Al(3+)-activated citrate efflux from root apices and greater tolerance to Al(3+) toxicity than nulls in hydroponic and short-term soil trials. Transgenic lines expressing FRD3 exhibited similar phenotypes except citrate release from roots occurred constitutively. The Al(3+) tolerance of these lines was compared with previously generated transgenic barley lines overexpressing the endogenous HvAACT1 gene and the TaALMT1 gene from wheat. Barley lines expressing TaALMT1 showed significantly greater Al(3+) tolerance than all lines expressing MATE genes. This study highlights the relative efficacy of different organic anion transport proteins for increasing the Al(3+) tolerance of an important crop species.

  8. Overexpression of cytokinin dehydrogenase genes in barley (Hordeum vulgare cv. Golden Promise fundamentally affects morphology and fertility.

    Directory of Open Access Journals (Sweden)

    Katarína Mrízová

    Full Text Available Barley is one of the most important cereal crops grown worldwide. It has numerous applications, but its utility could potentially be extended by genetically manipulating its hormonal balances. To explore some of this potential we identified gene families of cytokinin dehydrogenases (CKX and isopentenyl transferases, enzymes that respectively irreversibly degrade and synthesize cytokinin (CK plant hormones, in the raw sequenced barley genome. We then examined their spatial and temporal expression patterns by immunostaining and qPCR. Two CKX-specific antibodies, anti-HvCKX1 and anti-HvCKX9, predominantly detect proteins in the aleurone layer of maturing grains and leaf vasculature, respectively. In addition, two selected CKX genes were used for stable, Agrobacterium tumefaciens-mediated transformation of the barley cultivar Golden Promise. The results show that constitutive overexpression of CKX causes morphological changes in barley plants and prevents their transition to flowering. In all independent transgenic lines roots proliferated more rapidly and root-to-shoot ratios were higher than in wild-type plants. Only one transgenic line, overexpressing CKX under the control of a promoter from a phosphate transporter gene, which is expressed more strongly in root tissue than in aerial parts, yielded progeny. Analysis of several T1-generation plants indicates that plants tend to compensate for effects of the transgene and restore CK homeostasis later during development. Depleted CK levels during early phases of development are restored by down-regulation of endogenous CKX genes and reinforced de novo biosynthesis of CKs.

  9. Transcriptional regulatory programs underlying barley germination and regulatory functions of Gibberellin and abscisic acid

    Directory of Open Access Journals (Sweden)

    Lin Li

    2011-06-01

    Full Text Available Abstract Background Seed germination is a complex multi-stage developmental process, and mainly accomplished through concerted activities of many gene products and biological pathways that are often subjected to strict developmental regulation. Gibberellins (GA and abscisic acid (ABA are two key phytohormones regulating seed germination and seedling growth. However, transcriptional regulatory networks underlying seed germination and its associated biological pathways are largely unknown. Results The studies examined transcriptomes of barley representing six distinct and well characterized germination stages and revealed that the transcriptional regulatory program underlying barley germination was composed of early, late, and post-germination phases. Each phase was accompanied with transcriptional up-regulation of distinct biological pathways. Cell wall synthesis and regulatory components including transcription factors, signaling and post-translational modification components were specifically and transiently up-regulated in early germination phase while histone families and many metabolic pathways were up-regulated in late germination phase. Photosynthesis and seed reserve mobilization pathways were up-regulated in post-germination phase. However, stress related pathways and seed storage proteins were suppressed through the entire course of germination. A set of genes were transiently up-regulated within three hours of imbibition, and might play roles in initiating biological pathways involved in seed germination. However, highly abundant transcripts in dry barley and Arabidopsis seeds were significantly conserved. Comparison with transcriptomes of barley aleurone in response to GA and ABA identified three sets of germination responsive genes that were regulated coordinately by GA, antagonistically by ABA, and coordinately by GA but antagonistically by ABA. Major CHO metabolism, cell wall degradation and protein degradation pathways were up

  10. STUDIES ON SYNBIOTIC BARLEY GRAIN EXTRACT AGAINST SOME HUMAN PATHOGENS

    Directory of Open Access Journals (Sweden)

    T. Sheela

    2012-01-01

    Full Text Available This study evaluated that effect of prebiotic food containing oligosaccharide to enhance the growth and activity of probiotic strains. Barley grains probioticated using different strains of probiotics are Lactobacillus kefiranofaciens, Candida kefir,and saccharomyces boluradii. To select a suitable prebiotics like inulin for the development of Synbiotic barley and tested for antibacterial activity against diarrhoea causing pathogen such as Esherichia coli, Staphylococcus aureus, Salmonella paratyphi A, Shigella dysenteriae, Vibrio cholerae. Analysis of identified compound from synbiotic barley grain using GC-MS.

  11. Effect of simulated microgravity on nitric oxide synthase activity of osteocyte-like cell line MLO-Y4 in response to fluid shear stress

    Science.gov (United States)

    Sun, Lian-Wen; Yang, Xiao; Fan, Yu-Bo

    It is well known that microgravity could induce bone loss. However, the mechanism remains poorly understood. Osteocytes are extremely sensitive to fluid shear stress, even more than osteobleasts. The effect of simulated microgravity on osteocytes in response to fluid shear was investigated in this study in order to see if the mechanosensibility of osteocytes changed under simulated microgravity. The osteocyte-like cell line, MLO-Y4, was cultured and divided into four groups, including control (CON), control and shear (CONS), rotary (RT), rotary and shear (RTS). In RT and RTS, the cells were cultured in the rotary cell culture system to simulate microgravity condition. After 5 days, the cells in RTS and CONS were subjected to flow shear for 15 min. Then nitric oxide synthase (NOS) activity in the cells was measured using assay kit. The results showed that NOS activity in respond to fluid shear decreased significantly in RTS compared with CONS. In addition, there was significant difference in NOS activity between CONS and CON while no significant difference between RTS and RT. These indicates that the mechanosensibility of osteocytes decreased under simulated microgravity and this maybe the partly causes of the poor effect of exercise to counter microgravity-induced-bone loss. However, further research need to be done to support this finding.

  12. [Cold induced cDNA library construction of highland barley (Hordeum vulgare L. var. nudum Hk. f.) using suppression subtractive hybridization technology].

    Science.gov (United States)

    He, Tao; Jia, Jing Fen

    2008-12-01

    Cold-induced genes of highland barley (Hordeum vulgare L. var. nudum Hk. f.) were studied using suppression subtractive hybridization (SSH) technique. The cDNA from the materials treated with 4 degrees C was used as "tester", and that from the materials growing in green house (20+/-2 degrees C) as "driver". A subtractive library of highland barley including 640 cDNA clones was constructed in this study. Enzyme digestion of 32 clones chosen randomly from the library indicated that 87.5% of them contained inserts. The cDNA inserts of 16 clones were sequenced. Blast search analyses showed that these cDNAs were homologies to genes encoding the following proteins: metallothionein, protein kinase, ethylene signal transcription factor, bZIP transcription factor, zing finger transcription factor, ribulose-1,5-bisphosphate carboxylase, ribosomal protein, sodium: hydrogen antiporter, catalase, NADPH-cytochrome reductase, ascorbate peroxidase, DNA binding protein, and sugar transporter-like protein. These results indicated that the cDNA clones in the library were related to cold-induced genes, and suggested that the cold-tolerant mechanism of highland barley might be a complicated, interactive system involving multiple approaches and genes. Construction of subtractive cDNA library provided an advantage for further studies to isolate and clone cold-induced genes in highland barley.

  13. Effect of phytase supplementation to barley-canola meal and barley-soybean meal diets on phosphorus and calcium balance in growing pigs

    NARCIS (Netherlands)

    Sauer, W.C.; Cervantes, M.; He, J.M.M.; Schulze, H.

    2003-01-01

    Two metabolism experiments were carried out, to determine the effect of microbial phytase addition to barley-canola meal and barley-soybean meal diets on P and Ca balance in growing. pigs; In experiment 1, six barrows (29.6kg: initial LW) were fed a barley-canola meal diet, without or. with phytase

  14. Effects of feeding hull-less barley on production performance, milk fatty acid composition, and nutrient digestibility of lactating dairy cows.

    Science.gov (United States)

    Yang, Y; Ferreira, G; Teets, C L; Corl, B A; Thomason, W E; Griffey, C A

    2017-03-16

    The objectives of this study were to evaluate production performance, milk fatty acid composition, and nutrient digestibility in high-producing dairy cows consuming diets containing corn and hull-less barley (cultivar Amaze 10) in different proportions as the grain source. Eight primiparous and 16 multiparous Holstein cows were assigned to 1 of 4 diets in a replicated 4 × 4 Latin square design with 21-d periods. Cows were fed once daily (1200 h) by means of a Calan gate system (American Calan Inc., Northwood, NH). All diets contained ∼20% grain (dry matter basis). Treatments consisted of 100% corn (0B), 67% corn and 33% hull-less barley (33B), 33% corn and 67% hull-less barley (67B), and 100% hull-less barley (100B) as the grain sources. Total-tract nutrient digestibility was estimated using lanthanum chloride (LaCl3) as an external marker. Dry matter intake differed quadratically among treatments, being lowest for 67B and highest for 0B and 100B. Feeding hull-less barley did not affect milk yield, and milk fat concentration differed cubically among treatments. The cubic response was attributed to the higher milk fat concentration observed for the diet containing 67B. Neither the concentrations in milk of protein and lactose nor the yields of protein and lactose differed among treatments. The proportion of de novo synthesized fatty acids in milk did not differ among treatments. The apparent total-tract digestibility of dry matter, crude protein, and neutral detergent fiber did not differ among treatments. Although a quadratic effect was observed, starch digestibility was minimally affected by treatments. In conclusion, this study indicates that hull-less barley grain is as good as corn grain as an energy source when formulating diets for high-producing dairy cows.

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

    Science.gov (United States)

    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.

  16. Effects of nitrogen application rate on dry matter redistribution, grain yield, nitrogen use efficiency and photosynthesis in malting barley

    DEFF Research Database (Denmark)

    Cai, J; Jiang, D; Wollenweber, Bernd

    2012-01-01

    , and decreased with 300 kg N ha−1. Net photosynthetic rate (P N) and the amount of accumulated dry matter distributed into grains showed the same response to N application as grain yield. Grain protein content increased with increasing N application rates. Moreover, based on further analysis of these results......The harmonious combination of malting barley yield, quality and nitrogen (N) use-efficiency under nitrogen (N) rates applications was greatly conducive to production in China. The malting barley cultivar Supi 3 was planted during the growing seasons 2005 and 2006 at two contrasting sites in China....... Five nitrogen (N) application rates (0, 75, 150, 225 and 300 kg ha−1) were applied for research of effects of N rates application on grain yield, protein content and N use-efficiency. At both sites and in both years, grain yield increased with increasing N application rates up to 225 kg N ha−1...

  17. Barley starch bioengineering for high phosphate and amylose

    DEFF Research Database (Denmark)

    Blennow, Per Gunnar Andreas; Carciofi, Massimiliano; Shaik, Shahnoor Sultana;

    2011-01-01

    Starch is a biological polymer that can be industrially produced in massive amounts in a very pure form. Cereals is the main source for starch production and any improvement of the starch fraction can have a tremendous impact in food and feed applications. Barley ranks number four among cereal...... crops and barley is a genetically very well characterized. Aiming at producing new starch qualities in the cereal system, we used RNAi and overexpression strategies to produce pure amylose and high-phosphate starch, respectively, using the barley kernel as a polymer factory. By simultaneous silencing...... of the three genes encoding the starch-branching enzymes SBEI, SBEIIa, and SBEIIb using a triple RNAi chimeric hairpin construct we generated a virtually amylopectin-free barley. The grains of the transgenic lines were shrunken and had a yield of around 80% of the control line. The starch granules were...

  18. Barley peroxidase isozymes. Expression and post-translational modification in mature seeds as identified by two-dimensional gel electrophoresis and mass spectrometry

    DEFF Research Database (Denmark)

    Laugesen, Sabrina; Bak-Jensen, Kristian Sass; Hägglund, Per

    2007-01-01

    putative barley peroxidase. The presence of multiple spots for each of the isozymes reflected variations in post-translational glycosylation and protein truncation. Complete sequence coverage was achieved by using a series of proteases and chromatographic resins for sample preparation prior to mass....... In summary, a subset of three peroxidase isozymes was found to occur in the seed, whereas products of four other barley peroxidase genes were not detected. The present analysis documents the selective expression profiles and post-translational modifications of isozymes from a large plant gene family. (C...

  19. Glutathione S-transferase (GST) family in barley: identification of members, enzyme activity, and gene expression pattern.

    Science.gov (United States)

    Rezaei, Mohammad Kazem; Shobbar, Zahra-Sadat; Shahbazi, Maryam; Abedini, Raha; Zare, Sajjad

    2013-09-15

    Barley (Hordeum vulgare) is one of the most important cereals in many developing countries where drought stress considerably diminishes agricultural production. Glutathione S-transferases (GSTs EC 2.5.1.18) are multifunctional enzymes which play a crucial role in cellular detoxification and oxidative stress tolerance. In this study, 84 GST genes were identified in barley by a comprehensive in silico approach. Sequence alignment and phylogenetic analysis grouped these HvGST proteins in eight classes. The largest numbers of the HvGST genes (50) were included in the Tau class followed by 21 genes in Phi, five in Zeta, two in DHAR, two in EF1G, two in Lambda, and one each in TCHQD and Theta classes. Phylogenetic analysis of the putative GSTs from Arabidopsis, rice, and barley indicated that major functional diversification within the GST family predated the monocot/dicot divergence. However, intra-specious duplication seems to be common. Expression patterns of five GST genes from Phi and Tau classes were investigated in three barley genotypes (Yusof [drought-tolerant], Moroc9-75 [drought-sensitive], and HS1 [wild ecotype]) under control and drought-stressed conditions, during the vegetative stage. All investigated genes were up-regulated significantly under drought stress and/or showed a higher level of transcripts in the tolerant cultivar. Additionally, GST enzyme activity was superior in Yusof and induced in the extreme-drought-treated leaves, while it was not changed in Moroc9-75 under drought conditions. Moreover, the lowest and highest levels of lipid peroxidation were observed in the Yusof and Moroc9-75 cultivars, respectively. Based on the achieved results, detoxification and antioxidant activity of GSTs might be considered an important factor in the drought tolerance of barley genotypes for further investigations.

  20. Hydrothermal liquefaction of barley straw to bio-crude oil

    DEFF Research Database (Denmark)

    Zhu, Zhe; Rosendahl, Lasse; Toor, Saqib;

    2015-01-01

    Hydrothermal liquefaction (HTL) of barley straw with K2CO3 at different temperatures (280–400 C) was conducted and compared to optimize its process conditions; the aqueous phase as a co-product from this process was recycled to explore the feasibility of implementing wastewater reuse for bio...... a detailed insight into the HTL behavior of barley straw, and offers potential opportunities and benefits for bio-crude oil production through the reuse of aqueous phase....

  1. Androgenesis in anther culture of Lithuanian spring barley cultivars

    OpenAIRE

    Asakavičiūtė, Rita; Pašakinskienė, Izolda

    2006-01-01

    The method of anther culture was used for the production of doubled haploids in Lithuanian spring barley cultivars. Two methods, (i) regeneration from callus (Szarjeko’s method) and (ii) direct regeneration from embryoids (Caredda’s method) were applied to determine the androgenic potential according to the green regenerant yield and other morphogenetic factors. Green double haploid regenerants were obtained in four Lithuanian spring barley cultivars (‘Aura’, ‘Aidas’, ‘Alsa’ and ‘Auksiniai’) ...

  2. The spontaneous chlorophyll mutation frequency in barley

    DEFF Research Database (Denmark)

    Jørgensen, Jørgen Helms; Jensen, Hans Peter

    1986-01-01

    A total of 1866 barley plants were progeny tested in the greenhouse. Twenty-five plants segregated for newly arisen, spontaneous chlorophyll mutant genes. Among the total of 470,129 seedlings screened there were 79 mutants (1.7 .+-. 0.6 .times. 10-4). The data are added to data from three similar...... materials and the resulting estimate of the chlorophyll mutant frequency is 1.6 .times. 10-4 in about 1.43 million seedlings. The estimate of the chlorophyll mutation rate per generation is close to 67.3 .times. 10-4 per diploid genome or in the order of 6 .times. 10-7 per locus and haploid genome....

  3. The Barley Chromosome 5 Linkage Map

    DEFF Research Database (Denmark)

    Jensen, J.; Jørgensen, Jørgen Helms

    1975-01-01

    The literature is surveyed for data on recombination between loci on chromosome 5 of barley; 13 loci fall into the category “mapped” loci, more than 20 into the category “associated” loci and nine into the category “loci once suggested to be on chromosome 5”. A procedure was developed...... for estimating a linkage map; it involves (1) transformation by the Kosambi mapping function of the available recombination percentages to additive map distances, (2) calculations of a set of map distances from the transformed recombination percentages by a maximum likelihood method in which all the available...... data are utilized jointly, and (3) omission of inconsistent data and determination of the most likely order of the loci. This procedure was applied to the 42 recombination percentages available for the 13 “mapped” loci. Due to inconsistencies 14 of the recombination percentages and, therefore, two...

  4. Weed suppression ability of spring barley varieties

    DEFF Research Database (Denmark)

    Christensen, Svend

    1995-01-01

    Three years of experiments with spring barley showed significant differences in weed suppression ability among varieties. Weed dry matter in the most suppressive variety, Ida, was 48% lower than the mean weed dry matter of all varieties, whereas it was 31% higher in the least suppressive variety......, Grit. Ranking varietal responses to weed competition in terms of grain yield loss corresponded well to ranking weed dry matter produced in crop weed mixtures. There was no correspondence between the varietal grain yields in pure stands and their competitiveness, suggesting that breeding to optimize...... interception model was developed to describe the light interception profiles of the varieties. A study of the estimated parameters showed significant correlation between weed dry matter, rate of canopy height development and the light interception profile. However, when estimates were standardized to eliminate...

  5. Effects of sulfur nutritional level on cadmium toxicity in barley

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Yichang; Huerta, A.J. (Miami Univ., Oxford, OH (United States))

    1993-05-01

    The effects of S levels on Cd toxicity were studied in barley (Hordeum vulgare L.cv.UC 476). Barley was grown hydroponically in half-strength Hoagland's solution containing either 100% or 10% S in a growth chamber at constant 20 C, 290 umole M[sup [minus]2] s[sup [minus]1] light intensity, and a 16/18 hour light/dark period. Five days after the first true leaf appeared, 15 uM Cd was added to the nutrient solutions where appropriate. At 14 days after beginning of Cd treatment, plants were analyzed for photosynthetic characteristics. The photosynthetic characteristics measured were CO[sub 2] response curves (measured with a LICOR 6200 portable photosynthesis system), and fluorescence measurement system. At 21 days they were analyzed for morphological and biomass measurements. The CO[sub 2] response curves for leaves of plants treated with 10% S did not significantly differ from those of plants treated with 100% S. Treatment with Cd significantly reduced the CO[sup 2] saturated rates of photosynthesis and the reduction was more significant in the 10% S than in the 100% S plants. Photochemical efficiency of PSII (FV/FM) and fluorescence quenching capacity (FQ) were not affected by 10% S as compared to 100% S treatment. Interestingly, treatment with Cd significantly increased both FV/FM and FQ as compared to control., However, S level had no effect on the fluorescence parameters of Cd-treated plants. Leaf and root length, leaf area, root and shoot dry weight were only slightly affected (increased or decreased) by 10% S as compared to 100% S but very significantly reduced by treatment with Cd. Our results agree with the previous reports which show that S (an important component of glutathione and phytochelatins which are low molecular weight Cd binding proteins), is important in regulating Cd detoxification in plants. However, we are continuing to conduct experiments as even lower S concentrations and different Cd concentrations.

  6. Effects of toasting blue lupins, soybeans or barley as supplement for high-yielding organic dairy cows fed grass-clover silage ad libitum

    DEFF Research Database (Denmark)

    Mogensen, Lisbeth; Lund, Peter; Kristensen, Troels;

    2008-01-01

    . The supplement under investigation was: lupins in experiment 1, barley in experiment 2 and soybeans in experiment 3. The same forage mixture of grass-clover silage (84% of DM), grass pellets (11% of DM) and straw (5% of DM) was fed ad libitum in all the experiments. Toasting decreased effective rumen protein...

  7. A pathway-specific microarray analysis highlights the complex and co-ordinated transcriptional networks of the developing grain of field-grown barley

    DEFF Research Database (Denmark)

    Hansen, Michael; Friis, Carsten; Bowra, Steve

    2009-01-01

    The aim of the study was to describe the molecular and biochemical interactions associated with amino acid biosynthesis and storage protein accumulation in the developing grains of field-grown barley. Our strategy was to analyse the transcription of genes associated with the biosynthesis of stora...

  8. Characterization of recombinant high pI Barley α-Glucosidase

    DEFF Research Database (Denmark)

    Næsted, Henrik; Svensson, Birte

    α-glucosidase activity in barley seeds plays a crucial role in embryonic development. The concerted action of α-glucosidase, α-amylase, β-amylase, and limit dextrinase serve as the machinery responsible for the supply of glucose as energy source for the developing embryo in the germinating seed...... of the enzyme was grown under high cell-density fermentation conditions. This approach enabled a successful protein expression profile under the highly sensitive methanol utilization phase of the fermentation procedure. The enzyme was purified using a four step purification strategy. Interestingly, the purified...

  9. The effect of added enzymes on process potentials derived from different qualities of barley

    DEFF Research Database (Denmark)

    Shetty, Radhakrishna; Zhuang, Shiwen; Hansen, Preben Bøje;

    Barley sorting is an important step for picking up grain of desired quality. Whilst brewing with 100% sorted barley (picked high quality) has become realistic with the addition of exogenous enzymes, the effect of added enzymes on process potentials derived from un-sorted barley (mixed) and sorted...... filterability, the Ondea® Pro treatment resulted in significantly lower turbidity and smaller particle size compared to Cellic® CTec2; however, this effect was observed in sorted and un-sorted barley but not in sorted-out barley. Consequently the un-sorted barley demonstrated great potential in brewing process...

  10. Long-term reconstitution of dry barley increased phosphorus digestibility in pigs

    DEFF Research Database (Denmark)

    Ton Nu, Mai Anh; Blaabjerg, Karoline; Poulsen, Hanne Damgaard

    of reconstitution compared to dry stored barley on phosphorus (P) digestibility in pigs. Materials and Methods: Dry barley (13% moisture; phytate P, 1.7 g/kg DM) was rolled and stored directly or reconstituted with water to produce rolled barley with 35% moisture that was stored in air-tight conditions. After 49......: Reconstituted barley had higher soluble P (2.56 g/kg DM) and lower phytate P (0.93 g/ kg DM) compared with dry barley (0.78 and 1.7 g/kg DM, respectively). Pigs fed the reconstituted barley diet showed increased P absorption (52%) and decreased P excretion in feces (21%) (P

  11. Constructing the barley model for genetic transformation in Triticeae

    Institute of Scientific and Technical Information of China (English)

    LÜ Bo; WU Jia-jie; FU Dao-lin

    2015-01-01

    Barley (Hordeum vulgare L.) is one of the oldest domesticated crops, showing dramatic adaptation to various climate and environmental conditions. As a major cereal crop, barley ranks the 4th after wheat, maize and rice in terms of planting area and production al over the world. Due to its diploid nature, the cultivated barley is considered as an ideal model to study the polyploid wheat and other Triticeae species. Here, we reviewed the development, optimization, and application of transgenic approaches in barley. The most efifcient and robust genetic transformation has been built on the Agrobacterium-mediated transfer in conjunction with the immature embryo-based regeneration. We then discussed future considerations of using more practical technologies in barley transformation, such as the T-DNA/transposon tagging and the genome editing. As a cereal crop amenable to genetic transformation, barley wil serve as the most valuable carrier for global functional genomics in Triticeae and is becoming the most practical model for generating value-added products.

  12. Glycaemic response to barley porridge varying in dietary fibre content.

    Science.gov (United States)

    Thondre, Pariyarath S; Wang, Ke; Rosenthal, Andrew J; Henry, Christiani J K

    2012-03-01

    The interest in barley as a food is increasing worldwide because of its high dietary fibre (DF) content and low glycaemic index (GI). DF in cereals may prove beneficial in improving blood glucose response in the long term. However, a dose-dependent effect of insoluble fibre on reducing postprandial blood glucose levels is yet to be proven. The objective of the present study was to determine the glycaemic response to two barley porridges prepared from whole barley grains varying in fibre content. In two separate non-blind randomised crossover trials, ten human subjects consumed barley porridge with 16 g/100 g and 10 g/100 g fibre content provided in different serving sizes (equivalent to 25 and 50 g available carbohydrate). The glycaemic response to both barley porridges was significantly lower than the reference glucose (P porridges. We concluded that irrespective of the difference in total fibre content or serving size of barley porridges, their GI values did not differ significantly.

  13. Comparative Analysis of Nutritional Composition Between Black Barley and White Barley%黑大麦与白大麦营养成分对比分析

    Institute of Scientific and Technical Information of China (English)

    申晓蓉; 樊梦原; 陈莉; 李方敏; 任丽; 叶永丽; 韦紫玉

    2012-01-01

    以甘肃省永登县民乐乡黑大麦、白大麦为主要研究对象,采用国家标准方法,即灰化法、高锰酸钾滴定法、蒽酮比色法、索氏抽提法、凯氏定氮法等方法,测定黑大麦中水分、灰分、钙、磷、蛋白质、总糖、粗脂肪、淀粉等基本营养成分及纤维成分.本实验通过对比分析得出,黑大麦中灰分含量为2.06±0.04 g/100g,粗脂肪含量为1.66±0.42 g/100g,总糖含量为63.03±1.05 g/100g,粗纤维含量为6.12±0.34 g/100g,磷含量为371 mg/100g,分别比白大麦高出0.18%,0.22%,2.79%,1.47%和0.012%,相对于白大麦具有营养优越性.%The progress conditions of the analysis of nutritional composition in black barley and white barley from Yongdeng County Gansu Province were investigated using the national standards including permanganate titration method, Kjeldahl method, Soxhlet extraction method and other national standard methods. The water, ash, calcium, phosphorus, protein, total reducing sugar, fat, fiber and other basic nutrients were determined. The results showed that the contents of the ash, fat, total reducing sugar, fiber and phosphorus in the black barley were 2.06 ± 0.04 g/100g, 1.66 ± 0.42 g/100g, 63.03 ± 1.05 g/100g, 6.12 ± 0.34 g/100g, 371 ± 5.58 rng/100g respectively, which were 0.18 % ,0.22 % ,2.79 %, 1.47 % and 0. 012 % larger than those in the white barley.

  14. Purification, enzymatic characterization, and nucleotide sequence of a high-isoelectric-point alpha-glucosidase from barley malt

    DEFF Research Database (Denmark)

    Frandsen, T P; Lok, F; Mirgorodskaya, E;

    2000-01-01

    High-isoelectric-point (pI) alpha-glucosidase was purified 7, 300-fold from an extract of barley (Hordeum vulgare) malt by ammonium sulfate fractionation, ion-exchange, and butyl-Sepharose chromatography. The enzyme had high activity toward maltose (k(cat) = 25 s(-1)), with an optimum at pH 4...... in the transition state complex. Mass spectrometry of tryptic fragments assigned the 92-kD protein to a barley cDNA (GenBank accession no. U22450) that appears to encode an alpha-glucosidase. A corresponding sequence (HvAgl97; GenBank accession no. AF118226) was isolated from a genomic phage library using a c......DNA fragment from a barley cDNA library. HvAgl97 encodes a putative 96.6-kD protein of 879 amino acids with 93.8% identity to the protein deduced from U22450. The sequence contains two active site motifs of glycoside hydrolase family 31. Three introns of 86 to 4,286 bp interrupt the coding region. The four...

  15. Programmed cell death in barley aleurone cells is not directly stimulated by reactive oxygen species produced in response to gibberellin.

    Science.gov (United States)

    Aoki, Nozomi; Ishibashi, Yushi; Kai, Kyohei; Tomokiyo, Reisa; Yuasa, Takashi; Iwaya-Inoue, Mari

    2014-05-01

    The cereal aleurone layer is a secretory tissue that produces enzymes to hydrolyze the starchy endosperm during germination. We recently demonstrated that reactive oxygen species (ROS), produced in response to gibberellins (GA), promoted GAMyb expression, which induces α-amylase expression in barley aleurone cells. On the other hand, ROS levels increase during programmed cell death (PCD) in barley aleurone cells, and GAMyb is involved in PCD of these cells. In this study, we investigated whether the ROS produced in response to GA regulate PCD directly by using mutants of Slender1 (SLN1), a DELLA protein that negatively regulates GA signaling. The wild-type, the sln1c mutant (which exhibits gibberellin-type signaling even in the absence of GA), and the Sln1d mutant (which is gibberellin-insensitive with respect to α-amylase production) all produced ROS in response to GA, suggesting that ROS production in aleurone cells in response to GA is independent of GA signaling through this DELLA protein. Exogenous GA promoted PCD in the wild-type. PCD in sln1c was induced even without exogenous GA (and so without induction of ROS), whereas PCD in Sln1d was not induced in the presence of exogenous GA, even though the ROS content increased significantly in response to GA. These results suggest that PCD in barley aleurone cells is not directly stimulated by ROS produced in response to GA but is regulated by GA signaling through DELLA protein.

  16. Cloning and characterization of a gibberellin-induced RNase expressed in barley aleurone cells

    Energy Technology Data Exchange (ETDEWEB)

    Rogers, S.W.; Rogers, J.C. (Washington State Univ., Pullman, WA (United States). Inst. of Biological Chemistry)

    1999-04-01

    The authors cloned a cDNA for a gibberellin-induced ribonuclease (RNase) expressed in barley (Hordeum vulgare) aleurone and the gene for a second barley RNase expressed in leaf tissue. The protein encoded by the cDNA is unique among RNases described to date in that it contains a novel 23-amino acid insert between the C2 and C3 conserved sequences. Expression of the recombinant protein in tobacco (Ncotiana tabacum) suspension-cultured protoplasts gave an active RNase of the expected size, confirming the enzymatic activity of the protein. Analyses of hormone regulation of re-expression of mRNA for the aleurone RNase revealed that, like the pattern for [alpha]-amylase, mRNA levels increased in the presence of gibberellic acid, and its antagonist abscisic acid prevented this effect. Quantitative studies at early times demonstrated that cycloheximide treatment of aleurone layers increased mRNA levels 4-fold, whereas a combination of gibberellin plus cycloheximide treatment was required to increase [alpha]-amylase mRNA levels to the same extent. These results are consistent with loss of repression as an initial effect of gibberellic acid on transcription of those genes, although the regulatory pathways for the two genes may differ.

  17. From Proteomics to Structural Studies of Cytosolic/Mitochondrial-Type Thioredoxin Systems in Barley Seeds

    Institute of Scientific and Technical Information of China (English)

    Azar Shahpiri; Birte Svensson; Christine Finnie

    2009-01-01

    Thioredoxins (Trx) are ubiquitous proteins that participate in thiol disulfide reactions via two active site cysteine residues,allowing Trx to reduce disulfide bonds in target proteins.Recent progress in proteome analysis has resulted in identification of a wide range of potential target proteins for Trx,indicating that Trx plays a key role in several aspects of cell metabolism.In contrast to other organisms,plants contain multiple forms of Trx that are classified based on their primary structures and sub-cellular localization.The reduction of cytosolic and mitochondrial types of Trx is dependenl on NADPH and catalyzed by NADPH-dependent thioredoxin reductase (NTR).In barley,two isoforms each of Trx and NTR have been identified and investigated using proteomics,gene expression,and structural studies.This review outlines the diverse roles suggested for cytosolic/mitochondrial-type Trx systems in cereal seeds and summarizes the current knowl-edge of the barley system including recent data on function,regulation,interactions,and structure.Directions for future research are discussed.

  18. Differences in grain ultrastructure, phytochemical and proteomic profiles between the two contrasting grain Cd-accumulation barley genotypes.

    Directory of Open Access Journals (Sweden)

    Hongyan Sun

    Full Text Available 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.

  19. Competition and Facilitation in Hairy Vetch-Barley Intercrops

    Directory of Open Access Journals (Sweden)

    Giacomo Tosti

    2010-09-01

    Full Text Available Intercrops between legumes and non-legumes are widely used for fodder production and as cover crops, but little quantitative data are available on competition between species in the mixture. The objective of the present study was to assess the interaction between hairy vetch (Vicia villosa Roth. and barley (Hordeum vulgare L. grown as pure crops or intercrops with different proportions of seed rates at sowing. A 4-year field study was conducted using hairy vetch and barley as pure stands at full sowing density and as intercrops at different proportions of their own full seed rate according to the replacement principle. Interaction between species was evaluated on the basis of Land Equivalent Ratio (LER, Relative Neighbour Effect (RNE and Aggressivity (A calculated on biomass and nitrogen (N accumulation. The N accumulation of the mixed crops increased linearly with the legume proportion in the mixture. The mixtures were more efficient than the pure crops in terms of N use (LER > 1. Partial LER values indicated that the barley component benefited from the presence of the legume, while the hairy vetch partial LER decreased with increasing barley proportion in the mixture. The competitive response in terms of biomass accumulation was high for both species when their density in the mixture was high. Concerning N accumulation, barley benefited from an asymmetric interspecific facilitation while the vetch behaviour was similar to that observed for biomass accumulation. Barley dominance progressively increased reaching a maximum just before the last sampling date. At the last sampling date the competitive ability of hairy vetch showed a considerable increase in all mixtures (A ≈ 0. These findings indicate that the use of mixtures between hairy vetch and barley allows an increase in the use efficiency of N resource with respect to pure crops. Barley is the dominant component of the mixture and the hairy vetch is able to cope with the cereal

  20. HvALMT1 from barley is involved in the transport of organic anions.

    Science.gov (United States)

    Gruber, Benjamin D; Ryan, Peter R; Richardson, Alan E; Tyerman, Stephen D; Ramesh, Sunita; Hebb, Diane M; Howitt, Susan M; Delhaize, Emmanuel

    2010-03-01

    Members of the ALMT gene family contribute to the Al(3+) resistance of several plant species by facilitating malate efflux from root cells. The first member of this family to be cloned and characterized, TaALMT1, is responsible for most of the natural variation of Al(3+) resistance in wheat. The current study describes the isolation and characterization of HvALMT1, the barley gene with the greatest sequence similarity to TaALMT1. HvALMT1 is located on chromosome 2H which has not been associated with Al(3+) resistance in barley. The relatively low levels of HvALMT1 expression detected in root and shoot tissues were independent of external aluminium or phosphorus supply. Transgenic barley plants transformed with the HvALMT1 promoter fused to the green fluorescent protein (GFP) indicated that expression of HvALMT1 was relatively high in stomatal guard cells and in root tissues containing expanding cells. GFP fused to the C-terminus of the full HvALMT1 protein localized to the plasma membrane and motile vesicles within the cytoplasm. HvALMT1 conferred both inward and outward currents when expressed in Xenopus laevis oocytes that were bathed in a range of anions including malate. Both malate uptake and efflux were confirmed in oocyte assays using [(14)C]malate as a radiotracer. It is suggested that HvALMT1 functions as an anion channel to facilitate organic anion transport in stomatal function and expanding cells.

  1. Characterization of the entire cystatin gene family in barley and their target cathepsin L-like cysteine-proteases, partners in the hordein mobilization during seed germination.

    Science.gov (United States)

    Martinez, Manuel; Cambra, Ines; Carrillo, Laura; Diaz-Mendoza, Mercedes; Diaz, Isabel

    2009-11-01

    Plant cystatins are inhibitors of cysteine-proteases of the papain C1A and legumain C13 families. Cystatin data from multiple plant species have suggested that these inhibitors act as defense proteins against pests and pathogens and as regulators of protein turnover. In this study, we characterize the entire cystatin gene family from barley (Hordeum vulgare), which contain 13 nonredundant genes, and identify and characterize their target enzymes, the barley cathepsin L-like proteases. Cystatins and proteases were expressed and purified from Escherichia coli cultures. Each cystatin was found to have different inhibitory capability against barley cysteine-proteases in in vitro inhibitory assays using specific substrates. Real-time reverse transcription-polymerase chain reaction revealed that inhibitors and enzymes present a wide variation in their messenger RNA expression patterns. Their transcripts were mainly detected in developing and germinating seeds, and some of them were also expressed in leaves and roots. Subcellular localization of cystatins and cathepsin L-like proteases fused to green fluorescent protein demonstrated the presence of both protein families throughout the endoplasmic reticulum and the Golgi complex. Proteases and cystatins not only colocalized but also interacted in vivo in the plant cell, as revealed by bimolecular fluorescence complementation. The functional relationship between cystatins and cathepsin L-like proteases was inferred from their common implication as counterparts of mobilization of storage proteins upon barley seed germination. The opposite pattern of transcription expression in gibberellin-treated aleurones presented by inhibitors and enzymes allowed proteases to specifically degrade B, C, and D hordeins stored in the endosperm of barley seeds.

  2. Efficient production of tetraploid barley (Hordeum vulgare L. by colchicine treatment of diploid barley

    Directory of Open Access Journals (Sweden)

    Ayed Sourour

    2014-03-01

    Full Text Available An experiment was conducted to induce tetraploidy in three diploid barley varieties (Martin, Rihane and Manel through different colchicines treatments. Colchicine was added for three different concentrations at three different stages of plant development i.e. on seed (0.05% for 48 hours, on pre-germinated seeds (0.1% for 2 hours and on three leaf stage (0.1% for 16 hours. Colchicine application reduced significantly germination percentage and viability of plants. Seed germination was completely inhibited in Martin, while a reduction of 20% and 30% for germination percentage compared to control was recorded in varieties Manel and Rihane, respectively at 0.1% colchicine concentration. Ploidy evaluation showed no tetraploidy in all the three tested varieties by colchicine application of 0.05% for 48 hours on seeds and 0.1% for 2 hours on pre-germinated seeds. However, tetraploid plants were produced only by treatment with 0.1% for 16 hours of seedlings. The percentages of plants were 40%, 44% and 100% for Rihane, Manel and Martin, respectively. Cytological analyses showed the increase of chromosome numbers from 2n=2x=14 to 2n=4x=28. The increase of ploidy levels caused major changes in some morphological traits. In fact, the induced tetraploids in barley was accompanied by significant (P<0.01 decrease in plant height, tiller height, leaf number and leaf length compared to diploid control plants. colchicine treatment induce successfully the production of tetraploid barley plants and could be used in breeding programs.

  3. Molecular characterization of barley 3H semi-dwarf genes.

    Directory of Open Access Journals (Sweden)

    Haobing Li

    Full Text Available The barley chromosome 3H accommodates many semi-dwarfing genes. To characterize these genes, the two-rowed semi-dwarf Chinese barley landrace 'TX9425' was crossed with the Australian barley variety 'Franklin' to generate a doubled haploid (DH population, and major QTLs controlling plant height have been identified in our previous study. The major QTL derived from 'TX9425' was targeted to investigate the allelism of the semi-dwarf gene uzu in barley. Twelve sets of near-isogenic lines and a large NILF2 fine mapping population segregating only for the dwarfing gene from 'TX9425' were developed. The semi-dwarfing gene in 'TX9425' was located within a 2.8 cM region close to the centromere on chromosome 3H by fine mapping. Molecular cloning and sequence analyses showed that the 'TX9425'-derived allele contained a single nucleotide substitution from A to G at position 2612 of the HvBRI1 gene. This was apparently the same mutation as that reported in six-rowed uzu barley. Markers co-segregating with the QTL were developed from the sequence of the HvBRI1 gene and were validated in the 'TX9425'/'Franklin' DH population. The other major dwarfing QTL derived from the Franklin variety was distally located on chromosome 3HL and co-segregated with the sdw1 diagnostic marker hv20ox2. A third dwarfing gene, expressed only in winter-sown trials, was identified and located on chromosome 3HS. The effects and interactions of these dwarfing genes under different growing conditions are discussed. These results improve our understanding of the genetic mechanisms controlling semi-dwarf stature in barley and provide diagnostic markers for the selection of semi-dwarfness in barley breeding programs.

  4. Expression of the betaine aldehyde dehydrogenase gene in barley in response to osmotic stress and abscisic acid.

    Science.gov (United States)

    Ishitani, M; Nakamura, T; Han, S Y; Takabe, T

    1995-01-01

    When subjected to salt stress or drought, some vascular plants such as barley respond with an increased accumulation of the osmoprotectant glycine betaine (betaine), being the last step of betaine synthesis catalyzed by betaine aldehyde dehydrogenase (BADH). We report here cloning and characterization of BADH cDNA from barley, a monocot, and the expression pattern of a BADH transcript. An open reading frame of 1515 bp encoded a protein which showed high homology to BADH enzymes present in other plants (spinach and sugar-beet) and in Escherichia coli. Transgenic tobacco plants harboring the clone expressed high levels of both BADH protein and its enzymatic activity. Northern blot analyses indicated that BADH mRNA levels increased almost 8-fold and 2-fold, respectively, in leaves and roots of barley plants grown in high-salt conditions, and that these levels decreased upon release of the stress, whereas they did not decrease under continuous salt stress. BADH transcripts also accumulate in response to water stress or drought, indicating a common response of the plant to osmotic changes that affect its water status. The addition of abscisic acid (ABA) to plants during growth also increased the levels of BADH transcripts dramatically, although the response was delayed when compared to that found for salt-stressed plants. Removal of plant roots before transferring the plants to high-salt conditions reduced only slightly the accumulation of BADH transcripts in the leaves.

  5. Characterization of barley serpin Z7 that plays multiple roles in malt and beer.

    Science.gov (United States)

    Li, Xiaomin; Jin, Zhao; Gao, Fei; Lu, Jian; Cai, Guolin; Dong, Jianjun; Yu, Junhong; Yang, Mei

    2014-06-18

    Barley protein Z7 (BSZ7) is a well-known serine protease inhibitor that was regarded as a major effector of beer foam stability. Moreover, it has also been suggested to participate in haze formation and affect wort filterability. The present study purified BSZ7 from barley malt and characterized its secondary structure and modification, as well as its relationship with peroxidase, to elucidate the molecular base of BSZ7 that supports its multiple roles in malt and beer. It was found that after 30 min of heating, the secondary structure was not affected. BSZ7 has no inhibiting effect on nonspecific protease originated from malt, suggesting its negative role in wort filterability was accomplished by other means. Furthermore, the glycation of BSZ7 by the Maillard reaction may make some contribution to its survival during wort boiling. The interaction of BSZ7 with polysaccharides and polyphenols found by adding experiment may explain how it acts as a negative factor on wort filterability. Greater understanding of BSZ7 and other proteins of malts will lead to better improvements in brewing quality.

  6. Pearling barley to alter the composition of the raw material before brewing

    NARCIS (Netherlands)

    Donkelaar, van L.H.G.; Noordman, T.R.; Boom, R.M.; Goot, van der A.J.

    2015-01-01

    Partly replacing malt with unmalted barley is a trend in brewing. The use of unmalted barley, however, leads to issues such as haze and high mash viscosity, due to its higher content of undesired components. Pearling, an abrasive method to remove the outer layers of the barley kernels has been shown

  7. 7 CFR 457.102 - Wheat or barley winter coverage endorsement.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 6 2010-01-01 2010-01-01 false Wheat or barley winter coverage endorsement. 457.102... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.102 Wheat or barley... Wheat or Barley Winter Coverage Endorsement (This is a continuous endorsement) 1. In return for...

  8. Products based on a high fiber barley genotype, but not on common barley or oats, lower postprandial glucose and insulin responses in healthy humans.

    Science.gov (United States)

    Liljeberg, H G; Granfeldt, Y E; Björck, I M

    1996-02-01

    Postprandial blood glucose and insulin responses to cereal products made from common barley, oats or a barley genotype containing elevated levels of beta-glucans were evaluated in nine healthy subjects. Porridges were made from commercial Swedish whole-meal barley or oat flours, and a mixed whole-meal porridge using the high fiber barley genotype and commercial Swedish common barley (50:50). Also studied were two types of flour-based bread products composed of high fiber barley and common barley in ratios of 50:50 or 80:20, respectively. The common oat and barley porridges produced postprandial glucose and insulin responses similar to the white wheat bread reference, suggesting that the naturally occurring dietary fiber in these whole-meal flours has no impact on the glucose tolerance. In contrast, all high fiber barley products induced significantly lower responses than did the reference product, with the glycemic and insulin indices ranging from 57 to 72 or 42 to 72%, respectively. It is concluded that "lente" products of high sensory quality can be prepared from a barley genotype with an elevated content of soluble dietary fiber. The glycemic index of these products compares favorably with that of products made from common cereals, suggesting their use as a potential component of diets for patients with diabetes and hyperlipidemia, and for individuals predisposed to metabolic disease.

  9. The role of root hairs in cadmium acquisition by barley

    Energy Technology Data Exchange (ETDEWEB)

    Zheng Ruilun; Li Huafen [Key Laboratory of Plant-Soil Interactions of the Ministry of Education, College of Resources and Environmental Sciences, China Agricultural University, Beijing 100094 (China); Jiang Rongfeng, E-mail: rfjiang@cau.edu.c [Key Laboratory of Plant-Soil Interactions of the Ministry of Education, College of Resources and Environmental Sciences, China Agricultural University, Beijing 100094 (China); Roemheld, Volker [Institute of Plant Nutrition, University of Hohenheim, D-70593 Stuttgart (Germany); Zhang Fusuo [Key Laboratory of Plant-Soil Interactions of the Ministry of Education, College of Resources and Environmental Sciences, China Agricultural University, Beijing 100094 (China); Zhao Fangjie [Soil Science Department, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ (United Kingdom)

    2011-02-15

    The role of root hairs in Cd acquisition from soil was investigated in three pot experiments using a root hairless mutant (bald root barley, brb) and its wild-type (WT) cultivar of barley (Hordeum vulgare). brb had significantly lower concentrations and lower total amounts of Cd in shoots than WT. The Cd uptake efficiency based on total root length was 8-45% lower in brb than in WT. The difference between brb and WT increased with increasing extractable Cd in soil under the experimental conditions used. Additions of phosphate to soil decreased Cd extractability. Both soil and foliar additions of phosphate decreased root length, and root hair formation in WT. These effects resulted in decreased Cd uptake with increasing P supply. Cd uptake in WT correlated significantly with root length, root hair length and density, and soil extractable Cd. Root hairs contribute significantly to Cd uptake by barley. - Research highlights: The Cd uptake efficiency was significantly lower in brb than in WT. Additions of phosphate to soil decreased Cd extractability and Cd uptake. Both soil and foliar additions of phosphate decreased root length, and root hair formation in WT. Root hairs contribute significantly to Cd uptake by barley. - The Cd uptake efficiency based on total root length was 8-45% lower in a barley root hairless mutant than in its wild-type, indicating an important role of root hairs in Cd acquisition.

  10. Structure and expression of phosphoglucan phosphatase genes of Like Sex Four1 and Like Sex Four2 in barley.

    Science.gov (United States)

    Ma, Jian; Gao, Shang; Jiang, Qian-Tao; Yang, Qiang; Sun, Min; Wang, Ji-Rui; Qi, Peng-Fei; Liu, Ya-Xi; Li, Wei; Pu, Zhi-En; Lan, Xiu-Jin; Wei, Yu-Ming; Liu, Chunji; Zheng, You-Liang

    2016-06-01

    Phosphoglucan phosphatases (Like-SEX4 1 and 2; LSF1 and LSF2) were reported to play roles in starch metabolism in leaves of Arabidopsis. In this study, we identified and mapped the LSF1 and LSF2 genes in barley (HvLSF1 and HvLSF2), characterized their gene and protein structures, predicted the cis-elements of their promoters, and analysed their expression patterns. HvLSF1 and HvLSF2 were mapped on the long arm of chromosome 1H (1HL) and 5H (5HL), respectively. Our results revealed varied exon-intron structures and conserved exon-intron junctions in both LSF1 and LSF2 from a range of analysed species. Alignment of protein sequences indicated that cTP and CT domains are much less varied than the functional domains (PDZ, DPS and CBM48). LSF2 was mainly expressed in anthers of barley and rice, and in leaf of Arabidopsis. LSF1 was mainly expressed in endosperm of barley and leaf of Arabidopsis and rice. The expression of LSF1 exhibited a diurnal pattern in rice only and that of LSF2 in both rice and Arabidopsis. Of the investigated stresses, only cold stress significantly reduced expression level of LSF1 and LSF2 in barley and LSF2 in Arabidopsis at late stages of the treatments. While heat treatment significantly decreased expression levels of LSF1 at middle stage (4 h) of a treatment in Arabidopsis only. The strong relationships detected between LSF2 and starch excess4 (SEX4), glucan, water dikinases or phosphoglucan, water dikinases were identified and discussed. Taken together, these results provide information of genetic manipulation of LSF1 and LSF2, especially in monocotyledon and further elucidate their regulatory mechanism in plant development.

  11. Dynamics of phosphorus compounds in ripening and germinating cereal grains. Part II. Changes in phosphorus compounds content during germination of wheat, barley and rye grains

    Directory of Open Access Journals (Sweden)

    J. Tłuczkiewicz

    2015-01-01

    Full Text Available The dynamics of phosphorus compounds was investigated in the embryos and endosperm of wheat (Grana, barley (Kosmos and rye (Pancerne grain after 2, 4 and 6 days of germination. In germinating grain the phosphorus compounds transformations ran along the way of synthesis in the embryo and of breakdown in the endosperm. The level of phosphorus: inorganic, nucleotides, saccharides esters and their metabolites, lipid, RNA, DNA, phytin free and phytin bound with proteins and proteins rapidly increased in germinating seed embryos. The content of phosphorus free phytin and that bound with proteins, lipids, RNA and DNA in the endosperm rapidly decreased during germination. On the other hand, the saccharides phosphorus and that of their metabolites did not change, and inorganic, nuclotides and proteins-P slightly increased. Germinating rye grains showed a higher rate of synthesis in the embryos and of breakdown in the endosperm of the investigated phosphorus compounds than wheat and barley grains.

  12. Allelic barley MLA immune receptors recognize sequence-unrelated avirulence effectors of the powdery mildew pathogen.

    Science.gov (United States)

    Lu, Xunli; Kracher, Barbara; Saur, Isabel M L; Bauer, Saskia; Ellwood, Simon R; Wise, Roger; Yaeno, Takashi; Maekawa, Takaki; Schulze-Lefert, Paul

    2016-10-18

    Disease-resistance genes encoding intracellular nucleotide-binding domain and leucine-rich repeat proteins (NLRs) are key components of the plant innate immune system and typically detect the presence of isolate-specific avirulence (AVR) effectors from pathogens. NLR genes define the fastest-evolving gene family of flowering plants and are often arranged in gene clusters containing multiple paralogs, contributing to copy number and allele-specific NLR variation within a host species. Barley mildew resistance locus a (Mla) has been subject to extensive functional diversification, resulting in allelic resistance specificities each recognizing a cognate, but largely unidentified, AVRa gene of the powdery mildew fungus, Blumeria graminis f. sp. hordei (Bgh). We applied a transcriptome-wide association study among 17 Bgh isolates containing different AVRa genes and identified AVRa1 and AVRa13, encoding candidate-secreted effectors recognized by Mla1 and Mla13 alleles, respectively. Transient expression of the effector genes in barley leaves or protoplasts was sufficient to trigger Mla1 or Mla13 allele-specific cell death, a hallmark of NLR receptor-mediated immunity. AVRa1 and AVRa13 are phylogenetically unrelated, demonstrating that certain allelic MLA receptors evolved to recognize sequence-unrelated effectors. They are ancient effectors because corresponding loci are present in wheat powdery mildew. AVRA1 recognition by barley MLA1 is retained in transgenic Arabidopsis, indicating that AVRA1 directly binds MLA1 or that its recognition involves an evolutionarily conserved host target of AVRA1 Furthermore, analysis of transcriptome-wide sequence variation among the Bgh isolates provides evidence for Bgh population structure that is partially linked to geographic isolation.

  13. MARKER ASSISTED SELECTION (MAS FOR DEVELOPMENT OF BARLEY AND WHEAT LINES WITH REQUESTED TRAITS

    Directory of Open Access Journals (Sweden)

    M. Hudcovicová

    2008-09-01

    Full Text Available Molecular markers closely linked to interesting genes enable early, proper and fast detection of plant individuals with desired allele during backcross breeding, what can make plant breeding faster and cheaper. We are focused on molecular breeding of barley and wheat lines for disease resistance and some important quality traits. As acceptors of interesting genes we use especially elite Slovak and Czech cultivars and lines. After five backcross generations with the help of MAS new created lines carrying markers linked to desired genes undergo resistance, agronomic and technological tests. In breeding of winter barley for resistance to BaYMV/BaMMV viruses we use codominant STS and SSR markers linked to rym4 and rym11 resistance genes. Cultivar Romanze has been used as a donor of rym4 gene and landrace Russia57 as gene rym11 donor. In spring barley we are focused on transfer of Yd2 gene from landraces Shannon and Sutter resistant to BYDV by use of dominant ASPCR marker. We are also working on transfer of effective leaf rust resistance genes Lr19, Lr24 derived from Thinopyrum ponticum and gene Lr35 from Aegilops speltoides into hexaploid wheat by use of dominant STS and SCAR markers. Near isogenic lines with these genes are used in gene pyramiding to develop a single line with all three genes. By use of protein markers we develop near isogenic wheat lines for higher sedimentation values, higher dough strength and better breadmaking quality. These are lines with new combination of HMW glutenin subunits (21*, 7+8, 5+10 as well as wheat lines with new unknown HMW-GS and with new HMW-GS pair.

  14. A major barley allergen associated with baker's asthma disease is a glycosylated monomeric inhibitor of insect alpha-amylase: cDNA cloning and chromosomal location of the gene.

    Science.gov (United States)

    Mena, M; Sanchez-Monge, R; Gomez, L; Salcedo, G; Carbonero, P

    1992-11-01

    A 14.5 kDa barley endosperm protein that is a major allergen in baker's asthma disease, as previously shown by both in vitro (IgE binding) and in vivo tests, has been identified as a glycosylated monomeric member of the multigene family of inhibitors of alpha-amylase/trypsin from cereals. A cDNA encoding this allergen (renamed BMAI-1) has been isolated and characterized. The deduced sequence for the mature protein, which is 132 residues long, is identical in its N-terminal end to the 20 amino acid partial sequence previously determined from the purified allergen, and fully confirms that it is a member of the multigene family of cereal inhibitors. Southern-blot analysis of wheat/barley addition lines using the insert in the BMAI-1 cDNA clone as a probe, has led to the location of the allergen gene (Iam1) in barley chromosome 2, while another related member of this protein family, the barley dimeric alpha-amylase inhibitor BDAI-1 gene (Iad1) has been located in chromosome 6. Iam1 is the first member of this inhibitor family in cereals to be assigned to chromosome group 2, thus extending the dispersion of genes in the family to five out of the seven homology groups of chromosomes in wheat and barley (chromosomes 2, 3, 4, 6 and 7).

  15. 大麦发芽过程中蛋白质组的变化研究%Study on changes of barley proteome during the malting process

    Institute of Scientific and Technical Information of China (English)

    刘宝祥; 朴永哲; 翟明昌; 董亮; 郭敏利; 周华成; 赵长新

    2013-01-01

    通过双向电泳技术监测大麦发芽过程中水溶蛋白质组的变化,明确大麦发芽过程中蛋白质组的变化规律,为麦芽制造提供一定理论依据.以澳麦“schooner”为研究对象,提取其水溶蛋白进行双向电泳分析,结合分析软件监测大麦发芽过程中蛋白质组的变化.结果显示,本实验检测出大麦种子中大约有804种蛋白质,发芽过程中有424种蛋白质未发生变化,379种蛋白质降解消失或浓度降低,另有77种新的蛋白质产生.结果表明,进一步证实浸麦阶段主要是蛋白质的降解过程;发芽前期蛋白质的降解和合成转化较为缓和,发芽中期(48~72h)蛋白质的降解最为旺盛;发芽后期蛋白质的变化基本趋于平衡.%Study on the change of barley proteome during the malting process by two-dimensional electrophoresis,providing a theory of malting.In this study,Australian barley-" Schooner" was chosen.Two-dimensional electrophoresis and analysis software were used to survey the change of water-soluble barley proteome during the malting process.The results showed that during the malting process,only less than 424 kinds of protein retained,while more than 379 kinds of protein disappeared.And 77 kinds of new protein appeared,compared with the 804 kinds of protein in barley.The results showed that barley protein mainly resolved in steeping process.The solution and synthetic transformation of barley protein changed small at the start of germination.And protein resolved the most clearly in the middle of the germination(48~72h).So it was a gradual process of equilibrium.Later of it,protein mostly did not change.

  16. Effects of n-butanol on barley microspore embryogenesis

    DEFF Research Database (Denmark)

    Castillo, Ana Maria; Nielsen, Nanna; Jensen, Anni;

    2014-01-01

    Doubled haploid (DH) production is an efficient tool in barley breeding, but efficiency of DH methods is not consistent. Hence, the aim of this study was to study the effect of n-butanol application on DH barley plant production efficiency. Five elite cultivars of barley and thirteen breeding...... crosses with different microspore embryogenesis capacities were selected for n-butanol application in anther and isolated microspore cultures. Application of 0.1 % n-butanol after a mannitol stress treatment in anther culture significantly increased the number of embryos (up to almost twice) and green...... plants (from 1.7 to 3 times) in three low-responding cultivars: Albacete, Astoria and Majestic. No significant differences on microspore embryogenesis efficiency were observed in medium and high responding cultivars. The application of n-butanol treatment to isolated microspores from cold treated spikes...

  17. Oligosaccharide binding to barley alpha-amylase 1

    DEFF Research Database (Denmark)

    Robert, X.; Haser, R.; Mori, H.;

    2005-01-01

    Enzymatic subsite mapping earlier predicted 10 binding subsites in the active site substrate binding cleft of barley alpha-amylase isozymes. The three-dimensional structures of the oligosaccharide complexes with barley alpha-amylase isozyme 1 (AMY1) described here give for the first time a thorough...... insight into the substrate binding by describing residues defining 9 subsites, namely -7 through +2. These structures support that the pseudotetrasaccharide inhibitor acarbose is hydrolyzed by the active enzymes. Moreover, sugar binding was observed to the starch granule-binding site previously determined...... in barley alpha-amylase isozyme 2 (AMY2), and the sugar binding modes are compared between the two isozymes. The "sugar tongs" surface binding site discovered in the AMY1-thio-DP4 complex is confirmed in the present work. A site that putatively serves as an entrance for the substrate to the active site...

  18. Factors underlying restricted crossover localization in barley meiosis.

    Science.gov (United States)

    Higgins, James D; Osman, Kim; Jones, Gareth H; Franklin, F Chris H

    2014-01-01

    Meiotic recombination results in the formation of cytological structures known as chiasmata at the sites of genetic crossovers (COs). The formation of at least one chiasma/CO between homologous chromosome pairs is essential for accurate chromosome segregation at the first meiotic division as well as for generating genetic variation. Although DNA double-strand breaks, which initiate recombination, are widely distributed along the chromosomes, this is not necessarily reflected in the chiasma distribution. In many species there is a tendency for chiasmata to be distributed in favored regions along the chromosomes, whereas in others, such as barley and some other grasses, chiasma localization is extremely pronounced. Localization of chiasma to the distal regions of barley chromosomes restricts the genetic variation available to breeders. Studies reviewed herein are beginning to provide an explanation for chiasma localization in barley. Moreover, they suggest a potential route to manipulating chiasma distribution that could be of value to plant breeders.

  19. Gene Targeting Without DSB Induction Is Inefficient in Barley.

    Science.gov (United States)

    Horvath, Mihaly; Steinbiss, Hans-Henning; Reiss, Bernd

    2016-01-01

    Double strand-break (DSB) induction allowed efficient gene targeting in barley (Hordeum vulgare), but little is known about efficiencies in its absence. To obtain such data, an assay system based on the acetolactate synthase (ALS) gene was established, a target gene which had been used previously in rice and Arabidopsis thaliana. Expression of recombinases RAD51 and RAD54 had been shown to improve gene targeting in A. thaliana and positive-negative (P-N) selection allows the routine production of targeted mutants without DSB induction in rice. We implemented these approaches in barley and analysed gene targeting with the ALS gene in wild type and RAD51 and RAD54 transgenic lines. In addition, P-N selection was tested. In contrast to the high gene targeting efficiencies obtained in the absence of DSB induction in A. thaliana or rice, not one single gene targeting event was obtained in barley. These data suggest that gene targeting efficiencies are very low in barley and can substantially differ between different plants, even at the same target locus. They also suggest that the amount of labour and time would become unreasonably high to use these methods as a tool in routine applications. This is particularly true since DSB induction offers efficient alternatives. Barley, unlike rice and A. thaliana has a large, complex genome, suggesting that genome size or complexity could be the reason for the low efficiencies. We discuss to what extent transformation methods, genome size or genome complexity could contribute to the striking differences in the gene targeting efficiencies between barley, rice and A. thaliana.

  20. PHYSIOLOGICAL AND AGROECOLOGICAL ASPECTS OF CADMIUM INTERACTIONS WITH BARLEY PLANTS: AN OVERVIEW

    Directory of Open Access Journals (Sweden)

    A VASSILEV

    2003-07-01

    Full Text Available This work is a review of author’s previous publications, unpublished results as well as available literature on barley responses to Cd contamination. The physiological backgrounds of the acute Cd toxicity in barley plants are briefly described. Some data characterizing the chronic Cd toxicity in barley have been also provided in relation to its possible use for seed production and Cd phytoextraction on Cd-contaminated agricultural soils. Information about the main physiological factors limiting growth of Cd-exposed barley plants and grain yield, seedling quality as well as Cd phytoextraction capacity of barley grown in Cd-contaminated soils is presented.

  1. Genetics of Two Storage Protein Variants in Barley

    DEFF Research Database (Denmark)

    Oram, R. N.; Doll, Hans; Køie, B.

    1975-01-01

    The Bomi and Sultan varieties have different co-dominant alleles at one locus on chromosome 5 controlling the electrophoretic pattern of one group of hordeins. This locus is hypostatic to the recessive allele in Risø mutant 1508, which suppresses the formation of these and some other hordeins. Th...

  2. The transfer of {sup 137}Cs from barley to beer

    Energy Technology Data Exchange (ETDEWEB)

    Proehl, G.; Mueller, H.; Voigt, G. [Institut fuer Strahlenschutz, Oberschleibheim (Germany)] [and others

    1997-01-01

    Beer has been brewed from barley contaminated with {sup 137}Cs as a consequence of the Chernobyl accident. The {sup 137}Cs activity has been measured in all intermediate steps and in the by-products of the production process. About 35 % of the {sup 137}Cs in barley were recovered in beer. Processing factors defined as the concentration ratio of processed and raw products were determined to be 0.61, 3.3, 0.1 and 0.11 for malt, malt germs, spent grains and beer, respectively. 4 refs., 2 tabs.

  3. Implementation of biochemical screening to improve baking quality of Barley

    DEFF Research Database (Denmark)

    Aaslo, Per; Langkilde, Ane; Dionisio, Giuseppe

    2011-01-01

    Barley (Hordeum vulgare) is mostly used in feed and malt production but has the ability to provide humans nutritional benefits. The current wheat based “barley” breads can unfortunately not exceed more than 20% barley flour mixed into the dough due to poor leavening properties. Therefore...... the opportunity to give a forecast of the taste of the bread, as the AA composition is known to control certain aspects of the taste. We uses a MSE approach on a time of flight instrument coupled to a UPLC and in gel digestion to identify and characterize the different D-hordeins responsible for baking quality...

  4. Identification and characterization of barley RNA-directed RNA polymerases

    DEFF Research Database (Denmark)

    Madsen, Christian Toft; Stephens, Jennifer; Hornyik, Csaba

    2009-01-01

    in dicot species. In this report, we identi!ed and characterized HvRDR1, HvRDR2 and HvRDR6 genes in the monocot plant barley (Hordeum vulgare). We analysed their expression under various biotic and abiotic stresses including fungal and viral infections, salicylic acid treatment as well as during plant...... development. The different classes and subclasses of barley RDRs displayed contrasting expression patterns during pathogen challenge and development suggesting their involvement in speci!c regulatory pathways. Their response to heat and salicylic acid treatment suggests a conserved pattern of expression...

  5. 6-Plex microsphere immunoassay with imaging planar array detection for mycotoxins in barley.

    Science.gov (United States)

    Peters, Jeroen; Cardall, Alice; Haasnoot, Willem; Nielen, Michel W F

    2014-08-21

    Mycotoxins are produced by fungi as secondary metabolites. They often multi-contaminate food and feed commodities posing a health risk to humans and animals. A fast and easy to apply multiplex screening of these commodities could be useful to detect multi-contamination. For this, we developed a semi-quantitative 6-plex immunoassay using a suspension array of paramagnetic colour-coded microspheres combined with imaging planar array detection for the mycotoxins aflatoxin B1, ochratoxin A, zearalenone, deoxynivalenol, T2-toxin, HT-2 toxin and fumonisin B1. Mycotoxin specific monoclonal antibodies were coupled to different sets of microspheres and mycotoxins conjugated to the fluorescent protein R-phycoerythrin served as reporter molecules. Competition between free mycotoxins in the sample and mixed reporter molecules for antibody binding sites on mixed microspheres created a multiplex direct inhibition immunoassay. The reagents were selected for no or low cross-interactions between the assays and cross-reactions with metabolites and possible masked forms were determined. A within-laboratory validation was carried out using blank and spiked barley samples. Furthermore, the 6-plex was used to screen available barley, and malted barley, reference materials. The validation showed very high inter and intra-day precision for all samples with a maximum relative standard deviation value of 10%. The screening assay allows easy and rapid multiplex detection of the target mycotoxins in barley according to EU legislation. With a cut off factor of 50%, based on the EU maximum levels, we were able to screen at 2 μg kg(-1) for aflatoxin B1, 2.5 μg kg(-1) for ochratoxin A, 625 μg kg(-1) for deoxynivalenol, 50 μg kg(-1) for zearalenone, 1000 μg kg(-1) for fumonisin B1 and 25 μg kg(-1) for T-2 toxin. Thanks to the transportable planar array system, the developed 6-plex has potential for future on-site testing. Future implementation of this method as a pre-screening tool, prior to

  6. Overexpression, purification and enzymatic characterization of a recombinant plastidial glucose-6-phosphate dehydrogenase from barley (Hordeum vulgare cv. Nure) roots.

    Science.gov (United States)

    Cardi, Manuela; Chibani, Kamel; Castiglia, Daniela; Cafasso, Donata; Pizzo, Elio; Rouhier, Nicolas; Jacquot, Jean-Pierre; Esposito, Sergio

    2013-12-01

    In plant cells, the plastidial glucose 6-phosphate dehydrogenase (P2-G6PDH, EC 1.1.1.49) represents one of the most important sources of NADPH. However, previous studies revealed that both native and recombinant purified P2-G6PDHs show a great instability and a rapid loss of catalytic activity. Therefore it has been difficult to describe accurately the catalytic and physico-chemical properties of these isoforms. The plastidial G6PDH encoding sequence from barley roots (Hordeum vulgare cv. Nure), devoid of a long plastidial transit peptide, was expressed as recombinant protein in Escherichia coli, either untagged or with an N-terminal his-tag. After purification from both the soluble fraction and inclusion bodies, we have explored its kinetic parameters, as well as its sensitivity to reduction. The obtained results are consistent with values determined for other P2-G6PDHs previously purified from barley roots and from other land plants. Overall, these data shed light on the catalytic mechanism of plant P2-G6PDH, summarized in a proposed model in which the sequential mechanism is very similar to the mammalian cytosolic G6PDH. This study provides a rational basis to consider the recombinant barley root P2-G6PDH as a good model for further kinetic and structural studies.

  7. True-breeding targeted gene knock-out in barley using designer TALE-nuclease in haploid cells.

    Directory of Open Access Journals (Sweden)

    Maia Gurushidze

    Full Text Available Transcription activator-like effector nucleases (TALENs are customizable fusion proteins able to cleave virtually any genomic DNA sequence of choice, and thereby to generate site-directed genetic modifications in a wide range of cells and organisms. In the present study, we expressed TALENs in pollen-derived, regenerable cells to establish the generation of instantly true-breeding mutant plants. A gfp-specific TALEN pair was expressed via Agrobacterium-mediated transformation in embryogenic pollen of transgenic barley harboring a functional copy of gfp. Thanks to the haploid nature of the target cells, knock-out mutations were readily detected, and homozygous primary mutant plants obtained following genome duplication. In all, 22% of the TALEN transgenics proved knocked out with respect to gfp, and the loss of function could be ascribed to the deletions of between four and 36 nucleotides in length. The altered gfp alleles were transmitted normally through meiosis, and the knock-out phenotype was consistently shown by the offspring of two independent mutants. Thus, here we describe the efficient production of TALEN-mediated gene knock-outs in barley that are instantaneously homozygous and non-chimeric in regard to the site-directed mutations induced. This TALEN approach has broad applicability for both elucidating gene function and tailoring the phenotype of barley and other crop species.

  8. Isolation and characterization of Phi class glutathione transferase partial gene from Iranian barley

    Directory of Open Access Journals (Sweden)

    Hassan Mohabatkar

    2012-01-01

    Full Text Available Glutathione transferases are multifunctional proteins involved in several diverse intracellular events such as primary and secondary metabolisms, signaling and stress metabolism. These enzymes have been subdivided into eight classes in plants. The Phi class, being plant specific, is the most represented. In the present study, based on the sequences available at GenBank, different primers were designed for amplifying the Phi class of glutathione transferase gene in the genome and transcriptome of Iranian barley, Karoun cultivar. After extraction of DNA and total RNA, Phi class was amplified and sequenced. Bioinformatics analysis predicted that the deduced protein sequence has two ß-sheets, eight α-helices and some intermediate loops in its secondary structure. Consequently, the sequences were submitted to NCBI GenBank with GS262333 and GW342614 accession numbers. Phylogenic relationships of the sequences were compared with existing sequences in GenBank.

  9. Replacement of Dietary Barley Grain by Different Levels of Restaurant Waste and Its Effect on Hybrid Lambs Performance

    Directory of Open Access Journals (Sweden)

    M. Moradi

    2013-08-01

    Full Text Available This study was conducted to determine the nutritive value of restaurant waste (RW, substituted with dietary barley grain and its effects on the performance of finishing lambs. Dry matter, organic matter, crude protein, ether extract and ash content of RW were 33.4, 95.9, 15.1, 14.1 and 4 percent respectively. 36 male and female lambs, (initial weight of 33.4± 0.5 and 29.7± 0.5 kg respectively were used in the experiment. The experimental lambs were from three hybrid groups: Ghezel*Merino (n=12, Merino*Moghani (n=18 and Ghezel*Baluchi (n=6. Dietary barley grains at the levels of 50 and 100 percent were replaced with RW and along with control group (no RW compromised experimental treatments.. Experimental diets were offered three times daily at 6.00, 14.00 and 20.00 hours. Dry matter intake was not significantly different between the treatments. The average daily gain and feed conversion ratio were differ between sexes. Weight gain of male and female lambs during the fattening period was 250.3 and 171.6 g/day respectively. Replacement of barley grain with RW in the 3rd treatment significantly affected ruminal pH, N-NH3 and total volatile fatty acids content and fecal pH, as well as blood glucose and BUN comparing the control group. Replacement of RW with barley grain at levels of 50 and 100 percent reduced cost of the live weight gain up to 24 and 37.7 percent respectively versus control diet.

  10. The Quantum Mixed-Spin Heme State of Barley Peroxidase: A Paradigm for Class III Peroxidases

    Energy Technology Data Exchange (ETDEWEB)

    Howes, B.D.; Ma, J.; Marzocchi, M.P.; Schiodt, C.B.; Shelnutt, J.A.; Smulevich, G.; Welinder, K.G.; Zhang, J.

    1999-03-23

    Electronic absorption and resonance Raman (RR) spectra of the ferric form of barley grain peroxidase (BP 1) at various pH values both at room temperature and 20 K are . reported, together with EPR spectra at 10 K. The ferrous forms and the ferric complex with fluoride have also been studied. A quantum mechanically mixed-spin (QS) state has been identified. The QS heme species co-exists with 6- and 5-cHS heroes; the relative populations of these three spin states are found to be dependent on pH and temperature. However, the QS species remains in all cases the dominant heme spin species. Barley peroxidase appears to be further characterized by a splitting of the two vinyl stretching modes, indicating that the vinyl groups are differently conjugated with the porphyrin. An analysis of the presently available spectroscopic data for proteins from all three peroxidase classes suggests that the simultaneous occurrence of the QS heme state as well as the splitting of the two vinyl stretching modes is confined to class III enzymes. The former point is discussed in terms of the possible influences of heme deformations on heme spin state. It is found that moderate saddling alone is probably not enough to cause the QS state, although some saddling maybe necessary for the QS state.

  11. QTL analysis for Malt Quality in DH Lines of Barley (Steptoe × Morex grown in Iran

    Directory of Open Access Journals (Sweden)

    Mohammad Bahman Sadeghi

    2013-12-01

    Full Text Available In order to study the quantitative variability of malting quality-related traits and to determine the genomic locations which control these traits, an experiment was conducted using one hundred fifty double haploid (DH lines of barley and their two parents ‛Steptoe’ & ‛Morex’. Protein content (% was measured using Kjeldahl method, diastatic power calculated with Lintner formula, Malt extract was measured for each by special weight achieved and based on Malt Berix Charts. Transgressive segregation in both directions was observed for all traits. Genetic map is fairly saturated and comprising 327 RFLP markers with a total length of 1226.3cM with an average marker spacing of 3.75cM. Seventeen QTLs by LOD≤2(LRS≤9.21 controlling different studied traits were identified for all studied traits. Total phenotypic variance explained by these QTLs varies from 23.2 to 45.05%. Highest LOD scores were obtained for QTL’s controlling diastatic power (Qdip3Ha on chromosome 3H, and lowest LOD scores were obtained for QTL’s controlling seed yield per plant (Qsyp1Hb on chromosome 1H. Therefore gain through marker-assisted selection (MAS in this population would be limited and some of the “Steptoe× Morex” population was developed with the intention of isolating and advancing barley lines for release to the malting quality.

  12. Shining light on the differences in molecular structural chemical makeup and the cause of distinct degradation behavior between malting- and feed-type barley using synchrotron FTIR microspectroscopy: a novel approach.

    Science.gov (United States)

    Yu, Peiqiang; Doiron, Kevin; Liu, Dasen

    2008-05-14

    The objective of this study was to use advanced synchrotron-sourced FTIR microspectroscopy (SFTIRM) as a novel approach to identify the differences in protein and carbohydrate molecular structure (chemical makeup) between these two varieties of barley and illustrate the exact causes for their significantly different degradation kinetics. Items assessed included (1) molecular structural differences in protein amide I to amide II intensities and their ratio within cellular dimensions, (2) molecular structural differences in protein secondary structure profile and their ratios, and (3) molecular structural differences in carbohydrate component peak profile. Our hypothesis was that molecular structure (chemical makeup) affects barley quality, fermentation, and degradation behavior in both humans and animals. Using SFTIRM, the protein and carbohydrate molecular structural chemical makeup of barley was revealed and identified. The protein molecular structural chemical makeup differed significantly between the two varieties of barleys. No difference in carbohydrate molecular structural chemical makeup was detected. Harrington was lower than Valier in protein amide I, amide II, and protein amide I to amide II ratio, while Harrington was relatively higher in model-fitted protein alpha-helix and beta-sheet, but lower in the others (beta-turn and random coil). These results indicated that it is the molecular structure of protein (chemical makeup) that may play a major role in the different degradation kinetics between the two varieties of barleys (not the molecular structure of carbohydrate). It is believed that use of the advanced synchrotron technology will make a significant step and an important contribution to research in examining the molecular structure (chemical makeup) of plant, feed, and seeds.

  13. Shining Light on the Differences in Molecular Structural Chemical Makeup and the Cause of Distinct Degradation Behavior Between Malting- and Feed- Type Barley Using Synchrotorn FTIR Microspectroscopy: A Novel Approach

    Energy Technology Data Exchange (ETDEWEB)

    Yu,P.; Doiron, K.; Liu, D.

    2008-01-01

    The objective of this study was to use advanced synchrotron-sourced FTIR microspectroscopy (SFTIRM) as a novel approach to identify the differences in protein and carbohydrate molecular structure (chemical makeup) between these two varieties of barley and illustrate the exact causes for their significantly different degradation kinetics. Items assessed included (1) molecular structural differences in protein amide I to amide II intensities and their ratio within cellular dimensions, (2) molecular structural differences in protein secondary structure profile and their ratios, and (3) molecular structural differences in carbohydrate component peak profile. Our hypothesis was that molecular structure (chemical makeup) affects barley quality, fermentation, and degradation behavior in both humans and animals. Using SFTIRM, the protein and carbohydrate molecular structural chemical makeup of barley was revealed and identified. The protein molecular structural chemical makeup differed significantly between the two varieties of barleys. No difference in carbohydrate molecular structural chemical makeup was detected. Harrington was lower than Valier in protein amide I, amide II, and protein amide I to amide II ratio, while Harrington was relatively higher in model-fitted protein a-helix and {beta}-sheet, but lower in the others ({beta}-turn and random coil). These results indicated that it is the molecular structure of protein (chemical makeup) that may play a major role in the different degradation kinetics between the two varieties of barleys (not the molecular structure of carbohydrate). It is believed that use of the advanced synchrotron technology will make a significant step and an important contribution to research in examining the molecular structure (chemical makeup) of plant, feed, and seeds.

  14. A weed suppressive index for spring barley (Hordeum vulgare) varieties

    DEFF Research Database (Denmark)

    Hansen, P K; Kristensen, K; Willas, J

    2008-01-01

    A screening programme for crop variety competitiveness would ideally be based on only a few, non-destructive measurements of key growth traits. In this study we measured the weed suppressive ability of 79 varieties of spring barley in two ways: (i) directly, by weed coverage assessments under wee...

  15. Zinc biofortification of cereals: rice differs from wheat and barley

    NARCIS (Netherlands)

    Stomph, T.J.; Jiang, W.; Struik, P.C.

    2009-01-01

    In their review, mainly focused on bread wheat (Triticum aestivum), durum wheat (Triticum durum) and barley (Hordeum vulgare), Palmgren et al. 1 M.G. Palmgren et al., Zinc biofortification of cereals: problems and solutions, Trends Plant Sci. 13 (2008), pp. 464–473. Article | PDF (905 K) | View Reco

  16. Evaluation of a malting barley quality assessment system

    NARCIS (Netherlands)

    Lonkhuijsen, H.J. van; Douma, A.C.; Angelino, S.A.G.F.

    1998-01-01

    New malting barley varieties are annually tested for their malting and brewing potential according to a field trial set-up combined with quality evaluation on pilot scale. To assess the effects of trial year and location on quality evaluation data, a data base consisting of quality data from Dutch m

  17. Radiation hybrid map of barley chromosome 3H

    Science.gov (United States)

    Assembly of the barley genome is complicated by its large size (5.1 Gb) and proportion of repetitive elements (84%). This process is facilitated by high resolution maps for aligning BAC contigs along chromosomes. Available genetic maps; however, do not provide accurate information on the physical po...

  18. Registration of Harriman low-phytate, hulled spring barley

    Science.gov (United States)

    The Agricultural Research Service, U.S. Department of Agriculture (USDA-ARS), has released 'Harriman', (Hordeum vulgare L.) (Reg. No. xxxxxx, P.I. xxxxxx). Harriman is a hulled, low-phytate barley, the second to be developed and released by the USDA-ARS. Compared to the previously released hulled, l...

  19. Reclamation of Sodic-Saline Soils. Barley Crop Response

    Directory of Open Access Journals (Sweden)

    Giovanna Cucci

    2008-12-01

    Full Text Available The research was aimed at assessing the salinity and sodicity effects of two soil types submitted to correction on barley crop. The two soils, contained in cylindrical pots (0.40 m in size and 0.60 m h supplied with a bottom valve for the collection of drainage water and located under shed to prevent the leaching action of rainfall, were clay-textured and saline and sodic-saline at barley seeding, as they had been cultivated for 4 consecutive years with different herbaceous species irrigated with 9 types of brackish water. In 2002-2003 the 2 salinized and sodium-affected soils (ECe and ESP ranging respectively from 5.84-20.27 dSm-1 to 2.83-11.19%, submitted to correction, were cultivated with barley cv Micuccio, and irrigated with fresh water (ECw = 0.5 dS m-1 and SAR = 0.45 whenever 30% of the maximum soil available moisture was lost by evapotranspiration. Barley was shown to be a salt-tolerant species and did not experience any salt stress when grown in soils with an initial ECe up to 11 dS m-1. When it was grown in more saline soils (initial ECe of about 20 dS m-1, despite the correction, it showed a reduction in shoot biomass and kernel yield by 26% and 36% respectively, as compared to less saline soils.

  20. Microgeographic Edaphic Differentiation in Hordein Polymorphisms of Wild Barley

    DEFF Research Database (Denmark)

    Nevo, E.; Beiles, A.; Storch, N.

    1983-01-01

    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....

  1. Wheat and barley differently affect porcine intestinal microbiota

    DEFF Research Database (Denmark)

    Weiss, Eva; Aumiller, Tobias; Spindler, Hanns K

    2016-01-01

    Diet influences the porcine intestinal microbial ecosystem. Barrows were fitted with ileal T-cannulas to compare short-term effects of eight different wheat or barley genotypes and period-to-period effects on seven bacterial groups in ileal digesta and faeces by qPCR. Within genotypes of wheat an...

  2. Classification and salt tolerance analysis of barley varieties

    NARCIS (Netherlands)

    Katerji, N.; Hoorn, van J.W.; Hamdy, A.; Mastrorilli, M.; Fares, C.; Ceccarelli, S.; Grando, S.; Oweis, T.

    2006-01-01

    Six varieties of barley (Hordeum vulgare), five of which were provided by ICARDA, were tested in a green house experiment for their salt tolerance. Afterwards the ICARDA variety Melusine, selected from this experiment for its combination of high yield and salt tolerance, was compared in a lysimeter

  3. Durum wheat and barley productivity in saline-drought environments

    NARCIS (Netherlands)

    Katerji, N.; Mastrorilli, M.; Hoorn, van J.W.; Lahmer, F.Z.; Hamdy, A.; Oweis, T.

    2009-01-01

    In two Successive years, durum wheat (Triticum turgidum Desf.) and barley (Hodeum vulgare L.) were tested in a factorial salinity-drought experiment, combining three levels of salinity and two levels of drought. The two drought treatments were obtained by applying irrigation water when the pre-dawn

  4. Expression of lipoxygenase isoenzymes in developing barley grains

    NARCIS (Netherlands)

    Schmitt, N.F.; Mechelen, J.R. van

    1997-01-01

    Expression of lipoxygenase was studied in whole developing barley grains from 5 days after flowering (DAF) to full maturity. Lipoxygenase showed two distinct peaks of activity. The first peak of activity occurred in the early stages of grain development from 5 until 20 DAF, whereas the second peak o

  5. Biotic stress in barley: disease problems and solutions

    Science.gov (United States)

    Barley (Hordeum vulgare L.) is cultivated over a wider geographic range than almost any other major crop species. It can be found growing from the tropics to the high latitudes and from the seacoast to the highest arable mountaintops. On marginal lands where alkaline soils, drought, or cold summer t...

  6. Wheat and barley seed systems in Ethiopia and Syria

    NARCIS (Netherlands)

    Bishaw, Z.

    2004-01-01

    Keywords: Wheat,Triticumspp., Barley,Hordeumvulgare L., Seed Systems, Formal Seed Sector, Informal Seed Sector, National Seed Program, Seed Source, Seed Selection, Seed Management, Seed Quality,

  7. Wheat and barley exposure to nanoceria: Implications for agricultural productivity

    Science.gov (United States)

    The impacts of man-made nanomaterials on agricultural productivity are not yet well understood. A soil microcosm study was performed to assess the physiological, phenological, and yield responses of wheat (Triticum aestivum) and barley (Hordeum vulgare L.) exposed to nanoceria (n...

  8. Evaluation of Wheat and Barley Cultivars Tolerance to Metribuzine Application

    Directory of Open Access Journals (Sweden)

    E Izadi Darbandi

    2013-08-01

    Full Text Available In order to study of barely and wheat cultivars tolerance to metribuzin, a factorial experiment was conducted as a completely randomized design, with three replications in Greenhouse of Agricultural Research at Ferdowsi University of Mashhad. Treatments included wheat cultivars (Backcross roshan, Cross Arvand, Bahar, Sepahan, Gascosion, Sayonez, Bam garmsiry, Garmsiri, Ghods, Pishtaz, Chamran and Shoori 6, barely cultivars (Macouyi, Karoon and Bahman and metribuzin application rates ( 0, 175, 350, 700, 1050, 1400 and 2100 gr. ai.ha-1. Metribuzine was applied at 3-4 leaf stage and 3 weeks after herbicide spraying, plants survival and their biomass were determined. Results showed that metribuzin application had a significant effect (p≤0.01 on barley and wheat dry weight. Based on results, mertibuzin application did not affect on barley cultivars up to 30 g.a.i.ha-1 but in wheat varieties lead to significant reduction in their biomass and survival. Increasing of metribuzin rates reduced wheat and barley cultivars biomass (p≤0.01. Barely varieties were less sensitive than wheat cultivars to metribuzine. The highest and the lowest ED50 in wheat cultivars were observed in cross arvand (940 and shoori (25 varieties, respectively. In barley cultivars the highest and lowest ED50 were observed in Macouyi (614 and Karoon (396, respectively.

  9. Spatial aggregation of pathotypes of barley powdery mildew

    DEFF Research Database (Denmark)

    O'Hara, R.B.; Brown, J.K.M.

    1997-01-01

    Aggregation in the distribution of pathotypes of Erysiphe graminis f.sp. hordei, the barley powdery mildew pathogen, was investigated in field plots of 'Golden Promise', 'Proctor' and 'Tyra'. 'Golden Promise' and 'Proctor' have no effective mildew resistance alleles, whereas 'Tyra' has Mla1, which...

  10. Dormant barley aleurone shows heterogeneity and a specific cytodifferentiation

    NARCIS (Netherlands)

    Schuurink, R.C.; Bakhuizen, R.; Libbenga, K.R.; Boulanger, F.; Sinjorgo, K.M.C.

    1997-01-01

    In response to gibberellic acid, aleurone layers isolated from dormant barley (Hordeum distichum L. cv. Triumph) kernels produced significantly less alpha-amylase than aleurones from non-dormant kernels. Light microscopical investigations using the dye acridine orange as well as electron microscopic

  11. Allelopathic effects of barley straw on germination and seedling growth of corn, sugar beet and sunflower

    Directory of Open Access Journals (Sweden)

    mohamad taghi naseri poor yazdi

    2009-06-01

    Full Text Available Allelopathic effects of barley straw and root on germination and growth of maize, sugar beet, and sunflower were investigated under glasshouse and laboratory experiments in Faculty of Agriculture, Ferdowsi University of Mashhad in 2006. The glasshouse experiment was designed based on randomized complete block design with three replications, treatments included: 0, 200, 400, 600 g/m² of grounded barley straw and also 0 and 50 g/m2 barley root. A laboratory experiment was carried out in order to study the effect of different concentrations of barley water extracts on germination and seedling characteristics of corn, sugar beet and sunflower. Treatments in laboratory trial included 0, 33, 50 and 100 percent of barley extracts. Results showed that leaf area of corn was significantly affected by barley straw treatments. Shoot dry matter and seed weight per plant in corn , leaf and tuber weight in sugar beet and leaf , stem weights , plant per plant in corn , leaf and tuber weight in sugar beet and leaf, stem weights, plant height, head diameter, head weight and seed weight in sunflower were significantly higher in treatment of 50g/m² barley roots. Crop seed germination decreased with increasing the amount of barley straw. The best germination response to barley extract was observed in corn. Maize radicle weight was significantly decreased with increasing concentration of barley water extract.

  12. iTAG Barley: A 9-12 curriculum to explore inheritance of traits and genes using Oregon Wolfe barley

    Science.gov (United States)

    Segregating plants from the Informative & Spectacular Subset (ISS) of the Oregon Wolfe doubled haploid barley (OWB) population are easily grown on a lighted window bench in the classroom. These lines originate from a wide cross and have exceptionally diverse and dramatic phenotypes, making this an i...

  13. Tocopherols and tocotrienols in barley oil prepared from germ and other fractions from scarification and sieving of hulless barley

    Science.gov (United States)

    Two cultivars of hulless barley (Doyce and Merlin), were scarified to abrade the outer layers of the kernels (germ, pericarp, and aleurone). The resulting scarification fines fractions were then separated into four particle size subfractions using sieves. Each of the size subfractions was then extr...

  14. Dispersed gold nanoparticles potentially ruin gold barley yellow dwarf virus and eliminate virus infectivity hazards

    Science.gov (United States)

    Alkubaisi, Noorah A.; Aref, Nagwa M. A.

    2016-10-01

    Gold nanoparticles (AuNPs) application melted barley yellow dwarf virus-PAV (BYDV-PAV) spherical nanoparticle capsids. Synergistic therapeutic effects for plant virus resistance were induced by interaction with binding units of prepared AuNPs in a water solution which was characterized and evaluated by zeta sizer, zeta potential and transmission electron microscopy (TEM). The yield of purified nanoparticles of BYDV-PAV was obtained from Hordeum vulgare (Barley) cultivars, local and Giza 121/Justo. It was 0.62 mg/ml from 27.30 g of infected leaves at an A260/A280 ratio. Virus nanoparticle has a spherical shape 30 nm in size by TEM. BYDV-PAV combined with AuNPs to challenge virus function in vivo and in vitro. Dual AuNPs existence in vivo and in vitro affected compacted configuration of viral capsid protein in the interior surface of capsomers, the outer surface, or between the interface of coat protein subunits for 24 and 48 h incubation period in vitro at room temperature. The sizes of AuNPs that had a potentially dramatic deteriorated effect are 3.151 and 31.67 nm with a different intensity of 75.3% for the former and 24.7% for the latter, which enhances optical sensing applications to eliminate virus infectivity. Damages of capsid protein due to AuNPs on the surface of virus subunits caused variable performance in four different types of TEM named puffed, deteriorated and decorated, ruined and vanished. Viral yield showed remarkably high-intensity degree of particle symmetry and uniformity in the local cultivar greater than in Giza 121/Justo cultivar. A high yield of ruined VLPs in the local cultivar than Justo cultivar was noticed. AuNPs indicated complete lysed VLPs and some deteriorated VLPs at 48 h.

  15. Dispersed gold nanoparticles potentially ruin gold barley yellow dwarf virus and eliminate virus infectivity hazards

    Science.gov (United States)

    Alkubaisi, Noorah A.; Aref, Nagwa M. A.

    2017-02-01

    Gold nanoparticles (AuNPs) application melted barley yellow dwarf virus-PAV (BYDV-PAV) spherical nanoparticle capsids. Synergistic therapeutic effects for plant virus resistance were induced by interaction with binding units of prepared AuNPs in a water solution which was characterized and evaluated by zeta sizer, zeta potential and transmission electron microscopy (TEM). The yield of purified nanoparticles of BYDV-PAV was obtained from Hordeum vulgare (Barley) cultivars, local and Giza 121/Justo. It was 0.62 mg/ml from 27.30 g of infected leaves at an A260/A280 ratio. Virus nanoparticle has a spherical shape 30 nm in size by TEM. BYDV-PAV combined with AuNPs to challenge virus function in vivo and in vitro. Dual AuNPs existence in vivo and in vitro affected compacted configuration of viral capsid protein in the interior surface of capsomers, the outer surface, or between the interface of coat protein subunits for 24 and 48 h incubation period in vitro at room temperature. The sizes of AuNPs that had a potentially dramatic deteriorated effect are 3.151 and 31.67 nm with a different intensity of 75.3% for the former and 24.7% for the latter, which enhances optical sensing applications to eliminate virus infectivity. Damages of capsid protein due to AuNPs on the surface of virus subunits caused variable performance in four different types of TEM named puffed, deteriorated and decorated, ruined and vanished. Viral yield showed remarkably high-intensity degree of particle symmetry and uniformity in the local cultivar greater than in Giza 121/Justo cultivar. A high yield of ruined VLPs in the local cultivar than Justo cultivar was noticed. AuNPs indicated complete lysed VLPs and some deteriorated VLPs at 48 h.

  16. Diacylglycerol pyrophosphate binds and inhibits the glyceraldehyde-3-phosphate dehydrogenase in barley aleurone.

    Science.gov (United States)

    Astorquiza, Paula Luján; Usorach, Javier; Racagni, Graciela; Villasuso, Ana Laura

    2016-04-01

    The aleurona cell is a model that allows the study of the antagonistic effect of gibberellic acid (GA) and abscisic acid (ABA). Previous results of our laboratory demonstrated the involvement of phospholipids during the response to ABA and GA. ABA modulates the levels of diacylglycerol, phosphatidic acid and diacylglycerol pyrophosphate (DAG, PA, DGPP) through the activities of phosphatidate phosphatases, phospholipase D, diacylglycerol kinase and phosphatidate kinase (PAP, PLD, DGK and PAK). PA and DGPP are key phospholipids in the response to ABA, since both are capable of modifying the hydrolitic activity of the aleurona. Nevertheless, little is known about the mechanism of action of these phospholipids during the ABA signal. DGPP is an anionic phospholipid with a pyrophosphate group attached to diacylglycerol. The ionization of the pyrophosphate group may be important to allow electrostatic interactions between DGPP and proteins. To understand how DGPP mediates cell functions in barley aleurone, we used a DGPP affinity membrane assay to isolate DGPP-binding proteins from Hordeum vulgare, followed by mass spectrometric sequencing. A cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) was identified for being bound to DGPP. To validate our method, the relatively abundant GAPDH was characterized with respect to its lipid-binding properties, by fat western blot. GAPDH antibody interacts with proteins that only bind to DGPP and PA. We also observed that ABA treatment increased GAPDH abundance and enzyme activity. The presence of phospholipids during GAPDH reaction modulated the GAPDH activity in ABA treated aleurone. These data suggest that DGPP binds to GAPDH and this DGPP and GAPDH interaction provides new evidences in the study of DGPP-mediated ABA responses in barley aleurone.

  17. Structural basis for target protein recognition by the protein disulfide reductase thioredoxin

    DEFF Research Database (Denmark)

    Maeda, Kenji; Hägglund, Per; Finnie, Christine;

    2006-01-01

    Thioredoxin is ubiquitous and regulates various target proteins through disulfide bond reduction. We report the structure of thioredoxin (HvTrxh2 from barley) in a reaction intermediate complex with a protein substrate, barley alpha-amylase/subtilisin inhibitor (BASI). The crystal structure...... a major role in the specificity and protein disulfide reductase activity of thioredoxin. This novel insight into the function of thioredoxin constitutes a basis for comprehensive understanding of its biological role. Moreover, comparison with structurally related proteins shows that thioredoxin shares...

  18. GWAS of Barley Phenotypes Established Under Future Climate Conditions of Elevated Temperature, CO2, O3 and Elevated Temperature and CO2 Combined

    DEFF Research Database (Denmark)

    Ingvordsen, Cathrine Heinz; Backes, G.; Lyngkjær, M. F.

    2015-01-01

    Climate change is likely to decrease crop yields worldwide. Developing climate resilient cultivars is one way to combat this production scarcity, however, little is known of crop response to future climate conditions and in particular the variability within crops.In Scandinavia, barley is widely...... cultivated, but yields have stagnated since the start of this century. In this study we cultivated 138 spring barley accessions in a climate phytotron under four treatments mimicking forecasted levels of temperature, carbon dioxide concentration ([CO2]) and ozone ([O3]) at the end of the 21st century1...... yield, grain protein concentration, grain protein harvested, number of grains, number of ears, aboveground vegetative biomass and harvest index. In addition, stability of the production was calculated over the applied treatments for the assessed parameters.In the climate scenario of elevated temperature...

  19. Stress-inducible expression of barley Hva1 gene in transgenic mulberry displays enhanced tolerance against drought, salinity and cold stress.

    Science.gov (United States)

    Checker, Vibha G; Chhibbar, Anju K; Khurana, Paramjit

    2012-10-01

    Coping with different kinds of biotic and abiotic stresses is the foundation of sustainable agriculture. Although conventional breeding and marker-assisted selection are being employed in mulberry (Morus indica L.) to develop better varieties, nonetheless the longer time periods required for these approaches necessitates the use of precise biotechnological approaches for sustainable agriculture. In an attempt to improve stress tolerance of mulberry, an important plant of the sericulture industry, an encoding late embryogenesis abundant gene from barley (HVA1) was introduced into mulberry plants by Agrobacterium-mediated transformation. Transgenic mulberry with barley Hva1 under a constitutive promoter actin1 was shown to enhance drought and salinity tolerance. Here, we report that overexpression of barley Hva1 also confers cold tolerance in transgenic mulberry. Further, barley Hva1 gene under control of a stress-inducible promoter rd29A can effectively negate growth retardation under non-stress conditions and confer stress tolerance in transgenic mulberry. Transgenic lines display normal morphology to enhanced growth and an increased tolerance against drought, salt and cold conditions as measured by free proline, membrane stability index and PSII activity. Protein accumulation was detected under stress conditions confirming inductive expression of HVA1 in transgenics. Investigations to assess stress tolerance of these plants under field conditions revealed an overall better performance than the non-transgenic plants. Enhanced expression of stress responsive genes such as Mi dnaJ and Mi 2-cysperoxidin suggests that Hva1 can regulate downstream genes associated with providing abiotic stress tolerance. The investigation of transgenic lines presented here demonstrates the acquisition of tolerance against drought, salt and cold stress in plants overexpressing barley Hva1, indicating that Arabidopsis rd29A promoter can function in mulberry.

  20. Peroxidase profiling reveals genetic linkage between peroxidase gene clusters and basal host and non-host resistance to rusts and mildew in barley.

    Directory of Open Access Journals (Sweden)

    Ana M González

    Full Text Available BACKGROUND: Higher plants possess a large multigene family encoding secreted class III peroxidase (Prx proteins. Peroxidases appear to be associated with plant disease resistance based on observations of induction during disease challenge and the presence or absence of isozymes in resistant vs susceptible varieties. Despite these associations, there is no evidence that allelic variation of peroxidases directly determines levels of disease resistance. METHODOLOGY/PRINCIPAL FINDINGS: The current study introduces a new strategy called Prx-Profiling. We showed that with this strategy a large number of peroxidase genes can be mapped on the barley genome. In order to obtain an estimate of the total number of Prx clusters we followed a re-sampling procedure, which indicated that the barley genome contains about 40 peroxidase gene clusters. We examined the association between the Prxs mapped and the QTLs for resistance of barley to homologous and heterologous rusts, and to the barley powdery mildew fungus. We report that 61% of the QTLs for partial resistance to P. hordei, 61% of the QTLs for resistance to B. graminis and 47% of the QTLs for non-host resistance to other Puccinia species co-localize with Prx based markers. CONCLUSIONS/SIGNIFICANCE: We conclude that Prx-Profiling was effective in finding the genetic location of Prx genes on the barley genome. The finding that QTLs for basal resistance to rusts and powdery mildew fungi tend to co-locate with Prx clusters provides a base for exploring the functional role of Prx-related genes in determining natural differences in levels of basal resistance.

  1. Determination of mortality of different life stages of Tribolium castaneum (Coleoptera: Tenebrionidae) in stored barley using microwaves.

    Science.gov (United States)

    Vadivambal, R; Jayas, D S; White, N D G

    2008-06-01

    Barley, Hordeum vulgare L., one of the important crops in Canada, is used in malting, feed, and food industries. Disinfestation of barley using microwaves can be an alternative to chemical methods used to kill insects. A pilot-scale industrial microwave system operating at 2.45 MHz was used in this study to determine the mortality of life stages (egg, larva, pupa, and adult) of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). Barley samples of 50 g each at 14, 16, and 18% moisture content (MC; wet basis) were infested with various life stages of T. castaneum and exposed to microwave energy at different power levels and exposure times, and the mortality of the insects was determined. The average temperature of the 14% MC sample exposed to 28 s at 0, 200, 300, 400, and 500 W were 27.4, 42.6, 53.7, 66.9, and 73.0 degrees C, respectively, and those exposed to 56 s at 0, 200, 300, and 400 W were 27.4, 57.3, 75.5, and 91.2 degrees C, respectively. A similar range of temperature was observed for 16 and 18% MC barley. Complete mortality of all life stages of T. castaneum can be achieved at a power level of 400 W and an exposure time of 56 s or at 500 W for 28 s. Among the life stages of T. castaneum, eggs were the most susceptible to microwave energy and adults were the least susceptible. There was no significant difference in the mortality of larvae and adults at 14, 16, and 18% MC, but the mortality was significantly different for eggs and pupae at different MCs. There was a significant increase in the mortality with an increase in power level or exposure time or both. Germination capacity of the seeds was decreased with an increase in power level or exposure time or both. Quality characteristics such as alpha-amylase, diastatic power, soluble protein, viscosity, and density of the barley malt treated at 500 W for 28 s were same as the control sample, whereas the samples treated at 400 W for 56 s were significantly lower.

  2. Electrospray mass spectrometry characterization of post-translational modifications of barley alpha-amylase 1 produced in yeast

    DEFF Research Database (Denmark)

    Søgaard, M; Andersen, Jens S.; Roepstorff, P;

    1993-01-01

    We have used electrospray mass spectrometry (ESMS) in combination with protein chemistry and genetics to delineate post-translational modifications in yeast of barley alpha-amylase 1 (AMY1), a 45 kD enzyme crucial for production of malt, an important starting material in the manufacture of beer...... and whisky. In addition to signal peptide processing these modifications are: (1) removal of C-terminal Arg-Ser by Kex1p, (2) glutathionylation of Cys95, (3) O-glycosylation, and (4) additional degradation of the C-terminus....

  3. Interactions of barley alpha-amylase isozymes with Ca2+, substrates and proteinaceous inhibitors

    DEFF Research Database (Denmark)

    Abou Hachem, Maher; Bozonnet, Sophie; Willemoes, Martin

    2006-01-01

    , and proteinaceous inhibitors for alpha-amylases. Isozyme specific effects of Ca2+ on the 80% sequence identical barley alpha-amylases AMY1 and AMY2 are not obvious from the two crystal structures, containing three superimposable Ca2+ with identical ligands. A fully hydrated fourth Ca2+ at the interface of the AMY2...... has revealed that AMY1 has ten functional subsites which can be modified by means protein engineering to modulate the substrate specificity. Other mutational analyses show that surface carbohydrate binding sites are critical for interaction with polysaccharides. The conserved Tyr380 in the newly...... discovered 'sugar tongs' site in domain C of AMY1 is thus critical for binding to starch granules. Furthermore, mutations of binding sites mostly reduced the degree of multiple attack in amylose hydrolysis. AMY1 has higher substrate affinity than AMY2, but isozyme chimeras with AMY2 domain C and other...

  4. Proteome analysis of dissected barley seed tissue during germination and radicle elongation

    DEFF Research Database (Denmark)

    Bønsager, Birgit Christine

    2007-01-01

    Cereal grains are vital components of our diet, and therefore understanding of the biology of breakage of dormancy and initiation of germination very important. These processes are far from fully understood, despite extensive studies, and no specific markers for germination have been identified...... at the protein or the DNA level. In addition, germination of barley seeds is of interest for the brewing industry since this process corresponds to the steeping process that starts the industrial malting. In the present study a proteomics approach was employed to understand the initial changes in the water...... stress, storage breakdown, folding, and housekeeping were identified and tracked through 72 h PI. APX and the other enzymes involved in the ascorbate-glutathione cycle (dehydroascorbate reductase, monodehydroascorbate reductase, and glutathione reductase) were thought to have an important function during...

  5. Starch and Prolamin Level in Single and Double High-Lysine Barley Mutants

    DEFF Research Database (Denmark)

    Kreis, M.; Doll, Hans

    1980-01-01

    At maturity the high-lysine barley (Hordeum vulgare L.) Ris0 mutants 1508, 527 and 29 kernels contained about 20% less starch and twice as much free sugars as the parent varieties Bomi and Carlsberg II. An enhanched effect on starch reduction and free sugar accumulation was observed during kernel...... development when the single mutants 527 and 29 are combined with the mutant 1508. At maturity, kernels of the double mutants 527/1508 and 29/1508 contained, respectively, 68 and 43% less starch than Bomi. The double mutant 29/1508 kernel had a slightly lower prolamin content than mutant 1508 which is the most...... prolamin-deficient single mutant. In the double mutant 527/1508, however, an almost complete suppression of prolamin synthesis was observed during kernel development. The percentage of lysine in the seed proteins of the double mutants was about the same as in the most extreme single mutant 1508. Based...

  6. Determination of ergosterol levels in barley and malt varieties in the Czech Republic via HPLC.

    Science.gov (United States)

    Jedlicková, Lenka; Gadas, David; Havlová, Pavla; Havel, Josef

    2008-06-11

    Ergosterol is considered to be a suitable indicator of mold infestation in barley and malt. In this study ergosterol levels in different varieties of barley and malt produced in the Czech Republic were determined. A modified high-performance liquid chromatography (HPLC) method was statistically processed, validated (Effivalidation program), and applied to 124 samples of barley and malt. Ergosterol was isolated by extraction and saponification, and the quantification was performed using HPLC with diode array detection. The content of ergosterol ranged between the limit of detection (LOD) and 36.3 mg/kg in barley and between the LOD and 131.1 mg/kg in malt. Ergosterol is presumably connected with metabolites generated when barley grain is attacked by pathogens, and such barley often shows a high overfoaming (gushing) value. However, it was found that the content of ergosterol does not correlate with the degree of beer gushing.

  7. Protein

    Science.gov (United States)

    ... Food Service Resources Additional Resources About FAQ Contact Protein Protein is found throughout the body—in muscle, ... the heart and respiratory system, and death. All Protein Isn’t Alike Protein is built from building ...

  8. Barley (Hordeum vulgare L. yield with normal and coated urea in Pintag, Quito, Ecuador.

    Directory of Open Access Journals (Sweden)

    Andrea C. Lema-Aguirre

    2016-12-01

    Full Text Available The objective of this investigation was to evaluate the response of barley production to nitrogenized fertilization (NF and its impact on the protein content of the grain and the availability of nutrients on the soil. The study was carried out in two sites in Hacienda Valencia, in Pintag, Quito, Ecuador, between March 2014 and February 2015. A variety of INIAP Cañicapa 2003 was used, and the NF was done with normal urea and polymerized (urea + tiophosphate of N-nbutiltriamida. An experimental design of randomized full blocks in four repetitions of divided lots [the urea (normal and polymerized was used in the main lot and the NF (0, 30, 60, 90, 120, y 150 kg N/ha in the sub-lot]. 110 kg/ha of seed was used; 50% of the NF was applied three weeks after planting (broadcast and 50% eight weeks after. The type of urea did not affect the production (p>0,10, although the average was different between two sites (4,33 y 2,08 t/ha for sites 1 and 2, respectively. Excess of rain could have limit the effect on the urea type. On average a cuadratic response to NF was obtined, with an optimum dose of N (DON of 90 kg N/ha and a production of 3,41 t/ ha. This DON was similar to the dose of N recomended for barley in Ecuador. The NF augmented the protein content up to 14,50% (p<0,01 and acidi ed the ground, because the urea nitri cation produces H+.

  9. Cultivar and Environmental Variation of β-glucan Content in Chinese Barleys

    Institute of Scientific and Technical Information of China (English)

    CHEN Jin-xin; Zhang Guo-ping; QIANG Xiao-lin; WANG Jun-mei; DING Shou-ren

    2002-01-01

    β-glucan is a polysaccharide compound closely related to the quality of barley used as malting,feed and food. Low β-glucan content is expected for brewing and feed barley, while high β-glucan content is desirable for food barley. The β-glucan content of barley genotypes collected from various areas of China as well as from Canada and Australia were assayed. Meanwhile a multi-locations trial was conducted to determineβ-glucan content of 10 barley cultivars in 8 locations for two successive planting years. The results showed that barley genotypes from Tibet and Xinjiang had higher β-glucan content and the genotypes with higher than 8%of β-glucan content were detected in Tibet barleys, being valuable for use in the development of healthy food.Barley cultivars being planted now in winter-sowing areas of China had basically the same β-glucan content as those from Canada and Australia. Barley seeds produced in Hangzhou had lower β-glucan content than seeds from the original areas. There was a highly significant difference in β-glucan content among 10 barleys, 8locations and between years. On an average of two years, Xiumei 3 and Kongpei 1 had the highest and lowestβ-glucan content, respectively, and Taian and Hangzhou produced the highest and lowest β-glucan content barley seeds, respectively. Analysis of AMMI model showed that interaction effect between cultivar and environment was highly significant in both experimental years, and was dependent on cuitivar, suggesting that it is important to plant the suitable cultivars in a particular area in order to obtain barley seeds with reasonableβ-glucan content.

  10. Retention and growth performance of chicks given low-phytate conventional or hull-less barleys

    Science.gov (United States)

    Four low-phytate, hulled lines, M2 422 (now referred to as barley lpa1-1), M2 635 (now referred to as barley lpa3-1), M2 955 and M2 1070 (now referred to as barley lpa2-1), and a "hulless" version of M2 422, were evaluated in a chick feeding experiment. The diets were provided in meal form, with the...

  11. Effect of wide variation of the Waxy gene on starch properties in hull-less barley from Qinghai-Tibet plateau in China.

    Science.gov (United States)

    Li, Qiao; Pan, Zhifen; Deng, Guangbing; Long, Hai; Li, Zhongyi; Deng, Xiaoqing; Liang, JunJun; Tang, Yawei; Zeng, Xingquan; Tashi, Nyima; Yu, Maoqun

    2014-11-26

    Granule-bound starch synthase I (GBSS I) plays an important role in the synthesis of amylose and in the determination of starch properties in barley grains. Genomic DNAs for the Waxy gene encoding GBSS I protein were sequenced from 34 barley accessions or lines from Qinghai-Tibet plateau in China, to identify Waxy gene nucleotide variations and study the roles of polymorphic sites of the Waxy gene on expression levels of Waxy transcripts and GBSS I proteins and on resulting starch properties. A total of 116 DNA polymorphic sites were identified within the barley Waxy gene, which divided the studied accessions into 11 haplotypes. Among 33 nucleotide polymorphic sites in coding regions, 5 SNPs in three exons were found to play different roles on the expression level of the Waxy transcript and the GBSS I protein and on the amylose content and starch properties. One SNP G(3935)-to-T substitution in the 10th exon in the accession Z999 (HP II-2) caused a high expression level of the Waxy transcript and the GBSS I protein and the amylose free phenotype. The other SNP alteration was a C(2453)-to-T in the fifth exon in the accession Z1191 (HP I-5), which drastically reduced the expression level of the Waxy transcript and the GBSS I protein and, finally, produced the amylose free phenotype. Three SNPs in the seventh exon in the accession Z1337 (HP I-6) did not significantly change the level of Waxy transcript, the GBSS I protein, and starch properties, except obviously reducing the breakdown value of starch viscosity and extending the peak time. A total of 84 DNA polymorphic sites were found in the noncoding regions. A 403 bp deletion at 5'UTR in the accession Z1979 (HP I-3) had low transcript level, low GBSS I protein level, and low amylose content due to the deletion of cis-acting DNA regulatory elements. A 191 bp insertion and a 15 bp insertion in the first intron and second exons, respectively, may be closely related to a higher transcript level of the Waxy gene and

  12. Assessment of the Seedling Reactions of Some Hulless Barley Genotypes to Drechslera teres f. maculata

    OpenAIRE

    Gerlegiz, Emine Tuba; KARAKAYA, Aziz; Celik Oguz, Arzu; MERT, Zafer; Sayim, İsmail; Ergun, Namuk; Aydogan, Sinan

    2015-01-01

    The seedling reactions of three barley cultivars, one hulless barley cultivar, two candidate hulless barley lines and nine hulless barley genotypes were determined under greenhouse conditions to ten isolates of Drechslera teres f. maculata, the causal agent of spot form of net blotch. Isolates were obtained from Ankara, Çankırı, Eskişehir, Kayseri, Konya and Şanlıurfa provinces. The reactions of the cultivars and hulless cultivar ranged between suscepible-resistant. The reactions of the hulle...

  13. Barley Sprouts Extract Attenuates Alcoholic Fatty Liver Injury in Mice by Reducing Inflammatory Response

    Directory of Open Access Journals (Sweden)

    Yun-Hee Lee

    2016-07-01

    Full Text Available It has been reported that barley leaves possess beneficial properties such as antioxidant, hypolipidemic, antidepressant, and antidiabetic. Interestingly, barley sprouts contain a high content of saponarin, which showed both anti-inflammatory and antioxidant activities. In this study, we evaluated the effect of barley sprouts on alcohol-induced liver injury mediated by inflammation and oxidative stress. Raw barley sprouts were extracted, and quantitative and qualitative analyses of its components were performed. The mice were fed a liquid alcohol diet with or without barley sprouts for four weeks. Lipopolysaccharide (LPS-stimulated RAW 264.7 cells were used to study the effect of barley sprouts on inflammation. Alcohol intake for four weeks caused liver injury, evidenced by an increase in serum alanine aminotransferase and aspartate aminotransferase activities and tumor necrosis factor (TNF-α levels. The accumulation of lipid in the liver was also significantly induced, whereas the glutathione (GSH level was reduced. Moreover, the inflammation-related gene expression was dramatically increased. All these alcohol-induced changes were effectively prevented by barley sprouts treatment. In particular, pretreatment with barley sprouts significantly blocked inducible nitric oxide synthase (iNOS and cyclooxygenase (COX-2 expression in LPS-stimulated RAW 264.7. This study suggests that the protective effect of barley sprouts against alcohol-induced liver injury is potentially attributable to its inhibition of the inflammatory response induced by alcohol.

  14. Archaeogenetic evidence of ancient nubian barley evolution from six to two-row indicates local adaptation.

    Directory of Open Access Journals (Sweden)

    Sarah A Palmer

    Full Text Available BACKGROUND: Archaeobotanical samples of barley (Hordeum vulgare L. found at Qasr Ibrim display a two-row phenotype that is unique to the region of archaeological sites upriver of the first cataract of the Nile, characterised by the development of distinctive lateral bracts. The phenotype occurs throughout all strata at Qasr Ibrim, which range in age from 3000 to a few hundred years. METHODOLOGY AND FINDINGS: We extracted ancient DNA from barley samples from the entire range of occupancy of the site, and studied the Vrs1 gene responsible for row number in extant barley. Surprisingly, we found a discord between the genotype and phenotype in all samples; all the barley had a genotype consistent with the six-row condition. These results indicate a six-row ancestry for the Qasr Ibrim barley, followed by a reassertion of the two-row condition. Modelling demonstrates that this sequence of evolutionary events requires a strong selection pressure. CONCLUSIONS: The two-row phenotype at Qasr Ibrim is caused by a different mechanism to that in extant barley. The strength of selection required for this mechanism to prevail indicates that the barley became locally adapted in the region in response to a local selection pressure. The consistency of the genotype/phenotype discord over time supports a scenario of adoption of this barley type by successive cultures, rather than the importation of new barley varieties associated with individual cultures.

  15. Barley Sprouts Extract Attenuates Alcoholic Fatty Liver Injury in Mice by Reducing Inflammatory Response

    Science.gov (United States)

    Lee, Yun-Hee; Kim, Joung-Hee; Kim, Sou Hyun; Oh, Ji Youn; Seo, Woo Duck; Kim, Kyung-Mi; Jung, Jae-Chul; Jung, Young-Suk

    2016-01-01

    It has been reported that barley leaves possess beneficial properties such as antioxidant, hypolipidemic, antidepressant, and antidiabetic. Interestingly, barley sprouts contain a high content of saponarin, which showed both anti-inflammatory and antioxidant activities. In this study, we evaluated the effect of barley sprouts on alcohol-induced liver injury mediated by inflammation and oxidative stress. Raw barley sprouts were extracted, and quantitative and qualitative analyses of its components were performed. The mice were fed a liquid alcohol diet with or without barley sprouts for four weeks. Lipopolysaccharide (LPS)-stimulated RAW 264.7 cells were used to study the effect of barley sprouts on inflammation. Alcohol intake for four weeks caused liver injury, evidenced by an increase in serum alanine aminotransferase and aspartate aminotransferase activities and tumor necrosis factor (TNF)-α levels. The accumulation of lipid in the liver was also significantly induced, whereas the glutathione (GSH) level was reduced. Moreover, the inflammation-related gene expression was dramatically increased. All these alcohol-induced changes were effectively prevented by barley sprouts treatment. In particular, pretreatment with barley sprouts significantly blocked inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expression in LPS-stimulated RAW 264.7. This study suggests that the protective effect of barley sprouts against alcohol-induced liver injury is potentially attributable to its inhibition of the inflammatory response induced by alcohol. PMID:27455313

  16. Investigations of barley stripe mosaic virus as a gene silencing vector in barley roots and in Brachypodium distachyon and oat

    Directory of Open Access Journals (Sweden)

    Nilsson Lena

    2010-11-01

    Full Text Available Abstract Background Gene silencing vectors based on Barley stripe mosaic virus (BSMV are used extensively in cereals to study gene function, but nearly all studies have been limited to genes expressed in leaves of barley and wheat. However since many important aspects of plant biology are based on root-expressed genes we wanted to explore the potential of BSMV for silencing genes in root tissues. Furthermore, the newly completed genome sequence of the emerging cereal model species Brachypodium distachyon as well as the increasing amount of EST sequence information available for oat (Avena species have created a need for tools to study gene function in these species. Results Here we demonstrate the successful BSMV-mediated virus induced gene silencing (VIGS of three different genes in barley roots, i.e. the barley homologues of the IPS1, PHR1, and PHO2 genes known to participate in Pi uptake and reallocation in Arabidopsis. Attempts to silence two other genes, the Pi transporter gene HvPht1;1 and the endo-β-1,4-glucanase gene HvCel1, in barley roots were unsuccessful, probably due to instability of the plant gene inserts in the viral vector. In B. distachyon leaves, significant silencing of the PHYTOENE DESATURASE (BdPDS gene was obtained as shown by photobleaching as well as quantitative RT-PCR analysis. On the other hand, only very limited silencing of the oat AsPDS gene was observed in both hexaploid (A. sativa and diploid (A. strigosa oat. Finally, two modifications of the BSMV vector are presented, allowing ligation-free cloning of DNA fragments into the BSMV-γ component. Conclusions Our results show that BSMV can be used as a vector for gene silencing in barley roots and in B. distachyon leaves and possibly roots, opening up possibilities for using VIGS to study cereal root biology and to exploit the wealth of genome information in the new cereal model plant B. distachyon. On the other hand, the silencing induced by BSMV in oat seemed too

  17. Enzymatic hydrolysis of pretreated barley and wheat straw

    DEFF Research Database (Denmark)

    Rosgaard, Lisa

    2007-01-01

    pretreatment conditions; hot water extraction and acid- or water impregnation followed by steam explosion showed there were slight differences between the effect of pretreatment conditions in relation to the overall yield from enzymatic hydrolysis. The highest glucose concentration was found for barley straw...... subjected to acid impregnation followed by steam explosion; however when the glucose concentration was related to the glucose potential in the substrates, the highest yield was obtained with hot water extracted. Analysis of the supernatants from the pretreatments by mass spectrometry showed that the water...... feeding strategy to increase the substrate loading in the hydrolysis reaction. The substrate for the enzymatic hydrolysis was primarily steam pretreated wheat and barley straw since these substrates were the primary feedstocks for the Babilafuente Bioethanol process. The initial work showed...

  18. Effect of ozone pretreatment on hydrogen production from barley straw.

    Science.gov (United States)

    Wu, Jiangning; Ein-Mozaffari, Farhad; Upreti, Simant

    2013-09-01

    Application of ozone technology to lignocellulosic biohydrogen production was explored with a barley straw. Ozone pretreatment effectively degraded the straw lignin and increased reducing sugar yield. A simultaneous enzyme hydrolysis and dark fermentation experiment was conducted using a mixed anaerobic consortium together with saccharification enzymes. Both untreated and ozonated samples produced hydrogen. Compared to the untreated group, hydrogen produced by the groups ozonated for 15, 30, 45 and 90 min increased 99%, 133%, 166% and 94%, respectively. Some inhibitory effect on hydrogen production was observed with the samples ozonated for 90 min, and the inhibition was on the fermentative microorganisms, not the saccharification enzymes. These results demonstrate that production of biohydrogen from barley straw, a lignocellulosic biomass, can be significantly enhanced by ozone pretreatment.

  19. In vitro fermentation of ten cultivars of barley silage

    Directory of Open Access Journals (Sweden)

    Federico Infascelli

    2010-01-01

    Full Text Available The fermentation characteristics of whole-crop barley silages from ten different cultivars were evaluated by the in vitro gas production technique. The organic matter degradability of barley silage (62.9% in average was comparable to those reported in our previous trials for oat (59.7% and sorghum silages (65.5%; while the maximum gas production rate (5.38 ml/h in average was slightly lower respect to oat (6.71 ml/h and sorghum silage (6.74 ml/h. The mean nutritive value (4.00 MJ/kg DM calculated on the basis of both chemical composition and in vitro fermentation data was comparable to that (4.16 MJ/kg DM obtained in our previous research performed on corn silage, from crop sowed in the same area.

  20. Pollen grain of barley (Hordeum vulgare L. - pattern of development

    Directory of Open Access Journals (Sweden)

    Maria Charzyńska

    2014-01-01

    Full Text Available Pollen development in barley follows the general pattern established for other species of Poaceae: 1 microspore division occurs at the vacuolate microspore stage with polarly located nucleus; 2 microspore mitosis is immediately followed by phragmoplast and cell plate formation; 3 in consequence or unequal microspore division, the generative cell, at first attached to the pollen wall, is separated from the vegetative cell by a callosic wall; 4 during the postmitotic two-cell stage of development, the vegetative nucleus migrates to the aperture pole and is followed by the generative cell that is detached and free of callose wall. In this position the generative cell divides into two sperm cells. These data do not confirm the interpretation of pollen grain development in barley given by Cass and Karas in Can. J. Bot. 53: 1051-1062, 1975.

  1. Barley grain enrichement with essential elements by agronomic biofortification

    Directory of Open Access Journals (Sweden)

    Dragičević Vesna D.

    2016-01-01

    Full Text Available Barley grain is rich in mineral nutrients, but their bioavailability to humans depends on antinutrients that restrain bioavailability and promoters that promote bioavailability. The aim of this study was to examine composition of barley grain, including phytate and phenolics as antinutrients, carotenoids and glutathione as promoters and mineral elements, such as Ca, Mg, Fe, Si, Zn and Mn influenced by various non-standard foliar fertilizers (Zircon, Chitosan, Siliplant, Propikonazole, including some hormonal growth-stimulators (Epin Extra, Benzyladenine, as potential biofortification measure. Chitosan increased glutathione concentration in grain. Unfavorable meteorological conditions were partly mitigated by application of Benzyladenine and Siliplant, reflected through increased potential bioavailability of P, Mg, Ca and Fe. [Projekat Ministarstva nauke Republike Srbije, br. TR-31037

  2. Resistance in winter barley against Ramularia leaf spot

    DEFF Research Database (Denmark)

    Hjortshøj, Rasmus Lund

    Ramularia leaf spot is an emerging disease in barley caused by R. collo-cygni. At present little is known about the resistance mechanisms carried out by the host plant to avoid disease development. Nor is the lifecycle of the fungus or its populations structure fully understood. To gain insight...... into these aspects four experiments were set up; (1) a mapping experiment aiming at identifying QTL’s controlling disease levels under field conditions was conducted in two winterbarley populations. (2) a toxin assay testing the parental lines used in the mapping populations for response to Rubellin D was developed....... (3) microarray analysis of transcriptional response in barley to inoculation with R. collo-cygni was carried out, and finally the (4) population genetic structure of R. collo-cygni was assessed by AFLP and gene sequencing. Based on these experiments interaction was compared to the interactions of C...

  3. Suppression of Zn stress on barley by irradiated chitosan

    Energy Technology Data Exchange (ETDEWEB)

    Nagasawa, N.; Mitomo, H. [Gunma Univ., Faculty of Engineering, Department of Biological and Chemical Engineering, Kiryu, Gunma (Japan); Ha, P.T.L. [Nuclear Research Institute, Dalat (Viet Nam); Watanabe, S.; Ito, T.; Takeshita, H.; Yoshii, F.; Kume, T. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2001-03-01

    Chitosan was irradiated up to 1000 kGy in solid state. Irradiation of chitosan caused the reduction of molecular weight. The molecular weight of the chitosan reduced from ca. 4 x 10{sup 5} to ca. 6 x 10{sup 3} by irradiation at 1000 kGy. For the barley growth promotion, irradiated chitosan showed the significant effect and 1000 kGy irradiated chitosan improved 20% of growth. Using the positron emitting tracer imaging system (PETIS), the effect of chitosan on uptake and transportation of {sup 62}Zn in barley were investigated. It was found that the transportation of Zn from root to shoot and the damage of plant by Zn were suppressed with irradiated chitosan. (author)

  4. Drivers of phosphorus uptake by barley following secondary resource application

    Directory of Open Access Journals (Sweden)

    Eva eBrod

    2016-05-01

    Full Text Available Minable rock phosphate is a finite resource. Replacing mineral phosphorus (P fertilizer with P-rich secondary resources is one way to manage P more efficiently, but the importance of physicochemical and microbial soil processes induced by secondary resources for plant P uptake are still poorly understood. Using radioactive labelling techniques, the fertilization effects of dairy manure, fish sludge, meat bone meal and wood ash were studied as P uptake by barley after 44 days and compared with those of water-soluble mineral P (MinP and an unfertilized control (NoP in a pot experiment with an agricultural soil containing little available P at two soil pH levels, approximately pH 5.3 (unlimed soil and pH 6.2 (limed soil. In a parallel incubation experiment, the effects of the secondary resources on physicochemical and microbial soil processes were studied. The results showed that the relative agronomic efficiency compared with MinP decreased in the order: manure ≥ fish sludge ≥ wood ash ≥ meat bone meal. The solubility of inorganic P in secondary resources was the main driver for P uptake by barley (Hordeum vulgare. The effects of secondary resources on physicochemical and microbial soil processes were of little overall importance. Application of organic carbon with manure resulted in microbial P immobilisation and decreased uptake by barley of P derived from the soil. On both soils, P uptake by barley was best explained by a positive linear relationship with the H2O + NaHCO3-soluble inorganic P fraction in fertilizers, or by a linear negative relationship with the HCl-soluble inorganic P fraction in fertilizers.

  5. Ethylene production and peroxidase activity in aphid-infested barley.

    Science.gov (United States)

    Argandoña, V H; Chaman, M; Cardemil, L; Muñoz, O; Zúñiga, G E; Corcuera, L J

    2001-01-01

    The purpose of this work was to investigate whether ethylene is involved in the oxidative and defensive responses of barley to the aphids Schizaphis graminum (biotype C) and Rhopalophum padi. The effect of aphid infestation on ethylene production was measured in two barley cultivars (Frontera and Aramir) that differ in their susceptibility to aphids. Ethylene evolution was higher in plants infested for 16 hr than in plants infested for 4 hr in both cultivars. Under aphid infestation, the production of ethylene was higher in cv. Frontera than in Aramir, the more aphid susceptible cultivar. Ethylene production also increases with the degree of infestation. Maximum ethylene evolution was detected after 16 hr when plants were infested with 10 or more aphids. Comparing the two species of aphids, Schizaphis graminum induced more ethylene evolution than Rhopalosiphum padi. Infestation with S. graminum increased hydrogen peroxide content and total soluble peroxidase activity in cv. Frontera, with a maximum level of H2O2 observed after 20 min of infestation and the maximum in soluble peroxidase activity after 30 min of infestation. When noninfested barley seedlings from cv. Frontera were exposed to ethylene, an increase in hydrogen peroxide and in total peroxidase activity was detected at levels similar to those of infested plants from cv. Frontera. When noninfested plants were treated with 40 ppm of ethylene, the maximum levels of H2O2 and soluble peroxidase activity were at 10 and 40 min, respectively. Ethylene also increased the activity of both cell-wall-bound peroxidases types (ionically and covalently bound), comparable with infestation. These results suggest that ethylene is involved in the oxidative responses of barley plants induced by infestation.

  6. Modeling moisture absorption kinetics of barley grain using viscoelastic model and neural networks

    Directory of Open Access Journals (Sweden)

    M Kamali

    2015-09-01

    Full Text Available Introduction: Barley is one of the most important grains with high digestible starch making it a main source of energy in human nutrition as well as in livestock rations formulation and feeding. Starch is the main part of barley grain and it has an inverse relation with its protein. It has a digestible foodstuff of 80 to 84 percent of its dry matter content. Barley as livestock foodstuff should be processed and it is done in several ways. A customary method for processing barley in dairy farms is its size reduction by milling (Hunt, 1996. An alternative method of barley processing is steam rolling. However, because of the high cost of steam generators a method of soaking with heating has been considered as an alternative method for steam rolling (Yang et al., 2000. The rate of moisture absorption by grains during the soaking process varies considerably and depends on the size of the grain, water temperature and the length of soaking. High temperature water soaking is an ordinary way to reduce the time duration for reaching a high rate of moisture absorption during the soaking process (Kashaninejad et al., 2009. Various studies have shown that these models have adequate accuracy in analyzing drying and moisture absorption processes for most agricultural products (Abu-Ghannam and McKenna, 1997. Some researchers have modeled beans moisture absorption behavior using 14 mathematical models and found that the Weibull model had the most conformity with variations in experimental data (Shafaei and Masoumi, 2014c. Observations made by researchers indicate that the moisture absorption process in various materials encompasses a primary phase with a fast rate and a second phase with a lower rate. The second phase in moisture absorption is called the relaxation phase. The main problem with all the mathematical and experimental models is the lack of the model’s ability to evaluate the rate of moisture absorption in the secondary phase. Artificial Neural

  7. Effects of ethylene on root elongation in barley and rice

    Energy Technology Data Exchange (ETDEWEB)

    John, A.; Hall, M.A.; Crossett, R.N.

    1972-01-01

    Experiments were performed to determine the effects of rice and barley to growth inhibition by ethylene. The mechanism of growth inhibition was investigated at the cellular level and a detailed comparison was made between the responses of the two species. The following measurements were made on intact plants in short (up to 200 minutes), medium (up to 3 days) or long (up to 10 days) experiments: the rate of extension growth of main root axes; the final cell length and number of elongating cells produced; and the extensibility of the apical growing region. Results indicate that the effects of ethylene on the elongation of roots of rice and barley plants are different. In barley there is a rapid inhibition of root extension which persists with prolonged exposure to the gas but with little effect on the production of growing cells. However, rice roots exhibit no rapid growth inhibition response, but a reduction does occur after prolonged exposure. Low concentrations promote extension rice roots. The inhibition of root growth is reflected in a reduced extensibility of the apical growing region.

  8. Nitrate Uptake into Barley (Hordeum vulgare) Plants 1

    Science.gov (United States)

    Deane-Drummond, Celia E.; Glass, Anthony D. M.

    1982-01-01

    Evidence is presented that chlorate is an extremely good analog for nitrate during nitrate uptake by intact barley (Hordeum vulgare cv. Fergus) roots. The depletion of ClO3− or NO3− from uptake media over 2 to 6 hours by seedlings was found to be dependent on combined NO3− plus ClO3− concentrations, and total anion uptake was equivalent at different NO3−/ClO3− ratios. After loading barley seedlings with 36ClO3− for 6 hours, kinetic parameters were derived from the analysis of efflux of [36Cl] chlorate into unlabeled solution. On the basis of this analysis, the half times for exchange for the cytoplasmic and vacuolar phases were 17 minutes and 20 hours, respectively. Data pooled from a number of different experiments were used to calculate kinetic constants (Km and Vmax) for 36ClO3− influx into barley roots at different external ClO3−/NO3− ratios, using short (10 minutes) influx times. There appeared to be no discrimination by the root cells between ClO3− and NO3−. Lineweaver-Burk analysis of the interaction between nitrate and chlorate were characteristic of competitive inhibition at low nitrate concentrations (0-0.5 mm). At higher concentrations, in the range of >1 mm, similar interactions between these ions were evident. PMID:16662478

  9. Genetic analysis of aluminum tolerance in Brazilian barleys

    Directory of Open Access Journals (Sweden)

    Minella Euclydes

    2002-01-01

    Full Text Available Aluminum (Al toxicity is a major factor limiting barley growth in acid soils, and genotypes with adequate level of tolerance are needed for improving barley adaptation in Brazil. To study the inheritance of Al tolerance in Brazilian barleys, cultivars Antarctica 1, BR 1 and FM 404 were crossed to sensitive Kearney and PFC 8026, and intercrossed. Parental, F1, F2 and F6 generations were grown in nutrient solution containing 0.03, 0.05 and 0.07 mM of Al and classified for tolerance by the root tip hematoxylin staining assay. Tolerant by sensitive F2 progenies segregated three tolerant to one sensitive, fitting the 3:1 ratio expected for a single gene. The F6 populations segregated one tolerant to one sensitive also fitting a monogenic ratio. The F2 seedlings from crosses among tolerant genotypes scored the same as the parents. Since the population size used would allow detection of recombination as low as 7%, the complete absence of Al sensitive recombinants suggests that tolerance in these cultivars is most probably, controlled by the same gene. Thus, the potential for improving Al tolerance through recombination of these genotypes is very low and different gene sources should be evaluated.

  10. Alleviation of Al Toxicity in Barley by Addition of Calcium

    Institute of Scientific and Technical Information of China (English)

    GUO Tian-rong; CHEN Ying; ZHANG Yan-hua; JIN Ye-fei

    2006-01-01

    The potential mechanism by which Ca alleviates Al toxicity was investigated in barley seedlings. It was found that 100 μM Al-alone treatment inhibited barley plant growth and thereby reduced shoot height and root length, and dry weights of root, shoot and leaf; promoted Al accumulation but inhibited Ca absorption in plant tissues; and induced an increase in the activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) and in the level of lipid peroxidation (MDA content) in leaves. Except for the increase in Ca concentration in plant tissues, treatment with 0.5 mM Ca in the absence of Al had less effect on the above-mentioned parameters, compared with the control. Addition of Ca efficiently reduced Al toxicity, which is reflected by the promotion of plant growth, reduction in Al concentration and MDA content,increase in Ca concentration and in SOD, POD, and CAT activities compared with the Al-alone-treatment; with increase in Ca level (3.0 mM), the ameliorative effect became more dominant. This indicated that the alleviation of aluminum toxicity in barley seedlings with Ca supplementation could be associated with less absorption of Al and the enhancement of the protective ability of the cell because of increased activity of the antioxidative enzyme.

  11. Pysicochemical properties of Tibetan hull-less barley starch.

    Science.gov (United States)

    Yangcheng, Hanyu; Gong, Lingxiao; Zhang, Ying; Jane, Jay-lin

    2016-02-10

    Objectives of this study were to (1) determine the starch physicochemical properties of two commercial Tibetan hull-less barley varieties, Beiqing (BQ) and Kangqing (KQ); and (2) understand the relationship between unique properties of the starches, their structures, and impacts of growing conditions. The BQ barleys were grown at a location with lower temperature and less rainfall compared with the KQ barleys. The BQ starches showed significantly lower onset-gelatinization temperature (54.1-54.9 °C), larger gelatinization-temperature range (9.4-10.6 °C), and higher peak-viscosities (138.9-153.9RVU) than the KQ starches (55.1-56.1 °C, 7.4-8.8 °C, and 63.4-64.7RVU, respectively). After a treatment with 2% sodium-dodecyl-sulphate solution, the KQ starches showed substantially greater increases in peak viscosities than the BQ starches. Annealing of starch and enhanced amylose-lipid complex formation, resulting from higher growing temperature during the development of the KQ starches, likely contributed to the differences in thermal and pasting properties between the BQ and KQ starches.

  12. Screening of the aerodynamic and biophysical properties of barley malt

    Science.gov (United States)

    Ghodsvali, Alireza; Farzaneh, Vahid; Bakhshabadi, Hamid; Zare, Zahra; Karami, Zahra; Mokhtarian, Mohsen; Carvalho, Isabel. S.

    2016-10-01

    An understanding of the aerodynamic and biophysical properties of barley malt is necessary for the appropriate design of equipment for the handling, shipping, dehydration, grading, sorting and warehousing of this strategic crop. Malting is a complex biotechnological process that includes steeping; germination and finally, the dehydration of cereal grains under controlled temperature and humidity conditions. In this investigation, the biophysical properties of barley malt were predicted using two models of artificial neural networks as well as response surface methodology. Stepping time and germination time were selected as the independent variables and 1 000 kernel weight, kernel density and terminal velocity were selected as the dependent variables (responses). The obtained outcomes showed that the artificial neural network model, with a logarithmic sigmoid activation function, presents more precise results than the response surface model in the prediction of the aerodynamic and biophysical properties of produced barley malt. This model presented the best result with 8 nodes in the hidden layer and significant correlation coefficient values of 0.783, 0.767 and 0.991 were obtained for responses one thousand kernel weight, kernel density, and terminal velocity, respectively. The outcomes indicated that this novel technique could be successfully applied in quantitative and qualitative monitoring within the malting process.

  13. Selection of barley grain affects ruminal fermentation, starch digestibility, and productivity of lactating dairy cows.

    Science.gov (United States)

    Silveira, C; Oba, M; Yang, W Z; Beauchemin, K A

    2007-06-01

    The objective of this study was to evaluate the effects of 2 lots of barley grain cultivars differing in expected ruminal starch degradation on dry matter (DM) intake, ruminal fermentation, ruminal and total tract digestibility, and milk production of dairy cows when provided at 2 concentrations in the diet. Four primiparous ruminally cannulated (123 +/- 69 d in milk; mean +/- SD) and 4 multiparous ruminally and duodenally cannulated (46 +/- 14 d in milk) cows were used in a 4 x 4 Latin Square design with a 2 x 2 factorial arrangement of treatments with 16-d periods. Primiparous and multiparous cows were assigned to different squares. Treatments were 2 dietary starch concentrations (30 vs. 23% of dietary DM) and 2 lots of barley grain cultivars (Xena vs. Dillon) differing in expected ruminal starch degradation. Xena had higher starch concentration (58.7 vs. 50.0%) and greater in vitro 6-h starch digestibility (78.0 vs. 73.5%) compared with Dillon. All experimental diets were formulated to supply 18.3% crude protein and 20.0% forage neutral detergent fiber. Dry matter intake and milk yield were not affected by treatment. Milk fat concentration (3.55 vs. 3.29%) was greater for cows fed Dillon compared with Xena, but was not affected by dietary starch concentration. Ruminal starch digestion was greater for cows fed high-starch diets compared with those fed low-starch diets (4.55 vs. 2.49 kg/d), and tended to be greater for cows fed Xena compared with those fed Dillon (3.85 vs. 3.19 kg/d). Ruminal acetate concentration was lower, and propionate concentration was greater, for cows fed Xena or high-starch diets compared with cows fed Dillon or low-starch diets, respectively. Furthermore, cows fed Xena or high-starch diets had longer duration that ruminal pH was below 5.8 (6.6 vs. 4.0 and 6.4 vs. 4.2 h/d) and greater total tract starch digestibility (94.3 vs. 93.0 and 94.3 vs. 93.0%) compared with cows fed Dillon or low-starch diets, respectively. These results

  14. Influence of diurnal photosynthetic activity on the morphology, structure, and thermal properties of normal and waxy barley starch

    DEFF Research Database (Denmark)

    Goldstein, Avi; Annor, George; Vamadevan, Varatharajan

    2017-01-01

    This study investigated the influence of diurnal photosynthetic activity on the morphology, molecular composition, crystallinity, and gelatinization properties of normal barley starch (NBS) and waxy barley starch (WBS) granules from plants cultivated in a greenhouse under normal diurnal (16h light...... that normal barley starch is less influenced by the diurnal photosynthetic lighting regime than amylose-free barley starch suggesting a role of amylose to prevent structural disorder and increase starch granule robustness against environmental cues....

  15. The effects of salt stress cause a diversion of basal metabolism in barley roots: possible different roles for glucose-6-phosphate dehydrogenase isoforms.

    Science.gov (United States)

    Cardi, Manuela; Castiglia, Daniela; Ferrara, Myriam; Guerriero, Gea; Chiurazzi, Maurizio; Esposito, Sergio

    2015-01-01

    In this study the effects of salt stress and nitrogen assimilation have been investigated in roots of hydroponically-grown barley plants exposed to 150 mM NaCl, in presence or absence of ammonium as the sole nitrogen source. Salt stress determines a diversion of root metabolism towards the synthesis of osmolytes, such as glycine betaine and proline, and increased levels of reduced glutathione. The metabolic changes triggered by salt stress result in a decrease in both activities and protein abundance of key enzymes, namely GOGAT and PEP carboxylase, and in a slight increase in HSP70. These variations would enhance the requirement for reductants supplied by the OPPP, consistently with the observed increase in total G6PDH activity. The involvement and occurrence of the different G6PDH isoforms have been investigated, and the kinetic properties of partially purified cytosolic and plastidial G6PDHs determined. Bioinformatic analyses examining co-expression profiles of G6PDHs in Arabidopsis and barley corroborate the data presented. Moreover, the gene coding for the root P2-G6PDH isoform was fully sequenced; the biochemical properties of the corresponding protein were examined experimentally. The results are discussed in the light of the possible distinct roles and regulation of the different G6PDH isoforms during salt stress in barley roots.

  16. Wheat and barley allergens associated with baker's asthma. Glycosylated subunits of the alpha-amylase-inhibitor family have enhanced IgE-binding capacity.

    Science.gov (United States)

    Sanchez-Monge, R; Gomez, L; Barber, D; Lopez-Otin, C; Armentia, A; Salcedo, G

    1992-01-15

    A 16 kDa protein, designated CM16*, which strongly binds IgE from baker's-asthma patients has been identified as a glycosylated form of the previously reported WTAI-CM16, which is a subunit of the wheat tetrameric alpha-amylase inhibitor. A glycosylated form (CMb*) of BTAI-CMb, the equivalent inhibitor subunit from barley, has been also found to have significantly enhanced IgE-binding capacity. In all, 14 purified members of the alpha-amylase/trypsin-inhibitor family showed very different IgE-binding capacities when tested by a dot-blot assay. The glycosylated components CM16*, CMb* and the previously described non-glycosylated 14.5 kDa allergen from barley (renamed BMAI-1) were found to be the strongest allergens.

  17. Production of barley endoprotease B2 in Pichia pastoris and its proteolytic activity against native and recombinant hordeins

    DEFF Research Database (Denmark)

    Rosenkilde, Anne Lind; Dionisio, Giuseppe; Holm, Preben Bach

    2014-01-01

    Barley (Hordeum vulgare L.) cysteine proteases are of fundamental biological importance during germination but may also have a large potential as commercial enzyme. Barley cysteine endoprotease B2 (HvEPB2) was expressed in Pichia pastoris from a pPICZαA based construct encoding a HvEPB2 C......-terminal truncated version (HvEPB2ΔC) and a proteolytic resistant His6 tag. Maximum yield was obtained after 4 days of induction. Recombinant HvEPB2ΔC (r-HvEPB2ΔC) was purified using a single step of Ni2+-affinity chromatography. Purified protein was evaluated by SDS–PAGE, Western blotting and activity assays...

  18. Spatio-temporal changes in germination and radical elongation of barley seeds tracked by proteome analysis of dissected embryo, aleurone layer and endosperm tissues

    DEFF Research Database (Denmark)

    Bønsager, Birgit Christine; Finnie, Christine; Roepstorff, P.

    2007-01-01

    Germination of barley is accompanied by changes in water-soluble seed proteins. 2-DE was used to describe spatio-temporal proteome differences in dissected seed tissues associated with germination and the subsequent radicle elongation. Protein identification by MS enabled assignment of proteins...... protein and an ABA-induced protein, suggesting that these proteins are also involved in desiccation stress. Several redox-related proteins differed in spatio-temporal patterns at the end of germination and onset of radicle elongation. Notably, ascorbate peroxidase that was observed only in the embryo......, increased in abundance at 36 h PI. The surprisingly early changes seen in the protein profiles already 4 h after imbibition indicate that germination is programmed during seed maturation...

  19. Optimization of TaDREB3 gene expression in transgenic barley using cold-inducible promoters.

    Science.gov (United States)

    Kovalchuk, Nataliya; Jia, Wei; Eini, Omid; Morran, Sarah; Pyvovarenko, Tatiana; Fletcher, Stephen; Bazanova, Natalia; Harris, John; Beck-Oldach, Kontanze; Shavrukov, Yuri; Langridge, Peter; Lopato, Sergiy

    2013-08-01

    Constitutive over-expression of the TaDREB3 gene in barley improved frost tolerance of transgenic plants at the vegetative stage of plant development, but leads to stunted phenotypes and 3- to 6-week delays in flowering compared to control plants. In this work, two cold-inducible promoters with contrasting properties, the WRKY71 gene promoter from rice and the Cor39 gene promoter from durum wheat, were applied to optimize expression of TaDREB3. The aim of the work was to increase plant frost tolerance and to decrease or prevent negative developmental phenotypes observed during constitutive expression of TaDREB3. The OsWRKY71 and TdCor39 promoters had low-to-moderate basal activity and were activated by cold treatment in leaves, stems and developing spikes of transgenic barley and rice. Expression of the TaDREB3 gene, driven by either of the tested promoters, led to a significant improvement in frost tolerance. The presence of the functional TaDREB3 protein in transgenic plants was confirmed by the detection of strong up-regulation of cold-responsive target genes. The OsWRKY71 promoter-driven TaDREB3 provides stronger activation of the same target genes than the TdCor39 promoter. Analysis of the development of transgenic plants in the absence of stress revealed small or no differences in plant characteristics and grain yield compared with wild-type plants. The WRKY71-TaDREB3 promoter-transgene combination appears to be a promising tool for the enhancement of cold and frost tolerance in crop plants but field evaluation will be needed to confirm that negative development phenotypes have been controlled.

  20. Analysis of early events in the interaction between Fusarium graminearum and the susceptible barley (Hordeum vulgare) cultivar Scarlett

    DEFF Research Database (Denmark)

    Yang, Fen; Jensen, J.D.; Svensson, Birte;

    2010-01-01

    after inoculation. This coincided with the appearance of discrete F. graminearum-induced proteolytic fragments of beta-amylase. Based on these results, analysis of grain proteome changes prior to extensive proteolysis enabled identification of barley proteins responding early to infection by the fungus...... function was identified. Quantitative real-time RT-PCR analysis of selected genes showed a correlation between high gene expression and detection of the corresponding proteins. Fungal genes encoding alkaline protease and endothiapepsin were expressed during 1-3 days after inoculation, making them...... candidates for generation of the observed beta-amylase fragments. These fragments have potential to be developed as proteome-level markers for fungal infection that are also informative about grain protein quality....

  1. Antisense downregulation of the barley limit dextrinase inhibitor modulates starch granule size distribution, starch composition and amylopectin structure.

    Science.gov (United States)

    Stahl, Yvonne; Coates, Steve; Bryce, James H; Morris, Peter C

    2004-08-01

    The barley protein limit dextrinase inhibitor (LDI), structurally related to the alpha-amylase/trypsin inhibitor family, is an inhibitor of the starch debranching enzyme limit dextrinase (LD). In order to investigate the function of LDI, and the consequences for starch metabolism of reduced LDI activity, transgenic barley plants designed to downregulate LDI by antisense were generated. Homozygous antisense lines with reduced LDI protein level and activity were analysed and found to have enhanced free LD activity in both developing and germinating grains. In addition the antisense lines showed unpredicted pleiotropic effects on numerous enzyme activities, for example, alpha- and beta-amylases and starch synthases. Analysis of the starch showed much reduced numbers of the small B-type starch granules, as well as reduced amylose relative to amylopectin levels and reduced total starch. The chain length distribution of the amylopectin was modified with less of the longer chains (>25 units) and enhanced number of medium chains (10-15 units). These results suggest an important role for LDI and LD during starch synthesis as well as during starch breakdown.

  2. Phylogenetic and comparative gene expression analysis of barley (Hordeum vulgare)WRKY transcription factor family reveals putatively retained functions betweenmonocots and dicots

    Energy Technology Data Exchange (ETDEWEB)

    Mangelsen, Elke; Kilian, Joachim; Berendzen, Kenneth W.; Kolukisaoglu, Uner; Harter, Klaus; Jansson, Christer; Wanke, Dierk

    2008-02-01

    WRKY proteins belong to the WRKY-GCM1 superfamily of zinc finger transcription factors that have been subject to a large plant-specific diversification. For the cereal crop barley (Hordeum vulgare), three different WRKY proteins have been characterized so far, as regulators in sucrose signaling, in pathogen defense, and in response to cold and drought, respectively. However, their phylogenetic relationship remained unresolved. In this study, we used the available sequence information to identify a minimum number of 45 barley WRKY transcription factor (HvWRKY) genes. According to their structural features the HvWRKY factors were classified into the previously defined polyphyletic WRKY subgroups 1 to 3. Furthermore, we could assign putative orthologs of the HvWRKY proteins in Arabidopsis and rice. While in most cases clades of orthologous proteins were formed within each group or subgroup, other clades were composed of paralogous proteins for the grasses and Arabidopsis only, which is indicative of specific gene radiation events. To gain insight into their putative functions, we examined expression profiles of WRKY genes from publicly available microarray data resources and found group specific expression patterns. While putative orthologs of the HvWRKY transcription factors have been inferred from phylogenetic sequence analysis, we performed a comparative expression analysis of WRKY genes in Arabidopsis and barley. Indeed, highly correlative expression profiles were found between some of the putative orthologs. HvWRKY genes have not only undergone radiation in monocot or dicot species, but exhibit evolutionary traits specific to grasses. HvWRKY proteins exhibited not only sequence similarities between orthologs with Arabidopsis, but also relatedness in their expression patterns. This correlative expression is indicative for a putative conserved function of related WRKY proteins in mono- and dicot species.

  3. Removal and isolation of germ-rich fractions from hull-less barley using a fitzpatrick comminuting mill

    Science.gov (United States)

    A process was developed to produce a germ-enriched fraction from hull-less barley using a Fitzpatrick Comminuting Mill followed by sieving. Hulled and hull-less barleys contain 1.5-2.5% oil and, like wheat kernels which contain wheat germ oil, much of the oil in barley kernels is in the germ fracti...

  4. HvDep1 Is a Positive Regulator of Culm Elongation and Grain Size in Barley and Impacts Yield in an Environment-Dependent Manner

    Science.gov (United States)

    Wendt, Toni; Holme, Inger; Dockter, Christoph; Preuß, Aileen; Thomas, William; Waugh, Robbie; Braumann, Ilka

    2016-01-01

    Heterotrimeric G proteins are intracellular membrane-attached signal transducers involved in various cellular processes in both plants and animals. They consist of three subunits denoted as α, β and γ. The γ-subunits of the so-called AGG3 type, which comprise a transmembrane domain, are exclusively found in plants. In model species, these proteins have been shown to participate in the control of plant height, branching and seed size and could therefore impact the harvestable yield of various crop plants. Whether AGG3-type γ-subunits influence yield in temperate cereals like barley and wheat remains unknown. Using a transgenic complementation approach, we show here that the Scottish malting barley cultivar (cv.) Golden Promise carries a loss-of-function mutation in HvDep1, an AGG3-type subunit encoding gene that positively regulates culm elongation and seed size in barley. Somewhat intriguingly, agronomic field data collected over a 12-year period reveals that the HvDep1 loss-of-function mutation in cv. Golden Promise has the potential to confer either a significant increase or decrease in harvestable yield depending on the environment. Our results confirm the role of AGG3-type subunit-encoding genes in shaping plant architecture, but interestingly also indicate that the impact HvDep1 has on yield in barley is both genotypically and environmentally sensitive. This may explain why widespread exploitation of variation in AGG3-type subunit-encoding genes has not occurred in temperate cereals while in rice the DEP1 locus is widely exploited to improve harvestable yield. PMID:28005988

  5. The effect of free air carbon dioxide enrichment and nitrogen fertilisation on the chemical composition and nutritional value of wheat and barley grain.

    Science.gov (United States)

    Wroblewitz, Stefanie; Hüther, Liane; Manderscheid, Remy; Weigel, Hans-Joachim; Wätzig, Hermann; Dänicke, Sven

    2013-08-01

    A rising atmospheric CO2 concentration might influence the nutrient composition of feedstuffs and consequently the nutritional value for livestock. The present study investigates the effects of atmospheric CO2 enrichment on the chemical composition and nutritional value of winter wheat cv. "Batis" and winter barley cv. "Theresa". Both cereals were grown at two different atmospheric CO2 concentrations (ambient CO2 [AMBI]: 380 ppm and enriched CO2 [free air carbon dioxide enrichment, FACE]: 550 ppm) for two growing seasons. The influence of two different nitrogen (N) fertilisation levels (adequate N supply [N100] and nearly 50% of adequate N supply [N50]) were studied as well. A significant effect was observed for the crude protein content, which declined at FACE condition in a range of 8-16 g kg(-1) in wheat and of 10-20 g kg(-1) in barley. A reduced N fertilisation level resulted in a strong reduction of crude protein concentration in both cereal species. In wheat, a decrease in N supply significantly enhanced the concentration of starch and crude fibre. In barley, only the concentration of fructose increased under FACE condition and reduced N fertilisation. The FACE did not have major effects on the concentrations of minerals, while the influence of N fertilisation was different for both cereals. Whereas no effects could be observed for barley, a reduced N supply caused a significant reduction in concentrations of zinc, manganese and iron in wheat. Furthermore, an undirected effect of atmospheric CO2 and N fertilisation levels were found for the amino acid concentrations. Based on these results, future scenarios of climate change would have an impact on the nutritional value of cereal grains.

  6. Dryland malt barley yield and quality affected by tillage, cropping sequence, and nitrogen fertilization

    Science.gov (United States)

    Information is needed on the effects of management practices on dryland malt barley (Hordeum vulgaris L.) and pea (Pisum sativum L.) yields and quality. We evaluated the effects of tillage and cropping sequence combination and N fertilization on dryland malt barley and pea yields, grain characterist...

  7. Malt barley yield and quality affected by irrigation, tillage, crop rotation, and nitrogen fertilization

    Science.gov (United States)

    Little is known about the comparison of management practices on malt barley (Hordeum vulgare L.) yield and quality in irrigated and non-irrigated cropping systems. We evaluated the effects of irrigation, tillage, cropping system, and N fertilization on malt barley yield and quality in a sandy loam s...

  8. Influence of alkali catalyst on product yield and properties via hydrothermal liquefaction of barley straw

    DEFF Research Database (Denmark)

    Zhu, Z.; Toor, Saqib; Rosendahl, Lasse

    2015-01-01

    Barley straw was successfully converted to bio-crude by hydrothermal liquefaction at temperature of 280 e400 C using an alkali catalyst (K2CO3) in our previous work, and the maximum bio-crude yield was obtained at 300 C. This paper extends previous work on studying liquefaction behavior of barley...

  9. Regrowth in Barley (Hordeum vulgare L.) and Rye (Secale cereale L.)

    DEFF Research Database (Denmark)

    Christiansen, J L; Jørgensen, Johannes Ravn; Jørnsgård, B;

    1998-01-01

    Regrowth after cutting at four development stages, from heading to grain maturity, was investigated in a pot experiment containing three rye and four barley varieties (including 2 Hordeum spontaneum lines). Regrowth in the barley varieties decreased strongly from heading to grain maturity. Rye ge...

  10. Integration of weed management and tillage practices in spring barley production

    Science.gov (United States)

    Spring barley can be used to diversify and intensify winter wheat-based production systems in the U.S. Pacific Northwest, but the response of barley to conservation tillage systems, which are needed to reduce the risk of soil erosion, is not well documented. The objective of this study was to descri...

  11. Transformation of Barley (Hordeum vulgar L.) by Agrobacterium tumefasciens Infection of In Vitro Cultured Ovules

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Brinch-Pedersen, Henrik; Lange, Mette;

    2012-01-01

    Agrobacterium-mediated transformation of in vitro cultured barley ovules is an attractive alternative to well-established barley transformation methods of immature embryos. The ovule culture system can be used for transformation with and without selection and has successfully been used to transform...

  12. Compositional equivalence of barleys differing only in low and normal phytate levels

    Science.gov (United States)

    Recent breeding advances have led to the development of several barley lines with reduced levels of phytate. One of them was further developed and released as a hulless low phytate cultivar (Clearwater). Because barley oil contains high levels of tocotrienols and other functional lipids, we conduc...

  13. Determinants of barley grain yield in a wide range of Mediterranean environments

    NARCIS (Netherlands)

    Francia, E.; Tondelli, A.; Rizza, F.; Badeck, F.W.; Li Destri Nicosia, O.; Akar, T.; Grando, S.; Al-Yassin, A.; Benkelkacim, A.; Thomas, W.T.B.; Eeuwijk, van F.A.; Romagosa, I.; Stanca, A.M.; Pechionni, N.

    2011-01-01

    Barley grain yield in rainfed Mediterranean regions can be largely influenced by terminal drought events. In this study the ecophysiological performance of the ‘Nure’ (winter) × ‘Tremois’ (spring) barley mapping population (118 Doubled Haploids, DHs) was evaluated in a multi-environment trial of eig

  14. Dilute-acid pretreatment of barley straw for biological hydrogen production using Caldicellulosiruptor saccharolyticus

    NARCIS (Netherlands)

    Panagiotopoulos, I.A.; Bakker, R.R.C.; Vrije, de G.J.; Claassen, P.A.M.; Koukios, E.G.

    2012-01-01

    The main objective of this study was to use the fermentability test to investigate the feasibility of applying various dilute acids in the pretreatment of barley straw for biological hydrogen production. At a fixed acid loading of 1% (w/w dry matter) 28-32% of barley straw was converted to soluble m

  15. A comparison of energy use and productivity of wheat and barley (case study

    Directory of Open Access Journals (Sweden)

    S.M. Ziaei

    2015-01-01

    Full Text Available Comparison of energy productivity of different crops can be used as an effective tool to prioritize crops planting in each area. This study was conducted in order to compare wheat and barley farms of Sistan and Baluchestan province in Iran in relation to various aspects of energy consumption at 2009. 100 wheat and 100 barley fields were selected randomly from main 11 cities in the studied region. Input data and yield of wheat and barley fields were collected in the form of questionnaires in a face-to-face interview. Results showed that total energy inputs of wheat and barley fields were 32492.97 and 25655.81 MJ ha−1, respectively. Total energy outputs for wheat and barley fields were 48517.24 and 49800.87 MJ ha−1, respectively. Based on these results the amount of energy use efficiency for wheat and barley fields were 1.49 and 1.94, respectively, and the amount of energy productivity for mentioned fields were 0.056 and 0.066. The share of renewable energy as one of the sustainability indexes of agricultural systems was 19.60 for wheat and 14.60 for barley fields. Therefore, it seems that barley production is more efficient from various aspects of energy consumption rather than wheat in the studied region.

  16. Quantifying relationships between rooting traits and water uptake under drought in Mediterranean barley and durum wheat

    Institute of Scientific and Technical Information of China (English)

    Pedro Carvalho; Sayed AzamAli; M. John Foulkes

    2014-01-01

    In Mediterranean regions drought is the major factor limiting spring barley and durum wheat grain yields. This study aimed to compare spring barley and durum wheat root and shoot responses to drought and quantify relationships between root traits and water uptake under terminal drought. One spring barley (Hordeum vulgare L. cv. Rum) and two durum wheat Mediterranean cultivars (Triticum turgidum L. var durum cvs Hourani and Karim) were examined in soil-column experi-ments under wel watered and drought conditions. Root system architecture traits, water uptake, and plant growth were measured. Barley aerial biomass and grain yields were higher than for durum wheat cultivars in wel watered conditions. Drought decreased grain yield more for barley (47%) than durum wheat (30%, Hourani). Root-to-shoot dry matter ratio increased for durum wheat under drought but not for barley, and root weight increased for wheat in response to drought but decreased for barley. The critical root length density (RLD) and root volume density (RVD) for 90%available water capture for wheat were similar to (cv. Hourani) or lower than (cv. Karim) for barley depending on wheat cultivar. For both species, RVD accounted for a slightly higher proportion of phenotypic variation in water uptake under drought than RLD.

  17. High capacity of plant regeneration from callus of interspecific hybrids with cultivated barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Bagger Jørgensen, Rikke; Jensen, C. J.; Andersen, B.

    1986-01-01

    Callus was induced from hybrids between cultivated barley (Hordeum vulgare L. ssp. vulgare) and ten species of wild barley (Hordeum L.) as well as from one backcross line ((H. lechleri .times. H. vulgare) .times. H. vulgare). Successful callus induction and regeneration of plants were achieved from...

  18. A promising low beta-glucan barley mutation of m351 for better bioethanol production use

    Science.gov (United States)

    Bioethanol is an important liquid fuel complement. Barley is an alternative raw material for ethanol production and its byproduct is a nutritious feed. The barley m351mutant line, which has a mutation for low beta-glucan content, was tested for its ethanol production efficiency and feed fraction qua...

  19. Quantitative Analysis of the Early Powdery Mildew Infection Stages on Resistant Barley Genotypes

    DEFF Research Database (Denmark)

    Andersen, J. B.; Torp, J.

    1986-01-01

    A classification system was developed, that allowed quantification of the leaf surface development of the barley powdery mildew fungus on barley. An experiment with Manchuria and Pallas as susceptible controls and 4 resistance gene each represented by three lines with different gene backgrounds s...

  20. Differential levels of mite infestation of wheat and barley in Czech grain stores

    Institute of Scientific and Technical Information of China (English)

    Jan Hubert; Zuzana Kucerova; Radek Aulicky; Marta Nesvoma; Vaclav Stejskal

    2009-01-01

    While mites are able to utilize numerous food sources, the suitability of the food strongly influences population growth. The different suitabilities of various stored agricultural products will thus affect the level of infestation. In this study, we compared field mite infestation rates in two stored cereals: wheat and barley. We analyzed mite abundance, frequency and species composition in samples of grain obtained from 79 selected Czech grain stores. Stored barley seemed to be more vulnerable to mite attack than wheat, as we consistently found more infested samples, more species and higher mean and median mite abundance per sample in barley as compared to wheat. The mean mite abundance per sample were 55 and 506 individuals for wheat and barley, respectively. In barley, 10% of samples exceeded allergen risk threshold (i.e., 1 000 individuals per kg of grain). Altogether, 25 species were identified from approximately 35 000 individuals. The most frequently identified species were the same in wheat and barley, that is, Tydeus interruptus Sig Thor, Acarus siro L., Tarsonemus granarius Lindquist, Lepidoglyphus destructor (Schrank) and 1),rophagusputrescentiae (Schrank). Based on principal components analysis, we found a closer association of T. interruptus, T. putrescentiae, L. destructor and Cheyletus eruditus (Schrank) with barley samples, corresponding to the high frequency and abundance values of these mites. The probable reasons for the higher infestation, especially mite abundance in barley, are discussed in relation to the higher proportion of crushed parts, which may release favorable nutrient sources and amplify the abundance values.

  1. Immunomodulatory properties of oat and barley β-glucan populations on bone marrow derived dendritic cells

    NARCIS (Netherlands)

    Rosch, Christiane; Meijerink, Marjolein; Delahaije, Roy J.B.M.; Taverne, Nico; Gruppen, Harry; Wells, Jerry M.; Schols, Henk A.

    2016-01-01

    Specific structures of oat and barley β(1,3)(1,4)-glucans induced different in vitro immunomodulatory effects in bone marrow derived dendritic cells (BMDC) from TLR2/4 knock out mice. All barley β-glucan fractions induced larger amounts of cytokines in BMDCs than their oat equivalents. The particula

  2. Effect of different ascorbic acid levels (Vitamin C on eco-physiological properties of barley in soils contaminated with lead

    Directory of Open Access Journals (Sweden)

    Kamdin Akhavan Samimi

    2016-03-01

    Full Text Available This study was carried out to examine the effects of foliar application of ascorbic acid on barley in contaminated soil in a completely randomized factorial design with 2 factors, 9 treatments and 3 replications in Varamin in 1393.150 mg of lead nitrate per kg of soil were applied to infect the soil for all treatments. Superabsorbent was the first factor used in three levels (0, 3, 6 g per kg soil and ascorbic acid as the second factor was also used in three levels (0, 50 and 100 ppm. The results of this experiment showed that increase in superabsorbent and ascorbic acid concentrations in barley improved the morphological traits such as plant height and spike and grain number, grain weight, total weight of shoot, root dry weight and thousand grain weight and also improved physiological traits such as protein content and chlorophyll a, b and total chlorophyll in barley, moreover, increase in ascorbic acid in the plant resulted in reduction in antioxidant enzymes content such as superoxide dismutase andcatalase, and physiological traits such as proline, increased relative water content and reduced lead content in leaves and roots.So it can be concluded that, given that the country is located in arid and semiarid regions and considering Iran’s soils pollution with heavy metals,using effective treatments such as ascorbic acid can enhance crop water holding capacity and also reduce the effects of these elements toxicity. Therefore,the use of ascorbic acid seems essential.Due to the non-degradable and long life of heavy metals in soil, insoluble hydrophilic polymers with different amounts carboxylic groups are used. The surface carboxylic groups of the polymer (SAP due to exposure to pH are ionized and make strong bonds with soil pollutant metals, and eventually form a gel and are separated from soil.

  3. Effects of processing technologies combined with cell wall degrading enzymes on in vitro degradability of barley.

    Science.gov (United States)

    de Vries, S; Pustjens, A M; Schols, H A; Hendriks, W H; Gerrits, W J J

    2012-12-01

    Effects of processing technologies and cell wall degrading enzymes on in vitro degradation of barley were tested in a 5 × 2 factorial arrangement: 5 technologies (unprocessed, wet-milling, extrusion, autoclaving, and acid-autoclaving), with or without enzymes. Upper gastrointestinal tract digestion (Boisen incubation) and large intestinal fermentation (gas production technique) were simulated in duplicate. All technologies increased digestion of DM (13 to 43% units) and starch (22 to 51% units) during Boisen incubation, compared with the unprocessed control (P starch (≈ 20% units), and CP (≈ 10% units) in unprocessed and autoclaved barley (P starch present in the Boisen residues. In conclusion, wet-milling, extrusion, and acid-autoclaving improved in vitro starch and CP digestion in barley, which is related to the cell wall matrix disruption. Addition of xylanases and β-glucanases improved in vitro starch and CP digestion only in unprocessed barley or barley poorly affected by processing.

  4. Efficacy of imidacloprid for control of cereal leaf beetle (Coleoptera: Chrysomelidae) in barley.

    Science.gov (United States)

    Tharp, C; Blodgett, S L; Johnson, G D

    2000-02-01

    The toxicity of imidacloprid to the cereal leaf beetle, Oulema melanopus (L.), was measured under laboratory and field conditions. Insect mortality and plant damage were determined from artificial and natural infestations of O. melanopus applied to various growth stages of barley. All rates of imidacloprid formulated and applied as a seed treatment caused >90% mortality to cereal leaf beetle larvae when barley was infested with eggs at the 4-leaf stage, but were ineffective when barley was infested with eggs at the early tillering or flag-leaf stages of barley. This window of susceptibility influenced results obtained in field trials where peak larval emergence did not occur until the early tillering stage of barley. The resulting mortality in plants from treated seeds never exceeded 40% in the field. Foliar imidacloprid, however, caused >90% mortality in the field, and may be another option in the management of the cereal leaf beetle.

  5. 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.

    Directory of Open Access Journals (Sweden)

    Sachin Rustgi

    Full Text Available 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.

  6. Long-term reconstitution with water of dry barley before feeding increases phosphorus digestibility in pigs1

    DEFF Research Database (Denmark)

    Ton Nu, Mai Anh; Blaabjerg, Karoline; Poulsen, Hanne Damgaard

    2016-01-01

    compared to dry stored barley on apparent total tract digestibility (ATTD) of P in pigs. Dry barley (12% moisture; total P, 3.3 g/kg DM; phytate P, 1.7 g/kg DM) was rolled and stored directly or reconstituted with water to produce rolled barley with 35% moisture that was stored in air-tight conditions...... numerically more soluble P (2.56 g/kg DM) and less phytate P (0.93 g/kg DM) than dry barley (0.78 and 1.7 g/kg DM). Pigs fed the reconstituted barley diet showed an increase in P absorption (54%, P

  7. New starch phenotypes produced by TILLING in barley.

    Directory of Open Access Journals (Sweden)

    Francesca Sparla

    Full Text Available 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.

  8. Characterization of chloroplast proteins controlling manganese use efficiency by quantitative proteomics using 2D-LC-MS/MS

    DEFF Research Database (Denmark)

    Petersen, Jørgen; Sprenger, Richard Remko; Rogowska-Wrzesinska, Adelina;

    Manganese deficiency is a worldwide problem. We set up to identify and quantify key protein responsible for manganese use efficiency in different genotypes of barley, with the overall aim to make sustainable crops...

  9. Expression in Escherichia coli of cDNA encoding barley beta-amylase and properties of recombinant beta-amylase.

    Science.gov (United States)

    Yoshigi, N; Okada, Y; Sahara, H; Koshino, S

    1994-06-01

    To express the cloned beta-amylase cDNA in Escherichia coli under control of the tac promoter, a plasmid pBETA92 was constructed. The plasmid consisted of 6312 bp. An extract of E. coli JM109 harboring pBETA92 had beta-amylase activity that produced beta-maltose from soluble starch. The enzyme production started in the logarithmic phase, increased linearly, and reached a maximum after 12 h. The recombinant barley beta-amylase gave two major (pI 5.43 and 5.63) and four minor (pI 5.20, 5.36, 5.80, and 6.13) activity bands on isoelectric focusing, and their pIs didn't change throughout the incubation. But Western blot analysis found that one beta-amylase having a molecular weight of about 56,000 was synthesized. The recombinant beta-amylase was purified from the cells by consecutive column chromatography. The purified enzyme gave a single band of protein on SDS-PAGE but showed heterogeneity on isoelectric focusing. The N-terminal amino acid sequence showed that the recombinant beta-amylase lacked four amino acids at positions 2-5 (Glu-Val-Asn-Val) when compared with the presumed amino acid sequence of barley beta-amylase. Therefore, the recombinant beta-amylase consisted of 531 amino acids, and its molecular weight was calculated to be 59,169. The N-terminal amino acid sequence of the recombinant beta-amylase and the nucleotide sequence of the junction position in plasmid pBETA92 indicated that GTG (Val-5 in the case of barley beta-amylase) at positions 27-29 from the SD sequence (AGGA) was the translation initiation codon. The properties of the recombinant beta-amylase were almost the same as those of barley beta-amylase except for the pI and the Km values for maltohexaose and maltoheptaose.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. Modelling phytate degradation kinetics in soaked wheat and barley

    DEFF Research Database (Denmark)

    Blaabjerg, Karoline; Strathe, A B; Poulsen, Hanne Damgaard

    2012-01-01

    for describing the phytate degradation profiles of the soaked wheat and barley. The in vitro results showed that heat-treatment of the cereals increased the F0 and reduced the K although the plant phytase activity was reduced. This was possible due to induced structural changes of the grain that increased...... the contact between phytate and plant phytases. The lack of effect of additions of Phytase 1 or Phytase 2 on the phytate degradation in the cereals was possible due to a poor access of the microbial phytases to phytate. Therefore, more knowledge is needed about processing technologies that increase the access...... of plant and microbial phytases to phytate in the grains....

  11. Further molecular evidence for the Hordeum vulgare ssp. spontaneum in Tibet as ultimate progenitor of Chinese cultivated barley

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    106 accessions of Tibetan wild barley, including 50 accessions of the two-rowed wild barley Hordeum vulgare ssp. spontaneum(HS), 27 accessions of the six-rowed bottle-shaped wild barley H. lagunculiforme(HL) and 29 accessions of the six-rowed wild barley H. agriocrithon(HA) that separately represent different agrigeographical regions of Tibet, were used to study the genetic diversity and genetic differentiation using SSR markers selected from seven barley linkage groups. 229 allelic variants were identified with an average of 7.6 alleles/locus. The average of total number of alleles per locus in HA(6.4) is much higher than that in HS(3.9) and HL(3.4). The genetic diversity and its standard deviation among the three subspecies were in the order of HS>HL>HA. Very significant genetic differentiation was observed among the three subspecies of wild barley. Comparisons of the results from this and previous studies showed a strong Oriental-Occidental differentiation of barley, and that Shannan region of Tibet might be the center of origin of the Tibetan two-rowed wild barley, thus supporting not only the hypothesis of a mono-phyletic origin of cultivated barley but also the proposition that the Tibetan two-rowed wild barley as ultimate progenitor of Chinese cultivated barley.

  12. 大麦雄性不育的遗传研究%Genetic Study on Barley Male Sterility

    Institute of Scientific and Technical Information of China (English)

    张凤英; 刘志萍; 包海柱

    2009-01-01

    [Objective] The aim was to research the genetic mechanism of barley male sterility, and provide the theoretical reference for breeding strong heterosis hybrid of barley. [Method] Fertility segregation phenomenon, morphological characteristics and main agronomic characters of male sterility character of 4 kinds of barley male sterility materials 2001-17, 2001-37, 2001-84 and 2001-116 and their derived lines were observed and researched. [Result] Barley male sterility existed genetic phenomenon of single dominant nuclear gene caused by environmental stimulation, its sterility controlled by MS, the sterile genotype was MSms. There was stable nucleo-cytoplasmic inheritance on barley male sterility which was controlled by cytoplasmic male sterile gene S and nuclear gene rr, and its genotype was S(rr), this male sterility belonged to CMS type sterility. Temperature had no effect on fertility. [Conclusion] There was stable nucleo-cytoplasmic inheritance on barley male sterility, this result played a positively promoting role in barley practical production.

  13. Dynamics of phosphorus compounds in ripening and germinating cereal grains. Part I. Changes in phosphorus compounds content during ripening of wheat, barley and rye grains

    Directory of Open Access Journals (Sweden)

    J. Tłuczkiewicz

    2015-01-01

    Full Text Available The dynamics of phosphorus compounds was followed in ripening wheat (Grana, barley (Kosmos and rye (Pancerne seeds. In the investigated ontogenesis period the content in the seeds of total and acid-soluble phosphorus doubled. The phosphorus of free phytin and phytin that bound with proteins increased to the end of maturation. In dormant seeds the contribution of phosphorus bound with phytin was 62-70 per cent of total P. As the seeds ripened, the content of inorganic and nucleotides-P decreased, while that of saccharides-P and their metabolites started to decrease from the moment of wax ripeness. Later RNA-P and DNA-P level decreased only slight.ly and that of lipids-P markedly. Phosphoproteid content diminished at the beginning of ripening and further remained at almost the same level. Wheat, barley and rye seed exhibited similar dynamics of metabolically active (nucleotides, saccharide-P esters and their metabolites, functional (nucleic acids and structural (phospholipids phosphorus compounds. Accumulation of storage forms (phytin of phosphorus was higher in wheat and successively lower in rye and barley.

  14. Effects of lactic acid fermentation and gamma irradiation of barley on antinutrient contents and nutrient digestibility in mink (Mustela vison) with and without dietary enzyme supplement.

    Science.gov (United States)

    Skrede, Anders; Sahlstrøm, Stefan; Ahlstrøm, Oystein; Connor, Kirsti Hjelme; Skrede, Grete

    2007-06-01

    The experiment was conducted to study the effects of fermentation of barley, using two different strains of lactic acid bacteria, a Lactobacillus plantarum/pentosus strain isolated from spontaneously fermented rye sourdough (AD2) and a starch-degrading Lactobacillus plantarum (AM4), on contents of mixed-linked (1 --> 3) (1 --> 4)-beta-glucans, alpha-amylase inhibitor activity, inositol phosphates, and apparent digestibility of macronutrients in mink. Effects of fermentation were compared with effects of gamma irradiation (gamma-irradiation: 60Co gamma-rays at 25 kGy). The diets were fed to mink with and without a supplementary enzyme preparation. Both lactic acid fermentation and gamma-irradiation followed by soaking and incubation, reduced concentrations of soluble beta-glucans, phytate and alpha-amylase inhibitor activity. Dietary enzyme supplementation increased significantly digestibility of crude protein, fat, starch and crude carbohydrate (CHO). Fermentation of the barley increased digestibility of starch and CHO. Fermentation with lactic acid bacteria AD2 resulted in higher starch and CHO digestibility than strain AM4, and had greater effect than gamma-irradiation, soaking and incubation. The highest digestibility of starch and CHO was obtained after AD2 fermentation followed by enzyme supplementation. It is concluded that both lactic acid fermentation of barley and enzyme supplementation have positive nutritional implications in the mink by limiting the effects of antinutrients and improving digestibility and energy utilization.

  15. Endophytic colonization of barley (Hordeum vulgare) roots by the nematophagous fungus Pochonia chlamydosporia reveals plant growth promotion and a general defense and stress transcriptomic response.

    Science.gov (United States)

    Larriba, Eduardo; Jaime, María D L A; Nislow, Corey; Martín-Nieto, José; Lopez-Llorca, Luis Vicente

    2015-07-01

    Plant crop yields are negatively conditioned by a large set of biotic and abiotic factors. An alternative to mitigate these adverse effects is the use of fungal biological control agents and endophytes. The egg-parasitic fungus Pochonia chlamydosporia has been traditionally studied because of its potential as a biological control agent of plant-parasitic nematodes. This fungus can also act as an endophyte in monocot and dicot plants, and has been shown to promote plant growth in different agronomic crops. An Affymetrix 22K Barley GeneChip was used in this work to analyze the barley root transcriptomic response to P. chlamydosporia root colonization. Functional gene ontology (GO) and gene set enrichment analyses showed that genes involved in stress response were enriched in the barley transcriptome under endophytism. An 87.5% of the probesets identified within the abiotic stress response group encoded heat shock proteins. Additionally, we found in our transcriptomic analysis an up-regulation of genes implicated in the biosynthesis of plant hormones, such as auxin, ethylene and jasmonic acid. Along with these, we detected induction of brassinosteroid insensitive 1-associated receptor kinase 1 (BR1) and other genes related to effector-triggered immunity (ETI) and pattern-triggered immunity (PTI). Our study supports at the molecular level the growth-promoting effect observed in plants endophytically colonized by P. chlamydosporia, which opens the door to further studies addressing the capacity of this fungus to mitigate the negative effects of biotic and abiotic factors on plant crops.

  16. Nitrogen excretion of adult sheep fed silages made of a mixed sward or of pure unfertilised grass alone and in combination with barley.

    Science.gov (United States)

    Seip, Katharina; Breves, Gerhard; Isselstein, Johannes; Abel, Hansjörg

    2011-08-01

    Four adult rumen-fistulated wether sheep were fed silages combined with barley. The silages consisted of 48% grasses, 28% legumes and 24% other forbs (GCF) or of pure grass (G). The swards received no mineral fertiliser. The dry matter (DM) and fibre contents were lower in GCF than in G. Crude protein content of DM in GCF and G were 145 g/kg and 102 g/kg respectively. DM content as ash, lipids and non-fibrous carbohydrates were rather similar in both silages. About 40g DM were offered per kg BW0.75 and day either as silage alone or as a mixture of silage and barley (60:40). Faecal N excretion was greater with GCF than with G. The proportion of faecal bacterial and endogenous debris N reached 75 and 73% when GCF or G was fed, respectively. Undigested dietary N represented about 20%, and water soluble N accounted for 5-6% of faecal N. GCF caused more urinary N than G. Barley reduced urinary N excretion when supplemented to GCF. No dietary influence on urinary non-urea nitrogenous compounds was shown. GCF caused higher urinary urea N excretion than G and barley reduced this fraction when replacing part of GCF. Based on the urinary urea N proportions, it is concluded that N intake exceeded N requirement for any of the four diets fed. Dietary supplementation of ruminally fermentable carbohydrates can reduce urinary N excretion and this improves the efficiency of utilisation of N in N-unfertilised biodiverse grassland/ruminant farming systems.

  17. Identification of reference genes for quantitative expression analysis of microRNAs and mRNAs in barley under various stress conditions.

    Science.gov (United States)

    Ferdous, Jannatul; Li, Yuan; Reid, Nicolas; Langridge, Peter; Shi, Bu-Jun; Tricker, Penny J

    2015-01-01

    For accurate and reliable gene expression analysis using quantitative real-time reverse transcription PCR (qPCR), the selection of appropriate reference genes as an internal control for normalization is crucial. We hypothesized that non-coding, small nucleolar RNAs (snoRNAs)would be stably expressed in different barley varieties and under different experimental treatments,in different tissues and at different developmental stages of plant growth and therefore might prove to be suitable reference genes for expression analysis of both microRNAs (miRNAs)and mRNAs. In this study, we examined the expression stability of ten candidate reference genes in six barley genotypes under five experimental stresses, drought, fungal infection,boron toxicity, nutrient deficiency and salinity. We compared four commonly used housekeeping genes; Actin (ACT), alpha-Tubulin (α-TUB), Glycolytic glyceraldehyde-3-phosphate dehydrogenase(GAPDH), ADP-ribosylation factor 1-like protein (ADP), four snoRNAs; (U18,U61, snoR14 and snoR23) and two microRNAs (miR168, miR159) as candidate reference genes. We found that ADP, snoR14 and snoR23 were ranked as the best of these candidates across diverse samples. Additionally, we found that miR168 was a suitable reference gene for expression analysis in barley. Finally, we validated the performance of our stable and unstable candidate reference genes for both mRNA and miRNA qPCR data normalization under different stress conditions and demonstrated the superiority of the stable candidates. Our data demonstrate the suitability of barley snoRNAs and miRNAs as potential reference genes form iRNA and mRNA qPCR data normalization under different stress treatments [corrected].

  18. DNA polymorphisms and haplotype patterns of transcription factors involved in barley endosperm development are associated with key agronomic traits

    Directory of Open Access Journals (Sweden)

    Stracke Silke

    2010-01-01

    Full Text Available Abstract Background Association mapping is receiving considerable attention in plant genetics for its potential to fine map quantitative trait loci (QTL, validate candidate genes, and identify alleles of interest. In the present study association mapping in barley (Hordeum vulgare L. is investigated by associating DNA polymorphisms with variation in grain quality traits, plant height, and flowering time to gain further understanding of gene functions involved in the control of these traits. We focused on the four loci BLZ1, BLZ2, BPBF and HvGAMYB that play a role in the regulation of B-hordein expression, the major fraction of the barley storage protein. The association was tested in a collection of 224 spring barley accessions using a two-stage mixed model approach. Results Within the sequenced fragments of four candidate genes we observed different levels of nucleotide diversity. The effect of selection on the candidate genes was tested by Tajima's D which revealed significant values for BLZ1, BLZ2, and BPBF in the subset of two-rowed barleys. Pair-wise LD estimates between the detected SNPs within each candidate gene revealed different intra-genic linkage patterns. On the basis of a more extensive examination of genomic regions surrounding the four candidate genes we found a sharp decrease of LD (r2 Significant marker-trait associations between SNP sites within BLZ1 and flowering time, BPBF and crude protein content and BPBF and starch content were detected. Most haplotypes occurred at frequencies BPBF was associated to crude protein content and starch content, BLZ2 showed association to thousand-grain weight and BLZ1 was found to be associated with flowering time and plant height. Conclusions Differences in nucleotide diversity and LD pattern within the candidate genes BLZ1, BLZ2, BPBF, and HvGAMYB reflect the impact of selection on the nucleotide sequence of the four candidate loci. Despite significant associations, the analysed candidate

  19. Distribution and Molecular Characterization of β-Glucans from Hull-Less Barley Bran, Shorts and Flour

    Directory of Open Access Journals (Sweden)

    Qi Wang

    2011-02-01

    Full Text Available Six hull-less barley cultivars widely grown in China were roller-milled to produce bran, shorts and flour fractions. The distribution and molecular characteristics of β-glucans from the three roller-milled fractions were investigated. The β-glucan contents in the six hull-less barley cultivars varied from 4.96% to 7.62%. For all the six cultivars, the shorts fraction contained the highest concentration of β-glucan (8.12–13.01%, followed by bran (6.15–7.58% and flour (2.48–2.95%. Crude β-glucans were prepared from the three roller-milled fractions using aqueous sodium carbonate (pH 10. These preparations contained 45.38–71.41% β-glucan, 10.81–17.26% arabinoxylan, 2.6–9.6% protein, 2.7–9.0% starch, and 5.23–9.68% ash. Purification using a-amylase and β-xylanase in combination with pH adjustment and dialysis produced high purity β-glucan preparations (91–95%. The molecular weight (Mw of β-glucan preparations from roller-milled fractions ranged from 117,600 to 852,400 g/mol. β-Glucan from flour had higher Mw than those from shorts and bran within the same cultivar, and β-glucan preparations from bran had the lowest Mw.

  20. Expression analysis of abscisic acid (ABA) and metabolic signalling factors in developing endosperm and embryo of barley.

    Science.gov (United States)

    Chen, Zhiwei; Huang, Jianhua; Muttucumaru, Nira; Powers, Stephen J; Halford, Nigel G

    2013-09-01

    The expression of genes encoding components of ABA and metabolic signalling pathways in developing barley endosperm and embryo was investigated. The genes included HvRCAR35_47387 and HvRCAR35_2538 (encoding ABA receptors), HvABI1d (protein phosphatase 2C), HvSnRK2.4, HvSnRK2.6 and HvPKABA1 (SnRK2-type protein kinases) and HvABI5 (ABA response element binding protein; AREBP), as well as two genes encoding SnRK1-type protein kinases. Both SnRK1 and SnRK2 phosphorylate AREBPs, but SnRK2 is activated by ABA whereas SnRK1 may be broken down. Multiple cereal AREBPs with two conserved SnRK1/2 target sites and another class of BZIP transcription factors with SnRK1/2 binding sites, including HvBLZ1, were identified. Barley grain (cv. Triumph) was sampled at 15, 20, 25 and 30 days post-anthesis (dpa). HvRCAR35_47387, HvABI1d, HvSnRK2.4 and HvABI5 were expressed highly in the endosperm but at much lower levels in the embryo. Conversely, HvPKABA1 and HvRCAR35_2538 were expressed at higher levels in the embryo than the endosperm, while HvSnRK2.6 was expressed at similar levels in both. HvRCAR35_47387, HvABI1d, HvSnRK2.4 and HvABI5 all peaked in expression in the endosperm at 20 dpa. A model is proposed in which ABA brings about a transition from a SnRK1-dominated state in the endosperm during grain filling to a SnRK2-dominated state during maturation.

  1. Heat shock inhibits. alpha. -amylase synthesis in barley aleurone without inhibiting the activity of endoplasmic reticulum marker enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Sticher, L.; Biswas, A.K.; Bush, D.S.; Jones, R.L. (Univ. of California, Berkeley (USA))

    1990-02-01

    The effects of heat shock on the synthesis of {alpha}-amylase and on the membranes of the endoplasmic reticulum (ER) of barley (Hordeum vulgare) aleurone were studied. Heat shock, imposed by raising the temperature of incubation from 25{degree}C to 40{degree}C for 3 hours, inhibits the accumulation of {alpha}-amylase and other proteins in the incubation medium of barley aleurone layers treated with gibberellic acid and Ca{sup 2+}. When ER is isolated from heat-shocked aleurone layers, less newly synthesized {alpha}-amylase is found associated with this membrane system. ER membranes, as indicated by the activities of NADH cytochrome c reductase and ATP-dependent Ca{sup 2+} transport, are not destroyed by heat stress, however. Although heat shock did not reduce the activity of ER membrane marker enzymes, it altered the buoyant density of these membranes. Whereas ER from control tissue showed a peak of marker enzyme activity at 27% to 28% sucrose (1.113-1.120 grams per cubic centimeter), ER from heat-shocked tissue peaked at 30% to 32% sucrose (1.127-1.137 grams per cubic centimeter). The synthesis of a group of proteins designated as heat-shock proteins (HSPs) was stimulated by heat shock. These HSPs were localized to different compartments of the aleurone cell. Several proteins ranging from 15 to 30 kilodaltons were found in the ER and the mitochondrial/plasma membrane fractions of heat-shocked cells, but none of the HSPs accumulated in the incubation medium of heat-shocked aleurone layers.

  2. Barley metallothioneins: MT3 and MT4 are localized in the grain aleurone layer and show differential zinc binding.

    Science.gov (United States)

    Hegelund, Josefine Nymark; Schiller, Michaela; Kichey, Thomas; Hansen, Thomas Hesselhøj; Pedas, Pai; Husted, Søren; Schjoerring, Jan Kofod

    2012-07-01

    Metallothioneins (MTs) are low-molecular-weight, cysteine-rich proteins believed to play a role in cytosolic zinc (Zn) and copper (Cu) homeostasis. However, evidence for the functional properties of MTs has been hampered by methodological problems in the isolation and characterization of the proteins. Here, we document that barley (Hordeum vulgare) MT3 and MT4 proteins exist in planta and that they differ in tissue localization as well as in metal coordination chemistry. Combined transcriptional and histological analyses showed temporal and spatial correlations between transcript levels and protein abundance during grain development. MT3 was present in tissues of both maternal and filial origin throughout grain filling. In contrast, MT4 was confined to the embryo and aleurone layer, where it appeared during tissue specialization and remained until maturity. Using state-of-the-art speciation analysis by size-exclusion chromatography inductively coupled plasma mass spectrometry and electrospray ionization time-of-flight mass spectrometry on recombinant MT3 and MT4, their specificity and capacity for metal ion binding were quantified, showing a strong preferential Zn binding relative to Cu and cadmium (Cd) in MT4, which was not the case for MT3. When complementary DNAs from barley MTs were expressed in Cu- or Cd-sensitive yeast mutants, MT3 provided a much stronger complementation than did MT4. We conclude that MT3 may play a housekeeping role in metal homeostasis, while MT4 may function in Zn storage in developing and mature grains. The localization of MT4 and its discrimination against Cd make it an ideal candidate for future biofortification strategies directed toward increasing food and feed Zn concentrations.

  3. Effects of barley chromosome addition to wheat on behavior and development of Locusta migratoria nymphs.

    Science.gov (United States)

    Suematsu, Shunji; Harano, Ken-ichi; Tanaka, Seiji; Kawaura, Kanako; Ogihara, Yasunari; Watari, Yasuhiko; Saito, Osamu; Tokuda, Makoto

    2013-01-01

    Locusta migratoria feeds on various Poaceae plants but barley. Barley genes related to feeding deterrence may be useful for developing novel resistant crops. We investigated the effects of barley cultivar Betzes, wheat cultivar Chinese Spring (CS), and six barley chromosome disomic addition lines of wheat (2H-7H) on locomotor activity, feeding behavior, survival and development of L. migratoria nymphs. Locomotor activity was similar in nymphs kept with wheat and 2H-7H in an actograph, whereas it was generally high in those kept with barely. No-choice and choice feeding tests suggested that barley genes related to inhibition of feeding by L. migratoria are located on barley chromosomes 5H and 6H and those related to the palatability of plants on chromosomes 2H, 5H and 6H. Rearing experiments suggested the presence of barley genes negatively affecting the survival and growth of locust nymphs on chromosomes 5H and 2H, respectively, and the effects are phase-dependent.

  4. Genetic structure and linkage disequilibrium in landrace populations of barley in Sardinia.

    Science.gov (United States)

    Rodriguez, Monica; Rau, Domenico; O'Sullivan, Donal; Brown, Anthony H D; Papa, Roberto; Attene, Giovanna

    2012-06-01

    Multilocus digenic linkage disequilibria (LD) and their population structure were investigated in eleven landrace populations of barley (Hordeum vulgare ssp. vulgare L.) in Sardinia, using 134 dominant simple-sequence amplified polymorphism markers. The analysis of molecular variance for these markers indicated that the populations were partially differentiated (F(ST) = 0.18), and clustered into three geographic areas. Consistent with this population pattern, STRUCTURE analysis allocated individuals from a bulk of all populations into four genetic groups, and these groups also showed geographic patterns. In agreement with other molecular studies in barley, the general level of LD was low (13% of locus pairs, with P landrace populations, but that epistatic homogenising or diversifying selection was also present. Notably, the variance of the disequilibrium component was relatively high, which implies caution in the pooling of barley lines for association studies. Finally, we compared the analyses of multilocus structure in barley landrace populations with parallel analyses in both composite crosses of barley on the one hand and in natural populations of wild barley on the other. Neither of these serves as suitable mimics of landraces in barley, which require their own study. Overall, the results suggest that these populations can be exploited for LD mapping if population structure is controlled.

  5. Effect of supplementation with barley and calcium hydroxide on intake of Mediterranean shrubs

    Directory of Open Access Journals (Sweden)

    Dragan Skobic

    2011-04-01

    Full Text Available Maquis plant communities are one of the most varied vegetation types in the Mediterranean region and an important habitat for wild and domestic herbivores. Although the majority of these shrubs are nutritious, the secondary compounds are main impediments that reduce their forage value. In five experiments we determined the effect of supplementing goats with calcium hydroxide plus barley, and barley alone on intake of five dominant shrubs (Quercus ilex, Erica multiflora, Arbutus unedo, Viburnum tinus and Pistacia lentiscus of the Mediterranean maquis community. The combination of calcium hydroxide plus barley and barley alone increased utilization of all five investigated Mediterranean shrubs; therewith that intake of Arbutus unedo and Viburnum tinus was not statistically significant. Supplemented goats with calcium hydroxide plus barley or barley alone could be effective in controlling secondary compounds-rich Mediterranean shrubs where their abundance threatens biodiversity. This control can be facilitated by browsing dominant Mediterranean shrubs, which has been shown to be effective in managing Mediterranean maquis density. Calcium hydroxide and barley (energy enhance use of secondary compounds-containing plants, which may increase production of alternate forages and create a more diverse mix of plant species in the Mediterranean maquis plant community.

  6. Homeopathically prepared gibberellic acid and barley seed germination.

    Science.gov (United States)

    Hamman, B; Koning, G; Lok, K Him

    2003-07-01

    The potentisation process by which homeopathic preparations are produced raises the concern that these medicines have placebo effects only, since they theoretically no longer contain active molecules of the diluted substance. Plant models offer a method of examining the efficacy of homeopathically prepared solutions. This study examined the effects of homeopathically prepared gibberellic acid (HGA3) on the germination performance of barley (Hordeum vulgare L.) seeds. The effect of HGA3 (4-200 cH) on seed germination rate and seedling development was compared to that of the most commonly used form of gibberellic acid (GA3), 0.5 g l(-1), and control (distilled water). The extent and type of response was dependent on the vigour level of the seedlot. Treating seeds from three vigour groups in HGA3 consistently resulted in larger seedlings. High-vigour seeds treated with HGA3 4, 30 and 200 cH germinated faster, and roots of medium-vigour seedlots treated in HGA3 15 cH were longer. Biphasic effects of HGA3 were also demonstrated. As a plant model, germinating barley seeds successfully demonstrated the ability of HGA3 to produce a biological response.

  7. How barley growing conditions and its output change in Hungary

    Directory of Open Access Journals (Sweden)

    Éva Erdélyi

    2010-11-01

    Full Text Available There is no doubt that climate change has started. It is very important to make effort in developing impact analyses and adaptation strategies. First we were investigated how theproduction risk of winter barley is changing with time using the E,V efficiency criterion. Based on the regional yearly production data of the crop, we can conclude that beside other non-climatic effects, the changing climate has considerable impact on crops yield; its variability is increasing with the variability of meteorological parameters. We have used production data from 1951 to nowadays. Next, using comparison analyses for climate scenarios, we predict what we can expect in the future. For detecting the reasons of risk increase in the past, and forecasting the potential main points of future risk we have analysed statistically whether the climate needs of winter barley will be satisfied ornot in its important periods of growing. Frequency calculations were made based on the daily meteorological data. The situation doesn’t show big change, but It is no doubt that the anomalies of the indicators have been becoming more and more frequent. The morefrequent the extreme weather events are, the more we can be convinced of uncertainty.

  8. Effect of cultivar and year on phyllochron in winter barley

    Directory of Open Access Journals (Sweden)

    Pržulj Novo M.

    2013-01-01

    Full Text Available Development and growth of leaves in cereals significantly affects grain yield since dry matter accumulation depends on the leaf area that intercepts light. Phyllochron (PHY is defined as time interval between the emergences of successive leaves on the main stem. The aim of this study was to determine the effect of year and cultivar on phyllochron in winter barley. Twelve cultivars of winter barley differing in origin and time of anthesis were tested during six growing seasons (GS, from 2002/03 to 2007/08. The highest PHY across GSs was determined in the two-rowed cultivar Cordoba (81.6°Cd and the lowest in the two-rowed cultivar Novosadski 581 (71.0°Cd. The early cultivars had fast leaf development, the medium cultivars medium and the late cultivars slow development, 72.5°Cd, 75.6°Cd and 78.9°Cd, respectively. The tested cultivars showed significant variability in the PHY, which can be used for selecting most adaptable genotypes for specific growing conditions.

  9. Accumulation of Transcripts Abundance after Barley Inoculation with Cochliobolus sativus

    Directory of Open Access Journals (Sweden)

    Mohammad Imad Eddin Arabi

    2015-03-01

    Full Text Available Spot blotch caused by the hemibiotrophic pathogen Cochliobolus sativus has been the major yield-reducing factor for barley production during the last decade. Monitoring transcriptional reorganization triggered in response to this fungus is an essential first step for the functional analysis of genes involved in the process. To characterize the defense responses initiated by barley resistant and susceptible cultivars, a survey of transcript abundance at early time points of C. sativus inoculation was conducted. A notable number of transcripts exhibiting significant differential accumulations in the resistant and susceptible cultivars were detected compared to the non-inoculated controls. At the p-value of 0.0001, transcripts were divided into three general categories; defense, regulatory and unknown function, and the resistant cultivar had the greatest number of common transcripts at different time points. Quantities of differentially accumulated gene transcripts in both cultivars were identified at 24 h post infection, the approximate time when the pathogen changes trophic lifestyles. The unique and common accumulated transcripts might be of considerable interest for enhancing effective resistance to C. sativus.

  10. In Vitro Biochemical Characterization of All Barley Endosperm Starch Synthases

    Directory of Open Access Journals (Sweden)

    Jose Antonio Cuesta-Seijo

    2016-01-01

    Full Text Available Starch is the main storage polysaccharide in cereals and the major source of calories in the human diet. It is synthesized by a panel of enzymes including five classes of starch synthases (SSs. While the overall starch synthase (SS reaction is known, the functional differences between the five SS classes are poorly understood. Much of our knowledge comes from analyzing mutant plants with altered SS activities, but the resulting data are often difficult to interpret as a result of pleitropic effects, competition between enzymes, overlaps in enzyme activity and disruption of multi-enzyme complexes. Here we provide a detailed biochemical study of the activity of all five classes of SSs in barley endosperm. Each enzyme was produced recombinantly in E. coli and the properties and modes of action in vitro were studied in isolation from other SSs and other substrate modifying activities. Our results define the mode of action of each SS class in unprecedented detail; we analyze their substrate selection, temperature dependence and stability, substrate affinity and temporal abundance during barley development. Our results are at variance with some generally accepted ideas about starch biosynthesis and might lead to the reinterpretation of results obtained in planta. In particular, they indicate that granule bound SS is capable of processive action even in the absence of a starch matrix, that SSI has no elongation limit, and that SSIV, believed to be critical for the initiation of starch granules, has maltoligosaccharides and not polysaccharides as its preferred substrates.

  11. Nitric oxide and reactive oxygen species mediate metabolic changes in barley seed embryo during germination

    Directory of Open Access Journals (Sweden)

    Zhenguo eMa

    2016-02-01

    Full Text Available 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 postimbibition 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.

  12. Analysis on Interaction between Genotype of Four Main Flavonoids of Barley Grain and Environment

    Institute of Scientific and Technical Information of China (English)

    Tao YANG; Chengli DUAN; Yawen ZENG; Juan DU; Shuming YANG; Xiaoying PU; Shengchao YANG

    2012-01-01

    [Objective] This study aimed to analyze the interaction between genotype of flavonoids of barley grain and environment, to increase the flavonoid content of barley grain in cultivation and breeding. [Method] In this study, the content of cate- chin, myricetin, quercetin and kaempferol of barley grain planted in Kunming, Qujing and Baoshan were determined by HPLC, and the genotype, environment, genotype- environment interaction of the flavonoid content of barley grain were analyzed. [Result] According to the experimental results, the genotype variance, environmental variance and G x E interaction variance of catechin and kaempferol contents show the same trend: genotype variation 〉 environmental variation 〉 G × E interaction variation, which all reach a extremely significant level; the genotype variance, envi- ronmental variance and G × E interaction variance of quercetin and total flavonoid contents show the same trend: genetype variation 〉 G × E interaction variation 〉 environmental variation, which all reach a extremely significant level; the genotype variance and environmental variance of myricetin content both reach a extremely sig- nificant level, while the G × E interaction variance reaches a significant level, showing an order of genotype variation 〉 environmental variation 〉 G × E interaction variation; the genotype variance, environmental variance and G x E interaction vari- ance of total flavonoid content show an order of genotype variation 〉 environmental variation 〉 G × E interaction variation. Among different barley varieties, Ziguang- mangluoerling and Kuanyingdamai in Qujing, Kunming and Baoshan have relatively high content of quercetin, while other barley varieties barely contain any quercetin. The grains of Ziguangmangluoerling and Kuanyingdamai are purple, while the grains of other barley varieties are yellow. [Conclusion] Four main flavonoids and the total flavonoids of barley grain are mainly under genetic control and

  13. Extraction of starch from hulled and hull-less barley with papain and aqueous sodium hydroxide.

    Science.gov (United States)

    Sharma, Priyanka; Tejinder, S

    2014-12-01

    Starch was isolated from hulled (VJM 201) and hull-less (BL 134) barley with papain and aqueous sodium hydroxide treatments. For enzyme-assisted extraction, barley was steeped in water containing 0.2 % SO2 + 0.55 % lactic acid at 50° ± 2 °C for 4-5 h. The slurry was mixed with 0.4-2.0 g papain/kg barley and incubated at 50° ± 2 °C for 1-5 h. Aqueous sodium hydroxide (0.01-0.05 M) was added to the finely ground barley meal. The alkaline slurry was incubated at ambient temperature (25° ± 2 °C) for 15-60 min. The starch and grain fractions were isolated by screening and centrifugation. Increases in the time of treatment significantly affected the fiber, centrifugation and non-starch residue losses. Concentration of papain and sodium hydroxide had negligible effect on extraction losses. The enzyme-assisted extraction efficiency of starch was higher (80.7-84.6 %) than the alkaline method (70.9-83.7 %). The hulled barley showed higher extraction efficiency than the hull-less barley. The slurry treated with 0.4 g papain/kg barley for 5 h and 0.03 M sodium hydroxide for 60 min produced maximal yield of starch. Barley starch showed desirably high pasting temperature, water binding capacity and hold viscosity; and low final and setback viscosity compared with the commercial corn starch. The alkaline extracted hull-less barley starch showed exceptionally high peak and hold viscosities.

  14. Biotesting of radioactively contaminated forest soils using barley-based bioassay

    Science.gov (United States)

    Mel’nikova, T. V.; Polyakova, L. P.; Oudalova, A. A.

    2017-01-01

    Findings from radioactivity and phytotoxicity study are presented for soils from nine study-sites of the Klintsovsky Forestry located in the Bryansk region that were radioactively contaminated after the Chernobyl accident. According to the bioassay based on barley as test-species, stimulating effect of the soils analyzed is revealed for biological indexes of the length of barley roots and sprouts. From data on 137Cs specific activities in soils and plant biomass, the migration potential of radionuclide in the "soil-plant" system is assessed as a transfer factor. With correlation analysis, an impact of 137Cs in soil on the biological characteristics of barley is estimated.

  15. Distinct developmental defense activations in barley embryos identified by transcriptome profiling

    DEFF Research Database (Denmark)

    Nielsen, ME; Lok, F; Nielsen, Henrik Bjørn

    2006-01-01

    analyses of > 22,000 genes, which together with measurements of jasmonic acid and salicylic acid during embryo development provide new information on the initiation in the developing barley embryo of at least two distinct types of developmental defense activation (DDA). Early DDA is characterized by the up......-regulation of several PR genes is notable. Throughout barley embryo development, there are no indications of an increased biosynthesis of either jasmonic acid or salicylic acid. Collectively, the results help explain how the proposed DDA enables protection of the developing barley embryo and grain for purposes...

  16. Expression of Nudix hydrolase genes in barley under UV irradiation

    Science.gov (United States)

    Tanaka, Sayuri; Sugimoto, Manabu; Kihara, Makoto

    Seed storage and cultivation should be necessary to self-supply foods when astronauts would stay and investigate during long-term space travel and habitation in the bases on the Moon and Mars. Thought the sunlight is the most importance to plants, both as the ultimate energy source and as an environmental signal regulating growth and development, UV presenting the sunlight can damage many aspects of plant processes at the physiological and DNA level. Especially UV-C, which is eliminated by the stratospheric ozone layer, is suspected to be extremely harmful and give a deadly injury to plants in space. However, the defense mechanism against UV-C irradiation damage in plant cells has not been clear. In this study, we investigated the expression of Nudix hydrolases, which defense plants from biotic / abiotic stress, in barley under UV irradiation. The genes encoding the amino acid sequences, which show homology to those of 28 kinds of Nudix hydrolases in Arabidopsis thaliana, were identified in the barley full-length cDNA library. BLAST analysis showed 14 kinds of barley genes (HvNUDX1-14), which encode the Nudix motif sequence. A phylogenetic tree showed that HvNUDX1, HvNUDX7, HvNUDX9 and HvNUDX11 belonged to the ADP-ribose pyrophosphohydrolase, ADP-sugar pyrophosphohydrolase, NAD(P)H pyrophosphohydrolase and FAD pyrophosphohydrolase subfamilies, respectively, HvNUDX3, HvNUDX6, and HvNUDX8 belonged to the Ap _{n}A pyrophosphohydrolase subfamilies, HvNUDX5 and HvNUDX14 belonged to the coenzyme A pyrophosphohydrolase subfamilies, HvNUDX12 and HvNUDX13 belonged to the Ap _{4}A pyrophosphohydrolase subfamilies. Induction of HvNUDX genes by UV-A (340nm), UV-B (312nm), and UV-C (260nm) were analyzed by quantitative RT-PCR. The results showed that HvNUDX4 was induced by UV-A and UV-B, HvNUDX6 was induced by UV-B and UV-C, and HvNUDX7 and HvNUDX14 were induced by UV-C, significantly. Our results suggest that the response of HvNUDXs to UV irradiation is different by UV

  17. Differentially Expressed Genes between Two Barley Cultivars Contrasting in Drought Tolerance

    Institute of Scientific and Technical Information of China (English)

    P.G. Guo; M. Baum; R.H. Li; S. Grando; R.K. Varshney; J. Valkoun; S. Ceccarelli; A. Grane

    2007-01-01

    @@ Drought tolerance is a key trait for increasing and stabilizing barley productivity in dry areas. A number of genes have been described that respond to drought at the transcriptional level (Seki et al., 2002; Cheong et al.,2003).

  18. The effect of hydrolyzed Spirulina by malted barley on forced swimming test in ICR mice.

    Science.gov (United States)

    Kim, Na-Hyung; Jeong, Hyun-Ja; Lee, Ju-Young; Go, Hoyeon; Ko, Seong-Gyu; Hong, Seung-Heon; Kim, Hyung-Min; Um, Jae-Young

    2008-11-01

    Spirulina is a true puree of a filamentous, spiral-shaped blue alga and exerts the useful properties as a source of many biochemicals. This study investigated the antidepressant-like effect of hydrolyzed Spirulina by malted barley on the forced swimming test in mice. After the forced swimming test, we examined the levels of several blood biochemical parameters in mice. The effect of the hydrolyzed Spirulina by malted barley-treated group for 2 weeks on the immobility time was significantly reduced in comparison with the control group (p Spirulina by malted barley-treated group compared with the control group (p Spirulina by malted barley might be a candidate among antidepressant agents.

  19. Environmental impacts of barley cultivation under current and future climatic conditions

    DEFF Research Database (Denmark)

    Dijkman, Teunis Johannes; Birkved, Morten; Saxe, Henrik

    2017-01-01

    The purpose of this work is to compare the environmental impacts of spring barley cultivation in Denmark under current (year 2010) and future (year 2050) climatic conditions. Therefore, a Life Cycle Assessment was carried out for the production of 1 kg of spring barley in Denmark, at farm gate....... Both under 2010 and 2050 climatic conditions, four subscenarios were modelled, based on a combination of two soil types and two climates. Included in the assessment were seed production, soil preparation, fertilization, pesticide application, and harvest. When processes in the life cycle resulted in co...... categories, except human and freshwater eco-toxicity, are higher when the barley is produced under climatic circumstances representative for 2050. Comparison of the 2010 and 2050 climatic scenarios indicates that a predicted decrease in barley yields under the 2050 climatic conditions is the main driver...

  20. Extraction of starch from hulled and hull-less barley with papain and aqueous sodium hydroxide

    OpenAIRE

    2013-01-01

    Starch was isolated from hulled (VJM 201) and hull-less (BL 134) barley with papain and aqueous sodium hydroxide treatments. For enzyme-assisted extraction, barley was steeped in water containing 0.2 % SO2 + 0.55 % lactic acid at 50° ± 2 °C for 4–5 h. The slurry was mixed with 0.4–2.0 g papain/kg barley and incubated at 50° ± 2 °C for 1–5 h. Aqueous sodium hydroxide (0.01–0.05 M) was added to the finely ground barley meal. The alkaline slurry was incubated at ambient temperature (25° ± 2 °C) ...