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Sample records for barley endosperm development

  1. 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....

  2. Regulation of (1,3;1,4)-beta-D-glucan synthesis in developing endosperm of barley lys mutants

    DEFF Research Database (Denmark)

    Christensen, Ulla; Scheller, Henrik Vibe

    2012-01-01

    The mechanism behind altered content of (1,3:1,4)-beta-D-glucan was investigated in developing endosperm of barley lys3 and lys5 mutants. Both types of mutants are primarily affected in starch biosynthesis, and hence effects on (1,3;1,4)-beta-D-glucan are likely to be pleiotropic. The mutant alle...

  3. 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...

  4. Correlation-maximizing surrogate gene space for visual mining of gene expression patterns in developing barley endosperm tissue

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    Usadel Björn

    2007-05-01

    Full Text Available Abstract Background Micro- and macroarray technologies help acquire thousands of gene expression patterns covering important biological processes during plant ontogeny. Particularly, faithful visualization methods are beneficial for revealing interesting gene expression patterns and functional relationships of coexpressed genes. Such screening helps to gain deeper insights into regulatory behavior and cellular responses, as will be discussed for expression data of developing barley endosperm tissue. For that purpose, high-throughput multidimensional scaling (HiT-MDS, a recent method for similarity-preserving data embedding, is substantially refined and used for (a assessing the quality and reliability of centroid gene expression patterns, and for (b derivation of functional relationships of coexpressed genes of endosperm tissue during barley grain development (0–26 days after flowering. Results Temporal expression profiles of 4824 genes at 14 time points are faithfully embedded into two-dimensional displays. Thereby, similar shapes of coexpressed genes get closely grouped by a correlation-based similarity measure. As a main result, by using power transformation of correlation terms, a characteristic cloud of points with bipolar sandglass shape is obtained that is inherently connected to expression patterns of pre-storage, intermediate and storage phase of endosperm development. Conclusion The new HiT-MDS-2 method helps to create global views of expression patterns and to validate centroids obtained from clustering programs. Furthermore, functional gene annotation for developing endosperm barley tissue is successfully mapped to the visualization, making easy localization of major centroids of enriched functional categories possible.

  5. 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.

  6. 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.

  7. In Vitro Biochemical Characterization of All Barley Endosperm Starch Synthases

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    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.

  8. 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

  9. Endosperm Development in Autotetraploid Rice

    Institute of Scientific and Technical Information of China (English)

    WANG Lan; LIU Xiang-dong; LU Yong-gen; FENG Jiu-huan; XU Xue-bin; XU Shi-xiong (S.Y.Zee)

    2005-01-01

    By using the laser scanning confocal microscope and plastic (Leica 7022 historesin embedding kit) semi-thin sectioningtechnique, comparative studies on the development of endosperm were carried out between autotetraploid and diploid rices. Aboutone third of the ovaries in the autotetraploid showed normal endosperm development as those in the diploid. In these ovaries, one ofthe polar nuclei would fuse with the sperm nucleus, and the primary endosperm nucleus formed and underwent the first division in 4hours after pollination; the anticlinal wall began to grow centripetally between the free nuclei starting from the wall ingrowths of theembryo sac near the micropylar end, and some of the phragmoplasts formed transformed into periclinal walls. In addition, some of thecell wall situated in the middle of the endosperm appeared to originate from phragmoplasts, whereas others seemed to developrandomly without the obvious formation of phragmoplasts. Cellulose began to accumulate in the wall of aleurone cell layer at 6 daysafter pollination. The cellulose wall of the cells of the aleurone cell layer appeared to have completely formed within 7 to 8 days afterpollination. On the other hand, about two thirds of the ovaries in the autotetraploid showed abnormality in endosperm developmentwith various types, such as non-fertilization, abnormal fertilization, endosperm development-delay and non-synchronization in thedevelopment of cellulose wall of cells of the aleurone layer. These abnormalities usually resulted in decreased seed setting inautotetraploid rice.

  10. In vitro biochemical characterization of all barley endosperm starch synthases

    DEFF Research Database (Denmark)

    Cuesta-Seijo, Jose A.; Nielsen, Morten M.; Ruzanski, Christian;

    2016-01-01

    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...

  11. 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...

  12. 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.

  13. Cell wall degradation is required for normal starch mobilisation in barley endosperm.

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    Andriotis, Vasilios M E; Rejzek, Martin; Barclay, Elaine; Rugen, Michael D; Field, Robert A; Smith, Alison M

    2016-09-13

    Starch degradation in barley endosperm provides carbon for early seedling growth, but the control of this process is poorly understood. We investigated whether endosperm cell wall degradation is an important determinant of the rate of starch degradation. We identified iminosugar inhibitors of enzymes that degrade the cell wall component arabinoxylan. The iminosugar 1,4-dideoxy-1, 4-imino-l-arabinitol (LAB) inhibits arabinoxylan arabinofuranohydrolase (AXAH) but does not inhibit the main starch-degrading enzymes α- and β-amylase and limit dextrinase. AXAH activity in the endosperm appears soon after the onset of germination and resides in dimers putatively containing two isoforms, AXAH1 and AXAH2. Upon grain imbibition, mobilisation of arabinoxylan and starch spreads across the endosperm from the aleurone towards the crease. The front of arabinoxylan degradation precedes that of starch degradation. Incubation of grains with LAB decreases the rate of loss of both arabinoxylan and starch, and retards the spread of both degradation processes across the endosperm. We propose that starch degradation in the endosperm is dependent on cell wall degradation, which permeabilises the walls and thus permits rapid diffusion of amylolytic enzymes. AXAH may be of particular importance in this respect. These results provide new insights into the mobilization of endosperm reserves to support early seedling growth.

  14. Cell wall degradation is required for normal starch mobilisation in barley endosperm

    Science.gov (United States)

    Andriotis, Vasilios M. E.; Rejzek, Martin; Barclay, Elaine; Rugen, Michael D.; Field, Robert A.; Smith, Alison M.

    2016-01-01

    Starch degradation in barley endosperm provides carbon for early seedling growth, but the control of this process is poorly understood. We investigated whether endosperm cell wall degradation is an important determinant of the rate of starch degradation. We identified iminosugar inhibitors of enzymes that degrade the cell wall component arabinoxylan. The iminosugar 1,4-dideoxy-1, 4-imino-l-arabinitol (LAB) inhibits arabinoxylan arabinofuranohydrolase (AXAH) but does not inhibit the main starch-degrading enzymes α- and β-amylase and limit dextrinase. AXAH activity in the endosperm appears soon after the onset of germination and resides in dimers putatively containing two isoforms, AXAH1 and AXAH2. Upon grain imbibition, mobilisation of arabinoxylan and starch spreads across the endosperm from the aleurone towards the crease. The front of arabinoxylan degradation precedes that of starch degradation. Incubation of grains with LAB decreases the rate of loss of both arabinoxylan and starch, and retards the spread of both degradation processes across the endosperm. We propose that starch degradation in the endosperm is dependent on cell wall degradation, which permeabilises the walls and thus permits rapid diffusion of amylolytic enzymes. AXAH may be of particular importance in this respect. These results provide new insights into the mobilization of endosperm reserves to support early seedling growth. PMID:27622597

  15. Characterization of the microchemical structure of seed endosperm within a cellular dimension among six barley varieties with distinct degradation kinetics, using ultraspatially resolved synchrotron-based infrared microspectroscopy.

    Science.gov (United States)

    Liu, Na; Yu, Peiqiang

    2010-07-14

    Barley varieties have similar chemical composition but exhibit different rumen degradation kinetics and nutrient availability. These biological differences may be related to molecular, structural, and chemical makeup among the seed endosperm tissue. No detailed study was carried out. The objectives of this study were: (1) to use a molecular spectroscopy technique, synchrotron-based Fourier transform infrared microspectroscopy (SFTIRM), to determine the microchemical-structural features in seed endosperm tissue of six developed barley varieties; (2) to study the relationship among molecular-structural characteristics, degradation kinetics, and nutrient availability in six genotypes of barley. The results showed that inherent microchemical-structural differences in the endosperm among the six barley varieties were detected by the synchrotron-based analytical technique, SFTIRM, with the univariate molecular spectral analysis. The SFTIRM spectral profiles differed (P degradation). Weak correlations may indicate that limited variations of these six barley varieties might not be sufficient to interpret the relationship between spectroscopic information and the nutrient value of barley grain, although significant differences in biodegradation kinetics were observed. In conclusion, the studies demonstrated the potential of ultraspatially resolved synchrotron based technology (SFTIRM) to reveal the structural and chemical makeup within cellular and subcellular dimensions without destruction of the inherent structure of cereal grain tissue.

  16. Carotenoids of aleurone, germ, and endosperm fractions of barley, corn and wheat differentially inhibit oxidative stress.

    Science.gov (United States)

    Masisi, Kabo; Diehl-Jones, William L; Gordon, Joseph; Chapman, Donald; Moghadasian, Mohammed H; Beta, Trust

    2015-03-18

    The antioxidant potential of carotenoids from aleurone, germ, and endosperm fractions of barley, corn, and wheat has been evaluated. HPLC analysis confirmed the presence of lutein and zeaxanthin carotenoids (nd-15139 μg/kg) in extracts of cereal grain fractions. The antioxidant properties using 2,2-diphenyl-1-picrylhydrazyl, oxygen radical absorbance capacity, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assays revealed significantly higher (Paleurone and endosperm fractions. Using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay, 2,2'azobis (2-amidinopropane)dihydrochloride (AAPH)-induced cell loss was effectively reduced by preincubating Caco-2, HT-29, and FHs 74 Int cells with carotenoid extracts. Moreover, carotenoid extracts reduced (Paleurone, and endosperm fractions improved antioxidant capacity and thus have the potential to mitigate oxidative stress.

  17. 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...

  18. 454 Transcriptome sequencing suggests a role for two-component signalling in cellularization and differentiation of barley endosperm transfer cells.

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    Johannes Thiel

    Full Text Available BACKGROUND: Cell specification and differentiation in the endosperm of cereals starts at the maternal-filial boundary and generates the endosperm transfer cells (ETCs. Besides the importance in assimilate transfer, ETCs are proposed to play an essential role in the regulation of endosperm differentiation by affecting development of proximate endosperm tissues. We attempted to identify signalling elements involved in early endosperm differentiation by using a combination of laser-assisted microdissection and 454 transcriptome sequencing. PRINCIPAL FINDINGS: 454 sequencing of the differentiating ETC region from the syncytial state until functionality in transfer processes captured a high proportion of novel transcripts which are not available in existing barley EST databases. Intriguingly, the ETC-transcriptome showed a high abundance of elements of the two-component signalling (TCS system suggesting an outstanding role in ETC differentiation. All components and subfamilies of the TCS, including distinct kinds of membrane-bound receptors, have been identified to be expressed in ETCs. The TCS system represents an ancient signal transduction system firstly discovered in bacteria and has previously been shown to be co-opted by eukaryotes, like fungi and plants, whereas in animals and humans this signalling route does not exist. Transcript profiling of TCS elements by qRT-PCR suggested pivotal roles for specific phosphorelays activated in a coordinated time flow during ETC cellularization and differentiation. ETC-specificity of transcriptionally activated TCS phosphorelays was assessed for early differentiation and cellularization contrasting to an extension of expression to other grain tissues at the beginning of ETC maturation. Features of candidate genes of distinct phosphorelays and transcriptional activation of genes putatively implicated in hormone signalling pathways hint at a crosstalk of hormonal influences, putatively ABA and ethylene, and

  19. 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...

  20. 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.

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

    Science.gov (United States)

    Raboy, Victor; Cichy, Karen; Peterson, Kevin; Reichman, Sarah; Sompong, Utumporn; Srinives, Peerasak; Saneoka, Hirofumi

    2014-01-01

    Inositol hexaphosphate (Ins P6 or "phytic acid") typically accounts for 75 (± 10%) of seed total phosphorus (P). In some cases, genetic blocks in seed Ins P6 accumulation can also alter the distribution or total amount of seed P. In nonmutant barley (Hordeum vulgare L.) caryopses, ~80% of Ins P6 and total P accumulate in the aleurone layer, the outer layer of the endosperm, with the remainder in the germ. In barley low phytic acid 1-1 (Hvlpa1-1) seed, both endosperm Ins P6 and total P are reduced (~45% and ~25%, respectively), but germs are phenotypically wild type. This translates into a net reduction in whole-seed total P of ~15%. Nutrient culture studies demonstrate that the reduction in endosperm total P is not due to a reduction in the uptake of P into the maternal plant. Genetic tests (analyses of testcross and F2 seed) reveal that the Hvlpa1-1 genotype of the filial seed conditions the seed total P reduction; sibling seed in the same head of barley that differ in their Hvlpa1-1 genotype (heterozygous vs. homozygous recessive) differ in their total P (normal vs. reduced, respectively). Therefore, Hvlpa1 functions as a seed-specific or filial determinant of barley endosperm total P.

  2. Rice caryopsis development II:Dynamic changes in the endosperm

    Institute of Scientific and Technical Information of China (English)

    Xiaoba Wu; Jinxin Liu; Dongqi Li; Chun-Ming Liu

    2016-01-01

    The rice endosperm plays crucial roles in nourishing the embryo during embryogenesis and seed germination. Although previous studies have provided the general information about rice endosperm, a systematic investigation throughout the entire endosperm developmental process is still lacking. In this study, we examined in detail rice endosperm development on a daily basis throughout the 30-day period of post-fertilization development. We observed that coenocytic nuclear division occurred in the first 2 days after pollination (DAP), cellularization occurred between 3 and 5 DAP, differentiation of the aleurone and starchy endosperm occurred between 6 and 9 DAP, and accumulation of storage products occurred concurrently with the aleurone/starchy endosperm differentiation from 6 DAP onwards and was accomplished by 21 DAP. Changes in cytoplasmic membrane permeability, possibly caused by programmed cell death, were observed in the central region of the starchy endosperm at 8 DAP, and expanded to the whole starchy endosperm at 21 DAP when the aleurone is the only living component in the endosperm. Further, we observed that a distinct multi-layered dorsal aleurone formed near the dorsal vascular bundle, while the single- or occasionally two-cell layered aleurone was located in the lateral and ventral positions of endosperm. Our results provide in detail the dynamic changes in mitotic divisions, cellularization, cell differentiation, storage product accumulation, and programmed cell death that occur during rice endosperm development.

  3. Rice caryopsis development II: Dynamic changes in the endosperm.

    Science.gov (United States)

    Wu, Xiaoba; Liu, Jinxin; Li, Dongqi; Liu, Chun-Ming

    2016-09-01

    The rice endosperm plays crucial roles in nourishing the embryo during embryogenesis and seed germination. Although previous studies have provided the general information about rice endosperm, a systematic investigation throughout the entire endosperm developmental process is still lacking. In this study, we examined in detail rice endosperm development on a daily basis throughout the 30-day period of post-fertilization development. We observed that coenocytic nuclear division occurred in the first 2 days after pollination (DAP), cellularization occurred between 3 and 5 DAP, differentiation of the aleurone and starchy endosperm occurred between 6 and 9 DAP, and accumulation of storage products occurred concurrently with the aleurone/starchy endosperm differentiation from 6 DAP onwards and was accomplished by 21 DAP. Changes in cytoplasmic membrane permeability, possibly caused by programmed cell death, were observed in the central region of the starchy endosperm at 8 DAP, and expanded to the whole starchy endosperm at 21 DAP when the aleurone is the only living component in the endosperm. Further, we observed that a distinct multi-layered dorsal aleurone formed near the dorsal vascular bundle, while the single- or occasionally two-cell layered aleurone was located in the lateral and ventral positions of endosperm. Our results provide in detail the dynamic changes in mitotic divisions, cellularization, cell differentiation, storage product accumulation, and programmed cell death that occur during rice endosperm development.

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

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    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.

  5. The detection of QTLs in barley associated with endosperm hardness, grain density, grain size and malting quality using rapid phenotyping tools.

    Science.gov (United States)

    Walker, Cassandra K; Ford, Rebecca; Muñoz-Amatriaín, María; Panozzo, Joe F

    2013-10-01

    Using a barley mapping population, 'Vlamingh' × 'Buloke' (V × B), whole grain analyses were undertaken for physical seed traits and malting quality. Grain density and size were predicted by digital image analysis (DIA), while malt extract and protein content were predicted using near infrared (NIR) analysis. Validation of DIA and NIR algorithms confirmed that data for QTL analysis was highly correlated (R (2) > 0.82), with high RPD values (the ratio of the standard error of prediction to the standard deviation, 2.31-9.06). Endosperm hardness was measured on this mapping population using the single kernel characterisation system. Grain density and endosperm hardness were significantly inter-correlated in all three environments (r > 0.22, P hardness QTLs were found on 1H, 5H, and 7H. In this study, the majority of the genomic regions associated with grain texture were also coincident with QTLs for grain size, yield, flowering date and/or plant development genes. This study highlights the complexity of genomic regions associated with the variation of endosperm hardness and grain density, and their relationships with grain size traits, agronomic-related traits, and plant development loci.

  6. Isolation and Translation of Hordein Messenger RNA from Wild Type and Mutant Endosperms in Barley

    DEFF Research Database (Denmark)

    Brandt, Anders Bøving; Ingwersen, J.

    1978-01-01

    Nucleotide sequences of four cDNA clones coding for the carboxy-terminal portion of at least two different B1 hordein polypeptides are presented. The open reading frame in the nucleotide sequence of the the largest clone (pc hor2-4, 720 nucleotides) translates into the 181 carboxy-terminal amino...... was found to be a major constituent of the total messenger RNA population of the endosperm cell. Polyadenylated hordein messenger RNA sedimented at 11S in sucrose gradients and electrophoretic analysis reveals the presence of at least three RNA species with apparent molecular weights of 0.45, 0.36 and 0.......30 megadaltons. The 11S messenger RNA was translated in vitro into hordein precursor polypeptides which are 2–4 kilodaltons larger than the native hordein polypeptides. The endosperm cell of mutant No. 1508 contained twice as much RNA as the wild type endosperm cell but the same amount of polyadenylated 11S RNA...

  7. 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.

    2013-01-01

    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. PMID:24748715

  8. DNA endoreplication level in endosperm during seed development in three monocotyledonous species

    Directory of Open Access Journals (Sweden)

    Kazimierz Marciniak

    2014-02-01

    Full Text Available The DNA content after the Feulgen reaction in the endosperm of three monocotyledonous plant species (Asparagus officinalis, Muscari comosom, Haemanthus kurharinae differing in their 2C DNA content, was cytophotometrically measured. During endosperm development 1-6 endoreplication cycles take place, depending on the species. Differences in nuclear DNA endoreplication dynamics in the tested species are similar to those occurring in root parenchyma, but the endoreplication level in the endosperm is higher.

  9. Studies on the Programmed Cell Death in Rice During Starchy Endosperm Development

    Institute of Scientific and Technical Information of China (English)

    LI Rui; LAN Sheng-yin; XU Zhen-xiu

    2004-01-01

    Morphological variations of the nucleus in starchy endosperm cell were observed by the electron-transmisson microscope during endosperm development in rice. Along with the development of the starchy endosperm,the nuclei of the cells showed chromatin condensation,the typical feature of programmed cell death(PCD). The nuclei also showed nucleus deformation,disruption of nuclear envelope,nucleoplasm leaking into the cytoplasm and nucleus disintegration resulting in nuclear residue formation. From the nucleus deformation to the nucleus disintegration,the morphological changes of the nucleus were orderly progressive. This indicated that the cell death of starchy endosperm in rice was programmed cell death. Evans Blue staining observation showed that the cell death was initially detected in the central part of starchy endosperm in rice,then expanded outward. The activities of superoxide dismutase(SOD)and catalase(CAT)in rice starchy endosperm both descended continuously as development progressed. The analysis of DNA of rice starchy endosperm did not show the presence of DNA laddering. The above results showed that the cell death of starchy endosperm in rice was a special form of PCD.

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

    Directory of Open Access Journals (Sweden)

    Wobus Ulrich

    2009-01-01

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

  11. Positional cues specify and maintain aleurone cell fate in maize endosperm development.

    Science.gov (United States)

    Becraft, P W; Asuncion-Crabb, Y

    2000-09-01

    A genetic analysis of maize aleurone development was conducted. Cell lineage was examined by simultaneously marking cells with C1 for anthocyanin pigmentation in the aleurone and wx1 for amylose synthesis in the starchy endosperm. The aleurone and starchy endosperm share a common lineage throughout development indicating that positional cues specify aleurone fate. Mutants in dek1 block aleurone formation at an early stage and cause peripheral endosperm cells to develop as starchy endosperm. Revertant sectors of a transposon-induced dek1 allele showed that peripheral endosperm cells remain competent to differentiate as aleurone cells until late in development. Ds-induced chromosome breakage was used to generate Dek1 loss-of-function sectors. Events occurring until late development caused aleurone cells to switch fate to starchy endosperm indicating that cell fate is not fixed. Thus, positional cues are required to specify and maintain aleurone fate and Dek1 function is required to respond to these cues. An analysis of additional mutants that disrupt aleurone differentiation suggests a hierarchy of gene functions to specify aleurone cell fate and then control aleurone differentiation. These mutants disrupt aleurone differentiation in reproducible patterns suggesting a relationship to endosperm pattern formation.

  12. Expression of fatty acid and lipid biosynthetic genes in developing endosperm of Jatropha curcas

    Directory of Open Access Journals (Sweden)

    Gu Keyu

    2012-07-01

    Full Text Available Abstract Background Temporal and spatial expression of fatty acid and lipid biosynthetic genes are associated with the accumulation of storage lipids in the seeds of oil plants. In jatropha (Jatropha curcas L., a potential biofuel plant, the storage lipids are mainly synthesized and accumulated in the endosperm of seeds. Although the fatty acid and lipid biosynthetic genes in jatropha have been identified, the expression of these genes at different developing stages of endosperm has not been systemically investigated. Results Transmission electron microscopy study revealed that the oil body formation in developing endosperm of jatropha seeds initially appeared at 28 days after fertilization (DAF, was actively developed at 42 DAF and reached to the maximum number and size at 56 DAF. Sixty-eight genes that encode enzymes, proteins or their subunits involved in fatty acid and lipid biosynthesis were identified from a normalized cDNA library of jatropha developing endosperm. Gene expression with quantitative reverse-transcription polymerase chain reaction analysis demonstrated that the 68 genes could be collectively grouped into five categories based on the patterns of relative expression of the genes during endosperm development. Category I has 47 genes and they displayed a bell-shaped expression pattern with the peak expression at 28 or 42 DAF, but low expression at 14 and 56 DAF. Category II contains 8 genes and expression of the 8 genes was constantly increased from 14 to 56 DAF. Category III comprises of 2 genes and both genes were constitutively expressed throughout endosperm development. Category IV has 9 genes and they showed a high expression at 14 and 28 DAF, but a decreased expression from 42 to 56 DAF. Category V consists of 2 genes and both genes showed a medium expression at 14 DAF, the lowest expression at 28 or 42 DAF, and the highest expression at 56 DAF. In addition, genes encoding enzymes or proteins with similar function were

  13. Ultracytochemical Localization and Functional Analysis of ATPase During the Endosperm Development in Oryza sativa L.

    Institute of Scientific and Technical Information of China (English)

    WEI Cun-xu; LAN Sheng-yin; XU Zhen-xiu

    2003-01-01

    Ultracytochemical localization of ATPase during development of rice endosperm was performed using a lead phosphate precipitation technique. The results indicated that, at the coenocyte and ceilularization stages, active ATPase was mainly distributed in an embryo sac wall, nucleus, and plasma membrane. At the early stage of development and differentiation, active ATPase was observed in the plasma membrane. At the grain filling stage, ATPase was highly active in the plasma membrane, intercellular space, and plasmodesmata in aleurone, moderately active on the plasma membrane in subaleurone. In starchy endosperm, ATPase was localized in the plasma membrane and degenerated nucleus. ATPase activity also appeared around vacuole and protein body in endosperm cell. The relationships between the ultracytochemical localization of ATPase and its function during the development of rice endosperm were discussed. Overall, ATPase was involved in the process of nutrition absorption and protein synthesis.

  14. Diurnal oscillation of SBE expression in sorghum endosperm

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Chuanxin; Mutisya, J.; Rosenquist, S.; Baguma, Y.; Jansson, C.

    2009-01-15

    Spatial and temporal expression patterns of the sorghum SBEI, SBEIIA and SBEIIB genes, encoding, respectively, starch branching enzyme (SBE) I, IIA and IIB, in the developing endosperm of sorghum (Sorghum bicolor) were studied. Full-length genomic and cDNA clones for sorghum was cloned and the SBEIIA cDNA was used together with gene-specific probes for sorghum SBEIIB and SBEI. In contrast to sorghum SBEIIB, which was expressed primarily in endosperm and embryo, SBEIIA was expressed also in vegetative tissues. All three genes shared a similar temporal expression profile during endosperm development, with a maximum activity at 15-24 days after pollination. This is different from barley and maize where SBEI gene activity showed a significantly later onset compared to that of SBEIIA and SBEIIB. Expression of the three SBE genes in the sorghum endosperm exhibited a diurnal rhythm during a 24-h cycle.

  15. Dynamic expression of imprinted genes associates with maternally controlled nutrient allocation during maize endosperm development.

    Science.gov (United States)

    Xin, Mingming; Yang, Ruolin; Li, Guosheng; Chen, Hao; Laurie, John; Ma, Chuang; Wang, Dongfang; Yao, Yingyin; Larkins, Brian A; Sun, Qixin; Yadegari, Ramin; Wang, Xiangfeng; Ni, Zhongfu

    2013-09-01

    In angiosperms, the endosperm provides nutrients for embryogenesis and seed germination and is the primary tissue where gene imprinting occurs. To identify the imprintome of early developing maize (Zea mays) endosperm, we performed high-throughput transcriptome sequencing of whole kernels at 0, 3, and 5 d after pollination (DAP) and endosperms at 7, 10, and 15 DAP, using B73 by Mo17 reciprocal crosses. We observed gradually increased expression of paternal transcripts in 3- and 5-DAP kernels. In 7-DAP endosperm, the majority of the genes tested reached a 2:1 maternal versus paternal ratio, suggesting that paternal genes are nearly fully activated by 7 DAP. A total of 116, 234, and 63 genes exhibiting parent-specific expression were identified at 7, 10, and 15 DAP, respectively. The largest proportion of paternally expressed genes was at 7 DAP, mainly due to the significantly deviated parental allele expression ratio of these genes at this stage, while nearly 80% of the maternally expressed genes (MEGs) were specific to 10 DAP and were primarily attributed to sharply increased expression levels compared with the other stages. Gene ontology enrichment analysis of the imprinted genes suggested that 10-DAP endosperm-specific MEGs are involved in nutrient uptake and allocation and the auxin signaling pathway, coincident with the onset of starch and storage protein accumulation.

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

    DEFF Research Database (Denmark)

    Seefeldt, Helene Fast; van den Berg, Frans; Klöckenberger, Walter

    2007-01-01

    1H NMR imaging (MRI) was used as a noninvasive technique to study water distribution and mobility in hydrated barley (Hordeum vulgare L.) seeds of accessions with varying content of beta glucan (BG), a highly hygroscopic cell wall component. High contents of BG in barley are unfavorable in malting...... where it leads to clotting of filters and hazing of beer as well as in animal feed where it hinders the rapid uptake of energy. However, a high content of BG has a positive nutritional effect, as it lowers the cholesterol and the glycaemic index. It was studied whether water distribution and mobility...... were related to content and location of BG. Water mobility was investigated by following the rate and mode of desiccation in hydrated single seeds. In order to determine the different water components, a multispin echo experiment was set up to reveal the T2 transverse relaxation rates of water within...

  17. Two maize END-1 orthologs, BETL9 and BETL9like, are transcribed in a non-overlapping spatial pattern on the outer surface of the developing endosperm

    Directory of Open Access Journals (Sweden)

    Joaquín eRoyo

    2014-05-01

    Full Text Available In the course of a project aimed to isolate transfer cells-specific genes in maize endosperm we have identified the BETL9 gene. BETL9 encodes for a small protein very similar in sequence to the product of the barley transfer cell-specific gene end-1. Both BETL9 and END-1 proteins are lipid transfer proteins, but their function is currently unknown. In situ hybridization analysis confirms that the BETL9 gene is exclusively transcribed in the basal endosperm transfer cell layer during seed development since 10 days after pollination. However, immunolocalization data indicates that the BETL9 protein accumulates in the maternal placento-chalaza cells located just beside the transfer cell layer. This suggests that the BETL9 protein should be transported to the maternal side to exert its, still unknown, function. In addition, we have identified a second maize gene very similar in sequence to BETL9 and we have named it BETL9like. In situ hybridization shows that BETL9like is also specifically transcribed in the developing maize endosperm within the same time frame that BETL9, but in this case it is exclusively expressed in the aleurone cell layer. Consequently, the BETL9 and BETL9like genes are transcribed in a non-overlapping pattern on the outer surface of the maize endosperm. The BETL9 and BETL9like promoter sequences, fused to the GUS reporter gen, accurately reflected the expression pattern observed for the genes in maize. Finally, we have identified in the Arabidopsis genome a set of four genes orthologous to BETL9 and BETL9like and analysed the activity of their promoters in Arabidopsis transgenic plants carrying fusions of their promoter sequences to the GUS reporter. As in the case of the maize genes, the Arabidopsis orthologs showed highly complementary expression patterns.

  18. 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....

  19. DNA endoreplication level in endosperm and in cotyledons during seed development in three dicotyledonous species

    Directory of Open Access Journals (Sweden)

    Kazimierz Marciniak

    2014-01-01

    Full Text Available The DNA content of nuclear and cellular endosperm and in cotyledons in the course of seed development in Helianthus annuus, Pisum sativum and Vicia faba was cytophotometrically measured after Faulgen's reaction. Helianthus annuus is characterized by the lowest endoreplication dynamics attaining a maximum DNA content of 48C in nuclear endosperm, which corresponds to four endoreplication rounds, and 16C DNA (three rounds in cotyledons. In Pisum sativum cellular endosperm in the studied stages of seed development was absent and the highest DNA content in cotyledons was 512C DNA; this corresponds to eight endoreplication rounds. In Vicia faba, the maximum DNA endoreplication level was 1536C in nuclear endosperm, which corresponds to nine endoreplication rounds; in cotyledons the maximum DNA content was 64C (five rounds. In the course of seed development, differences in the dynamics of DNA endoreplication in throphic tissues of the examined species seem to be similar to differences in the endoreplication level in root and leaf parenchyma, but the DNA endoreplication level is much higher in throphic tissues. Therefore, the endoreplication pattern might be species-specific and genetically conditioned.

  20. Entwicklung transgener Gerste (Hordeum vulgare L.) mit dem Ziel der Lysin- und Threoninanreicherung im Endosperm

    OpenAIRE

    Ibrahim, Ahmed Shawky Ahmed

    2006-01-01

    An efficient Agrobacterium-mediated barley transformation system was established with a transformation rate of 13.4 % on average. Towards improving the nutritional value of barley, a set of novel transformation vectors was developed including the dapA and lysC genes encoding the feed-back-inhibition insensitive form of the dihydrodipicolinate synthase (DHDPS) and aspartate kinase (AK) respectively. Both genes under the control of the endosperm-specific D-hordein promoter or the constitutive u...

  1. A roadmap for zinc transport into the developing barley grain. Gene expression profiling using laser microdissection

    DEFF Research Database (Denmark)

    Borg, Søren; Tauris, Birgitte

    2011-01-01

    Nutrients destined for the developing cereal grain encounter several restricting barriers on their path towards their final storage sites in the grain. In order to identify transporters and chelating agents that may be involved in transport and deposition of zinc in the barley grain, expression...... profiles have been generated of four different tissue types: the transfer cells, the aleurone layer, the endosperm, and the embryo. Cells from these tissues were isolated with the 'laser capture microdissection' technology and the extracted RNA was subjected to three rounds of T7-based amplification...... were taken into account. On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain....

  2. Caspase-like activities accompany programmed cell death events in developing barley grains.

    Directory of Open Access Journals (Sweden)

    Van Tran

    Full Text Available Programmed cell death is essential part of development and cell homeostasis of any multicellular organism. We have analyzed programmed cell death in developing barley caryopsis at histological, biochemical and molecular level. Caspase-1, -3, -4, -6 and -8-like activities increased with aging of pericarp coinciding with abundance of TUNEL positive nuclei and expression of HvVPE4 and HvPhS2 genes in the tissue. TUNEL-positive nuclei were also detected in nucellus and nucellar projection as well as in embryo surrounding region during early caryopsis development. Quantitative RT-PCR analysis of micro-dissected grain tissues revealed the expression of HvVPE2a, HvVPE2b, HvVPE2d, HvPhS2 and HvPhS3 genes exclusively in the nucellus/nucellar projection. The first increase in cascade of caspase-1, -3, -4, -6 and -8-like activities in the endosperm fraction may be related to programmed cell death in the nucellus and nucellar projection. The second increase of all above caspase-like activities including of caspase-9-like was detected in the maturating endosperm and coincided with expression of HvVPE1 and HvPhS1 genes as well as with degeneration of nuclei in starchy endosperm and transfer cells. The distribution of the TUNEL-positive nuclei, tissues-specific expression of genes encoding proteases with potential caspase activities and cascades of caspase-like activities suggest that each seed tissue follows individual pattern of development and disintegration, which however harmonizes with growth of the other tissues in order to achieve proper caryopsis development.

  3. Transcriptome Analysis of High-Temperature Stress in Developing Barley Caryopses :Early Stress Responses and Effects on Storage Compound Biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Elke Mangelsen; Joachim Kilian; Klaus Harter; Christer Jansson; Dierk Wanke; Eva Sundberg

    2011-01-01

    High-temperature stress,like any abiotic stress,impairs the physiology and development of plants,including the stages of seed setting and ripening.We used the Aflymetrix 22K Barley1 GeneChip microarray to investigate the response of developing barley(Hordeum vulgare)seeds,termed caryopses,after 0.5,3,and 6 h of heat stress exposure;958 induced and 1122 repressed genes exhibited spatial and temporal expression patterns that provide a detailed insight into the caryopses'early heat stress responses.Down-regulation of genes related to storage compound biosynthesis and cell growth provides evidence for a rapid impairment of the caryopsis' development.Increased levels of sugars and amino acids were indicative for both production of compatible solutes and feedback-induced accumulation of substrates for storage compound biosynthesis.Metadata analysis identified embryo and endosperm as primary locations of heat stress responses,indicating a strong impact of short-term heat stress on central developmental functions of the caryopsis.A comparison with heat stress responses in Arabidopsis shoots and drought stress responses in barley caryopses identified both conserved and presumably heat-and caryopsis-specific stress-responsive genes.Summarized,our data provide an important basis for further investigation of gene functions in order to aid an improved heat tolerance and reduced losses of yield in barley as a model for cereal crops.

  4. 5种谷物胚乳表层细胞的比较研究%Comparison of peripheral endosperm cell development in five cereals

    Institute of Scientific and Technical Information of China (English)

    王慧慧; 熊飞; 王峰; 刘智; 顾蕴洁; 王忠

    2011-01-01

    Light microscopy, transmission and scanning electron microscopy were employed to investigate the structure of peripheral endosperm cells (PECs) of different cereals. Results showed that the differentiation of PECs was determined by their surface position, and the development extent of PECs from different cereals was various. PECs in cereals with large grain weight were generally more developed than those in cereals with small grain weight. All wall ingrowths in endosperm transfer cells of different cereals belonged to the flange-type wall ingrowths with their long axes consistent with that of the predicted direction of the solute flow. Wall ingrowths in endosperm transfer cells of maize and sorghum were more developed than those of barley and wheat. Aleurone cells of different cereals had the similar structure with thick walls. According to the position and structure features, PECs may play an important role in apoplastic and symplas-tic transport of nutrients. The structure of endosperm transfer cells is more suitable for the transport of grain filling nutrients than that of aleurone cells.%通过光学显微镜及透射和扫描电镜研究水稻、大麦、小麦、高粱、玉米5种谷物胚乳表层细胞的结构.结果表明:胚乳表层细胞的分化具有位置效应,且不同谷物胚乳表层细胞的发育程度不同.一般而言,粒重较大的谷物其胚乳表层细胞比粒重较小的谷物发达.谷物胚乳传递细胞中壁内突皆为肋状内突,其长轴方向与溶质流运输方向一致.玉米、高粱胚乳传递细胞中的肋状内突较大麦、小麦发达.不同谷物糊粉层细胞结构类似,其细胞壁较厚.根据胚乳表层细胞分布位置和结构特点可推测,胚乳表层细胞在养分的质外体和共质体运输过程中起着重要作用,且胚乳传递细胞的结构比糊粉层细胞更适合灌浆物质的运输.

  5. Dynamic Expression of Imprinted Genes Associates with Maternally Controlled Nutrient Allocation during Maize Endosperm Development[W][OPEN

    Science.gov (United States)

    Xin, Mingming; Yang, Ruolin; Li, Guosheng; Chen, Hao; Laurie, John; Ma, Chuang; Wang, Dongfang; Yao, Yingyin; Larkins, Brian A.; Sun, Qixin; Yadegari, Ramin; Wang, Xiangfeng; Ni, Zhongfu

    2013-01-01

    In angiosperms, the endosperm provides nutrients for embryogenesis and seed germination and is the primary tissue where gene imprinting occurs. To identify the imprintome of early developing maize (Zea mays) endosperm, we performed high-throughput transcriptome sequencing of whole kernels at 0, 3, and 5 d after pollination (DAP) and endosperms at 7, 10, and 15 DAP, using B73 by Mo17 reciprocal crosses. We observed gradually increased expression of paternal transcripts in 3- and 5-DAP kernels. In 7-DAP endosperm, the majority of the genes tested reached a 2:1 maternal versus paternal ratio, suggesting that paternal genes are nearly fully activated by 7 DAP. A total of 116, 234, and 63 genes exhibiting parent-specific expression were identified at 7, 10, and 15 DAP, respectively. The largest proportion of paternally expressed genes was at 7 DAP, mainly due to the significantly deviated parental allele expression ratio of these genes at this stage, while nearly 80% of the maternally expressed genes (MEGs) were specific to 10 DAP and were primarily attributed to sharply increased expression levels compared with the other stages. Gene ontology enrichment analysis of the imprinted genes suggested that 10-DAP endosperm-specific MEGs are involved in nutrient uptake and allocation and the auxin signaling pathway, coincident with the onset of starch and storage protein accumulation. PMID:24058158

  6. A roadmap for zinc trafficking in the developing barley grain based on laser capture microdissection and gene expression profiling

    DEFF Research Database (Denmark)

    Tauris, Birgitte; Borg, Søren; Gregersen, Per L;

    2009-01-01

    Nutrients destined for the developing cereal grain encounter several restricting barriers on their path towards their final storage sites in the grain. In order to identify transporters and chelating agents that may be involved in transport and deposition of zinc in the barley grain, expression...... profiles have been generated of four different tissue types: the transfer cells, the aleurone layer, the endosperm, and the embryo. Cells from these tissues were isolated with the ‘laser capture microdissection' technology and the extracted RNA was subjected to three rounds of T7-based amplification...... were taken into account. On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain....

  7. 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

  8. 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...

  9. Endosperm degradation during seed development of Echinocystis lobata (Cucurbitaceae) as a manifestation of programmed cell death (PCD) in plants.

    Science.gov (United States)

    Wojciechowska, Marzena; Olszewska, Maria J

    2003-01-01

    Programmed cell death (PCD) is an active, genetically controlled process that ultimately leads to elimination of unnecessary or damaged cells from multicellular organism. It occurs during normal growth and development or in response to a variety of environmental triggers and is indispensable for survival of the organism. In Echinocystis lobata the endosperm, an ephemeral tissue in angiosperm plants, undergoes distinct cytological, physiological and molecular changes during seed development and maturation. As a result, mature seeds are deprived of this tissue. The endosperm was analyzed at the consecutive stages of seed development. The morphological changes of cells were studied at light and electron microscope levels. In this paper we report that endosperm cells undergo morphological and biochemical changes characteristic of apoptosis, a particular type of PCD, i.e. cell shrinkage, chromatin condensation, nuclear fragmentation, and cytoplasm degradation, while the ultrastructure of mitochondria seems to be less changed. Furthermore, the progression of DNA degradation has been shown by agarose gel electrophoresis (ladder pattern of DNA fragmentseparation), TUNEL and comet assay. It isconcluded that during seed maturation, endosperm degradation process is accompanied by typical PCD-related changes of cell morphology and internucleosomal DNA cleavage.

  10. Functions of heteromeric and homomeric isoamylase-type starch-debranching enzymes in developing maize endosperm.

    Science.gov (United States)

    Kubo, Akiko; Colleoni, Christophe; Dinges, Jason R; Lin, Qiaohui; Lappe, Ryan R; Rivenbark, Joshua G; Meyer, Alexander J; Ball, Steven G; James, Martha G; Hennen-Bierwagen, Tracie A; Myers, Alan M

    2010-07-01

    Functions of isoamylase-type starch-debranching enzyme (ISA) proteins and complexes in maize (Zea mays) endosperm were characterized. Wild-type endosperm contained three high molecular mass ISA complexes resolved by gel permeation chromatography and native-polyacrylamide gel electrophoresis. Two complexes of approximately 400 kD contained both ISA1 and ISA2, and an approximately 300-kD complex contained ISA1 but not ISA2. Novel mutations of sugary1 (su1) and isa2, coding for ISA1 and ISA2, respectively, were used to develop one maize line with ISA1 homomer but lacking heteromeric ISA and a second line with one form of ISA1/ISA2 heteromer but no homomeric enzyme. The mutations were su1-P, which caused an amino acid substitution in ISA1, and isa2-339, which was caused by transposon insertion and conditioned loss of ISA2. In agreement with the protein compositions, all three ISA complexes were missing in an ISA1-null line, whereas only the two higher molecular mass forms were absent in the ISA2-null line. Both su1-P and isa2-339 conditioned near-normal starch characteristics, in contrast to ISA-null lines, indicating that either homomeric or heteromeric ISA is competent for starch biosynthesis. The homomer-only line had smaller, more numerous granules. Thus, a function of heteromeric ISA not compensated for by homomeric enzyme affects granule initiation or growth, which may explain evolutionary selection for ISA2. ISA1 was required for the accumulation of ISA2, which is regulated posttranscriptionally. Quantitative polymerase chain reaction showed that the ISA1 transcript level was elevated in tissues where starch is synthesized and low during starch degradation, whereas ISA2 transcript was relatively abundant during periods of either starch biosynthesis or catabolism.

  11. Near infrared spectra indicate specific mutant endosperm genes and reveal a new mechanism for substituting starch with (1-->3,1-->4)-[beta]-glucan in barley

    DEFF Research Database (Denmark)

    Munck, L.; Møller, B.; Jacobsen, Susanne

    2004-01-01

    Near Infrared Reflectance spectroscopy was tested as a screening method to characterise high lysine mutants from a barley collection by classification through Principal Component Analysis (PCA). Mean spectra of the samples within each cluster identified gene-specific patterns in the 2270-2360 nm ...

  12. Preferred carbon precursors for lipid labelling in the heterotrophic endosperm of developing oat (Avena sativa L.) grains.

    Science.gov (United States)

    Grimberg, Åsa

    2014-10-01

    Oat (Avena sativa L.) is unusual among the cereal grains in storing high amounts of oil in the endosperm; up to 90% of total grain oil. By using oat as a model species for oil metabolism in the cereal endosperm, we can learn how to develop strategies to redirect carbon from starch to achieve high-oil yielding cereal crops. Carbon precursors for lipid synthesis were compared in two genetically close oat cultivars with different endosperm oil content (about 6% and 10% of grain dw, medium-oil; MO, and high-oil; HO cultivar, respectively) by supplying a variety of (14)C-labelled substrates to the grain from both up- and downstream parts of glycolysis, either through detached oat panicles in vitro or by direct injection in planta. When supplied by direct injection, (14)C from acetate was identified to label the lipid fraction of the grain to the highest extent among substrates tested; 46% of net accumulated (14)C, demonstrating its applicability as a marker for lipids in the endosperm. Time course analyses of injected (14)C acetate during grain development suggested a more efficient transfer of fatty acids from polar lipids to triacylglycerol in the HO as compared to the MO cultivar, and turnover of triacylglycerol was suggested to not play a major role for the final oil content of oat grain endosperm despite the low amount of protective oleosins in this tissue. Moreover, availability of light was shown to drastically affect grain net carbon accumulation from (14)C-sucrose when supplied through detached panicles for the HO cultivar.

  13. 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.

  14. Drought stress delays endosperm development and misregulates genes associated with cytoskeleton organization and grain quality proteins in developing wheat seeds.

    Science.gov (United States)

    Begcy, Kevin; Walia, Harkamal

    2015-11-01

    Drought stress is a major yield-limiting factor for wheat. Wheat yields are particularly sensitive to drought stress during reproductive development. Early seed development stage is an important determinant of seed size, one of the yield components. We specifically examined the impact of drought stress imposed during postzygotic early seed development in wheat. We imposed a short-term drought stress on plants with day-old seeds and observed that even a short-duration drought stress significantly reduced the size of developing seeds as well as mature seeds. Drought stress delayed the developmental transition from syncytial to cellularized stage of endosperm. Coincident with reduced seed size and delayed endosperm development, a subset of genes associated with cytoskeleton organization was misregulated in developing seeds under drought-stressed. Several genes linked to hormone pathways were also differentially regulated in response to drought stress in early seeds. Notably, drought stress strongly repressed the expression of wheat storage protein genes such as gliadins, glutenins and avenins as early as 3 days after pollination. Our results provide new insights on how some of the early seed developmental events are impacted by water stress, and the underlying molecular pathways that can possibly impact both grain size and quality in wheat.

  15. Embryo and endosperm development in caryopses of hybrids from crosses between tetraploid wheats and their alloplasmic lines with rye

    Directory of Open Access Journals (Sweden)

    Wijciech Sodkiewicz

    2014-01-01

    Full Text Available Data concerning the embryo and endosperm development in twenty-day-old caryopses of hybrids obtained as the result of pollination with rye pollen of tetraploid wheats (Triticum dicoccoides, T. dicoccum, T. durum and T. polonicum, their alloplasmic lines with T. timopheevi plasma and aIlaplasmic T. timopheevi lines with cytoplasma of the above mentioned tetraploid wheats and hexaploid wheat (T. macha were analysed. A high variability was noted between the tetraploid wheats as regards the degree of development of the embryo and of the endosperm in the hybrid caryopses and a decisive influence of the wheat genotype on these characters. The data for alloplasmic lines showed that the cytoplasm may have a modifying effect on the expression of these genotype characters.

  16. Gene expression in the developing aleurone and starchy endosperm of wheat.

    Science.gov (United States)

    Gillies, Susan A; Futardo, Agnelo; Henry, Robert J

    2012-08-01

    Wheat is a critical food source globally. Food security is an increasing concern; current production levels are not expected to keep pace with global demand. New technologies have provided a vast array of wheat genetic data; however, best use of this data requires placing it within a framework in which the various genes, pathways and interactions can be examined. Here we present the first systematic comparison of the global transcriptomes of the aleurone and starchy endosperm of the developing wheat seed (Triticum aestivum), at time points critical to the development of the aleurone layer; 6-, 9- and 14-day post-anthesis. Illumina sequencing gave 25-55 million sequence reads per tissue, of the trimmed reads, 70%-81% mapped to reference expressed sequence transcripts. Transcript abundance was analysed by performing RNA-Seq normalization to generate reads per kilobase of exon model per million mapped reads values, and these were used in comparative analyses between the tissues at each time point using Kal's Z-test. This identified 9414-13 202 highly differentially expressed transcripts that were categorized on the basis of tissue and time point expression and functionally analysed revealing two very distinct tissues. The results demonstrate the fundamental biological reprogramming of the two major biologically and economically significant tissues of the wheat seed over this time course. Understanding these changes in gene expression profiles is essential to mining the potential these tissues hold for human nutrition and contributing to the systems biology of this important crop plant.

  17. Isolation of Promoters and Fragments of Genes Controlling Endosperm Development Without Fertilization in Arabidopsis and Engineering of the Antisense Constructions

    Directory of Open Access Journals (Sweden)

    Grigory A. Gerashchenkov

    2015-06-01

    Full Text Available Apomixis is asexual seed reproduction without both meiosis and fertilization based on the complex developmental processes such as apomeiosis, parthenogenesis and specific endosperm development. This investigation is aimed at engineering of apomixis in Arabidopsis thaliana with sexual seed reproduction. The fragments of known genes of endosperm formation MEA, FIE, FIS2 and gene of apomeiosis DYAD (as control were isolated using Q5 high fidelity DNA polymerase. These gene fragments of interest at the antisense orientation were fused with isolated constitutive and meiosis specific promoters of Arabidopsis at NcoI sites. The fused promoter-gene fragment modules were cloned in pCambia1301 at SalI cites. The engineered constructions will be used for the floral dip transformation of Arabidopsis and down regulation of these genes at engineering of apomixis.

  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. Cadmium translocation and accumulation in developing barley grains

    DEFF Research Database (Denmark)

    Chen, Fei; Wu, Feibo; Dong, Jing

    2007-01-01

    nutrient solution containing the markers for phloem (rubidium) and xylem (strontium) transport. Cd concentration in each part of detached spikes increased with external Cd levels, and Cd concentration in various organs over the three Cd levels of 0.5, 2, 8 μM Cd on 15-day Cd exposure was in the order: awn...... Soil cadmium (Cd) contamination has posed a serious problem for safe food production and become a potential agricultural and environmental hazard worldwide. In order to study the transport of Cd into the developing grains, detached ears of two-rowed barley cv. ZAU 3 were cultured in Cd stressed...... dramatic reduction in Cd transport to developing grains. The results indicated that awn, rachis and glume may involve in Cd transport into developing grains, and suggested that Cd redistribution in maturing cereals be considered as an important physiological process influencing the quality of harvested...

  20. The regulation of gene expression in transformed maize aleurone and endosperm protoplasts. Analysis of promoter activity, intron enhancement, and mRNA untranslated regions on expression.

    Science.gov (United States)

    Gallie, D R; Young, T E

    1994-11-01

    Gene expression in the aleurone and endosperm is highly regulated during both seed development and germination. Studies of alpha-amylase expression in the aleurone of barley (Hordeum vulgare) have generated the current paradigm for hormonal control of gene expression in germinating cereal grain. Gene expression studies in both the aleurone and endosperm tissues of maize (Zea mays) seed have been hampered because of a lack of an efficient transformation system. We report here the rapid isolation of protoplasts from maize aleurone and endosperm tissue, their transformation using polyethylene glycol or electroporation, and the regulation of gene expression in these cells. Adh1 promoter activity was reduced relative to the 35S promoter in aleurone and endosperm protoplasts compared to Black Mexican Sweet suspension cells in which it was nearly as strong as the 35S promoter. Intron-mediated stimulation of expression was substantially higher in transformed aleurone or endosperm protoplasts than in cell-suspension culture protoplasts, and the data suggest that the effect of an intron may be affected by cell type. To examine cytoplasmic regulation, the 5' and 3' untranslated regions from a barley alpha-amylase were fused to the firefly luciferase-coding region, and their effect on translation and mRNA stability was examined following the delivery of in vitro synthesized mRNA to aleurone and endosperm protoplasts. The alpha-amylase untranslated regions regulated translational efficiency in a tissue-specific manner, increasing translation in aleurone or endosperm protoplasts but not in maize or carrot cell-suspension protoplasts, in animal cells, or in in vitro translation lysates.

  1. Development and Meiosis of Three Interspecific Hybrids with Cultivated Barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Von Bothmer, R.; Flink, J.; Linde-Laursen, Ib

    1986-01-01

    The development and meiosis of three interspecific hybrids between cultivated barley (Hordeum vulgare L.) and H. secalinum Schreb., H. tetraploidum Covas, and H. parodii Covas, respectively, were studied. All three hybrid combinations developed very slowly vegetatively. Meiosis of the hybrids...

  2. 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.

  3. 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.

  4. 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.

  5. An analysis of expressed sequence tags of developing castor endosperm using a full-length cDNA library

    Directory of Open Access Journals (Sweden)

    Wallis James G

    2007-07-01

    Full Text Available Abstract Background Castor seeds are a major source for ricinoleate, an important industrial raw material. Genomics studies of castor plant will provide critical information for understanding seed metabolism, for effectively engineering ricinoleate production in transgenic oilseeds, or for genetically improving castor plants by eliminating toxic and allergic proteins in seeds. Results Full-length cDNAs are useful resources in annotating genes and in providing functional analysis of genes and their products. We constructed a full-length cDNA library from developing castor endosperm, and obtained 4,720 ESTs from 5'-ends of the cDNA clones representing 1,908 unique sequences. The most abundant transcripts are genes encoding storage proteins, ricin, agglutinin and oleosins. Several other sequences are also very numerous, including two acidic triacylglycerol lipases, and the oleate hydroxylase (FAH12 gene that is responsible for ricinoleate biosynthesis. The role(s of the lipases in developing castor seeds are not clear, and co-expressing of a lipase and the FAH12 did not result in significant changes in hydroxy fatty acid accumulation in transgenic Arabidopsis seeds. Only one oleate desaturase (FAD2 gene was identified in our cDNA sequences. Sequence and functional analyses of the castor FAD2 were carried out since it had not been characterized previously. Overexpression of castor FAD2 in a FAH12-expressing Arabidopsis line resulted in decreased accumulation of hydroxy fatty acids in transgenic seeds. Conclusion Our results suggest that transcriptional regulation of FAD2 and FAH12 genes maybe one of the mechanisms that contribute to a high level of ricinoleate accumulation in castor endosperm. The full-length cDNA library will be used to search for additional genes that affect ricinoleate accumulation in seed oils. Our EST sequences will also be useful to annotate the castor genome, which whole sequence is being generated by shotgun sequencing at

  6. A barley PHD finger transcription factor that confers male sterility by affecting tapetal development.

    Science.gov (United States)

    Fernández Gómez, José; Wilson, Zoe A

    2014-08-01

    Controlling pollen development is of major commercial importance in generating hybrid crops and selective breeding, but characterized genes for male sterility in crops are rare, with no current examples in barley. However, translation of knowledge from model species is now providing opportunities to understand and manipulate such processes in economically important crops. We have used information from regulatory networks in Arabidopsis to identify and functionally characterize a barley PHD transcription factor MALE STERTILITY1 (MS1), which expresses in the anther tapetum and plays a critical role during pollen development. Comparative analysis of Arabidopsis, rice and Brachypodium genomes was used to identify conserved regions in MS1 for primer design to amplify the barley MS1 gene; RACE-PCR was subsequently used to generate the full-length sequence. This gene shows anther-specific tapetal expression, between late tetrad stage and early microspore release. HvMS1 silencing and overexpression in barley resulted in male sterility. Additionally, HvMS1 cDNA, controlled by the native Arabidopsis MS1 promoter, successfully complemented the homozygous ms1 Arabidopsis mutant. These results confirm the conservation of MS1 function in higher plants and in particular in temperate cereals. This has provided the first example of a characterized male sterility gene in barley, which presents a valuable tool for the future control of male fertility in barley for hybrid development.

  7. 植物胚乳发育的表观遗传学调控%Epigenetic Regulation in Plant Endosperm Development

    Institute of Scientific and Technical Information of China (English)

    张美善; 刘宝

    2012-01-01

    In angiosperms, seed development initiates from double fertilization, which produces a diploid embryo and a triploid endosperm. The endosperm, a terminally differentiated tissue that nourishes the embryo during seed development and germination, is the prominent tissue of imprinting in plants. Proper endosperm development is crucial for normal embryo and seed development. The expression of imprinted genes is regulated by epigenetic mechanisms, including DNA methylation, H3K27 trimethylation and PollV-dependent siRNA (p4-siRNA). Such epigenetic regulation of imprinting is vital to proper endosperm development and seed viability. Recent studies show that endosperm DNA methylation is reduced genome-wide, which likely originates from demethylation in the central cell nucleus of the female gametophyte. This review focuses on the latest research advances in the epigenetic regulation in plant seed development, including the mechanism of plant genomic imprinting and dynamics of the genome-wide demethylation in endosperm.%被子植物的种子发育从双受精开始,产生二倍体的胚和三倍体的胚乳.在种子发育和萌发过程中,胚乳向胚组织提供营养物质,因此胚乳对胚和种子的正常生长发育至关重要.开花植物发生基因组印迹的主要器官是胚乳.印迹基因的表达受表观遗传学机制的调控,包括DNA甲基化和组蛋白H3K27甲基化修饰以及依赖于PollV的siRNAs(p4-siRNAs)调控.基因组印迹的表观遗传学调控对胚乳的正常发育和种子育性具有不可或缺的重要作用.最新研究显示,胚乳的整个基因组DNA甲基化水平降低,而且去甲基化作用可能源于雌配子体的中央细胞.该文综述了种子发育的表观遗传学调控机制,包括基因组印迹机制以及胚乳基因组DNA甲基化变化研究的最新进展.

  8. Effects of the activities of key enzymes involved in starch biosynthesis on the fine structure of amylopectin in developing rice (Oryza sativa L.) endosperms

    Institute of Scientific and Technical Information of China (English)

    L(U) Bing; GUO ZhiGang; LIANG JianSheng

    2008-01-01

    The dynamic changes of the activities of enzymes involving in starch biosynthesis, including ADP-glucose pyrophosphorylase (AGPase), soluble starch synthases (SSS), starch branching enzyme (SBE) and starch debranching enzymes (DBE) were studied, and changes of fine structure of amy-Iopectin were characterized by isoamylase treatment during rice grain development, using trans anti-waxy gene rice plants. The relationships between the activities of those key enzymes were also analyzed. The amylose synthesis was significantly inhibited in transgenic Wanjing 9522, but the total starch content and final grain weight were less affected as compared with those of non-transgenic Wanjing 9522 rice cultivar. Analyses on the changes of activities of enzymes involving in starch bio-synthesis showed that different enzyme activities were expressed differently during rice endosperm development. Soluble starch synthase is relatively highly expressed in earlier stage of endosperm de-velopment, whilst maximal expression of granule-bound starch synthase (GBSS) occurred in mid-stage of endosperm development. No obvious differences in changes of the activities of AGPase and SBE between two rice cultivars investigated, except the DBEs. Distribution patterns of branches of amy-Iopectin changed continually during the development of rice grains and varied between two rice culti-vars. It was suggested that amylopectin synthesis be prior to the synthesis of amylose and different enzymes have different roles in controlling syntheses of branches of amylopectin.

  9. Effects of the activities of key enzymes involved in starch biosynthesis on the fine structure of amylopectin in developing rice (Oryza sativa L.) endosperms.

    Science.gov (United States)

    Lü, Bing; Guo, ZhiGang; Liang, JianSheng

    2008-10-01

    The dynamic changes of the activities of enzymes involving in starch biosynthesis, including ADP-glucose pyrophosphorylase (AGPase), soluble starch synthases (SSS), starch branching enzyme (SBE) and starch debranching enzymes (DBE) were studied, and changes of fine structure of amylopectin were characterized by isoamylase treatment during rice grain development, using trans anti-waxy gene rice plants. The relationships between the activities of those key enzymes were also analyzed. The amylose synthesis was significantly inhibited in transgenic Wanjing 9522, but the total starch content and final grain weight were less affected as compared with those of non-transgenic Wanjing 9522 rice cultivar. Analyses on the changes of activities of enzymes involving in starch biosynthesis showed that different enzyme activities were expressed differently during rice endosperm development. Soluble starch synthase is relatively highly expressed in earlier stage of endosperm development, whilst maximal expression of granule-bound starch synthase (GBSS) occurred in mid-stage of endosperm development. No obvious differences in changes of the activities of AGPase and SBE between two rice cultivars investigated, except the DBEs. Distribution patterns of branches of amylopectin changed continually during the development of rice grains and varied between two rice cultivars. It was suggested that amylopectin synthesis be prior to the synthesis of amylose and different enzymes have different roles in controlling syntheses of branches of amylopectin.

  10. Effects of the activities of key enzymes involved in starch biosynthesis on the fine structure of amylopectin in developing rice (Oryza sativa L.) endosperms

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The dynamic changes of the activities of enzymes involving in starch biosynthesis, including ADP-glucose pyrophosphorylase (AGPase), soluble starch synthases (SSS), starch branching enzyme (SBE) and starch debranching enzymes (DBE) were studied, and changes of fine structure of amy- lopectin were characterized by isoamylase treatment during rice grain development, using trans anti-waxy gene rice plants. The relationships between the activities of those key enzymes were also analyzed. The amylose synthesis was significantly inhibited in transgenic Wanjing 9522, but the total starch content and final grain weight were less affected as compared with those of non-transgenic Wanjing 9522 rice cultivar. Analyses on the changes of activities of enzymes involving in starch bio- synthesis showed that different enzyme activities were expressed differently during rice endosperm development. Soluble starch synthase is relatively highly expressed in earlier stage of endosperm de- velopment, whilst maximal expression of granule-bound starch synthase (GBSS) occurred in mid-stage of endosperm development. No obvious differences in changes of the activities of AGPase and SBE between two rice cultivars investigated, except the DBEs. Distribution patterns of branches of amy- lopectin changed continually during the development of rice grains and varied between two rice culti- vars. It was suggested that amylopectin synthesis be prior to the synthesis of amylose and different enzymes have different roles in controlling syntheses of branches of amylopectin.

  11. A comparative glycoproteome study of developing endosperm in the hexose-deficient miniature1 (mn1 seed mutant and its wild type Mn1 in maize

    Directory of Open Access Journals (Sweden)

    Cecilia eSilva-Sanchez

    2014-02-01

    Full Text Available In maize developing seeds, transfer cells are prominently located at the basal endosperm transfer layer (BETL. As the first filial cell layer, BETL is a gateway to sugars, nutrients and water from mother plant; and anchor of numerous functions such as sucrose turnover, auxin and cytokinin biosynthesis/accumulation, energy metabolism, defense response, and signaling between maternal and filial generations. Previous studies showed that basal developing endosperms of miniature1 (mn1 mutant seeds lacking the Mn1-encoded cell wall invertase II, are also deficient for hexose. Given the role of glucose as one of the key sugars in protein glycosylation and proper protein folding; we performed a comparative large scale glycoproteome profiling of total proteins of these two genotypes (mn1 mutant vs Mn1 wild type using 2D gel electrophoresis and glycosylation/total protein staining, followed by image analysis. Protein identification was done by LC-MS/MS. A total of 413 spots were detected; from which, 113 spots matched between the two genotypes. Of these, 45 showed > 20% decrease/increase in glycosylation level and were selected for protein identification. A large number of identified proteins showed decreased glycosylation levels in mn1 developing endosperms as compared to the Mn1. Functional classification of proteins, showed mainly of post-translational modification, protein turnover, chaperone activities, carbohydrate and amino acid biosynthesis / transport, and cell wall biosynthesis. These proteins and activities were related to endoplasmic reticulum (ER stress and unfolded protein response (UPR as a result of the low glycolsylation levels of the mutant proteins. Overall, these results provide for the first time a global glycoproteome profile of maize BETL-enriched basal endosperm to better understand their role in seed development in maize.

  12. Roles of the haustorium and endosperm during the development of seedlings of Acrocomia aculeata (Arecaceae): dynamics of reserve mobilization and accumulation.

    Science.gov (United States)

    Mazzottini-Dos-Santos, Hellen Cássia; Ribeiro, Leonardo Monteiro; Oliveira, Denise Maria Trombert

    2016-11-24

    The mobilization of palm seed reserves is a complex process because of the abundance and diversity of stored compounds and results from the development of a highly specialized haustorium. This work focused on the important Neotropical oleaginous palm Acrocomia aculeata, with the aim of defining phases of seedling development associated with mobilization of reserves and elucidating the role of haustorium and endosperm in this process. Standard methods were performed, including biometric, anatomical, and histochemical analyses, as well as the evaluation of the activities of the enzymes endo-β-mannanase and lipase, throughout the reserve mobilization in seeds during germination and in seedlings. Seeds of A. aculeata stored large quantities of proteins, lipids, and polysaccharides in the embryo and endosperm. The mobilization of reserves initiated in the haustorium during germination and subsequently occurred in the endosperm adjacent to the haustorium, forming a gradually increasing zone of digestion. Proteins and polysaccharides were the first to be mobilized, followed by lipids and cell wall constituents. The haustorium activates and controls the mobilization, forming transitory reserves and translocating them to the vegetative axis, while the endosperm, which also has an active role, serves as a site of intense enzymatic activity associated with protein bodies. Seedling development can be described as occurring in six phases over a long period (approximately 150 days) due to the large amount of seed reserves. This process exhibits an alternation between stages of accumulation and translocation of protein, lipid, and carbohydrate reserves in the haustorium, which favors the seedling establishment and the reproductive success of the species.

  13. Defining the SUMO System in Maize: SUMOylation Is Up-Regulated during Endosperm Development and Rapidly Induced by Stress.

    Science.gov (United States)

    Augustine, Robert C; York, Samuel L; Rytz, Thérèse C; Vierstra, Richard D

    2016-07-01

    In response to abiotic and biotic challenges, plants rapidly attach small ubiquitin-related modifier (SUMO) to a large collection of nuclear proteins, with studies in Arabidopsis (Arabidopsis thaliana) linking SUMOylation to stress tolerance via its modification of factors involved in chromatin and RNA dynamics. Despite this importance, little is known about SUMOylation in crop species. Here, we describe the plant SUMO system at the phylogenetic, biochemical, and transcriptional levels with a focus on maize (Zea mays). In addition to canonical SUMOs, land plants encode a loosely constrained noncanonical isoform and a variant containing a long extension upstream of the signature β-grasp fold, with cereals also expressing a novel diSUMO polypeptide bearing two SUMO β-grasp domains in tandem. Maize and other cereals also synthesize a unique SUMO-conjugating enzyme variant with more restricted expression patterns that is enzymatically active despite a distinct electrostatic surface. Maize SUMOylation primarily impacts nuclear substrates, is strongly induced by high temperatures, and displays a memory that suppresses subsequent conjugation. Both in-depth transcript and conjugate profiles in various maize organs point to tissue/cell-specific functions for SUMOylation, with potentially significant roles during embryo and endosperm maturation. Collectively, these studies define the organization of the maize SUMO system and imply important functions during seed development and stress defense.

  14. 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.

  15. 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

  16. Molecular Genetic Traits Influencing Maize Endosperm Development and Value: Closeout Report for DOE Grant DE-FG02-96ER20242

    Energy Technology Data Exchange (ETDEWEB)

    Brian A. Larkins

    2012-09-12

    Development of the endosperm in cereal grasses entails different phases characterized by cell division, endoreduplication, accumulation of storage metabolites and cell death, which need to be carried out in an orderly fashion. While correct regulation of the cell cycle plays an essential role in endosperm development, the key regulatory factors and how the cell cycle interfaces with other pathways in this developmental context are largely unknown. We investigated the cyclin-dependent kinase (CDK)-retinoblastoma pathway and how it controls the cell cycle and coordinates it with other processes during maize endosperm development. Retinoblastoma-related (RBR) proteins may be inactivated through CDK-mediated phosphorylation, but the identity of the responsible kinase in maize is unknown. We have previously shown that down-regulation of CDKA;1 severely inhibits the endoreduplication cell cycle and suggested that CDK may be an up-stream regulator of the retinoblastoma pathway. We discovered two types of maize RBR genes, RBR1 and RBR3, which differ in terms of structure, regulation and function. Phylogenetic analyses indicate that these genes may be distinctive features of the Poaceae. We found that RBR3 plays a positive rather than a negative role in DNA replication, cell transformation, and the expression of the minichromosome maintenance (MCM)2-7 family of DNA replication factors. These features are a paradigm shift in RBR gene function and appear to be unique within the RBR gene family. They suggest the existence in maize and related cereal crops of specific RBR/E2F-dependent pathways impinging on the cell cycle and development. RBR1 was down-regulated in transgenic endosperm using RNAi approaches. This resulted in the de-repression of a number of down-stream E2F targets, including RBR3, the MCM2-7 gene family, DNA methyltransferase (MET)1, CDKB;1, and the recently identified RBR4 gene. It also increased endosperm ploidy levels, stimulated the production of a larger

  17. Root border cell development is a temperature-insensitive and Al-sensitive process in barley.

    Science.gov (United States)

    Pan, Jian-Wei; Ye, Dan; Wang, Li-Ling; Hua, Jing; Zhao, Gu-Feng; Pan, Wei-Huai; Han, Ning; Zhu, Mu-Yuan

    2004-06-01

    In vivo and in vitro experiments showed that border cell (BC) survival was dependent on root tip mucigel in barley (Hordeum vulgare L. cv. Hang 981). In aeroponic culture, BC development was an induced process in barley, whereas in hydroponic culture, it was a kinetic equilibrium process during which 300-400 BCs were released into water daily. The response of root elongation to temperatures (10-35 degrees C) was very sensitive but temperature changes had no great effect on barley BC development. At 35 degrees C, the root elongation ceased whereas BC production still continued, indicating that the two processes might be regulated independently under high temperature (35 degrees C) stress. Fifty microM Al could inhibit significantly BC development by inhibiting pectin methylesterase activity in the root cap of cv. 2000-2 (Al-sensitive) and cv. Humai 16 (Al-tolerant), but 20 microM Al could not block BC development in cv. Humai 16. BCs and their mucigel of barley had a limited role in the protection of Al-induced inhibition of root elongation, but played a significant role in the prevention of Al from diffusing into the meristems of the root tip and the root cap. Together, these results suggested that BC development was a temperature-insensitive but Al-sensitive process, and that BCs and their mucigel played an important role in the protection of root tip and root cap meristems from Al toxicity.

  18. Temporal small RNA transcriptome profiling unraveled partitioned miRNA expression in developing maize endosperms between reciprocal crosses

    Directory of Open Access Journals (Sweden)

    Mingming eXin

    2015-09-01

    Full Text Available In angiosperms, the endosperm nurtures embryo and provides nutrients for seed germination. We have performed high-throughput small RNA transcriptome sequencing of kernels at 0, 3, and 5 days after pollination (DAP and endosperms at 7, 10, and 15 DAP by using B73 and Mo17 reciprocal crosses in previous study. Here, we further explored these small RNA-seq data to investigate the potential roles of miRNAs in regulating gene expression process. In total, 57 conserved miRNAs and 18 novel miRNAs were observed highly expressed in maize endosperm. Temporal expression profiling indicates these miRNAs exhibited dynamic and partitioned expression patterns at different developmental stages between maize reciprocal crosses, and qRT-PCR results further confirmed our observation. In addition, we found a subset of distinct tandem miRNAs are generated from a single stem-loop structure in maize which might be conserved in monocots. Furthermore, a SNP variation of Zma-miR408-5p at 11th base position was characterized between B73 and Mo17 which leads to completely different functions in repressing targets. More interestingly, Zma-miR408-5p exhibited B73-biased expression pattern in the B73 and Mo17 reciprocal hybrid endosperms at 7, 10 and 15 DAP according to the reads abundance with SNPs and CAPS experiment.

  19. Vacuolar H+-translocating inorganic pyrophosphatase (Vpp1) marks partial aleurone cell fate in cereal endosperm development.

    Science.gov (United States)

    Wisniewski, Jean-Pierre; Rogowsky, Peter M

    2004-10-01

    Cereal endosperm is a model system for cell fate determination in plants. In wild-type plants the outermost endosperm cells adopt aleurone cell fate, while all underlying cells display starchy endosperm cell fate. Mutant analysis showed that cell fate is determined by position rather than lineage. To further characterise the precise cell fate of the outermost cells, we performed a differential screen and isolated the novel marker gene Vpp1 . It encodes a vacuolar H+-translocating inorganic pyrophosphatase (V-PPase) and is mainly expressed in kernels, leaves and tassels. In kernels, its expression is restricted to the aleurone layer with the maximum of expression shifting from the adaxial to the abaxial side during early stages. Together with three other marker genes Vpp1 was then used to analyse the cell fate of the outermost cells in Dap3 , Dap7 , cr4 and dek1 mutants, all of which have aberrant aleurone layers. In the Dap3 and Dap7 mutants the Vpp1 and Ltp2 markers but not the A1 and Zein markers were expressed in patches without aleurone indicating that the outermost cells had some but not all features of aleurone cells and did not simply adopt starchy endosperm cell fate. A similar result was obtained in the cr4 mutant, although Ltp2 expression was less generalised. In other Dap7 patches characterised by multiple aleurone-like cell layers the expression of Vpp1 and Ltp2 confirmed the aleurone cell fate of the cells in the additional cell layers. The analysis of dek1 mutants confirmed the starchy endosperm cell fate of the majority but not all outermost cells. Based on these data we propose a model suggesting a stepwise commitment to aleurone cell fate. Sequential steps are marked by the expression of Vpp1 , the expression of Ltp2 , the acquisition of a regular shape and thick walls and finally pigmentation coupled with A1 expression.

  20. A chemometric evaluation of the underlying physical and chemical patterns that support near infrared spectroscopy of barley seeds as a tool for explorative classification of endosperm genes and gene combinations

    DEFF Research Database (Denmark)

    Jacobsen, Susanne; Søndergaard, Ib; Møller, Birthe

    2005-01-01

    Near infrared spectroscopic (NIR; 1100-2500 nm), chemical and genetic data were combined to study the pleiotropic secondary effects of mutant genes on milled samples in a barley seed model. NIR and chemical data were both effective in classifying gene and gene combinations by Principal Component ...

  1. 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.

  2. Global Identification of Significantly Expressed Genes in Developing Endosperm of Rice by Expression Sequence Tags and cDNA Array Approaches

    Institute of Scientific and Technical Information of China (English)

    Qichao Tu; Haitao Dong; Haigen Yao; Yongqi Fang; Cheng'en Dai; Hongmei Luo; Jian Yao; Dong Zhao; Debao Li

    2008-01-01

    Rice endosperm plays a very important role in seedling germination and determines the qualities of fice grain.Although studies on specific gene categories in endosperm have been carried out,global view of gene expression at a transcription level in rice endosperm is still limited.To gain a better understanding of the global and tissue-specific gene expression profiles in rice endosperm,a cDNA library from rice endosperm of immature seeds was sequenced.A cDNA array was constructed based on the tentative unique transcripts derived from expression sequence tag (EST) assembling results and then hybridized with cONAs from five different tissues or organs including endosperm,embryo,leaf,stem and root of rice.Significant redundancy was found for genes encoding prolamin,glutelin,allergen,and starch synthesis proteins,accounting for~34% of the total ESTs obtained.The cDNA array revealed 87 significantly expressed genes in endosperm compared with the other four organs or tissues.These genes included 13 prolamin family proteins,17 glutelin family proteins,12 binding proteins,nine catalytic proteins and four ribosomal proteins,indicating a complicated biological processing in rice endosperm.In addition,Northern verification of 1,4-alpha-glucan branching enzyme detected two isoforms in rice endosperm,the larger one of which only existed in endosperm.

  3. Structure and Development of Endosperm Transfer Cells in Wheat%小麦胚乳传递细胞发育的结构观察

    Institute of Scientific and Technical Information of China (English)

    王慧慧; 王峰; 刘大同; 顾蕴洁; 王忠

    2011-01-01

    In developing wheat caryopsis,grain filling nutrients transport into the endosperm via endosperm transfer cells(ETCs).The developmental extent of ETCs is in close relation to yield and qualities of wheat.In this paper we investigated the developmental process and structural features of ETCs in wheat systematically by the combination of light microscopy and transmission electron microscopy.Main results were as follows:(1) ETCs occured in the endosperm epidermis bordering the endosperm cavity and were the first cell type histologically differentiated during endosperm development.ETCs could be divided into two subtypes: outer 1~2 layer of aleurone transfer cells and inner 1~2 layer of starchy endosperm transfer cells.(2) As caryopsis matured,the nuclei of starchy endosperm transfer cells disappeared,while the nuclei of aleurone transfer cells remained intact.(3) The development of ETCs showed obvious polarity and the pattern of temporal and spatial gradient.(4) Aleurone transfer cells had dense cytoplasm rich in rough endoplasmic reticulum,mitochondria,dictyosomes and lipid bodies;their plasma membrane wrinkled and in some region evaginated to form numerous plasmatubules.(5) Starchy endosperm transfer cells had sparse cytoplasm rich in amyloplasts and were highly vacuolated.(6) Lots of plasmodesmata penetrated into adjoining primary walls of ETCs where thickening and wall ingrowths didn't occur.(7) Mitochondria in ETCs showed polarized distribution,and most of them lied in the proximity of plasma membrane.The structural features of ETCs suggested that they may play a role in nutrients transportation into the endosperm both via apoplastic and symplastic pathway.%为从细胞学方面了解小麦产量和品质的形成机制,以扬麦5号为材料,利用光镜和透射电镜观察了小麦颖果发育过程中胚乳传递细胞的结构变化,并探讨了胚乳传递细胞的生理功能。结果表明:(1)胚乳传递细胞

  4. The functions of the endosperm during seed germination.

    Science.gov (United States)

    Yan, Dawei; Duermeyer, Lisza; Leoveanu, Catalina; Nambara, Eiji

    2014-09-01

    In angiosperms, a double fertilization event initiates the development of two distinct structures, the embryo and endosperm. The endosperm plays an important role in supporting embryonic growth by supplying nutrients, protecting the embryo and controlling embryo growth by acting as a mechanical barrier during seed development and germination. Its structure and function in the mature dry seed is divergent and specialized among different plant species. A subset of endospermic tissues are composed of living cells even after seed maturation, and play an active role in the regulation of seed germination. Transcriptome analysis has provided new insights into the regulatory functions of the endosperm during seed germination. It is well known that the embryo secretes signals to the endosperm to induce the degradation of the seed reserve and to promote endosperm weakening during germination. Recent advances in seed biology have shown that the endosperm is capable of sensing environmental signals, and can produce and secrete signals to regulate the growth of the embryo. Thus, germination is a systemic response that involves bidirectional interactions between the embryo and endosperm.

  5. Aberrant splicing in maize rough endosperm3 reveals a conserved role for U12 splicing in eukaryotic multicellular development

    Science.gov (United States)

    Barbazuk, W. Brad

    2017-01-01

    RNA splicing of U12-type introns functions in human cell differentiation, but it is not known whether this class of introns has a similar role in plants. The maize ROUGH ENDOSPERM3 (RGH3) protein is orthologous to the human splicing factor, ZRSR2. ZRSR2 mutations are associated with myelodysplastic syndrome (MDS) and cause U12 splicing defects. Maize rgh3 mutants have aberrant endosperm cell differentiation and proliferation. We found that most U12-type introns are retained or misspliced in rgh3. Genes affected in rgh3 and ZRSR2 mutants identify cell cycle and protein glycosylation as common pathways disrupted. Transcripts with retained U12-type introns can be found in polysomes, suggesting that splicing efficiency can alter protein isoforms. The rgh3 mutant protein disrupts colocalization with a known ZRSR2-interacting protein, U2AF2. These results indicate conserved function for RGH3/ZRSR2 in U12 splicing and a deeply conserved role for the minor spliceosome to promote cell differentiation from stem cells to terminal fates. PMID:28242684

  6. Enzymic hydrolysis and industrial importance of barley. beta. -glucans and wheat flour pentosans

    Energy Technology Data Exchange (ETDEWEB)

    McCleary, B.V.; Gibson, T.S.; Allen, H.; Gams, T.C.

    1986-12-01

    Mixed linkage ..beta..-glucane and pentosanes (mainly arabinoxylanes) are the major endosperm cell-wall polysaccharides of barley and wheat respectively. These polysaccharides, although minor components of the whole grain, significantly affect the industrial utilization of these cereals. The modification of barley corns during malting requires the dissolution of the ..beta..-glucane in the cell-wall of the starch endosperm. High ..beta..-glucane concentration in wort and beer effect the rate of filtration and can also lead to precipitate or gel formation in the final product. In a similar manner, pentosane is thought to cause filtration problems with wheat starch hydrolysates by increasing viscosity and by producing gelatinous precipitate which blocks filters. Ironically, it is this same viscosity building and water binding capacity which is considered to render pentosanes of considerable value in dough development and bread storage (anti-staling functions). In the current paper, some aspects of the beneficial and detrimental effects of pentosanes and ..beta..-glucane in the industrial utilization of wheat and barley are discussed. More specifically, enzymic methods for the preparation, analysis and identification of these polysaccharides and for the removal of their functional properties, are described in detail.

  7. Investigation of Endosperm Cell Development of Different Rice Varieties%不同类型水稻品种胚乳发育的研究

    Institute of Scientific and Technical Information of China (English)

    李栋梁; 李小刚; 顾蕴洁; 王忠

    2014-01-01

    不同部位的分布上,4种水稻品种各自呈现了一定程度的差异与联系。粳稻2个品种与籼稻2个品种间差异更多体现在颖果发育进程上;而糯稻与非糯稻的区别则更多体现在淀粉体的形态与灌浆的充实程度上。%The objective of this research is to clarify the difference of rice endosperm of different varieties during the development process.[Method]Four rice varieties, Nipponbare, Yangdao 6, Wuyunuo16 and Yangfunuo 4 were used as the experimental materials, and their caryopsis development days precisely were precisely recorded and their caryopsis development was closely observed. Starch accumulation and physiological activities were observed by I2-KI and TTC staining. Structure changes of rice starch endosperm cells were observed by applying spur resin embedding and semi-thin sectioning, and light microscopy. The ultrastructure and element analysis of full ripe rice caryopsis were observed under scanning electron microscope and EDS. [Result]Four stages occurred in caryopses development of all the rice varieties tested: formation stage, milky stage, dough stage, and full maturity stage. Compared to development stages of the endosperm, formation stage runs in parallel to the coenocyte and cellurization stages, which are the initial stage of endosperm formation; milky stage corresponds to the differentiation stage of endosperm cells, while the last two stages, dough and full maturity stages, are equivalent to that of the endosperm maturity stage. The shape of two rice sub subspecies, japonica and indica, varies significantly. Of the four rice varieties tested, two indica varieties showed a relatively faster growth rate than two japonica cultivars; Wuyunuo16 and Yangfunuo 4 had almost identical growth rate of dry matter accumulation. Nuclei of starch endosperm cells degenerated with the extension of amyloplasts, while amyloplasts were still enlarging and growing. Amyloplasts in the endosperm cells were compound granules, and their

  8. 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.

  9. Cuticular permeance in relation to wax and cutin development along the growing barley (Hordeum vulgare) leaf.

    Science.gov (United States)

    Richardson, Andrew; Wojciechowski, Tobias; Franke, Rochus; Schreiber, Lukas; Kerstiens, Gerhard; Jarvis, Mike; Fricke, Wieland

    2007-05-01

    The developing leaf three of barley provides an excellent model system for the direct determination of relationships between amounts of waxes and cutin and cuticular permeance. Permeance of the cuticle was assessed via the time-course of uptake of either toluidine blue or (14)C-labelled benzoic acid ([(14)C] BA) along the length of the developing leaf. Toluidine blue uptake only occurred within the region 0-25 mm from the point of leaf insertion (POLI). Resistance--the inverse of permeance--to uptake of [(14)C] BA was determined for four leaf regions and was lowest in the region 10-20 mm above POLI. At 20-30 and 50-60 mm above POLI, it increased by factors of 6 and a further 32, respectively. Above the point of emergence of leaf three from the sheath of leaf two, which was 76-80 mm above POLI, resistance was as high as at 50-60 mm above POLI. GC-FID/MS analyses of wax and cutin showed that: (1) the initial seven fold increase in cuticular resistance coincided with increase in cutin coverage and appearance of waxes; (2) the second, larger and final increase in cuticle resistance was accompanied by an increase in wax coverage, whereas cutin coverage remained unchanged; (3) cutin deposition in barley leaf epidermis occurred in parallel with cell elongation, whereas deposition of significant amounts of wax commenced as cells ceased to elongate.

  10. 水稻胚乳细胞发育的结构观察及其矿质元素分析%Structure Observation of Rice Endosperm Cell Development and Its Mineral Element Analysis

    Institute of Scientific and Technical Information of China (English)

    王忠; 顾蕴洁; 郑彦坤; 王慧慧

    2012-01-01

    In order to prove up mechanism of rice endosperm development and aleurone layer formation, structure changes of rice aleurone cells and inner endosperm cells during caryopsis development were observed under light microscope and electron microscope. Kinds and relative content of endosperm cell elements were analyzed with energy spectrometer. The results show that aleurone cells are differentiated from endosperm epidermis. The contents of P,K, Mg and Ca mineral elements in aleurone cells are obviously higher than those in inner endosperm cells. During the early development stage, aleurone cells contain many mitochondria, spherosomes and small vacuoles; during the middle and late development stages, small vacuoles accumulate protein and mineral elements to form aleurone granules. During middle and late development stages of endosperm, nuclei of endosperm cells distort and degenerate when cells accumulate starch; however, nuclei of aleurone cells do not degenerate during the development. The formation of aleurone layers is related to accumulation of mineral elements, lipids and other "filling wastes" (not endosperm cell reserve substance things). Therefore,cell layers of the aleurone layer in dorsal caryopsis through which more nutrients are transported overnumber those in ventral and side caryopsis. Development of cereal endosperm should be divided into four stages: coenocyte stage,cellularizalion stage,differentiation stage and maturation stage.%为了探明水稻胚乳发育的过程和糊粉层形成的机理,用光学显微镜和电子显微镜观察了水稻糊粉层细胞和内胚乳细胞在颖果发育过程中的结构变化,用能谱仪分析了胚乳细胞中元素的种类和相对含量.结果表明,糊粉层细胞是由胚乳表层细胞转化而来的.糊粉层细胞中的P、K、Mg和Ca等矿质元素含量要明显高于内胚乳细胞.发育初期糊粉层细胞中富含线粒体、圆球体和小液泡;发育中后期小液泡积累蛋白质和

  11. The 5' flanking region of a barley B hordein gene controls tissue and developmental specific CAT expression in tobacco plants.

    Science.gov (United States)

    Marris, C; Gallois, P; Copley, J; Kreis, M

    1988-07-01

    The 549 base pairs of the 5' flanking region of a barley seed storage protein (B1 hordein) gene were linked to the reporter gene encoding chloramphenicol acetyl transferase (CAT). The chimaeric gene was transferred into tobacco plants using Agrobacterium tumefaciens. CAT enzyme activity was detected in the seeds, but not in the leaves, of the transgenic plants. Furthermore, enzyme activity was found only in the endosperm, and only from fifteen days after pollination. In contrast, the constitutive 19S promoter from cauliflower mosaic virus (CaMV) directed the expression of the CAT gene in the leaves as well as in both the endosperm and embryo and at all stages in seed development.

  12. The MADS Box Genes ABS, SHP1, and SHP2 Are Essential for the Coordination of Cell Divisions in Ovule and Seed Coat Development and for Endosperm Formation in Arabidopsis thaliana.

    Science.gov (United States)

    Ehlers, Katrin; Bhide, Amey S; Tekleyohans, Dawit G; Wittkop, Benjamin; Snowdon, Rod J; Becker, Annette

    2016-01-01

    Seed formation is a pivotal process in plant reproduction and dispersal. It begins with megagametophyte development in the ovule, followed by fertilization and subsequently coordinated development of embryo, endosperm, and maternal seed coat. Two closely related MADS-box genes, SHATTERPROOF 1 and 2 (SHP1 and SHP2) are involved in specifying ovule integument identity in Arabidopsis thaliana. The MADS box gene ARABIDOPSIS BSISTER (ABS or TT16) is required, together with SEEDSTICK (STK) for the formation of endothelium, part of the seed coat and innermost tissue layer formed by the maternal plant. Little is known about the genetic interaction of SHP1 and SHP2 with ABS and the coordination of endosperm and seed coat development. In this work, mutant and expression analysis shed light on this aspect of concerted development. Triple tt16 shp1 shp2 mutants produce malformed seedlings, seed coat formation defects, fewer seeds, and mucilage reduction. While shp1 shp2 mutants fail to coordinate the timely development of ovules, tt16 mutants show less peripheral endosperm after fertilization. Failure in coordinated division of the innermost integument layer in early ovule stages leads to inner seed coat defects in tt16 and tt16 shp1 shp2 triple mutant seeds. An antagonistic action of ABS and SHP1/SHP2 is observed in inner seed coat layer formation. Expression analysis also indicates that ABS represses SHP1, SHP2, and FRUITFUL expression. Our work shows that the evolutionary conserved Bsister genes are required not only for endothelium but also for endosperm development and genetically interact with SHP1 and SHP2 in a partially antagonistic manner.

  13. Lipid partitioning in maize (Zea mays L.) endosperm highlights relationships among starch lipids, amylose, and vitreousness.

    Science.gov (United States)

    Gayral, Mathieu; Bakan, Bénédicte; Dalgalarrondo, Michele; Elmorjani, Khalil; Delluc, Caroline; Brunet, Sylvie; Linossier, Laurent; Morel, Marie-Hélène; Marion, Didier

    2015-04-08

    Content and composition of maize endosperm lipids and their partition in the floury and vitreous regions were determined for a set of inbred lines. Neutral lipids, i.e., triglycerides and free fatty acids, accounted for more than 80% of endosperm lipids and are almost 2 times higher in the floury than in the vitreous regions. The composition of endosperm lipids, including their fatty acid unsaturation levels, as well as their distribution may be related to metabolic specificities of the floury and vitreous regions in carbon and nitrogen storage and to the management of stress responses during endosperm cell development. Remarkably, the highest contents of starch lipids were observed systematically within the vitreous endosperm. These high amounts of starch lipids were mainly due to lysophosphatidylcholine and were tightly linked to the highest amylose content. Consequently, the formation of amylose-lysophosphatidylcholine complexes has to be considered as an outstanding mechanism affecting endosperm vitreousness.

  14. Protein accumulation in aleurone cells, sub-aleurone cells and the center starch endosperm of cereals.

    Science.gov (United States)

    Zheng, Yankun; Wang, Zhong

    2014-10-01

    There are mainly three endosperm storage tissues in the cereal endosperm: aleurone cells, sub-aleurone cells and the center starch endosperm. The protein accumulation is very different in the three endosperm storage tissues. The aleurone cells accumulate protein in aleurone granules. The sub-aleurone cells and the center starch endosperm accumulate protein in endoplasmic reticulum-derived protein bodies and vacuolar protein bodies. Proteins are deposited in different patterns within different endosperm storage tissues probably because of the special storage properties of these tissues. There are several special genes and other molecular factors to mediate the protein accumulation in these tissues. Different proteins have distinct functions in the protein body formation and the protein interactions determine protein body assembly. There are both cooperation and competition relationships between protein, starch and lipid in the cereal endosperm. This paper reviews the latest investigations on protein accumulation in aleurone cells, sub-aleurone cells and the center starch endosperm. Useful information will be supplied for future investigations on the cereal endosperm development.

  15. The Maize Viviparous8 locus, encoding a putative ALTERED MERISTEM PROGRAM1-like peptidase, regulates abscisic acid accumulation and coordinates embryo and endosperm development.

    Science.gov (United States)

    Suzuki, Masaharu; Latshaw, Susan; Sato, Yutaka; Settles, A Mark; Koch, Karen E; Hannah, L Curtis; Kojima, Mikiko; Sakakibara, Hitoshi; McCarty, Donald R

    2008-03-01

    We describe a mutant of Zea mays isolated from a W22 inbred transposon population, widow's peak mutant1 (wpk1), with an altered pattern of anthocyanin synthesis and aleurone cell differentiation in endosperm. In addition, a failure of the developing mutant embryo to form leaf initials is associated with decreased expression of a subset of meristem regulatory genes that includes Abphyl1 and Td1. We show that the viviparous8 (vp8) mutant has a similar pleiotropic phenotype in the W22 inbred background in contrast to the viviparous embryo phenotype exhibited in the standard genetic background, and we confirmed that wpk1 is allelic to vp8. Further genetic analysis revealed that the standard vp8 stock contains an unlinked, partially dominant suppressor of the vp8 mutation that is not present in W22. Consistent with the early-onset viviparous phenotype of vp8, expression of several embryonic regulators, including LEC1/B3 domain transcription factors, was reduced in the mutant embryo. Moreover, reduced abscisic acid (ABA) content of vp8/wpk1 embryos was correlated with altered regulation of ABA biosynthesis, as well as ABA catabolic pathways. The ABA biosynthetic gene Vp14 was down-regulated in the nonsuppressed background, whereas the ZmABA8'oxA1a ABA 8'-hydroxylase gene was strongly up-regulated in both genetic backgrounds. Molecular analysis revealed that Vp8 encodes a putative peptidase closely related to Arabidopsis thaliana ALTERED MERISTEM PROGRAM1. Because the Vp8 regulates meristem development as well as seed maturation processes, including ABA accumulation, we propose that VP8 is required for synthesis of an unidentified signal that integrates meristem and embryo formation in seeds.

  16. The Maize Viviparous8 Locus, Encoding a Putative ALTERED MERISTEM PROGRAM1-Like Peptidase, Regulates Abscisic Acid Accumulation and Coordinates Embryo and Endosperm Development1[W

    Science.gov (United States)

    Suzuki, Masaharu; Latshaw, Susan; Sato, Yutaka; Settles, A. Mark; Koch, Karen E.; Hannah, L. Curtis; Kojima, Mikiko; Sakakibara, Hitoshi; McCarty, Donald R.

    2008-01-01

    We describe a mutant of Zea mays isolated from a W22 inbred transposon population, widow's peak mutant1 (wpk1), with an altered pattern of anthocyanin synthesis and aleurone cell differentiation in endosperm. In addition, a failure of the developing mutant embryo to form leaf initials is associated with decreased expression of a subset of meristem regulatory genes that includes Abphyl1 and Td1. We show that the viviparous8 (vp8) mutant has a similar pleiotropic phenotype in the W22 inbred background in contrast to the viviparous embryo phenotype exhibited in the standard genetic background, and we confirmed that wpk1 is allelic to vp8. Further genetic analysis revealed that the standard vp8 stock contains an unlinked, partially dominant suppressor of the vp8 mutation that is not present in W22. Consistent with the early-onset viviparous phenotype of vp8, expression of several embryonic regulators, including LEC1/B3 domain transcription factors, was reduced in the mutant embryo. Moreover, reduced abscisic acid (ABA) content of vp8/wpk1 embryos was correlated with altered regulation of ABA biosynthesis, as well as ABA catabolic pathways. The ABA biosynthetic gene Vp14 was down-regulated in the nonsuppressed background, whereas the ZmABA8′oxA1a ABA 8′-hydroxylase gene was strongly up-regulated in both genetic backgrounds. Molecular analysis revealed that Vp8 encodes a putative peptidase closely related to Arabidopsis thaliana ALTERED MERISTEM PROGRAM1. Because the Vp8 regulates meristem development as well as seed maturation processes, including ABA accumulation, we propose that VP8 is required for synthesis of an unidentified signal that integrates meristem and embryo formation in seeds. PMID:18203869

  17. Arabidopsis haiku mutants reveal new controls of seed size by endosperm.

    Science.gov (United States)

    Garcia, Damien; Saingery, Virginie; Chambrier, Pierre; Mayer, Ulrike; Jürgens, Gerd; Berger, Frédéric

    2003-04-01

    In flowering plants, maternal seed integument encloses the embryo and the endosperm, which are both derived from double fertilization. Although the development of these three components must be coordinated, we have limited knowledge of mechanisms involved in such coordination. The endosperm may play a central role in these mechanisms as epigenetic modifications of endosperm development, via imbalance of dosage between maternal and paternal genomes, affecting both the embryo and the integument. To identify targets of such epigenetic controls, we designed a genetic screen in Arabidopsis for mutants that phenocopy the effects of dosage imbalance in the endosperm. The two mutants haiku 1 and haiku 2 produce seed of reduced size that resemble seed with maternal excess in the maternal/paternal dosage. Homozygous haiku seed develop into plants indistinguishable from wild type. Each mutation is sporophytic recessive, and double-mutant analysis suggests that both mutations affect the same genetic pathway. The endosperm of haiku mutants shows a premature arrest of increase in size that causes precocious cellularization of the syncytial endosperm. Reduction of seed size in haiku results from coordinated reduction of endosperm size, embryo proliferation, and cell elongation of the maternally derived integument. We present further evidence for a control of integument development mediated by endosperm-derived signals.

  18. In vitro biochemical characterization of all barley endosperm starch synthases

    DEFF Research Database (Denmark)

    Cuesta-Seijo, Jose A.; Nielsen, Morten M.; Ruzanski, Christian

    2016-01-01

    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...... maltoligosaccharides and not polysaccharides as its preferred substrates....

  19. Transgenic rice endosperm as a bioreactor for molecular pharming.

    Science.gov (United States)

    Ou, Jiquan; Guo, Zhibin; Shi, Jingni; Wang, Xianghong; Liu, Jingru; Shi, Bo; Guo, Fengli; Zhang, Chufu; Yang, Daichnag

    2014-04-01

    Plants provide a promising expression platform for producing recombinant proteins with several advantages in terms of high expression level, lower production cost, scalability, and safety and environment-friendly. Molecular pharming has been recognized as an emerging industry with strategic importance that could play an important role in economic development and healthcare in China. Here, this review represents the significant advances using transgenic rice endosperm as bioreactor to produce various therapeutic recombinant proteins in transgenic rice endosperm and large-scale production of OsrHSA, and discusses the challenges to develop molecular pharming as an emerging industry with strategic importance in China.

  20. Vertical gradient in soil temperature stimulates development and increases biomass accumulation in barley.

    Science.gov (United States)

    Füllner, K; Temperton, V M; Rascher, U; Jahnke, S; Rist, R; Schurr, U; Kuhn, A J

    2012-05-01

    We have detailed knowledge from controlled environment studies on the influence of root temperature on plant performance, growth and morphology. However, in all studies root temperature was kept spatially uniform, which motivated us to test whether a vertical gradient in soil temperature affected development and biomass production. Roots of barley seedlings were exposed to three uniform temperature treatments (10, 15 or 20°C) or to a vertical gradient (20-10°C from top to bottom). Substantial differences in plant performance, biomass production and root architecture occurred in the 30-day-old plants. Shoot and root biomass of plants exposed to vertical temperature gradient increased by 144 respectively, 297%, compared with plants grown at uniform root temperature of 20°C. Additionally the root system was concentrated in the upper 10cm of the soil substrate (98% of total root biomass) in contrast to plants grown at uniform soil temperature of 20°C (86% of total root biomass). N and C concentrations in plant roots grown in the gradient were significantly lower than under uniform growth conditions. These results are important for the transferability of 'normal' greenhouse experiments where generally soil temperature is not controlled or monitored and open a new path to better understand and experimentally assess root-shoot interactions.

  1. 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....

  2. Characterization of a gene from chromosome 1B encoding the large subunit of ADPglucose pyrophosphorylase from wheat: evolutionary divergence and differential expression of Agp2 genes between leaves and developing endosperm.

    Science.gov (United States)

    Thorneycroft, David; Hosein, Felicia; Thangavelu, Madan; Clark, Joanna; Vizir, Igor; Burrell, Michael M; Ainsworth, Charles

    2003-07-01

    A full-length genomic clone containing the gene encoding the large subunit of the ADPglucose pyrophosphorylase (Agp2), was isolated from a genomic library prepared from etiolated shoots of hexaploid wheat (Triticum aestivum L., cv, Chinese Spring). The coding region of this gene is identical to one of the cDNA clones previously isolated from a developing wheat grain cDNA library and is therefore an actively transcribed gene. The sequence represented by the cDNA spans 4.8 kb of the genomic clone and contains 15 introns. 2852 bp of DNA flanking the transcription start site of the gene was cloned upstream of the GUS (beta-glucuronidase) reporter gene. This Agp2::GUS construct and promoter deletions were used to study the pattern of reporter gene expression in both transgenic tobacco and wheat plants. Histochemical analysis of GUS expression in transgenic tobacco demonstrated that the reporter gene was expressed in guard cells of leaves and throughout the seed. In transgenic wheat, reporter gene expression was confined to the endosperm and aleurone with no expression in leaves. The cloned Agp2 gene was located to chromosome 1B by gene-specific PCR with nullisomic-tetrasomic lines. Northern analysis demonstrated that the Agp2 genes are differentially expressed in leaves and developing endosperm; while all three classes of Agp2 genes are transcribed in developing wheat grain endosperm, only one is transcribed in leaves. The differences between the Agp2 genes are discussed in relation to the evolution of hexaploid wheat.

  3. A PCR-based forward genetics screening, using expression domain-specific markers, identifies mutants in endosperm transfer cell development

    Directory of Open Access Journals (Sweden)

    Luis Miguel Muñiz

    2014-04-01

    Full Text Available Mutant collections are an invaluable source of material on which forward genetic approaches allow the identification of genes affecting a wide variety of biological processes. However, some particular developmental stages and morphological structures may resist analysis due to their physical inaccessibility or to deleterious effects associated to their modification. Furthermore, lethal mutations acting early in development may escape detection. We have approached the characterisation of 101 maize seed mutants, selected from a collection of 27500 visually screened Mu-insertion lines, using a molecular marker approach based on a set of genes previously ascribed to different tissue compartments within the early developing kernel. A streamlined combination of qRT-PCR assays has allowed us to preliminary pinpoint the affected compartment, establish developmental comparisons to WT siblings and select mutant lines with alterations in the different compartments. Furthermore, clusters of markers co-affected by the underlying mutation were identified. We have analysed more extensively a set of lines presenting significant variation in transfer cell-associated expression markers, and have performed morphological observations, and immunolocalization experiments to confirm the results, validating this approach as an efficient mutant description tool.

  4. The Role of alpha-Glucosidase in Germinating Barley Grains

    DEFF Research Database (Denmark)

    Stanley, Duncan; Rejzek, Martin; Næsted, Henrik;

    2011-01-01

    in the endosperm 10 d after imbibition, implying inhibition of maltase activity. Three of the four inhibitors also reduced starch degradation and seedling growth, but the fourth did not affect these parameters. Inhibition of starch degradation was apparently not due to inhibition of amylases. Inhibition......The importance of alpha-glucosidase in the endosperm starch metabolism of barley (Hordeum vulgare) seedlings is poorly understood. The enzyme converts maltose to glucose (Glc), but in vitro studies indicate that it can also attack starch granules. To discover its role in vivo, we took complementary...... silencing cassette for HvAgl97, alpha-glucosidase activity was reduced by up to 50%. There was a large decrease in the Glc-to-maltose ratio in these lines but no effect on starch degradation or seedling growth. Our results suggest that the alpha-glucosidase HvAGL97 is the major endosperm enzyme catalyzing...

  5. The embryo and the endosperm contribute equally to argan seed oil yield but confer distinct lipid features to argan oil.

    Science.gov (United States)

    Errouane, Kheira; Doulbeau, Sylvie; Vaissayre, Virginie; Leblanc, Olivier; Collin, Myriam; Kaid-Harche, Meriem; Dussert, Stéphane

    2015-08-15

    In the perspective of studying lipid biosynthesis in the argan seed, the anatomy, ploidy level and lipid composition of mature seed tissues were investigated using an experimental design including two locations in Algeria and four years of study. Using flow cytometry, we determined that mature argan seeds consist of two well-developed tissues, the embryo and the endosperm. The lipid content of the embryo was higher than that of the endosperm, but the dry weight of the endosperm was higher. Consequently, both tissues contribute equally to seed oil yield. Considerable differences in fatty acid composition were observed between the two tissues. In particular, the endosperm 18:2 percentage was twofold higher than that of the embryo. The tocopherol content of the endosperm was also markedly higher than that of the embryo. In contrast, the endosperm and the embryo had similar sterol and triterpene alcohol contents and compositions.

  6. Genotyping of endosperms to determine seed dormancy genes regulating germination through embryonic, endospermic, or maternal tissues in rice.

    Science.gov (United States)

    Gu, Xing-You; Zhang, Jinfeng; Ye, Heng; Zhang, Lihua; Feng, Jiuhuan

    2015-02-01

    Seed dormancy is imposed by one or more of the embryo, endosperm, and maternal tissues that belong to two generations and represent two ploidy levels. Many quantitative trait loci (QTL) have been identified for seed dormancy as measured by gross effects on reduced germination rate or delayed germination in crop or model plants. This research developed an endosperm genotype-based genetic approach to determine specific tissues through which a mapped QTL regulates germination using rice as a model. This approach involves testing germination velocity for partially after-ripened seeds harvested from single plants heterozygous for a tested QTL and genotyping endosperms from individual germinated and nongerminated seeds with a codominant DNA marker located on the QTL peak region. Information collected about the QTL includes genotypic frequencies in germinated and/or nongerminated subpopulations; allelic frequency distributions during a germination period; endosperm or embryo genotypic differences in germination velocity; and genotypic frequencies for gametes involved in the double fertilization to form the sampled seeds. Using this approach, the seed dormancy loci SD12, SD1-2, and SD7-1 were determined to regulate germination through the embryo, endosperm, and maternal tissues, respectively; SD12 and SD1-2 acted additively on germination velocity in the offspring tissues; and SD12 also was associated with the preferential fertilization of male gametes in rice. This new genetic approach can be used to characterize mapped genes/QTL for tissue-specific functions in endospermic seeds and for marker-assisted selection of QTL alleles before or immediately after germination in crop breeding.

  7. 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...

  8. The role of alpha-glucosidase in germinating barley grains.

    Science.gov (United States)

    Stanley, Duncan; Rejzek, Martin; Naested, Henrik; Smedley, Mark; Otero, Sofía; Fahy, Brendan; Thorpe, Frazer; Nash, Robert J; Harwood, Wendy; Svensson, Birte; Denyer, Kay; Field, Robert A; Smith, Alison M

    2011-02-01

    The importance of α-glucosidase in the endosperm starch metabolism of barley (Hordeum vulgare) seedlings is poorly understood. The enzyme converts maltose to glucose (Glc), but in vitro studies indicate that it can also attack starch granules. To discover its role in vivo, we took complementary chemical-genetic and reverse-genetic approaches. We identified iminosugar inhibitors of a recombinant form of an α-glucosidase previously discovered in barley endosperm (ALPHA-GLUCOSIDASE97 [HvAGL97]), and applied four of them to germinating grains. All four decreased the Glc-to-maltose ratio in the endosperm 10 d after imbibition, implying inhibition of maltase activity. Three of the four inhibitors also reduced starch degradation and seedling growth, but the fourth did not affect these parameters. Inhibition of starch degradation was apparently not due to inhibition of amylases. Inhibition of seedling growth was primarily a direct effect of the inhibitors on roots and coleoptiles rather than an indirect effect of the inhibition of endosperm metabolism. It may reflect inhibition of glycoprotein-processing glucosidases in these organs. In transgenic seedlings carrying an RNA interference silencing cassette for HvAgl97, α-glucosidase activity was reduced by up to 50%. There was a large decrease in the Glc-to-maltose ratio in these lines but no effect on starch degradation or seedling growth. Our results suggest that the α-glucosidase HvAGL97 is the major endosperm enzyme catalyzing the conversion of maltose to Glc but is not required for starch degradation. However, the effects of three glucosidase inhibitors on starch degradation in the endosperm indicate the existence of unidentified glucosidase(s) required for this process.

  9. 21 CFR 73.315 - Corn endosperm oil.

    Science.gov (United States)

    2010-04-01

    ... corn endosperm oil is a reddish-brown liquid composed chiefly of glycerides, fatty acids, sitosterols.... (b) Specifications. Corn endosperm oil conforms to the following specifications: Total fatty...

  10. Impaired auxin biosynthesis in the defective endosperm18 mutant is due to mutational loss of expression in the ZmYuc1 gene encoding endosperm-specific YUCCA1 protein in maize.

    Science.gov (United States)

    A seed-specific maize mutant, defective endosperm18 (de18), accumulates approximately 40% less dry mass and 10- to 15- fold less auxin (IAA) as compared to the De18; however, a causal basis of these changes is not known. Cellular analyses here showed that the de18 developing endosperm had lower tota...

  11. 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.

  12. Development and characterization of a new barley yellow dwarf virus (BYDV)-resistant wheat germplasm

    Institute of Scientific and Technical Information of China (English)

    聂道泰; 贾旭; 胡适全; 朱立煌; 胡含; 胡含; 周广和; 钱幼亭; 庄家骏

    1996-01-01

    A barley yellow dwarf virus (BYDV)-resistant line HG295 was selected from a cross between cv. 77-5433 and Zhong 5 after extensive investigation in field, greenhouse and ELISA. Cytological analysis revealed that it was an euploid line and genetically stable. The existence of alien DNA in HG295 was identified by RAPD and Southern hybridization analyses showed that the alien DNAs came from Zhong 5 or Th. intermedium. The differences of BYDV resistance between L1 and HG295 are discussed.

  13. 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.

  14. Distribution of carotenoids in endosperm, germ, and aleurone fractions of cereal grain kernels.

    Science.gov (United States)

    Ndolo, Victoria U; Beta, Trust

    2013-08-15

    To compare the distribution of carotenoids across the grain, non-corn and corn cereals were hand dissected into endosperm, germ and aleurone fractions. Total carotenoid content (TCC) and carotenoid composition were analysed using spectrophotometry and HPLC. Cereal carotenoid composition was similar; however, concentrations varied significantly (paleurone layer had zeaxanthin levels 2- to 5-fold higher than lutein among the cereals. Positive significant correlations (paleurone layer. Our findings suggest that the aleurone of wheat, oat, corn and germ of barley have significantly enhanced carotenoid levels.

  15. 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...

  16. 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...

  17. Shoot apex development date of anthesis and grain yield of autumn-sown spring and winter barley hordeum-vulgare after different sowing times

    DEFF Research Database (Denmark)

    Knopp, E.

    1985-01-01

    One spring and eight winter barley cultivars were sown at intervals from the middle of September (normal) until the end of October (late). The apical meristem of the spring cultivar had developed furthest until the onset of winter. It had started ear differentiation after normal sowing. Among the...

  18. Possible Roles for Polycomb Repressive Complex 2 in Cereal Endosperm

    Directory of Open Access Journals (Sweden)

    Kaoru eTonosaki

    2015-03-01

    Full Text Available The Polycomb Repressive Complex 2 (PRC2 is an evolutionarily conserved multimeric protein complex in both plants and animals. In contrast to animals, plants have evolved a range of different components of PRC2 and form diverse complexes that act in the control of key regulatory genes at many stages of development during the life cycle. A number of studies, particularly in the model species Arabidopsis thaliana, have highlighted the role of PRC2 and of epigenetic controls via parent-of-origin specific gene expression for endosperm development. However, recent research in cereal plants has revealed that although some components of PRC2 show evolutionary conservation with respect to parent-of-origin specific gene expression patterns, the identity of the imprinted genes encoding PRC2 components is not conserved. This disparity may reflect the facts that cereal plant genomes have undergone different patterns of duplication during evolution compared to Arabidopsis thaliana and that the endosperm development program is not identical in monocots and eudicots. In this context, we focus this review on the expression of imprinted PRC2 genes and their roles in endosperm development in cereals.

  19. 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...

  20. Low-temperature acclimation of barley cultivars used as parents in mapping populations: response to photoperiod, vernalization and phenological development.

    Science.gov (United States)

    Limin, Allen; Corey, Ann; Hayes, Patrick; Fowler, D Brian

    2007-06-01

    Six barley (Hordeum vulgare L.) accessions, previously used as parents of mapping populations, were evaluated for characters potentially affecting the location of low-temperature (LT) tolerance QTLs. Three were of winter growth habit (Kompolti Korai, Nure, and Strider), one was facultative (Dicktoo) and two were spring (Morex and Tremois). Final leaf number (FLN) and LT(50 )were determined at weekly intervals from 0 to 98 days of LT acclimation/vernalization under both long day (LD) and short day (SD) photoperiods. The point of vegetative/reproductive transition was determined from measurements of double ridge (DR) formation and FLN. With the exception of Nure, SD delayed development by increasing leaf production. Dicktoo was extremely SD sensitive lengthening its vegetative phase by more than 63 days relative to the LD photoperiod. SD had the opposite effect on Nure, causing an accelerating of flowering exhibiting the characteristic of 'short day vernalization'. All accessions except Dicktoo and Kompolti Korai acclimated rapidly in the first 7 days of LT exposure, approaching their maximum LT tolerance in 14-21 days. Dicktoo and Kompolti Korai continued to slowly acclimate until reproductive transition. The results emphasize two important points: (1) the location of QTLs for LT tolerance, and as a consequence the identification of putative candidate genes, will be a function of the genotypes sampled, the experimental conditions used, and the quality of the phenotypic data and (2) the barley LT tolerance pathway reaches an early impediment relative to closely related more hardy members of the Triticeae such as wheat and rye.

  1. Cloning of a 14-3-3 Gene from Developing Wheat Endosperm and Expression of its Recombinant Protein in Escherichia coli%小麦胚乳14-3-3基因的克隆及其重组蛋白的原核表达

    Institute of Scientific and Technical Information of China (English)

    戴双; 李豪圣; 程敦公; 刘爱峰; 曹新有; 刘建军; 宋健民

    2012-01-01

    [Objective] This research was conducted to clone 14-3-3 genes from developing wheat endosperm and express their recombinant proteins in Escherichia coli aiming to investigate their functions in wheat development [ Method 1 Specific primers with restriction enzymes cut site were designed according to conserved sequence of homologous genes registered in NCBI. The target genes were cloned by RT-PCR from filling wheat grain. Then the cloned genes were inserted into expressing vectors after confirmation by sequencing and multiple alignments. The recombinant protein was expressed in E. coli and purified for further research. [Result] A 14-3-3 gene was amplified from developing endosperm of 13-15 d after anthesis of bread wheat cultivar Jimai22 and inserted into Top plasmid vector then transformed into E. coli strain DH5α by heat shock. The cloned gene was sequenced after the recombinant plasmid was extracted. The results of sequencing analysis showed that the gene belongs to nonε group and contained an open reading frame of 777 bp in length encoding a protein with 259 aa with predicted molecular weight about 29 kD. According to multiple alignments using DNAMAN program, the gene was highly homologous to other 14-3-3 genes from main crops such as wheat rice, maize, barley, soybean and model plant Arabidopsis with the maximum homology of 98%, as well as their encoding proteins. Furthermore, the heterologous protein with molecular weight about 30 kD expressed in E. coli was coincident with predicted size based on deduced amino acid sequence. All results suggested that the cloned gene is a 14-3-3 gene and it was correctly inserted into the vector and expressed heterologously. The cloned gene was inserted into expressing vector pET29c possessing a S-tag specific bounding to S-protein agarose. The recombinant vector was transformed into E. coli strain BL21-CodonPlus (DE3)-RP supplied with additional rare codes to improve heterologous expression. The recombinant protein was

  2. Proteomic Comparison of Basal Endosperm in Maize miniature1 Mutant and its Wild-type Mn1

    Directory of Open Access Journals (Sweden)

    Cecilia eSilva-Sanchez

    2013-06-01

    Full Text Available Developing endosperm in maize seed is a major site for biosynthesis and storage of starch and proteins, and of immense economic importance for its role in food, feed and biofuel production. The basal part of endosperm performs a major role in solute, water and nutrition acquisition from mother plant to sustain these functions. The miniature1 (mn1 mutation is a loss-of-function mutation of the Mn1-encoded cell wall invertase that is entirely expressed in the basal endosperm and is essential for many of the metabolic and signaling functions associated with metabolically released hexose sugars in developing endosperm. Here we report a comparative proteomic study between Mn1 and mn1 basal endosperm to better understand basis of pleiotropic effects on many diverse traits in the mutant. Specifically, we used iTRAQ based quantitative proteomics combined with Gene Ontology and bioinformatics to understand functional basis of the proteomic information. A total of 2518 proteins were identified from soluble and cell wall associated protein fractions; of these 131 proteins were observed to be differentially expressed in the two genotypes. The main functional groups of proteins that were significantly different were those involved in the carbohydrate metabolic and catabolic process, and cell homeostasis. The study constitutes the first proteomic analysis of basal endosperm cell layers in relation to endosperm growth and development in maize.

  3. 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.

  4. 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.

  5. 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

  6. The trafficking pathway of a wheat storage protein in transgenic rice endosperm

    Science.gov (United States)

    Oszvald, Maria; Tamas, Laszlo; Shewry, Peter R.; Tosi, Paola

    2014-01-01

    Background and Aims The trafficking of proteins in the endoplasmic reticulum (ER) of plant cells is a topic of considerable interest since this organelle serves as an entry point for proteins destined for other organelles, as well as for the ER itself. In the current work, transgenic rice was used to study the pattern and pathway of deposition of the wheat high molecular weight (HMW) glutenin sub-unit (GS) 1Dx5 within the rice endosperm using specific antibodies to determine whether it is deposited in the same or different protein bodies from the rice storage proteins, and whether it is located in the same or separate phases within these. Methods The protein distribution and the expression pattern of HMW sub-unit 1Dx5 in transgenic rice endosperm at different stages of development were determined using light and electron microscopy after labelling with antibodies. Key results The use of HMW-GS-specific antibodies showed that sub-unit 1Dx5 was expressed mainly in the sub-aleurone cells of the endosperm and that it was deposited in both types of protein body present in the rice endosperm: derived from the ER and containing prolamins, and derived from the vacuole and containing glutelins. In addition, new types of protein bodies were also formed within the endosperm cells. Conclusions The results suggest that the HMW 1Dx5 protein could be trafficked by either the ER or vacuolar pathway, possibly depending on the stage of development, and that its accumulation in the rice endosperm could compromise the structural integrity of protein bodies and their segregation into two distinct populations in the mature endosperm. PMID:24603605

  7. Effects of sulphur dioxide on the development of fungal diseases of winter barley in an open-air fumigation system

    Energy Technology Data Exchange (ETDEWEB)

    Mansfield, P.J.; Bell, J.N.B.; Wheeler, B.E.J. (Imperial Coll., Ascot (UK)); McLeod, A.R. (National Power Technology and Environmental Centre, Leatherhead (UK))

    1991-01-01

    The development of fungal diseases was monitored through three consecutive growing seasons (1984-1985, 1985-1986 and 1986-1987) on field plots of winter barley (Hordeum vulgare L.) cv. 'Igri' exposed to sulfur dioxide (SO{sub 2}) concentrations in the range 0.005-0.048 microliter l{sup {minus}1} using an open-air fumigation system. In each season the incidence and severity of powdery mildew (Erysiphe graminis D.C. ex Merat f.sp. hordei Em. Marchal) increased and leaf blotch (Rhynchosporium secalis Oudem J.J. Davis) decreased in plots exposed to elevated levels of SO{sub 2} (0.014-0.048 microliter l{sup {minus}1} SO{sub 2}) compared with control plots exposed to background levels of 0.005-0.009 microliter l{sup {minus}1} SO{sub 2}. The apparent infection rate of powdery mildew was also greater in SO{sub 2}-treated plots. The effects of SO{sub 2} exposure on eyespot (Pseudocercosporella herpotrichoides (Fron) Deighton) and black ear moulds (Cladosporium and Alternaria spp.) were variable. Sharp eyespot (Rhizoctonia cerealis van der Hoeven), Fusarium foot rot, brown rust (Puccinia hordei Otth.), Septoria nodorum Berk., net blotch (Pyrenophora teres Sacc.) and Botrytis spp. infection on the ear were all apparently unaffected by SO{sub 2}. 3 figs., 36 refs., 4 tabs.

  8. Variability of barley aleurone layer induced by X-rays

    Directory of Open Access Journals (Sweden)

    Romuald Kosina

    2015-05-01

    Full Text Available A series of Hordeum vulgare cultivars was irradiated by X-rays to induce mutations in endosperm. Many structural defects of endosperm were revealed in plants irradiated 8 DAF. Change of a cell cycle was especially frequent and this was visible in the form of clones of small or large cells in the aleurone layer. X-irradiation appeared as a successful tool in the study of development.

  9. 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.

  10. Development of a monoclonal antibody-based enzyme-linked immunosorbent assay to quantify soluble beta-glucans in oats and barley.

    Science.gov (United States)

    Rampitsch, Christof; Ames, Nancy; Storsley, Joanne; Marien, Lindsay

    2003-09-24

    A set of 31 murine monoclonal antibodies was produced against (1-->3,1-->4)beta-d-glucan from oats (Avena sativa L.) chemically cross-linked to keyhole limpet hemocyanin. Monoclonal antibodies were tested for their cross-reactivity to related and unrelated polysaccharides. The antibodies reacted strongly to unmodified beta-glucan from oats and barley (Hordeum vulgare L.) and to lichenan from Icelandic moss, a polysaccharide with a structure similar to that of beta-glucan but which is not encountered in cereals. Cross-reaction to other polysaccharides tested was minimal at physiological levels. An enzyme-linked immunosorbent assay (ELISA) that could routinely detect and quantify nanogram levels of soluble beta-glucan extracted from the flour of oats or barley was designed with one of these monoclonal antibodies. The beta-glucan extraction procedure from ground oat and barley samples and the ELISA were both optimized for reproducibility, accuracy, and throughput, and results were compared to values obtained from an established, commercially available enzyme-based assay. Correlations between the two assays were consistently high (r (2) > 0.9), indicating that the ELISA presented in this paper is a valuable alternative for assaying beta-glucan levels in cereals and cereal products, both routinely and in preparations in which beta-glucans are present in nanogram amounts. Development of the extraction procedure for ELISA is discussed.

  11. Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

    Institute of Scientific and Technical Information of China (English)

    CHEN Jun-ying; YUE Run-qing; XU Hai-xian; CHEN Xin-jian

    2006-01-01

    To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars (Triticum aestivum L. cv.) were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm (endosperm-supported culture, ES), the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm (non-endosperm-supported culture, NES). This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.

  12. Contrast observation and investigation of wheat endosperm transfer cells and nucellar projection transfer cells.

    Science.gov (United States)

    Zheng, Yankun; Wang, Zhong

    2011-07-01

    In cereal seed, there are no symplastic connections between the maternal tissues and the endosperm. In order to facilitate solute transport, both the nucellar projection and its opposite endosperm epithelial cells in wheat caryopsis differentiate into transfer cells. In this paper, we did contrast observation and investigation of wheat endosperm transfer cells (ETC) and nucellar projection transfer cells (NPTC). The experimental results showed that there were some similarities and differences between ETC and NPTC. ETC and NPTC almost developed synchronously. Wall ingrowths of ETC and NPTC formed firstly in the first layer nearest to the endosperm cavity, and formed later in the inner layer further from the endosperm cavity. The mature ETC were mainly three layers and the mature NPTC were mainly four layers. Wall ingrowths of ETC were flange type and wall ingrowths of NPTC were reticulate type. NPTC were not nutrient-storing cells, but the first layer of ETC had aleurone cell features, and the second layer and third layer of ETC accumulated starch granules and protein bodies.

  13. Spatio-Temporal Metabolite Profiling of the Barley Germination Process by MALDI MS Imaging.

    Directory of Open Access Journals (Sweden)

    Karin Gorzolka

    Full Text Available MALDI mass spectrometry imaging was performed to localize metabolites during the first seven days of the barley germination. Up to 100 mass signals were detected of which 85 signals were identified as 48 different metabolites with highly tissue-specific localizations. Oligosaccharides were observed in the endosperm and in parts of the developed embryo. Lipids in the endosperm co-localized in dependency on their fatty acid compositions with changes in the distributions of diacyl phosphatidylcholines during germination. 26 potentially antifungal hordatines were detected in the embryo with tissue-specific localizations of their glycosylated, hydroxylated, and O-methylated derivates. In order to reveal spatio-temporal patterns in local metabolite compositions, multiple MSI data sets from a time series were analyzed in one batch. This requires a new preprocessing strategy to achieve comparability between data sets as well as a new strategy for unsupervised clustering. The resulting spatial segmentation for each time point sample is visualized in an interactive cluster map and enables simultaneous interactive exploration of all time points. Using this new analysis approach and visualization tool germination-dependent developments of metabolite patterns with single MS position accuracy were discovered. This is the first study that presents metabolite profiling of a cereals' germination process over time by MALDI MSI with the identification of a large number of peaks of agronomically and industrially important compounds such as oligosaccharides, lipids and antifungal agents. Their detailed localization as well as the MS cluster analyses for on-tissue metabolite profile mapping revealed important information for the understanding of the germination process, which is of high scientific interest.

  14. 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...

  15. Starch and Free Sugars during Kernel Development of Bomi Barley and its High-Lysine Mutant 1508

    DEFF Research Database (Denmark)

    Kreis, Michael

    1978-01-01

    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...

  16. Characterization of the imprinting and expression patterns of ZAG2 in maize endosperm and embryo

    Directory of Open Access Journals (Sweden)

    Chaoxian Liu

    2015-02-01

    Full Text Available ZAG2 has been identified as a maternally expressed imprinted gene in maize endosperm. Our study revealed that paternally inherited ZAG2 alleles were imprinted in maize endosperm and embryo at 14 days after pollination (DAP, and consistently imprinted in endosperm at 10, 12, 16, 18, 20, 22, 24, 26, and 28 DAP in reciprocal crosses between B73 and Mo17. ZAG2 alleles were also imprinted in reciprocal crosses between Zheng 58 and Chang 7-2 and between Huang C and 178. ZAG2 alleles exhibited differential imprinting in hybrids of 178 × Huang C and B73 × Mo17, while in other hybrids ZAG2 alleles exhibited binary imprinting. The tissue-specific expression pattern of ZAG2 showed that ZAG2 was expressed at a high level in immature ears, suggesting that ZAG2 plays important roles in not only kernel but ear development.

  17. Development of high-density genetic maps for barley and wheat using a novel two-enzyme genotyping-by-sequencing approach.

    Directory of Open Access Journals (Sweden)

    Jesse A Poland

    Full Text Available Advancements in next-generation sequencing technology have enabled whole genome re-sequencing in many species providing unprecedented discovery and characterization of molecular polymorphisms. There are limitations, however, to next-generation sequencing approaches for species with large complex genomes such as barley and wheat. Genotyping-by-sequencing (GBS has been developed as a tool for association studies and genomics-assisted breeding in a range of species including those with complex genomes. GBS uses restriction enzymes for targeted complexity reduction followed by multiplex sequencing to produce high-quality polymorphism data at a relatively low per sample cost. Here we present a GBS approach for species that currently lack a reference genome sequence. We developed a novel two-enzyme GBS protocol and genotyped bi-parental barley and wheat populations to develop a genetically anchored reference map of identified SNPs and tags. We were able to map over 34,000 SNPs and 240,000 tags onto the Oregon Wolfe Barley reference map, and 20,000 SNPs and 367,000 tags on the Synthetic W9784 × Opata85 (SynOpDH wheat reference map. To further evaluate GBS in wheat, we also constructed a de novo genetic map using only SNP markers from the GBS data. The GBS approach presented here provides a powerful method of developing high-density markers in species without a sequenced genome while providing valuable tools for anchoring and ordering physical maps and whole-genome shotgun sequence. Development of the sequenced reference genome(s will in turn increase the utility of GBS data enabling physical mapping of genes and haplotype imputation of missing data. Finally, as a result of low per-sample costs, GBS will have broad application in genomics-assisted plant breeding programs.

  18. Immature Seed Endosperm and Embryo Proteomics of the Lotus (Nelumbo Nucifera Gaertn. by One-Dimensional Gel-Based Tandem Mass Spectrometry and a Comparison with the Mature Endosperm Proteome

    Directory of Open Access Journals (Sweden)

    Carlo F. Moro

    2015-08-01

    Full Text Available Lotus (Nelumbo nucifera Gaertn. seed proteome has been the focus of our studies, and we have recently established the first proteome dataset for its mature seed endosperm. The current study unravels the immature endosperm, as well as the embryo proteome, to provide a comprehensive dataset of the lotus seed proteins and a comparison between the mature and immature endosperm tissues across the seed’s development. One-dimensional gel electrophoresis (SDS-PAGE linked with tandem mass spectrometry provided a protein inventory of the immature endosperm (122 non-redundant proteins and embryo (141 non-redundant proteins tissues. Comparing with the previous mature endosperm dataset (66 non-redundant proteins, a total of 206 non-redundant proteins were identified across all three tissues of the lotus seed. Results revealed some significant differences in proteome composition between the three lotus seed tissues, most notably between the mature endosperm and its immature developmental stage shifting the proteins from nutrient production to nutrient storage.

  19. 高粱胚乳细胞与母体组织发育关系的研究%Investigation of Sorghum Endosperm Cell Development and the Relationship with Its Maternal Tissue

    Institute of Scientific and Technical Information of China (English)

    李栋梁; 荆彦平; 李小刚; 顾蕴洁; 王忠

    2014-01-01

    势,中果皮发育后期细胞内观察到淀粉体存在特殊的二次增长,构成从前期复粒淀粉体为主转为单粒;“种皮”来源于珠心、珠被细胞依次降解后的胞壁残留堆叠而成,中间含有透明角质层。【结论】白高粱KS-304胚乳发育与玉米颖果胚乳类似,最终形成角质与粉质胚乳;颖果胚乳淀粉体发育存在独特的“发生中心”;中果皮发育后期其细胞可能行使积累同化产物“库”的作用。%Objective]The objective of this research was to clarify the relationship between endosperm and its maternal tissue during development process of the sorghum caryopsis.[Method]Sorghum KS-304 was used as the experimental material with its caryopsis development days precisely recorded and its caryopsis development closely observed. Structural changes of endosperm cells and relation with their maternal tissue were observed through semi-thin sections under light microscopy by applying Spurr resin tissue embedding. The ultrastructure of cells from different tissues and the starch granules wherein in developing and full ripe caryopsis were observed under SEM. The relationships of testa and aleurone layer of full ripe caryopsis were studied through fluorescent microscopy using cryosectioning.[Result]Four stages could be found in caryopsis development, they were formation stage, milky stage, dough stage, and full maturity stage. Accordingly, endosperm development was divided into five stages, i.e. coenocyte stage, cellurization stage, differentiation stage, developmental stage and the final maturation stage, and the first three of which are equal to that of the formation stage of caryopsis; while the last maturation stage corresponds to the last two stages of caryopsis development. It took as long as 15 days for the nucellar epidermis to be fully degraded. The outer peripheral endosperm cells started to accumulate lipid bodies as early as 7 DPA (days after pollination) and, turned

  20. 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.

  1. Biochemical and Molecular Characterization of a Barley Seed ß-Glucosidase

    DEFF Research Database (Denmark)

    Leah, R.; Kigel, J.; Svendsen, I.

    1995-01-01

    A 60-kDa ß-glucosidase (BGQ60) was purified and characterized from seeds of barley (Hordeum vulgare L.). BGQ60 catalytic activity was restricted to the cleavage of short-chain oligosaccharides composed of(1, 2) -,(1, 2, 3) -, and/or(1, 2, 3, 4) -ß-linked glucose or mannose units. These oligosacch......A 60-kDa ß-glucosidase (BGQ60) was purified and characterized from seeds of barley (Hordeum vulgare L.). BGQ60 catalytic activity was restricted to the cleavage of short-chain oligosaccharides composed of(1, 2) -,(1, 2, 3) -, and/or(1, 2, 3, 4) -ß-linked glucose or mannose units....... These oligosaccharides are the primary products of endosperm cell wall polysaccharide hydrolysis by other enzymes. In keeping with this, complete hydrolysis of the major polysaccharide of barley starchy endosperm cell wall, (1-3,1-4)-ß-glucan, to free glucose was shown to require the concerted action of endo-(1-3,1-4)-ß...... interrupted by 9 introns. The complete nucleotide sequence of the bgq60 structural gene represents the first characterized plant gene encoding a ß-glucosidase. The barley BGQ60 is a novel plant ß-glucosidase with a hitherto undescribed specific enzymatic activity. The possible biological functions of BGQ60...

  2. Barley plants over-expressing the NAC transcription factor gene HvNAC005 show stunting and delay in development combined with early senescence

    DEFF Research Database (Denmark)

    Christiansen, Michael W.; Matthewman, Colette; Podzimska-Sroka, Dagmara Agata;

    2016-01-01

    -expressing plants showed up-regulation of genes involved with secondary metabolism, hormone metabolism, stress, signalling, development, and transport. Up-regulation of senescence markers and hormone metabolism and signalling genes supports a role of HvNAC005 in the cross field of different hormone and signalling......The plant-specific NAC transcription factors have attracted particular attention because of their involvement in stress responses, senescence, and nutrient remobilization. The HvNAC005 gene of barley encodes a protein belonging to subgroup NAC-a6 of the NAC family. This study shows that HvNAC005...

  3. Cereal bioengineering: Amylopectin-free and hyper-phosphorylated barley starch

    DEFF Research Database (Denmark)

    Carciofi, Massimiliano; Shaik, Shahnoor Sultana; Jensen, Susanne Langgård;

    2011-01-01

    Barley lines producing grains with either amylopectin-free or hyper-phosphorylated starches were made by transgenic methods. Cereals producing these kind of starches have not been reported before. Amylopectin-free barley was generated by simultaneously silencing the three genes encoding the starch...... branching enzymes SBEIIa, SBEIIb and SBEI by a chimeric hairpin. The construct was inherited as a single locus with a distinct 1:3 segregation in consecutive generations. The transgenic grains were shrunken and the yield was around 80% of that found in wildtype. The starch granules were irregularly......, elongated and globose shaped. Transgenic grains also had a higher beta-glucan content. In order to increase barley starch phosphorylation, endosperm specific overexpression of glucan water dikinase from potato (StGWD) was conducted. The content of phosphate esters in starch from consecutive generations (T0...

  4. Programmed cell death or desiccation tolerance : two possible routes for wheat endosperm cells

    NARCIS (Netherlands)

    Golovina, E.A.; Hoekstra, F.A.; Aelst, van A.C.

    2000-01-01

    The fate of cells in the endosperm of developing wheat kernels was investigated under normal conditions and upon premature slow drying on the cut ear. To follow the changes in membrane integrity and cellular ultrastructure, an electron spin resonance (ESR) spin probe technique and low temperature sc

  5. 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....

  6. Cyclin-Dependent Kinase Inhibitors in Maize Endosperm and Their Potential Role in Endoreduplication1

    Science.gov (United States)

    Coelho, Cintia M.; Dante, Ricardo A.; Sabelli, Paolo A.; Sun, Yuejin; Dilkes, Brian P.; Gordon-Kamm, William J.; Larkins, Brian A.

    2005-01-01

    Two maize (Zea mays) cyclin-dependent kinase (CDK) inhibitors, Zeama;KRP;1 and Zeama;KRP;2, were characterized and shown to be expressed in developing endosperm. Similar to the CDK inhibitors in Arabidopsis (Arabidopsis thaliana) and tobacco (Nicotiana tabacum), the maize proteins contain a carboxy-terminal region related to the inhibitory domain of the mammalian Cip/Kip inhibitors. Zeama;KRP;1 is present in the endosperm between 7 and 21 d after pollination, a period that encompasses the onset of endoreduplication, while the Zeama;KRP;2 protein declines during this time. Nevertheless, Zeama;KRP;1 accounts for only part of the CDK inhibitory activity that peaks coincident with the endoreduplication phase of endosperm development. In vitro assays showed that Zeama;KRP;1 and Zeama;KRP;2 are able to inhibit endosperm Cdc2-related CKD activity that associates with p13Suc1. They were also shown to specifically inhibit cyclin A1;3- and cyclin D5;1-associated CDK activities, but not cyclin B1;3/CDK. Overexpression of Zeama;KRP;1 in maize embryonic calli that ectopically expressed the wheat dwarf virus RepA protein, which counteracts retinoblastoma-related protein function, led to an additional round of DNA replication without nuclear division. PMID:16055680

  7. Evidence for osmotic regulation of hydrolytic enzyme production in germinating barley seeds.

    Science.gov (United States)

    Jones, R L; Armstrong, J E

    1971-08-01

    alpha-Amylase levels in intact seeds of barley (Hordeum vulgare L. cv. Himalaya) reach a maximum at 3 to 4 days of germination while gibberellin levels continue to increase beyond 6 days of germination. In contrast to its effect on half seeds, gibberellic acid does not increase the total amount of alpha-amylase produced in germinating seeds. The inability of gibberellic acid to stimulate alpha-amylase production is not related to its availability; rather, evidence suggests that a factor(s) in whole seeds prevents further enhancement of alpha-amylase formation and accumulation. Hydrolysis products accumulate in the subaleurone space of the endosperm of germinating seeds up to concentrations of 570 milliosmolar. Chromatography of these hydrolysis products indicate the presence of maltose and glucose. Calculations based on reducing sugar determinations show that glucose accounts for as much as 57% of the solutes present in the endosperm fluid. Both maltose and glucose in the range of 0.2 to 0.4 M effectively inhibit the production of alpha-amylase by isolated barley aleurone layers. This inhibition is quantitatively similar to that brought about by solutions of polyethylene glycol and mannitol. On the basis of these data we propose that hydrolysis products which accumulate in the starchy endosperm of germinating seeds function to regulate the production of hydrolytic enzymes by the aleurone layer.

  8. Protein disulfide isomerase-like protein 1-1 controls endosperm development through regulation of the amount and composition of seed proteins in rice.

    Directory of Open Access Journals (Sweden)

    Yeon Jeong Kim

    Full Text Available Protein disulfide isomerase (PDI is a chaperone protein involved in oxidative protein folding by acting as a catalyst and assisting folding in the endoplasmic reticulum (ER. A genome database search showed that rice contains 19 PDI-like genes. However, their functions are not clearly identified. This paper shows possible functions of rice PDI-like protein 1-1 (PDIL1-1 during seed development. Seeds of the T-DNA insertion PDIL1-1 mutant, PDIL1-1Δ, identified by genomic DNA PCR and western blot analysis, display a chalky phenotype and a thick aleurone layer. Protein content per seed was significantly lower and free sugar content higher in PDIL1-1Δ mutant seeds than in the wild type. Proteomic analysis of PDIL1-1Δ mutant seeds showed that PDIL1-1 is post-translationally regulated, and its loss causes accumulation of many types of seed proteins including glucose/starch metabolism- and ROS (reactive oxygen species scavenging-related proteins. In addition, PDIL1-1 strongly interacts with the cysteine protease OsCP1. Our data indicate that the opaque phenotype of PDIL1-1Δ mutant seeds results from production of irregular starch granules and protein body through loss of regulatory activity for various proteins involved in the synthesis of seed components.

  9. A new statistical method for mapping QTLs underlying endosperm traits

    Institute of Scientific and Technical Information of China (English)

    HU Zhiqiu; XU Chenwu

    2005-01-01

    Genetic expression for an endosperm trait in seeds of cereal crops may be controlled simultaneously by the triploid endosperm genotypes and the diploid maternal genotypes. However, current statistical methods for mapping quantitative trait loci (QTLs) underlying endosperm traits have not been effective in dealing with the putative maternal genetic effects. Combining the quantitative genetic model for diploid maternal traits with triploid endosperm traits, here we propose a new statistical method for mapping QTLs controlling endosperm traits with maternal genetic effects. This method applies the data set of both DNA molecular marker genotypes of each plant in segregation population and the quantitative observations of single endosperms in each plant to map QTL. The maximum likelihood method implemented via the expectation-maximization algorithm was used to the estimate parameters of a putative QTL. Since this method involves the maternal effect that may contribute to endosperm traits, it might be more congruent with the genetics of endosperm traits and more helpful to increasing the precision of QTL mapping. The simulation results show the proposed method provides accurate estimates of the QTL effects and locations with high statistical power.

  10. Understanding the differences in molecular conformation of carbohydrate and protein in endosperm tissues of grains with different biodegradation kinetics using advanced synchrotron technology

    Science.gov (United States)

    Yu, P.; Block, H. C.; Doiron, K.

    2009-01-01

    Conventional "wet" chemical analyses rely heavily on the use of harsh chemicals and derivatization, thereby altering native seed structures leaving them unable to detect any original inherent structures within an intact tissue sample. A synchrotron is a giant particle accelerator that turns electrons into light (million times brighter than sunlight) which can be used to study the structure of materials at the molecular level. Synchrotron radiation-based Fourier transform IR microspectroscopy (SR-FTIRM) has been developed as a rapid, direct, non-destructive and bioanalytical technique. This technique, taking advantage of the brightness of synchrotron light and a small effective source size, is capable of exploring the molecular chemistry within the microstructures of a biological tissue without the destruction of inherent structures at ultraspatial resolutions within cellular dimensions. This is in contrast to traditional 'wet' chemical methods, which, during processing for analysis, often result in the destruction of the intrinsic structures of feeds. To date there has been very little application of this technique to the study of plant seed tissue in relation to nutrient utilization. The objective of this study was to use novel synchrotron radiation-based technology (SR-FTIRM) to identify the differences in the molecular chemistry and conformation of carbohydrate and protein in various plant seed endosperms within intact tissues at cellular and subcellular level from grains with different biodegradation kinetics. Barley grain (cv. Harrington) with a high rate (31.3%/h) and extent (78%), corn grain (cv. Pioneer) with a low rate (9.6%/h) and extent of (57%), and wheat grain (cv. AC Barrie) with an intermediate rate (23%/h) and extent (72%) of ruminal DM degradation were selected for evaluation. SR-FTIRM evaluations were performed at the National Synchrotron Light Source at the Brookhaven National Laboratory (Brookhaven, NY). The molecular structure spectral analysis

  11. Structural genes of wheat and barley 5-methylcytosine DNA glycosylases and their potential applications for human health.

    Science.gov (United States)

    Wen, Shanshan; Wen, Nuan; Pang, Jinsong; Langen, Gregor; Brew-Appiah, Rhoda A T; Mejias, Jaime H; Osorio, Claudia; Yang, Mingming; Gemini, Richa; Moehs, Charles P; Zemetra, Robert S; Kogel, Karl-Heinz; Liu, Bao; Wang, Xingzhi; von Wettstein, Diter; Rustgi, Sachin

    2012-12-11

    Wheat supplies about 20% of the total food calories consumed worldwide and is a national staple in many countries. Besides being a key source of plant proteins, it is also a major cause of many diet-induced health issues, especially celiac disease. The only effective treatment for this disease is a total gluten-free diet. The present report describes an effort to develop a natural dietary therapy for this disorder by transcriptional suppression of wheat DEMETER (DME) homeologs using RNA interference. DME encodes a 5-methylcytosine DNA glycosylase responsible for transcriptional derepression of gliadins and low-molecular-weight glutenins (LMWgs) by active demethylation of their promoters in the wheat endosperm. Previous research has demonstrated these proteins to be the major source of immunogenic epitopes. In this research, barley and wheat DME genes were cloned and localized on the syntenous chromosomes. Nucleotide diversity among DME homeologs was studied and used for their virtual transcript profiling. Functional conservation of DME enzyme was confirmed by comparing the motif and domain structure within and across the plant kingdom. Presence and absence of CpG islands in prolamin gene sequences was studied as a hallmark of hypo- and hypermethylation, respectively. Finally the epigenetic influence of DME silencing on accumulation of LMWgs and gliadins was studied using 20 transformants expressing hairpin RNA in their endosperm. These transformants showed up to 85.6% suppression in DME transcript abundance and up to 76.4% reduction in the amount of immunogenic prolamins, demonstrating the possibility of developing wheat varieties compatible for the celiac patients.

  12. Gibberellins regulate seed germination in tomato by endosperm weakening: a study with gibberellin-deficient mutants.

    Science.gov (United States)

    Groot, S P; Karssen, C M

    1987-08-01

    The germination of seeds of tomato [Lycopersicon esculentum (L.) Mill.] cv. Moneymaker has been compared with that of seeds of the gibberellin-deficient dwarf-mutant line ga-1, induced in the same genetic background. Germination of tomato seeds was absolutely dependent on the presence of either endogenous or exogenous gibberellins (GAs). Gibberellin A4+7 was 1000-fold more active than commercial gibberellic acid in inducing germination of the ga-1 seeds. Red light, a preincubation at 2°C, and ethylene did not stimulate germination of ga-1 seeds in the absence of GA4+7; however, fusicoccin did stimulate germination independently. Removal of the endosperm and testa layers opposite the radicle tip caused germination of ga-1 seeds in water. The seedlings and plants that develop from the detipped ga-1 seeds exhibited the extreme dwarfy phenotype that is normal to this genotype. Measurements of the mechanical resistance of the surrounding layers showed that the major action of GAs was directed to the weakening of the endosperm cells around the radicle tip. In wild-type seeds this weakening occurred in water before radicle protrusion. In ga-1 seeds a similar event was dependent on GA4+7, while fusicoccin also had some activity. Simultaneous incubation of de-embryonated endosperms and isolated axes showed that wild-type embryos contain and endosperm-weakening factor that is absent in ga-1 axes and is probably a GA. Thus, an endogenous GA facilitates germination in tomato seeds by weakening the mechanical restraint of the endosperm cells to permit radicle protrusion.

  13. Field studies on the regulation of abscisic acid content and germinability during grain development of barley: molecular and chemical analysis of pre-harvest sprouting.

    Science.gov (United States)

    Chono, Makiko; Honda, Ichiro; Shinoda, Shoko; Kushiro, Tetsuo; Kamiya, Yuji; Nambara, Eiji; Kawakami, Naoto; Kaneko, Shigenobu; Watanabe, Yoshiaki

    2006-01-01

    To investigate whether the regulation of abscisic acid (ABA) content was related to germinability during grain development, two cDNAs for 9-cis-epoxycarotenoid dioxygenase (HvNCED1 and HvNCED2) and one cDNA for ABA 8'-hydroxylase (HvCYP707A1), which are enzymes thought to catalyse key regulatory steps in ABA biosynthesis and catabolism, respectively, were cloned from barley (Hordeum vulgare L.). Expression and ABA-quantification analysis in embryo revealed that HvNCED2 is responsible for a significant increase in ABA levels during the early to middle stages of grain development, and HvCYP707A1 is responsible for a rapid decrease in ABA level thereafter. The change in the embryonic ABA content of imbibing grains following dormancy release is likely to reflect changes in the expression patterns of HvNCEDs and HvCYP707A1. A major change between dormant and after-ripened grains occurred in HvCYP707A1; the increased expression of HvCYP707A1 in response to imbibition, followed by a rapid ABA decrease and a high germination percentage, was observed in the after-ripened grains, but not in the dormant grains. Under field conditions, HvNCED2 showed the same expression level and pattern during grain development in 2002, 2003, and 2004, indicating that HvNCED2 expression is regulated in a growth-dependent manner in the grains. By contrast, HvNCED1 and HvCYP707A1 showed a different expression pattern in each year, indicating that the expression of these genes is affected by environmental conditions during grain development. The varied expression levels of these genes during grain development and imbibition, which would have effects on the activity of ABA biosynthesis and catabolism, might be reflected, in part, in the germinability in field-grown barley.

  14. Starch Bioengineering in Barley

    DEFF Research Database (Denmark)

    Shaik, Shahnoor Sultana

    the wild-type cultivar. These exciting results may provide a potential clean technological approach to starch modification by in-planta bioengineering and avoid environmental hazards resulting from post-harvest treatments by chemical modifications. The third study was to investigate the effects...... involved in this process, has enabled the genetic modification f crops in a rational manner to produce novel designer starches with improved functionality. The hypothesis of the present study is that the hyper-phosphorylation of cereal endosperm starch makes it easily accessible and degradable...... by the amylolytic enzymes while the amylose-only endosperm starch exhibits high resistance to degradation and hence less available for degradation. With the aim to investigate the hypothesis, starch molecular structures were modulated with the above mentioned modifications and were studied for the effects...

  15. Potassium channels in barley: cloning, functional characterization and expression analyses in relation to leaf growth and development.

    Science.gov (United States)

    Boscari, Alexandre; Clément, Mathilde; Volkov, Vadim; Golldack, Dortje; Hybiak, Jolanta; Miller, Anthony J; Amtmann, Anna; Fricke, Wieland

    2009-12-01

    It is not known how the uptake and retention of the key osmolyte K(+) in cells are mediated in growing leaf tissue. In the present study on the growing leaf 3 of barley, we have cloned the full-length coding sequence of three genes which encode putative K(+) channels (HvAKT1, HvAKT2, HvKCO1/HvTPK1), and of one gene which encodes a putative K(+) transporter (HvHAK4). The functionality of the gene products of HvAKT1 and HvAKT2 was tested through expression in Xenopus laevis oocytes. Both are inward-rectifying K(+) channels which are inhibited by Cs(+). Function of HvAKT1 in oocytes requires co-expression of a calcineurin-interacting protein kinase (AtCIPK23) and a calcineurin B-like protein (AtCBL9) from Arabidopsis, showing cross-species complementation of function. In planta, HvAKT1 is expressed primarily in roots, but is also expressed in leaf tissue. HvAKT2 is expressed particularly in leaf tissue, and HvHAK4 is expressed particularly in growing leaf tissue. Within leaves, HvAKT1 and HvAKT2 are expressed predominantly in mesophyll. Expression of genes changes little in response to low external K(+) or salinity, despite major changes in K(+) concentrations and osmolality of cells. Possible contributions of HvAKT1, HvAKT2, HvKCO1 and HvHAK4 to regulation of K(+) relations of growing barley leaf cells are discussed.

  16. Control of cell proliferation, endoreduplication, cell size, and cell death by the retinoblastoma-related pathway in maize endosperm

    KAUST Repository

    Sabelli, Paolo A.

    2013-04-22

    The endospermof cereal grains is one of the most valuable products of modern agriculture. Cereal endosperm development comprises different phases characterized by mitotic cell proliferation, endoreduplication, the accumulation of storage compounds, and programmed cell death. Although manipulation of these processes could maximize grain yield, how they are regulated and integrated is poorly understood. We show that the Retinoblastoma-related (RBR) pathway controls key aspects of endosperm development in maize. Down-regulation of RBR1 by RNAi resulted in up-regulation of RBR3-type genes, as well as the MINICHROMOSOME MAINTENANCE 2-7 gene family and PROLIFERATING CELL NUCLEAR ANTIGEN, which encode essential DNA replication factors. Both the mitotic and endoreduplication cell cycles were stimulated. Developing transgenic endosperm contained 42-58% more cells and ~70% more DNA than wild type, whereas there was a reduction in cell and nuclear sizes. In addition, cell death was enhanced. The DNA content of mature endosperm increased 43% upon RBR1 downregulation, whereas storage protein content and kernel weight were essentially not affected. Down-regulation of both RBR1 and CYCLIN DEPENDENT KINASE A (CDKA);1 indicated that CDKA;1 is epistatic to RBR1 and controls endoreduplication through an RBR1- dependent pathway. However, the repressive activity of RBR1 on downstream targets was independent from CDKA;1, suggesting diversification of RBR1 activities. Furthermore, RBR1 negatively regulated CDK activity, suggesting the presence of a feedback loop. These results indicate that the RBR1 pathway plays a major role in regulation of different processes during maize endosperm development and suggest the presence of tissue/organlevel regulation of endosperm/seed homeostasis.

  17. Alterations in barley ribulose-1,5-bisphosphate carboxylase/oxygenase activase gene expression during development and in response to illumination.

    Science.gov (United States)

    Rundle, S J; Zielinski, R E

    1991-08-05

    Two genes encode Rbu-P2-carboxylase activase in barley (RcaA and RcaB): RcaA encodes polypeptides of 46 and 42 kDa, which are generated by the alternatively spliced RcaA1 and RcaA2 mRNAs, respectively; RcaB encodes a 42-kDa polypeptide (Rundle, S. J., and Zielinski, R. E. (1991) J. Biol. Chem. 266, 4677-4685). In the cellular differentiation gradient of the first leaf of barley, the three Rca mRNAs accumulate differentially. RcaA1 and A2 mRNAs accumulate predominantly in the mature, most photosynthetically active regions of the leaf in a pattern that parallels accumulation of total Rbu-P2-carboxylase activase protein. However, the kinetics of accumulation of RcaA1 and RcaA2 mRNA differ slightly, indicating that either changes in RcaA pre-mRNA splicing or mRNA turnover occur during development. RcaB mRNA, in contrast, accumulates in the youngest and oldest cell populations at the base and tip of the leaf, respectively. In the mid-region of the leaf, the difference in accumulation between RcaA and RcaB mRNAs is largely attributable to differences in the rates of transcription of the two Rca genes. In this region of the leaf, the three Rca mRNAs accumulate differentially throughout the course of the diurnal cycle. Steady state levels of the three Rca mRNA species increase in parallel in response to increasing irradiance; these changes were accompanied by increased Rbu-P2-carboxylase activase protein accumulation.

  18. 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.

  19. 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.

  20. Transgenic multivitamin corn through biofortification of endosperm with three vitamins representing three distinct metabolic pathways

    OpenAIRE

    2009-01-01

    Vitamin deficiency affects up to 50% of the world's population, disproportionately impacting on developing countries where populations endure monotonous, cereal-rich diets. Transgenic plants offer an effective way to increase the vitamin content of staple crops, but thus far it has only been possible to enhance individual vitamins. We created elite inbred South African transgenic corn plants in which the levels of 3 vitamins were increased specifically in the endosperm through the simultaneou...

  1. Understanding the Differences in Molecular Conformation of Carbohydrate and Protein in Endosperm Tissues of Grains with Different Biodegradation Kinetics Using Advanced Synchrotron Technology

    Energy Technology Data Exchange (ETDEWEB)

    Yu, P.; Block, H; Doiron, K

    2009-01-01

    Conventional 'wet' chemical analyses rely heavily on the use of harsh chemicals and derivatization, thereby altering native seed structures leaving them unable to detect any original inherent structures within an intact tissue sample. A synchrotron is a giant particle accelerator that turns electrons into light (million times brighter than sunlight) which can be used to study the structure of materials at the molecular level. Synchrotron radiation-based Fourier transform IR microspectroscopy (SR-FTIRM) has been developed as a rapid, direct, non-destructive and bioanalytical technique. This technique, taking advantage of the brightness of synchrotron light and a small effective source size, is capable of exploring the molecular chemistry within the microstructures of a biological tissue without the destruction of inherent structures at ultraspatial resolutions within cellular dimensions. This is in contrast to traditional 'wet' chemical methods, which, during processing for analysis, often result in the destruction of the intrinsic structures of feeds. To date there has been very little application of this technique to the study of plant seed tissue in relation to nutrient utilization. The objective of this study was to use novel synchrotron radiation-based technology (SR-FTIRM) to identify the differences in the molecular chemistry and conformation of carbohydrate and protein in various plant seed endosperms within intact tissues at cellular and subcellular level from grains with different biodegradation kinetics. Barley grain (cv. Harrington) with a high rate (31.3%/h) and extent (78%), corn grain (cv. Pioneer) with a low rate (9.6%/h) and extent of (57%), and wheat grain (cv. AC Barrie) with an intermediate rate (23%/h) and extent (72%) of ruminal DM degradation were selected for evaluation. SR-FTIRM evaluations were performed at the National Synchrotron Light Source at the Brookhaven National Laboratory (Brookhaven, NY). These results suggest

  2. 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.

  3. 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...

  4. 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.

  5. Mass Spectrometric Imaging of Wheat (Triticum spp.) and Barley (Hordeum vulgare L.) Cultivars: Distribution of Major Cell Wall Polysaccharides According to Their Main Structural Features.

    Science.gov (United States)

    Veličković, Dušan; Saulnier, Luc; Lhomme, Margot; Damond, Aurélie; Guillon, Fabienne; Rogniaux, Hélène

    2016-08-17

    Arabinoxylans (AX) and (1→3),(1→4)-β-glucans (BG) are the main components of cereal cell walls and influence many aspects of their end uses. Important variations in the composition and structure of these polysaccharides have been reported among cereals and cultivars of a given species. In this work, the spatial distribution of AX and BG in the endosperm of mature grains was established for nine wheat varieties and eight barley varieties using enzymatically assisted mass spectrometry imaging (MSI). Important structural features of the AX and BG polymers that were previously shown to influence their physicochemical properties were assessed. Differences in the distribution of AX and BG structures were observed, both within the endosperm of a given cultivar and between wheat and barley cultivars. This study provides a unique picture of the structural heterogeneity of AX and BG polysaccharides at the scale of the whole endosperm in a series of wheat and barley cultivars. Thus, it can participate meaningfully in a strategy aiming at understanding the structure-function relationships of these two polymers.

  6. Barley callus: a model system for bioengineering of starch in cereals

    Directory of Open Access Journals (Sweden)

    Carciofi Massimiliano

    2012-09-01

    Full Text Available Abstract Background Starch is the most important source of calories for human nutrition and the majority of it is produced by cereal farming. Starch is also used as a renewable raw material in a range of industrial sectors. It can be chemically modified to introduce new physicochemical properties. In this way starch is adapted to a variety of specific end-uses. Recombinant DNA technologies offers an alternative to starch industrial processing. The plant biosynthetic pathway can be manipulated to design starches with novel structure and improved technological properties. In the future this may reduce or eliminate the economical and environmental costs of industrial modification. Recently, many advances have been achieved to clarify the genetic mechanism that controls starch biosynthesis. Several genes involved in the synthesis and modification of complex carbohydrates in many organisms have been identified and cloned. This knowledge suggests a number of strategies and a series of candidate genes for genetic transformation of crops to generate new types of starch-based polymers. However transformation of cereals is a slow process and there is no easy model system available to test the efficiency of candidate genes in planta. Results We explored the possibility to use transgenic barley callus generated from immature embryo for a fast test of transgenic modification strategies of starch biosynthesis. We found that this callus contains 4% (w/w dw starch granules, which we could modify by generating fully transgenic calli by Agrobacterium-transformation. A Green Fluorescent Protein reporter protein tag was used to identify and propagate only fully transgenic callus explants. Around 1 – 1.5 g dry weight of fully transgenic callus could be produced in 9 weeks. Callus starch granules were smaller than endosperm starch granules and contained less amylose. Similarly the expression profile of starch biosynthesis genes were slightly different in callus

  7. 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...

  8. 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.

  9. Inherited phenotype instability of inflorescence and floral organ development in homeotic barley double mutants and its specific modification by auxin inhibitors and 2,4-D

    Science.gov (United States)

    Šiukšta, Raimondas; Vaitkūnienė, Virginija; Kaselytė, Greta; Okockytė, Vaiva; Žukauskaitė, Justina; Žvingila, Donatas; Rančelis, Vytautas

    2015-01-01

    on the phenotypic instability observed, definite trends in the development of ectopic flower structures may be detected, from insignificant outgrowths on awns to flowers with sterile organs. Phenotypically unstable barley double mutants provide a highly promising genetic system for the investigation of gene expression modules and trend orders. PMID:25660346

  10. 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.

  11. Mapping Quantitative Trait Loci Controlling Endosperm Traits with Molecular Marker

    Institute of Scientific and Technical Information of China (English)

    XU Chen-wu; LI Tao; SUN Chang-sen; GU Shi-liang

    2002-01-01

    Based on the genetic models for triploid endosperm traits and on the methods for mapping diploid quantitative traits loci (QTLs), the genetic constitutions, components of means and genetic variances of QTL controlling endosperm traits under flanking marker genotypes of different generations were presented. From these results, a multiple linear regression method for mapping QTL underlying endosperm traits in cereals was proposed, which used the means of endosperm traits under flanking marker genotypes as a dependent variable, the coefficient of additive effect ( d ) and dominance effect ( h 1 and/or h2 ) of a putative QTL in a given interval as independent variables. This method can work at any position in a genome covered by markers and increase the estimation precision of QTL location and their effects by eliminating the interference of other relative QTLs. This method can also be easily used in other uneven data such as markers and quantitative traits detected or measured in plants and tissues different either in generations or at chromosomal ploidy levels, and in endosperm traits controlled by complicated genetic models considering the effects produced by genotypes of both maternal plants and seeds on them.

  12. The seeds of Lotus japonicus lines transformed with sense, antisense, and sense/antisense galactomannan galactosyltransferase constructs have structurally altered galactomannans in their endosperm cell walls.

    Science.gov (United States)

    Edwards, Mary E; Choo, Tze-Siang; Dickson, Cathryn A; Scott, Catherine; Gidley, Michael J; Reid, J S Grant

    2004-03-01

    Galactomannan biosynthesis in legume seed endosperms involves two Golgi membrane-bound glycosyltransferases, mannan synthase and galactomannan galactosyltransferase (GMGT). GMGT specificity is an important factor regulating the distribution and amount of (1-->6)-alpha-galactose (Gal) substitution of the (1-->4)-beta-linked mannan backbone. The model legume Lotus japonicus is shown now to have endospermic seeds with endosperm cell walls that contain a high-Gal galactomannan (mannose [Man]/Gal = 1.2-1.3). Galactomannan biosynthesis in developing L. japonicus endosperms has been mapped, and a cDNA encoding a functional GMGT has been obtained from L. japonicus endosperms during galactomannan deposition. L. japonicus has been transformed with sense, antisense, and sense/antisense ("hairpin loop") constructs of the GMGT cDNA. Some of the sense, antisense, and sense/antisense transgenic lines exhibited galactomannans with altered (higher) Man/Gal values in their (T(1) generation) seeds, at frequencies that were consistent with posttranscriptional silencing of GMGT. For T(1) generation individuals, transgene inheritance was correlated with galactomannan composition and amount in the endosperm. All the azygous individuals had unchanged galactomannans, whereas those that had inherited a GMGT transgene exhibited a range of Man/Gal values, up to about 6 in some lines. For Man/Gal values up to 4, the results were consistent with lowered Gal substitution of a constant amount of mannan backbone. Further lowering of Gal substitution was accompanied by a slight decrease in the amount of mannan backbone. Microsomal membranes prepared from the developing T(2) generation endosperms of transgenic lines showed reduced GMGT activity relative to mannan synthase. The results demonstrate structural modification of a plant cell wall polysaccharide by designed regulation of a Golgi-bound glycosyltransferase.

  13. Iron-biofortification in rice by the introduction of three barley genes participated in mugineic acid biosynthesis with soybean ferritin gene

    Directory of Open Access Journals (Sweden)

    Hiroshi eMasuda

    2013-05-01

    Full Text Available Iron deficiency is a serious problem around the world, especially in developing countries. The production of iron-biofortified rice will help ameliorate this problem. Previously, expression of the iron storage protein, ferritin, in rice using an endosperm-specific promoter resulted in a two-fold increase in iron concentration in the resultant transgenic seeds. However, further overexpression of ferritin did not produce an additional increase in the seed iron concentration, and symptoms of iron deficiency were noted in the leaves of the transgenic plants. In the present study, we aimed to further increase the iron concentration in rice seeds without increasing the sensitivity to iron deficiency by enhancing the uptake and transport of iron via a ferric iron chelator, mugineic acid. To this end, we introduced the soybean ferritin gene (SoyferH2 driven by two endosperm-specific promoters, along with the barley nicotianamine synthase gene (HvNAS1, two nicotianamine aminotransferase genes (HvNAAT-A and -B, and a mugineic acid synthase gene (IDS3 to enhance mugineic acid production in rice plants. A marker-free vector was utilized as a means of increasing public acceptance. Representative lines were selected from 102 transformants based on the iron concentration in polished seeds and ferritin accumulation in the seeds. These lines were grown in both commercially supplied soil (iron-sufficient conditions and calcareous soil (iron-deficient conditions. Lines expressing both ferritin and mugineic acid biosynthetic genes showed signs of iron-deficiency tolerance in calcareous soil. The iron concentration in polished T3 seeds was increased by 4 and 2.5 times, as compared to that in non-transgenic lines grown in normal and calcareous soil, respectively. These results indicate that the concomitant introduction of the ferritin gene and mugineic acid biosynthetic genes effectively increased the seed iron level without causing iron sensitivity under iron

  14. 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.

  15. 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...

  16. 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.

  17. Binding of ABI4 to a CACCG motif mediates the ABA-induced expression of the ZmSSI gene in maize (Zea mays L.) endosperm.

    Science.gov (United States)

    Hu, Yu-Feng; Li, Yang-Ping; Zhang, Junjie; Liu, Hanmei; Tian, Mengliang; Huang, Yubi

    2012-10-01

    Starch synthase I (SSI) contributes the majority of the starch synthase activity in developing maize endosperm. In this work, the effects of various plant hormones and sugars on the expression of the starch synthase I gene (ZmSSI) in developing maize endosperms were examined. The accumulation of ZmSSI mRNA was induced using abscisic acid (ABA) but not with glucose, sucrose, or gibberellin treatment. To investigate the molecular mechanism underlying this effect, the ZmSSI promoter region (-1537 to +51) was isolated and analysed. A transient expression assay in maize endosperm tissue showed that the full-length ZmSSI promoter is activated by ABA. The results of deletion and mutation assays demonstrated that a CACCG motif in the ZmSSI promoter is responsible for the ABA inducibility. The results of binding shift assays indicated that this CACCG motif interacts with the maize ABI4 protein in vitro. The overexpression of ABI4 in endosperm tissue enhanced the activity of a promoter containing the CACCG motif in the absence of ABA treatment. Expression pattern analysis indicated that the transcription pattern of ABI4 in the developing maize endosperm was induced by ABA treatment but was only slightly affected by glucose or sucrose treatment. Taken together, these data indicate that ABI4 binds to the CACCG motif in the ZmSSI promoter and mediates its ABA inducibility.

  18. 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...

  19. 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.

  20. EFFECT OF ENDOSPERM HARDNESS ON AN ETHANOL PROCESS USING A GRANULAR STARCH HYDROLYZING ENZYME

    Energy Technology Data Exchange (ETDEWEB)

    Wang, P; W Liu, D B; Johnston, K D; Rausch, S J; Schmidt, M E; Tumbleson, V Singh

    2010-01-01

    Granular starch hydrolyzing enzymes (GSHE) can hydrolyze starch at low temperature (32°C). The dry grind process using GSHE (GSH process) has fewer unit operations and no changes in process conditions (pH 4.0 and 32°C) compared to the conventional process because it dispenses with the cooking and liquefaction step. In this study, the effects of endosperm hardness, protease, urea, and GSHE levels on GSH process were evaluated. Ground corn, soft endosperm, and hard endosperm were processed using two GSHE levels (0.1 and 0.4 mL per 100 g ground material) and four treatments of protease and urea addition. Soft and hard endosperm materials were obtained by grinding and sifting flaking grits from a dry milling pilot plant; classifications were confirmed using scanning electron microscopy. During 72 h of simultaneous granular starch hydrolysis and fermentation (GSHF), ethanol and glucose profiles were determined using HPLC. Soft endosperm resulted in higher final ethanol concentrations compared to ground corn or hard endosperm. Addition of urea increased final ethanol concentrations for soft and hard endosperm. Protease addition increased ethanol concentrations and fermentation rates for soft endosperm, hard endosperm, and ground corn. The effect of protease addition on ethanol concentrations and fermentation rates was most predominant for soft endosperm, less for hard endosperm, and least for ground corn. Samples (soft endosperm, hard endosperm, or corn) with protease resulted in higher (1.0% to 10.5% v/v) ethanol concentration compared to samples with urea. The GSH process with protease requires little or no urea addition. For fermentation of soft endosperm, GSHE dose can be reduced. Due to nutrients (lipids, minerals, and soluble proteins) present in corn that enhance yeast growth, ground corn fermented faster at the beginning than hard and soft endosperm.

  1. 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.

  2. Methods for mapping QTLs underlying endosperm traits based on random hybridization design

    Institute of Scientific and Technical Information of China (English)

    WEN Yongxian; WU Weiren

    2006-01-01

    Several methods of interval mapping of QTLs underlying endosperm traits based on random hybridization designs and the triploid genetic model are proposed. The basic idea is: plants (or lines) from a population with known marker genotype information are randomly hybridized to generate a population of hybrid lines for endosperm QTL mapping; a mixture of seeds of each hybrid line is measured for the endosperm trait to get the mean of the line; then endosperm QTL mapping and effect estimation is performed using the endosperm trait means of hybrid lines and the marker genotype information of parental plants (or lines). The feasibility and efficiency of the methods are examined by computer simulations. Results show that the methods can precisely map endosperm QTLs and unbiasedly and efficiently estimate the three effects (additive effect, first dominant effect, second dominant effect) of endosperm QTLs.

  3. 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...

  4. 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...

  5. 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.

  6. 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.

  7. Genomic imprinting in plants: what makes the functions of paternal and maternal genes different in endosperm formation?

    Science.gov (United States)

    Ohnishi, Takayuki; Sekine, Daisuke; Kinoshita, Tetsu

    2014-01-01

    Genomic imprinting refers to the unequal expression of maternal and paternal alleles according to the parent of origin. This phenomenon is regulated by epigenetic controls and has been reported in placental mammals and flowering plants. Although conserved characteristics can be identified across a wide variety of taxa, it is believed that genomic imprinting evolved independently in animal and plant lineages. Plant genomic imprinting occurs most obviously in the endosperm, a terminally differentiated embryo-nourishing tissue that is required for seed development. Recent studies have demonstrated a close relationship between genomic imprinting and the development of elaborate defense mechanisms against parasitic elements during plant sexual reproduction. In this chapter, we provide an introductory description of genomic imprinting in plants, and focus on recent advances in our understanding of its role in endosperm development, the frontline of maternal and paternal epigenomes.

  8. FRAKSINASI ENZIM LIPASE DARI ENDOSPERM KELAPA DENGAN METODE SALTING OUT (Lipase fractionation of Coconut Endosperm by Salting out Method

    Directory of Open Access Journals (Sweden)

    Moh. Su'i

    2014-02-01

    Full Text Available This research learns about fractionation of lipases activity from coconut endosperm by using ammonium sulphate of 0–15%; 15-30 %, 30–45 %, 45–60 %, 60–75 % and 75–90 %. The results showed that the fractions of 0–15% ; 30–45 %, 45–60 % and 60–75 % have lipase activity. Meanwhile, the highest activity was fractions of 60-75%. fractions of 15-30% and 75-90%  have no lipase enzym activity. Molecule weigh of lipase enzyme was 72 kDa. Keywords: Lipases, endosperm, coconut, fractionation, ammonium sulphate   ABSTRAK Penelitian ini mempelajari fraksinasi enzim lipase dari endosperm kelapa menggunakan ammonium sulfat. fraksinasi dilakukan dengan variasi konsentrasi ammonium sulfat 0–15% ; 15-30%; 30–45 %, 45–60 %, 60–75 % dan 75–90 %. Hasil penelitian menunjukkan bahwa enzim lipase terdapat pada fraksi 0–15% ; 30–45 %, 45–60 % dan fraksi 60–75 % dengan aktivitas enzim tertinggi pada fraksi 60-75%. Sedangkan fraksi 15-30% dan 75-90% tidak ada enzim lipase. Berat molekul enzim lipase pada semua fraksi 72 kDa. Kata kunci: Lipase, endosperm, fraksinasi, ammonium sulfat

  9. 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.

  10. 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...

  11. 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.

  12. 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...

  13. 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....

  14. The induction of enzyme activity in the endosperm of germinating castor-bean seeds.

    Science.gov (United States)

    Marriott, K M; Northcote, D H

    1975-10-01

    Endosperm extracts were prepared at various times during germination from intact castor-bean seeds and from seeds from which the embryos had been removed. The sterilized seeds were incubated either on solid water agar or on agar containing 0.3 mM-gibberellic acid. 2. Isocitrate lyase and 3-hydroxyacyl-CoA dehydrogenase had very low activities in the mature seeds, but increased 44-fold and 27-fold respectively during germination. In contrast, the extracts of mature seeds had considerable acid and alkaline lipase activity and this only increased two- to three-fold during the incubation period. 3. Incubation of the seeds with gibberellic acid accelerated the rate of appearance of isocitrate lyase and 3-hydroxyacyl-CoA dehydrogenase. It also increased the total activity attained. However, the application of hormone had, in comparison, little effect on the development of lipase activity. 4. The removal of the embryo had little influence on the development of enzyme activity in the endosperm tissue; only with isocitrate lyase was a decrease in activity observed in the absence of the embryo.

  15. Use of the Endosperm Balance Number (EBN) in Potato Genetics and Breeding

    Institute of Scientific and Technical Information of China (English)

    Carputo Domenico; Jin Liping

    2006-01-01

    Solanum species have a wide geographic distribution and show a large range of ecological adaptation, and thus have developed strong resistances to a wide range of pests, diseases and abiotic stresses. They represent unique material as a source of useful genes and allelic diversity for the tetraploid cultivated potato (Solanum tuberosum). Knowledge of the endosperm balance number (EBN) incompatibility system may be extremely useful for breeders in designing specific crosses schemes for potato breeding. EBN has been experimentally assigned to Solanum species after crosses with standard species whose EBN was arbitrarily established, and assuming the 2: 1 ratio as a prerequisite for normal endosperm development. The role of the EBN incompatibility system in potato breeding is strongly complemented by 2n gametes. Only 2n gametes can allow inter-EBN crosses and thus gene flow between sexually isolated species. This complementary role of EBN and 2n gametes not only facilitates gene introgression between inter-EBN species, but also maintains the ploidy integrity of the two parental species. Examples are given to illustrate how to use EBN in crossing schemes designed for 2x (2EBN) and 2x(1EBN) species. As increasing in the interest in exploiting exotic germplasm for breeding purposes, it is strongly believed that an understanding of EBN can be used for interspecific gene transfer in any crop species where an EBN-like system operates.

  16. 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...

  17. Circadian oscillation of starch branching enzyme gene expression in the sorghum endosperm

    Energy Technology Data Exchange (ETDEWEB)

    Mutisya, J.; Sun, C.; Jansson, C.

    2009-08-31

    Expression of the three SBE genes, encoding starch branching enzymes, in the sorghum endosperm exhibited a diurnal rhythm during a 24-h cycle. Remarkably, the oscillation in SBE expression was maintained in cultured spikes after a 48-h dark treatment, also when fed a continuous solution of sucrose or abscisic acid. Our findings suggest that the rhythmicity in SBE expression in the endosperm is independent of cues from the photosynthetic source and that the oscillator resides within the endosperm itself.

  18. Effects of Different Comsteep Liquids on Structure of Starch and Protein Binding in Corn Endosperm

    OpenAIRE

    Rong Yan; Xinhua Li; Xiaojun Qi

    2015-01-01

    The changes in the structures of corn endosperms immersed in different reagents were studied, in order to preliminarily determine the mode of binding of starch and protein in the endosperm. The corn endosperm was immersed in Sulfurous acid, NaOH, ethanol, L-cysteine and morel berry, respectively. The dissociated starch content was analyzed using water as the reference. The results of the treatment were analyzed by the dissociated starch content, optical microscopy and scanning electron micros...

  19. Effect of Flowering Time of Spikelets Within a Rice Panicle on Endosperm Deve lopment and Its Physiological Mechanism

    Institute of Scientific and Technical Information of China (English)

    Yang Jianchang; Liu Lijun; Wang Zhiqin; Lang Youzhong; Zhu Qingsen

    2000-01-01

    The flowering duration of spikelets within a panicle of the rice cultivars of Wuyujing 3 (Japonica), Yangdao 4 (Indica), Shanyou 63 (Indica hybrid rice) and PC311/Zaoxiandang 18 (Indica/japonica hybrid rice) was 5d, 7d, 7d and 8d, respectively. The spikelets flowered on the 2nd day produced the highest endosperm weight (EW) and the most endosperm cells (ECs), and followed by the spikelets flowered on the 1st day. ECs decreased with the delay of flowering of the spikelets flowered from the 3rd day. Within a variety or a hybrid combination, the difference in endosperm cell weight (ECW) was not significant among the grains flowered on different dates. EW and grain-filled percentage (G-FP) were very significantly correlated with ECs, but not with ECW. The earlier the spikelets flowered (except those flowered on the 1st day), the greater the initial proliferation power (R°), the higher the maximum proliferation rate (Vmax) and the higher the mean proliferation rate (V) of ECs, and the shorter the time reaching Vmax, and vice versa. R°, Vmax,V and ECs were significantly correlated with the physiological activities (ATPase activity and the content of spermidine and spermine) of the grains at early grain filling stage. The physiological activities of the grains, R°,Vmax,V, ECs and EW significantly increased after removing 1/2 plants at booting stage and spraying 6-BA 〔N6(benzyl) adenine〕 at early heading stage, and the results were reversed after cutting 1/2 leaves at booting stage. These results suggest that the difference in ECs results in the difference in the grain weight, while the low physiological activities of late-flowered grains are the important reasons for their poor endosperm development.

  20. Identification and characterization of paternal-preferentially expressed gene NF-YC8 in maize endosperm.

    Science.gov (United States)

    Mei, Xiupeng; Liu, Chaoxian; Yu, Tingting; Liu, Xiaoli; Xu, De; Wang, Jiuguang; Wang, Guoqiang; Cai, Yilin

    2015-10-01

    Gene imprinting describes an epigenetic phenomenon, whereby genetically identical alleles are differentially expressed dependent on parent-of-origin. Some imprinted genes belonged to NUCLEAR FACTOR Y (NF-Y) transcription factors, which were involved in many important metabolic processes in plant. The characterizations of imprinted genes are of great importance for their function exploration. In this paper, 15 non-redundant NF-YC genes were identified in the maize genome and the paternally expressed gene NF-YC8 was further analyzed. NF-YC8 primarily expressed in maize immature ear and tassel and phylogenetic analysis showed that NF-YC8 was highly homologous with Arabidopsis thaliana NF-YC2 genes which function in regulation of the flowering processes, ER stress response. Furthermore, NF-YC8 was a differential, gene-specific imprinted gene at 14 DAP and persistently imprinted throughout later endosperm development in the B73/Mo17 genetic background. Bisulfite sequencing for NF-YC8 in maize endosperm showed that the paternal alleles were higher methylated (CG, CHG and CHH contexts) than maternal alleles in the 5' upstream region, and the coding region was highly methylated in CG context. Additionally, TE (CG, CHG and CHH contexts) and repetitive region (CG and CHG contexts) were all highly methylated. These results are the first description of evolution and molecular characterization of maize NF-YC8 and will provide new references for maize NF-YC genetic analysis.

  1. Growth performance, behaviour, forestomach development and meat quality of veal calves provided with barley grain or ground wheat straw for welfare purpose

    Directory of Open Access Journals (Sweden)

    Igino Andrighetto

    2010-01-01

    Full Text Available Two different feeding plans for veal calves were compared in the study: a traditional liquid diet supplemented with 250  g/calf/d of barley grain or with 250 g/calf/d of ground wheat straw. The two solid feeds had different chemical composi-  tion but a similar particle size obtained by grinding the straw in a mill with an 8-mm mesh screen. Twenty-four Polish  Friesian male calves were used in the study and they were housed in individual wooden stalls (0.83 x 1.80 m. The health  status of all the calves was satisfactory for the entire fattening period and no specific medical treatment was required  during the trial. Calves fed wheat straw showed a greater intake of solid feed (196 vs. 139 g/d; P  average daily gain (1288 vs. 1203 g/d; P  not affected by the type of solid feed and no milk refusal episodes were detected. The haemoglobin concentration was  similar in calves receiving the two feeding treatments despite the higher iron intake provided by the wheat straw through-  out the fattening period (2.12 vs. 1.15 g; P  calves’ metabolism. Feeding behaviour was affected by the provision of solid feeds. Eating and chewing were prolonged  in calves receiving ground wheat straw and the same solid feed reduced the frequency of oral stereotypies at the end of  the fattening period. At the slaughterhouse, no differences were observed between the feeding treatments as regards  carcass weight and dressing percentage. The calves fed ground wheat straw had a heavier weight of the empty omasum  (518 vs. 341 g; P  fed barley grain. The incidence of abomasal erosions, ulcers and scars was similar in both treatments; however the index  of abomasal damage, which considers the number and the seriousness of different type of lesions, was higher in calves  receiving barley grain. Therefore, the grinding of straw particles, as opposed to barley grain, can reduce the abrasive-  ness of roughage at the abomasum level. Visual evaluation of the

  2. Temperature Stress at Grain Filling Stage Mediates Expression of Three Isoform Genes Encoding Starch Branching Enzymes in Rice Endosperm

    Institute of Scientific and Technical Information of China (English)

    WEI Ke-su; CHENG Fang-min; ZHANG Qi-fang; LIU Kui-gang

    2009-01-01

    An early-maturity indica rice variety Zhefu 49, whose grain quality and starch structure are sensitive to environmental temperature, was subjected to different temperatures (32oC for high temperature and 22oC for optimum temperature) at the grain filling stage in plant growth chambers, and the different expressions of three isoform genes (SBEI, SBEIII and SBEIV) encoding starch branching enzyme (SBE) in the endosperms were studied by the real-time fluorescence quantitative PCR (FQ-PCR) method. Effects of high temperature on the SBE expression in developing rice endosperms were isoform-dependent. High temperature significantly down-regulated the expressions of SBEI and SBEIII, while up-regulated the expression of SBEIV. Compared with SBEIV and SBEIII, the expression of SBEI gene in Zhefu 49 rice endosperms was more sensitive to temperature variation at the grain filling stage. This study indicates that changes in weather/climate conditions especially temperature stress influence rice grain formation and its quality as evidenced by isoform expression.

  3. 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...

  4. Spatio-temporal profiling and degradation of α-amylase isozymes during barley seed germination

    DEFF Research Database (Denmark)

    Bak-Jensen, K.S.; Laugesen, S.; Østergaard, O.

    2007-01-01

    Ten genes from two multigene families encode barley alpha-amylases. To gain insight into the occurrence and fate of individual isoforms during seed germination, the alpha-amylase repertoire was mapped by using a proteomics approach consisting of 2D gel electrophoresis, western blotting, and mass...... identified only by immunostaining. Mass spectrometry identified 12 full-length forms and 12 fragments from the cultivar Barke. Products of both alpha-amylase 2 entries co-migrated in five full-length and one fragment spot. The alpha-amylase abundance and the number of fragments increased during germination...... and near the C-terminus. Only two shorter fragments were identified of the other alpha-amylase 2 (gi|166985). The 2D gels of dissected tissues showed alpha-amylase degradation to be confined to endosperm. In contrast, the aleurone layer contained essentially only full-length alpha-amylase forms. While only...

  5. Spatio-temporal profiling and degradation of alpha-amylase isozymes during barley seed germination

    DEFF Research Database (Denmark)

    Bak-Jensen, K.S.; Laugesen, Sabrina; Østergaard, Ole

    2007-01-01

    Ten genes from two multigene families encode barley alpha-amylases. To gain insight into the occurrence and fate of individual isoforms during seed germination, the alpha-amylase repertoire was mapped by using a proteomics approach consisting of 2D gel electrophoresis, western blotting, and mass...... increased during germination. Assessing the fragment minimum chain length by peptide mass fingerprinting suggested that alpha-amylase 2 ( gi vertical bar 4699831) initially was cleaved just prior to domain B that protrudes from the (beta alpha)(8)-barrel between beta-strand 3 and alpha-helix 3, followed...... by cleavage on the C-terminal side of domain B and near the C-terminus. Only two shorter fragments were identified of the other alpha-amylase 2 (gi vertical bar 166985). The 2D gels of dissected tissues showed alpha-amylase degradation to be confined to endosperm. In contrast, the aleurone layer contained...

  6. Concerted suppression of all starch branching enzyme genes in barley produces amylose-only starch granules

    DEFF Research Database (Denmark)

    Carciofi, Massimiliano; Blennow, Per Gunnar Andreas; Jensen, Susanne Langgård;

    2012-01-01

    Background Starch is stored in higher plants as granules composed of semi-crystalline amylopectin and amorphous amylose. Starch granules provide energy for the plant during dark periods and for germination of seeds and tubers. Dietary starch is also a highly glycemic carbohydrate being degraded...... is preferentially derived from amylose, which can be increased by suppressing amylopectin synthesis by silencing of starch branching enzymes (SBEs). However all the previous works attempting the production of high RS crops resulted in only partly increased amylose-content and/or significant yield loss. Results...... In this study we invented a new method for silencing of multiple genes. Using a chimeric RNAi hairpin we simultaneously suppressed all genes coding for starch branching enzymes (SBE I, SBE IIa, SBE IIb) in barley (Hordeum vulgare L.), resulting in production of amylose-only starch granules in the endosperm...

  7. 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

  8. 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.

  9. 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...

  10. 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...

  11. 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...

  12. Effect of nitrogen fertilizer on distribution of starch granules in different regions of wheat endosperm

    Directory of Open Access Journals (Sweden)

    Fei Xiong

    2014-02-01

    Full Text Available This study provided visual evidence of a nitrogen effect on starch granules (SGs in wheat endosperm. Winter wheat (Titicum aestivum L. cultivar Xumai 30 was cultured under no nitrogen (control and 240 kg ha− 1 of nitrogen applied at the booting stage. The number, morphology, and size of A- and B-type SGs in subaleurone of dorsal endosperm (SDE, center of dorsal endosperm (CDE, modified aleurone (MA, subaleurone of ventral endosperm (SVE, and center of ventral endosperm (CVE were observed under light and electron microscopes. (1 The distribution of SGs in SDE was similar to that in SVE, the distributions of SGs in CDE and CVE were similar, but the distribution of SGs in MA was different from those in the other four endosperm regions. The number of SGs in the five endosperm regions was in the order SDE > CDE > SVE > CVE > MA. (2 Nitrogen increased the number of A- and B-type SGs in SDE and SVE. Nitrogen also increased the number of B-type SGs but decreased the number of A-type SGs in CDE and CVE. Nitrogen decreased the numbers of A-type and B-type SGs in MA. The results suggest that increased N fertilizer application mainly increased the numbers of small SGs and decreased the numbers of large SGs, but that the results varied in different regions of the wheat endosperm.

  13. Effect of nitrogen fertilizer on distribution of starch granules in different regions of wheat endosperm

    Institute of Scientific and Technical Information of China (English)

    Fei; Xiong; Xurun; Yu; Liang; Zhou; Jing; Zhang; Yanping; Jin; Dongliang; Li; Zhong; Wang

    2014-01-01

    This study provided visual evidence of a nitrogen effect on starch granules(SGs) in wheat endosperm. Winter wheat(Titicum aestivum L.) cultivar Xumai 30 was cultured under no nitrogen(control) and 240 kg ha-1of nitrogen applied at the booting stage. The number, morphology, and size of Aand B-type SGs in subaleurone of dorsal endosperm(SDE), center of dorsal endosperm(CDE), modified aleurone(MA), subaleurone of ventral endosperm(SVE), and center of ventral endosperm(CVE) were observed under light and electron microscopes.(1) The distribution of SGs in SDE was similar to that in SVE, the distributions of SGs in CDE and CVE were similar, but the distribution of SGs in MA was different from those in the other four endosperm regions. The number of SGs in the five endosperm regions was in the order SDE > CDE > SVE > CVE > MA.(2) Nitrogen increased the number of Aand B-type SGs in SDE and SVE. Nitrogen also increased the number of B-type SGs but decreased the number of A-type SGs in CDE and CVE. Nitrogen decreased the numbers of A-type and B-type SGs in MA. The results suggest that increased N fertilizer application mainly increased the numbers of small SGs and decreased the numbers of large SGs, but that the results varied in different regions of the wheat endosperm.

  14. Division of Free Nuclei in Rice Endosperm and Its Influencing Factors

    Institute of Scientific and Technical Information of China (English)

    LIU Man-xi; WANG Zhong; WU Xiao-mei; CHEN Yi-fang; GU Yun-jie

    2009-01-01

    The division process of endosperm in rice and its influencing factors were investigated. Total dissection, resin embedding and sectioning under light and transmission electron microscopes were used to observe the division patterns of free nuclei and cytological characters of an endosperm. The various division patterns of free nuclei in rice endosperm included mitosis, amitosis, and the mitosis to amitosis change without an interphase. The division velocity of free nuclei in rice endosperm varied with the internucleus distance. Comparing to a cluster, the division velocity increased when the free nuclei were separated from each other. Moderately higher temperatures also enhanced the proliferation and shortened the division cycle of free nuclei in rice endosperm. To a certain extent, nitrogenous fertilizer applied at the booting stage together with a moderate drought treatment accelerated the division of free nuclei as well.

  15. Chemical genetics and cereal starch metabolism: structural basis of the non-covalent and covalent inhibition of barley β-amylase.

    Science.gov (United States)

    Rejzek, Martin; Stevenson, Clare E; Southard, Andrew M; Stanley, Duncan; Denyer, Kay; Smith, Alison M; Naldrett, Mike J; Lawson, David M; Field, Robert A

    2011-03-01

    There are major issues regarding the proposed pathway for starch degradation in germinating cereal grain. Given the commercial importance but genetic intractability of the problem, we have embarked on a program of chemical genetics studies to identify and dissect the pathway and regulation of starch degradation in germinating barley grains. As a precursor to in vivo studies, here we report systematic analysis of the reversible and irreversible inhibition of the major β-amylase of the grain endosperm (BMY1). The molecular basis of inhibitor action was defined through high resolution X-ray crystallography studies of unliganded barley β-amylase, as well as its complexes with glycone site binder disaccharide iminosugar G1M, irreversible inhibitors α-epoxypropyl and α-epoxybutyl glucosides, which target the enzyme's catalytic residues, and the aglycone site binders acarbose and α-cyclodextrin.

  16. 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

  17. Cutin plays a role in differentiation of endosperm-derived callus of kiwifruit.

    Science.gov (United States)

    Popielarska-Konieczna, Marzena; Kozieradzka-Kiszkurno, Małgorzata; Bohdanowicz, Jerzy

    2011-11-01

    Cutin fluorescence, after auramine O treatment, was detected on the surface of organogenic areas (protuberances) of endosperm derived callus induced on Murashige and Skoog medium with thidiazuron (0.5 mg l(-1)) in darkness. Electron micrographs of the protuberances revealed cuticle, visible as a dark-staining layer, and amorphous waxes on the cell wall. In some cases the cells of the epidermis-like layer and shoot buds at early stages of development showed thick and characteristically wavy cutin. This waviness corresponds with the wrinkled appearance of the cell wall as observed by scanning electron microscopy. The role of multivesicular bodies in cutin production and transfer to the plasma membrane is discussed.

  18. 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.

  19. Sequencing of 15,622 gene-bearing BACs clarifies the gene-dense regions of the barley genome

    Science.gov (United States)

    Barley (Hordeum vulgare L.) possesses a large and highly repetitive genome of 5.1 Gb that has hindered the development of a complete sequence. In 2012, the International Barley Sequencing Consortium released a resource integrating whole-genome shotgun sequences with a physical and genetic framework....

  20. 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

  1. 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.

  2. Dormancy of the barley grain is correlated with gibberellic Acid responsiveness of the isolated aleurone layer.

    Science.gov (United States)

    Schuurink, R C; Sedee, N J; Wang, M

    1992-12-01

    The relationship between barley grain dormancy and gibberellic acid (GA(3)) responsiveness of aleurone layers has been investigated. Barley (Hordeum distichum L. cvs Triumph and Kristina) grains were matured under defined conditions in a phytotron. Grains of Triumph plants grown under long-day/warm conditions had lower dormancy levels than grains of plants grown under short-day/cool conditions. Aleurone layers isolated from grains of long-day Triumph plants secreted more alpha-amylase and had a higher responsiveness to GA(3) as measured by alpha-amylase secretion. Storage of the grains increased both the percentage of germination and the responsiveness of the aleurone to GA(3). Use of different sterilization methods to break dormancy confirmed the correlation between germination percentage and aleurone layer GA(3) responsiveness. The response of embryoless Triumph grains to GA(3) was lower than that of the isolated aleurone layers, suggesting a role of the starchy endosperm in regulating the GA(3) response of the aleurone layer. Grains of the cultivar Kristina harvested from short day- and long day-grown plants lacked dormancy, and their isolated aleurone layers had a similar responsiveness to GA(3) as measured by alpha-amylase secretion. The data indicate that the physiological state of the aleurone layers contributes to the percentage germination of the grains.

  3. 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...

  4. Targeted expression of HvHMA2 increases the mineral content of the inner endosperm in barley

    DEFF Research Database (Denmark)

    Noeparvar, Shahin; Darbani, Behrooz; Tauris, Birgitte

    2017-01-01

    Cereals are a major source of dietary energy and protein but are nutritionally poor in micronutrients. Zinc (Zn) biofortification of staple crops has been proposed as a promising strategy to combat the global challenge of human Zn-deficiency. The aim of this study was to improve the Zn content in...

  5. 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...

  6. 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

  7. 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.

  8. 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).

  9. 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.

  10. Bioacumulación de arsénico en las etapas de desarrollo de la cebada maltera (Hordeum distichon L. Arsenic bioaccumulation in the stages of development of malt barley (Hordeum distichon L.

    Directory of Open Access Journals (Sweden)

    Francisco Prieto García

    cultivo para estimar niveles de contaminación por arsénico.The contamination by arsenic in water and ground of Zimapán, Hidalgo, Mexico, causes that malt barley (Hordeum distichon L. grown in this region, accumulates this element. The objective was to measure the levels of arsenic accumulated in organs and tissues of barley under controlled conditions, at different stages of development. Malt barley cv. Esmeralda was grown in 25 pots, five pots per differentiated trial. Trials include negative (not contaminated ground and water and positive controls. Five plants by pot were used and the accumulation of arsenic in each one was evaluated in five organs: root, stem, leaf, glean and grain, at each of five stages of development (coleoptiles formation or first leaf, macollo or true leaves from an armpit first knot that becomes present separated from the ground and forms the first spike, formation of spikes and seed filling. A random block design with three treatments was used: with arsenic, positive control and negative control. Malt barley presented high accumulation of arsenic at different stages of development, directly proportional to the time of exposition. Roots presented the highest accumulation of arsenic (100 and 300 mg kg-1, followed by leaves (80 to 180 mg kg-1 and stem (70 to 110 mg kg-1. The level of accumulation of arsenic in grains was between 40-90 mg kg-1, amount that according to Argentinean norms can be considered as highly contaminated. For the bioaccumulation index (BI, the leaves presented the high values, classified as medium values (0.1-1 for green plants. This organ can be indicative of contamination level in the ground. The largest values for the BI were observed in the stem, this organ can be a malt barley specific indicator of the level of arsenic contamination.

  11. Abscisic acid controls embryo growth potential and endosperm cap weakening during coffee (Coffea arabica cv. Rubi) seed germination.

    Science.gov (United States)

    da Silva, E A Amaral; Toorop, Peter E; van Aelst, Adriaan C; Hilhorst, Henk W M

    2004-12-01

    The mechanism and regulation of coffee seed germination were studied in Coffea arabica L. cv. Rubi. The coffee embryo grew inside the endosperm prior to radicle protrusion and abscisic acid (ABA) inhibited the increase in its pressure potential. There were two steps of endosperm cap weakening. An increase in cellulase activity coincided with the first step and an increase in endo-beta-mannanase (EBM) activity with the second step. ABA inhibited the second step of endosperm cap weakening, presumably by inhibiting the activities of at least two EBM isoforms and/or, indirectly, by inhibiting the pressure force of the radicle. The increase in the activities of EBM and cellulase coincided with the decrease in the force required to puncture the endosperm and with the appearance of porosity in the cell walls as observed by low-temperature scanning electronic microscopy. Tissue printing showed that EBM activity was spatially regulated in the endosperm. Activity was initiated in the endosperm cap whereas later during germination it could also be detected in the remainder of the endosperm. Tissue printing revealed that ABA inhibited most of the EBM activity in the endosperm cap, but not in the remainder of the endosperm. ABA did not inhibit cellulase activity. There was a transient rise in ABA content in the embryo during imbibition, which was likely to be responsible for slow germination, suggesting that endogenous ABA also may control embryo growth potential and the second step of endosperm cap weakening during coffee seed germination.

  12. 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...

  13. Novas observações citológicas no endosperma de café Cytological observations on coffee endosperm

    Directory of Open Access Journals (Sweden)

    Dixier M. Medina

    1965-01-01

    Full Text Available Contagens de cromossomos foram efetuadas em endosperma de algumas espécies de Coffea, empregando-se a técnica da orceína acética, já referida em trabalho anterior. Os resultados encontrados para C. arabica (variedades typica, semperflorens, 2n = 44, e plantas monossômicas, 2n = 43 ; C. canephora, 2n = 22, C. dewevrei, 2n = 22, C. eugenioides x C. kapakata, 2n = 22 e C. eugenioides duplicado, 2n = 44, são discutidos em cada caso. Em todo o material estudado foi constatada a endomitose. O endosperma é, no início, um tecido de constituição 3x, transformando-se depois num tecido quimérico com células polissomáticas de diferentes graus. As numerosas figuras conseguidas, que permitiram contagens ou microfotografias, confirmam que, no cafeeiro, o endosperma é um material favorável para pesquisas citológicas.Chromosome countings have been rnade on lhe endosperm of Coffea using the same technique of acetic oreein already referred to in previous work of the author. Endosperm from the varieties typica and semperflorens (2n = 44 and monosomic plants (2n = 43 of C. arabica as well as of other species as C. canephora (2n = 22, C. dewevrei (2n = 22, C. eugenioides x C. kapakata (2n = 22 and duplicated C. eugenioides (2n = 44 were studied. Gametic number of chromosomes are discussed leading in some cases to the conclusion that gametes with missing chromosomes and gametes with supernumerary chromosomes were functional. Endomitosis was observed in all material studied. Endosperm tissue is constituted, therefore, by polysomatic cells (6x, 12x in addition to the monosomatic ones (3x initially formed through fertilization. The possible relation of endomitosis and embryo development is also discussed. The great number of photomicrographies obtained indicates once more that coffee endosperm is a favourable material for cytological investigations.

  14. 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...

  15. Ethnobotany, diverse food uses, claimed health benefits and implications on conservation of barley landraces in North Eastern Ethiopia highlands

    Directory of Open Access Journals (Sweden)

    Sopade Peter A

    2011-06-01

    Full Text Available Abstract Background Barley is the number one food crop in the highland parts of North Eastern Ethiopia produced by subsistence farmers grown as landraces. Information on the ethnobotany, food utilization and maintenance of barley landraces is valuable to design and plan germplasm conservation strategies as well as to improve food utilization of barley. Methods A study, involving field visits and household interviews, was conducted in three administrative zones. Eleven districts from the three zones, five kebeles in each district and five households from each kebele were visited to gather information on the ethnobotany, the utilization of barley and how barley end-uses influence the maintenance of landrace diversity. Results According to farmers, barley is the "king of crops" and it is put for diverse uses with more than 20 types of barley dishes and beverages reportedly prepared in the study area. The products are prepared from either boiled/roasted whole grain, raw- and roasted-milled grain, or cracked grain as main, side, ceremonial, and recuperating dishes. The various barley traditional foods have perceived qualities and health benefits by the farmers. Fifteen diverse barley landraces were reported by farmers, and the ethnobotany of the landraces reflects key quantitative and qualitative traits. Some landraces that are preferred for their culinary qualities are being marginalized due to moisture shortage and soil degradation. Conclusions Farmers' preference of different landraces for various end-use qualities is one of the important factors that affect the decision process of landraces maintenance, which in turn affect genetic diversity. Further studies on improving maintenance of landraces, developing suitable varieties and improving the food utilization of barley including processing techniques could contribute to food security of the area.

  16. In situ histochemical localisation of alkaloids and acetogenins in the endosperm and embryonic axis of Annona macroprophyllata Donn. Sm. seeds during germination.

    Science.gov (United States)

    Brechú-Franco, A E; Laguna-Hernández, G; De la Cruz-Chacón, I; González-Esquinca, A R

    2016-02-01

    Currently, the Annonaceae family is characterised by the production of acetogenins (ACGs), and also by the biosynthesis of alkaloids, primarily benzylisoquinolines derived from tyrosine. The objective of this study was to confirm the presence of alkaloids and acetogenins in the idioblasts of the endosperm and the embryonic axis of A. macroprophyllata seeds in germination. The Dragendorff, Dittmar, Ellram, and Lugol reagents were used to test for alkaloids, and Kedde's reagent was used to determine the presence of acetogenins in fresh sections of the endosperm and embryonic axis of seeds after twelve days of germination. A positive reaction was observed for all the reagents, and the presence of alkaloids and acetogenins was confirmed in the idioblasts of the endosperm and those involved in the differentiation of the embryonic axis of the developing seedling. We concluded that the idioblasts store both metabolites, acetogenins and alkaloids. Beginning at differentiation, the idioblasts of the embryonic axis simultaneously biosynthesise acetogenins and alkaloids that are characteristic of the species during the development of the seedling. The method used here can be applied to histochemically confirm the presence of acetogenins and alkaloids in tissues and structures of the plant in different stages of its life cycle.

  17. In situ histochemical localisation of alkaloids and acetogenins in the endosperm and embryonic axis of Annona macroprophyllata Donn. Sm. seeds during germination

    Directory of Open Access Journals (Sweden)

    A.E. Brechú-Franco

    2016-02-01

    Full Text Available Currently, the Annonaceae family is characterised by the production of acetogenins (ACGs, and also by the biosynthesis of alkaloids, primarily benzylisoquinolines derived from tyrosine. The objective of this study was to confirm the presence of alkaloids and acetogenins in the idioblasts of the endosperm and the embryonic axis of A. macroprophyllata seeds in germination. The Dragendorff, Dittmar, Ellram, and Lugol reagents were used to test for alkaloids, and Kedde’s reagent was used to determine the presence of acetogenins in fresh sections of the endosperm and embryonic axis of seeds after twelve days of germination. A positive reaction was observed for all the reagents, and the presence of alkaloids and acetogenins was confirmed in the idioblasts of the endosperm and those involved in the differentiation of the embryonic axis of the developing seedling. We concluded that the idioblasts store both metabolites, acetogenins and alkaloids. Beginning at differentiation, the idioblasts of the embryonic axis simultaneously biosynthesise acetogenins and alkaloids that are characteristic of the species during the development of the seedling. The method used here can be applied to histochemically confirm the presence of acetogenins and alkaloids in tissues and structures of the plant in different stages of its life cycle.

  18. Effect of salicylhydroxamic acid on endosperm strength and embryo growth of Lactuca sativa L. cv Waldmann's Green seeds

    Science.gov (United States)

    Brooks, C. A.; Mitchell, C. A.

    1988-01-01

    Salicylhydroxamic acid (SHAM) stimulated germination of photosensitive lettuce (Lactuca sativa L. cv Waldmann's Green) seeds in darkness. To determine whether SHAM acts on the embryo or the endosperm, we investigated separately effects of SHAM on growth potential of isolated embryos as well as on endosperm strength. Embryo growth potential was quantified by incubating decoated embryos in various concentrations of osmoticum and measuring subsequent radicle elongation. Growth potential of embryos isolated from seeds pretreated with 4 millimolar SHAM was equal to that of untreated controls. Rupture strength of endosperm tissue excised from seeds pretreated with SHAM was 33% less than that of controls in the micropylar region. To determine if the embryo must be in contact with the endosperm of SHAM to weaken the endosperm, some endosperms were incubated with SHAM only after dissection from seeds. Rupture strength of SHAM-treated, isolated endosperms in the micropylar region was 25% less than that of untreated controls. There was no difference in rupture strength in the cotyledonary region of endosperm isolated from seeds treated with SHAM in buffer or buffer alone. SHAM therefore stimulates germination not by enhancing embryo growth potential, but by weakening the micropylar region of the endosperm enclosing the embryo.

  19. Thioredoxin reduction alters the solubility of proteins of wheat starchy endosperm: an early event in cereal germination.

    Science.gov (United States)

    Wong, Joshua H; Cai, Nick; Tanaka, Charlene K; Vensel, William H; Hurkman, William J; Buchanan, Bob B

    2004-04-01

    A KCl-soluble, albumin/globulin fraction of wheat (Triticum aestivum L.) starchy endosperm was further separated into a methanol-insoluble fraction that contained metabolic proteins and a methanol-soluble fraction that contained "chloroform-methanol" or CM-like proteins. Reduction of the disulfide bonds of the CM proteins with thioredoxin or dithiothreitol altered their properties so that, like the metabolic proteins, they were insoluble in methanol. Glutathione had little effect, indicating dithiol specificity. Proteomic analysis of the CM protein fraction revealed the presence of isoforms of low molecular weight disulfide proteins (alpha-amylase, alpha-amylase/trypsin and WCI proteinase inhibitors, lipid transfer proteins, gamma-thionins), stress enzymes (Cu-Zn superoxide dismutase and peroxidase), storage proteins (alpha-, gamma- and omega-gliadins, low molecular weight glutenin subunits and globulins of the avenin N9 type), and a component of protein degradation (polyubiquitin). These findings support the view that, in addition to modifying activity and increasing protease sensitivity, reduction by thioredoxin alters protein solubility, thereby promoting processes of the grain starchy endosperm, notably the mobilization of reserves during germination and seedling development.

  20. Proteomic Analysis Reveals Different Involvement of Embryo and Endosperm Proteins during Aging of Yliangyou 2 Hybrid Rice Seeds

    Science.gov (United States)

    Zhang, Ying-Xue; Xu, Heng-Heng; Liu, Shu-Jun; Li, Ni; Wang, Wei-Qing; Møller, Ian M.; Song, Song-Quan

    2016-01-01

    Seed aging is a process that results in a delayed germination, a decreased germination percentage, and finally a total loss of seed viability. However, the mechanism of seed aging is poorly understood. In the present study, Yliangyou 2 hybrid rice (Oryza sativa L.) seeds were artificially aged at 100% relative humidity and 40°C, and the effect of artificial aging on germination, germination time course and the change in protein profiles of embryo and endosperm was studied to understand the molecular mechanism behind seed aging. With an increasing duration of artificial aging, the germination percentage and germination rate of hybrid rice seeds decreased. By comparing the protein profiles from the seeds aged for 0, 10 and 25 days, a total of 91 and 100 protein spots were found to show a significant change of more than 2-fold (P cell defense and rescue (28%), and with storage protein (18%). In endosperms, most of the identified proteins were involved in metabolism (37%), in energy (27%), and in protein synthesis and destination (11%). The most marked change was the increased abundance of many glycolytic enzymes together with the two fermentation enzymes pyruvate decarboxylase and alcohol dehydrogenase in the embryos during aging. We hypothesize that the decreased viability of hybrid rice seeds during artificial aging is caused by the development of hypoxic conditions in the embryos followed by ethanol accumulation.

  1. Quantification of amylose, amylopectin and β-glucan in the search for genes controlling the three major quality traits in barley using genome-wide association studies

    Directory of Open Access Journals (Sweden)

    Soren K Rasmussen

    2014-05-01

    Full Text Available Genome-wide association studies (GWAS for amylose, amylopectin and β-glucan concentration in a collection of 254 European spring barley varieties allowed to identify 20, 17 and 21 single nucleotide polymorphic (SNP markers, respectively, associated with these important grain quality traits. Negative correlations between the content of amylose and β-glucan (R=-0.62, P<0.01 and amylopectin and β-glucan (R= -0.487, P<0.01 were found in this large collection of spring barley varieties. Besides HvCslF6, amo1 and AGPL2, sex6 and waxy were identified among the major genes responsible for β-glucan, amylose and amylopectin content, respectively. Several minor genes like HvGSL4, HvGSL3 and HvCesA6, PWD were also detected by GWAS for the first time. Furthermore, the gene encoding β-fructofuranosidase, located on the short arm of chromosome 7H at 1.49cM, and SRF6, encoding ‘leucine-rich repeat receptor kinase protein’ on chromosome 2H, are proposed to be new candidate genes for amylopectin formation in barley endosperm. Several of the associated SNPs on chromosome 1H, 5H, 6H and 7H mapped to overlapping regions containing QTLs and genes controlling the three grain constituents. In particular chromosomes 5H and 7H carry many QTLs controlling barley grain quality. Amylose, amylopectin and β-glucan were interacted among each other through a metabolic network connected by UDP showing pleiotropic effects. Taken together, these results showed that cereal quality traits related each other and regulated through an interaction network, the identified major genes and genetic regions for amylose, amylopectin and β-glucan is a helpful for further research on carbohydrates and barley breeding.

  2. Sequencing of 15 622 gene-bearing BACs clarifies the gene-dense regions of the barley genome.

    Science.gov (United States)

    Muñoz-Amatriaín, María; Lonardi, Stefano; Luo, MingCheng; Madishetty, Kavitha; Svensson, Jan T; Moscou, Matthew J; Wanamaker, Steve; Jiang, Tao; Kleinhofs, Andris; Muehlbauer, Gary J; Wise, Roger P; Stein, Nils; Ma, Yaqin; Rodriguez, Edmundo; Kudrna, Dave; Bhat, Prasanna R; Chao, Shiaoman; Condamine, Pascal; Heinen, Shane; Resnik, Josh; Wing, Rod; Witt, Heather N; Alpert, Matthew; Beccuti, Marco; Bozdag, Serdar; Cordero, Francesca; Mirebrahim, Hamid; Ounit, Rachid; Wu, Yonghui; You, Frank; Zheng, Jie; Simková, Hana; Dolezel, Jaroslav; Grimwood, Jane; Schmutz, Jeremy; Duma, Denisa; Altschmied, Lothar; Blake, Tom; Bregitzer, Phil; Cooper, Laurel; Dilbirligi, Muharrem; Falk, Anders; Feiz, Leila; Graner, Andreas; Gustafson, Perry; Hayes, Patrick M; Lemaux, Peggy; Mammadov, Jafar; Close, Timothy J

    2015-10-01

    Barley (Hordeum vulgare L.) possesses a large and highly repetitive genome of 5.1 Gb that has hindered the development of a complete sequence. In 2012, the International Barley Sequencing Consortium released a resource integrating whole-genome shotgun sequences with a physical and genetic framework. However, because only 6278 bacterial artificial chromosome (BACs) in the physical map were sequenced, fine structure was limited. To gain access to the gene-containing portion of the barley genome at high resolution, we identified and sequenced 15 622 BACs representing the minimal tiling path of 72 052 physical-mapped gene-bearing BACs. This generated ~1.7 Gb of genomic sequence containing an estimated 2/3 of all Morex barley genes. Exploration of these sequenced BACs revealed that although distal ends of chromosomes contain most of the gene-enriched BACs and are characterized by high recombination rates, there are also gene-dense regions with suppressed recombination. We made use of published map-anchored sequence data from Aegilops tauschii to develop a synteny viewer between barley and the ancestor of the wheat D-genome. Except for some notable inversions, there is a high level of collinearity between the two species. The software HarvEST:Barley provides facile access to BAC sequences and their annotations, along with the barley-Ae. tauschii synteny viewer. These BAC sequences constitute a resource to improve the efficiency of marker development, map-based cloning, and comparative genomics in barley and related crops. Additional knowledge about regions of the barley genome that are gene-dense but low recombination is particularly relevant.

  3. Barley Leaf Area and Leaf Growth Rates Are Maximized during the Pre-Anthesis Phase

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    Ahmad M. Alqudah

    2015-04-01

    Full Text Available Leaf developmental traits are an important component of crop breeding in small-grain cereals. Surprisingly, little is known about the genetic basis for the differences in barley (Hordeum vulgare L. leaf development. The two barley row-type classes, i.e., two- and six-rowed, show clear-cut differences in leaf development. To quantify these differences and to measure the genetic component of the phenotypic variance for the leaf developmental differences in both row-type classes we investigated 32 representative spring barley accessions (14 two- and 18 six-rowed accessions under three independent growth conditions. Leaf mass area is lower in plants grown under greenhouse (GH conditions due to fewer, smaller, and lighter leaf blades per main culm compared to pot- and soil-grown field plants. Larger and heavier leaf blades of six-rowed barley correlate with higher main culm spike grain yield, spike dry weight, and harvest index; however, smaller leaf area (LA in two-rowed barley can be attributed to more spikes, tillers, and biological yield (aboveground parts. In general, leaf growth rate was significantly higher between awn primordium and tipping stages. Moderate to very high broad-sense heritabilities (0.67–0.90 were found under all growth conditions, indicating that these traits are predominantly genetically controlled. In addition, our data suggests that GH conditions are suitable for studying leaf developmental traits. Our results also demonstrated that LA impacts single plant yield and can be reconsidered in future breeding programs. Six-rowed spike 1 (Vrs1 is the major determinate of barley row-types, the differences in leaf development between two- and six-rowed barleys may be attributed to the regulation of Vrs1 in these two classes, which needs further testing.

  4. Lipid composition of organelles from germinating castor bean endosperm

    Energy Technology Data Exchange (ETDEWEB)

    Donaldson, R.P.; Beevers, H.

    1977-02-01

    Glyoxysome, endoplasmic reticulum, mitochondria, and proplastid fractions were isolated from endosperm of castor beans (Ricinus communis) germinated for 5 days at 30 C. Samples from sucrose density gradients were diluted with 0.15 M KCl and the membranes pelleted. Lipid extracts of these membranes were analyzed for phosphoglyceride, acyl lipid, and sterol content. The endoplasmic reticulum contains 1.24 ..mu..mol of phosphoglyceride per mg of protein; the mitochondria, 0.65 ..mu..mol/mg; and the glyoxysome membranes, 0.55 ..mu..mol/mg. Phosphatidyl choline and phosphatidyl ethanolamine are the most abundant lipids in all membranes studied, accounting for 70% or more of the lipid phosphorus and 50% or more of the fatty acid. Glyoxysome membranes and endoplasmic reticulum also contain phosphatidyl inositol (respectively, 9 and 17% of the lipid phosphorus) and free fatty acids (13% of the total fatty acid in each). Compared with other organelles, mitochondrial membranes have more phosphatidyl ethanolamine relative to phosphatidyl choline and are characterized by the presence of cardiolipin, in which 80% of the fatty acid is linoleate. The relative amounts of linoleate, palmitate, oleate, stearate, and linolenate in each of the phosphotoglycerides are constant regardless of the membrane source. Stimasgasterol and ..beta..-sitosterol are present in the membranes (1 to 9 nmol each/mg protein). The data provide further evidence that glyoxysome membranes are derived from the endoplasmic reticulum but at the same time indicate some differentiation.

  5. Distribution of soluble amino acids in maize endosperm mutants

    Directory of Open Access Journals (Sweden)

    Toro Alejandro Alberto

    2003-01-01

    Full Text Available For human nutrition the main source of vegetable proteins are cereal and legume seeds. The content of total soluble amino acids in mature endosperm of wild-type, opaque and floury maize (Zea mays L. mutants were determined by HPLC. The total absolute concentration of soluble amino acids among the mutants varied depending on the mutant. The o11 and o13 mutants exhibited the highest average content, whereas o10, fl3 and fl1 exhibited the lowest average content. In general, the mutants exhibited similar concentrations of total soluble amino acids when compared to the wild-type lines, with the clear exception of mutants o11 and fl1, with the o11 mutant exhibiting a higher concentration of total soluble amino acids when compared to its wild-type counterpart W22 and the fl1 mutant a lower concentration when compared to its wild-type counterpart Oh43. For methionine, the mutants o2 and o11 and wild-type Oh43 exhibited the highest concentrations of this amino acid. Significant differences were not observed between mutants for other amino acids such as lysine and threonine. The high lysine concentrations obtained originally for these mutants may be due to the amino acids incorporated into storage proteins, but not those present in the soluble form.

  6. 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.

  7. 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.

  8. 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...

  9. 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...

  10. 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..

  11. Abscisic acid controls embryo growth potential and endosperm cap weakening during coffee (Coffea arabica cv. Rubi) seed germination

    NARCIS (Netherlands)

    Silva, da E.A.A.; Toorop, P.E.; Aelst, van A.C.; Hilhorst, H.W.M.

    2004-01-01

    The mechanism and regulation of coffee seed germination were studied in Coffea arabica L. cv. Rubi. The coffee embryo grew inside the endosperm prior to radicle protrusion and abscisic acid (ABA) inhibited the increase in its pressure potential. There were two steps of endosperm cap weakening. An in

  12. Fermentation of the endosperm cell walls of monocotyledon and dicotyledon plant species: The relationship between cell wall characteristics and fermentability

    NARCIS (Netherlands)

    Laar, van H.; Tamminga, S.; Williams, B.A.; Verstegen, M.W.A.

    2000-01-01

    Cell walls from the endosperm of four monocotyledons (maize, wheat, rye, and rice) and four dicotyledons (soya bean, lupin, faba bean, and pea) seeds were studied to relate cell wall composition and structure with fermentation characteristics. Cell wall material was isolated from the endosperm of th

  13. Block-surface staining for differentiation of starch and cell walls in wheat endosperm.

    Science.gov (United States)

    Glenn, G M; Pitts, M J; Liao, K; Irving, D W

    1992-03-01

    A staining technique for differentiating starch granules and cell walls was developed for computer-assisted studies of starch granule distribution in cells of wheat (Triticum aestivum L.) caryopses. Blocks of embedded caryopses were sectioned, exposing the endosperm tissue, and stained with iodine potassium iodide (IKI) and Calcofluor White. Excessive tissue hydration during staining was avoided by using stains prepared in 80% ethanol and using short staining times. The IKI quenched background fluorescence which facilitated the use of higher concentrations of Calcofluor White. Cell wall definition was improved with the IKI-Calcofluor staining combination compared to Calcofluor alone. The high contrast between darkly stained starch granules and fluorescent cell walls permitted computer assisted analysis of data from selected hard and soft wheat varieties. The ratio of starch granule area to cell area was similar for both wheat classes. The starch granule sizes ranged from 2.1 microns 3 to 22,000 microns 3 with approximately 90% of the granules measuring less than 752 microns 3 (ca. 11 microns in diameter). Hard wheat samples had a greater number of small starch granules and a lower mean starch granule area compared to the soft wheat varieties tested. The starch size distribution curve was bimodal for both the hard and soft wheat varieties. Three-dimensional starch size distribution was measured for four cells near the central cheek region of a single caryopsis. The percentage of small granules was higher at the ends than at the mid-section of the cells.

  14. Mapping genes in barley for resistance to Puccinia coronata from couch grass and to P. striiformis from brome, wheat and barley

    NARCIS (Netherlands)

    Niks, R.E.; Alemu, Sisay K.; Marcel, T.C.; Heyzen, van Skye

    2015-01-01

    Barley (Hordeum vulgare L.) mapping populations have been developed that are useful to study the inheritance of quantitative resistance to adapted and unadapted rust fungi. In a recent host range study, we found that the parents of those mapping populations also differed in their resistance to th

  15. 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...

  16. 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.

  17. Rice endosperm is cost-effective for the production of recombinant griffithsin with potent activity against HIV.

    Science.gov (United States)

    Vamvaka, Evangelia; Arcalis, Elsa; Ramessar, Koreen; Evans, Abbey; O'Keefe, Barry R; Shattock, Robin J; Medina, Vicente; Stöger, Eva; Christou, Paul; Capell, Teresa

    2016-06-01

    Protein microbicides containing neutralizing antibodies and antiviral lectins may help to reduce the rate of infection with human immunodeficiency virus (HIV) if it is possible to manufacture the components in large quantities at a cost affordable in HIV-endemic regions such as sub-Saharan Africa. We expressed the antiviral lectin griffithsin (GRFT), which shows potent neutralizing activity against HIV, in the endosperm of transgenic rice plants (Oryza sativa), to determine whether rice can be used to produce inexpensive GRFT as a microbicide ingredient. The yield of (OS) GRFT in the best-performing plants was 223 μg/g dry seed weight. We also established a one-step purification protocol, achieving a recovery of 74% and a purity of 80%, which potentially could be developed into a larger-scale process to facilitate inexpensive downstream processing. (OS) GRFT bound to HIV glycans with similar efficiency to GRFT produced in Escherichia coli. Whole-cell assays using purified (OS) GRFT and infectivity assays using crude extracts of transgenic rice endosperm confirmed that both crude and pure (OS) GRFT showed potent activity against HIV and the crude extracts were not toxic towards human cell lines, suggesting they could be administered as a microbicide with only minimal processing. A freedom-to-operate analysis confirmed that GRFT produced in rice is suitable for commercial development, and an economic evaluation suggested that 1.8 kg/ha of pure GRFT could be produced from rice seeds. Our data therefore indicate that rice could be developed as an inexpensive production platform for GRFT as a microbicide component.

  18. Purification and characterisation of arabinoxylan arabinofuranohydrolase I responsible for the filterability of barley malt.

    Science.gov (United States)

    Li, Xiaomin; Gao, Fei; Cai, Guolin; Jin, Zhao; Lu, Jian; Dong, Jianjun; Yin, Hua; Yu, Junhong; Yang, Mei

    2015-05-01

    Dan'er is a widely grown malt barley cultivar in China, but its filterability defects have severely impeded its application in beer brewing. Previous investigations have suggested that we should identify the malt filterability correlated proteins, one of which was postulated to be arabinoxylan arabinofuranohydrolase I (AXAH-I). To verify this hypothesis, we purified AXAH-I from Dan'er malt, characterised its enzyme performance, and investigated its influence on filterability by adding different amounts of purified enzyme to the mash. With 6 mU g(-1) malt AXAH-I supplemented, the wort separation rate increased by 31.8%, viscosity decreased by 3.6%, and the endosperm reserve contents declined concomitantly. Unexpectedly, the wort turbidity increased with increasing AXAH-I. We also tried to optimise the use of currently available commercial enzyme products for filterability improvement in beer brewing, by supplementing them with purified AXAH-I and β-amylase. AXAH-I could be a functional component for novel commercial enzyme products in the beer industry.

  19. 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)....

  20. Estimation of Barley (Hordeum Vulgare L. Crop Water Requirements Using Cropwat Software in Ksar-Chellala Region, Algeria

    Directory of Open Access Journals (Sweden)

    M. B. Laouisset

    2016-09-01

    Full Text Available This paper estimates the reference Evapotranspiration (ET0 and Water requirements of barley (Hordeum vulgare L. in Ksar-Chellala region, Algeria, for one dry year by using CROPWAT software. Determination of Evapotranspiration ( ET is important in application such as irrigation design, irrigation scheduling, water resource management, hydrology and cropping systems modeling. Estimation of crop water requirements of barley ( CWR b respected the methodology adopted by the service of development and management service of FAO, based on the use of software CROPWAT 8.0. The total water requirements for barley depend on a variety of target yields and crops management. The period of climatic data used is 23 years (1990-2012, the average rain in this period is 254 mm. The total rain of the dry year is 190 mm. The results of this study show, during the vegetative cycle of barley which is 6 months, the calculation of ET 0 is 453 mm, the potential water which was used by the crop barley is estimated at 281.4 mm, the efficiency of rainfall is 69 mm and a total water requirements of barley ( CWR b equals to 211 mm, this amount distributed on three months coincided with important stages of development in barley. The supplementary irrigation in these conditions with optimal contents equals water requirements estimated by CROPWAT software that increases significantly grain yield of barely. Consequently, the gross irrigation water requirements ( GIWR of 1250000 ha which project to grow barley in the Algerian steppes regions are estimated at 3.77 billion and this for a dry year and a irrigation efficiency of 70%.

  1. 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...

  2. Impact of Low Concentration of Cadmium on Photosynthesis and Growth of Pea and Barley

    Directory of Open Access Journals (Sweden)

    Irena Januškaitienė

    2010-10-01

    Full Text Available Photosynthetic gas exchange and growth characteristics were examined in pea and barley plants using 1 mM Cd treatment. Plants were sown into neutral peat substrate and at a leaf development stage were treated with 1 mM cadmium concentration solution. Gas exchange parameters (photosynthetic rate; intercellular CO2 concentration; transpiration rate; water use efficiency were measured with portable photosynthesis system LI-6400 on the fifth day after Cd treatment. Under Cd stress the photosynthetic rate of pea and barley plants decreased by 16.7 % (p 2 concentration decreased by 27.4 % (p 2 reduction processes of Cd treated pea leaves increased (because intercellular CO2 concentration decreased, but that had no positive effect on a photosynthetic rate, and the photosynthetic rate of pea decreased by 4 % more than that of barley. The changes of dry biomass of cadmium treated plants were weak and statistically insignificant.

  3. Effect of Soil Erosion on Spring Barley Growth in East Anglia,England:Preliminary Results

    Institute of Scientific and Technical Information of China (English)

    LUXI-XI; Y.BIOT

    1994-01-01

    The effect of soil erosion on spring barley growth was studied on a deep loamy soil in East Anglia,England,in 1992,Soil erosion was simulated by three levels of soil desurfacing,7.5,18and 30cm with three replicates.Significant differences in crop height,ground cover and crop yield were observed between the three levels of desurfacing.Soil desurfacing also has a singnificant effect on soil moisture at the 20cm depth.The interaction between soil removal and crop performance affected soil moisture at the depths of 50 and 100cm,No significant differences were found in runoff and sediment etween the three topsoil removals due to very dry growing season.Regression equations were developed between spring barley yield and soil desurfacing Spring barley grain yield declined by 97.6kg/ha per cm soil desurfacing.

  4. 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...

  5. The development and evaluation of single cell suspension from wheat and barley as a model system; a first step towards functional genomics application

    DEFF Research Database (Denmark)

    Dong, Jing; Bowra, Steve; Vincze, Éva

    2010-01-01

    chose to study the impact of selected abiotic stress elicitors at the physiological, biochemical and molecular level. We report the results of osmotic stress imposed by NaCl and PEG. As proline is an important osmoprotectant of the cereal cells, colorimetric assay for proline detection was developed...

  6. NAC Transcription Factors of Barley (Hordeum vulgare L.) and their Involvement in Leaf Senescence

    DEFF Research Database (Denmark)

    Wagner, Michael

    yielding cereal crops are generated. In cereals, the process of leaf senescence is of utmost relevance when discussing yield. It is during the senescence process that all nutrients are transported from the withering leaf to the developing grains. Furthermore, the timing of senescence determines...... parts of the senescence process. The specific aims of this study were therefore (1) to establish and characterise the NAC transcription factors of the model cereal crop barley (Hordeum vulgare L.) (2) to identify and study putative barley NAC transcription factors involved in the regulation of leaf...

  7. 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....

  8. 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....

  9. Influence of instrument rigidity and specimen geometry on calculations of compressive strength properties of wheat endosperm

    Science.gov (United States)

    Endosperm texture is one of the most important quality features in wheat that defines milling energy requirements and the suitability of flour or semolina for the various food products such as pan breads, crackers, cakes, and pastas. Rooted in low molecular weight proteins known as puroindolines a a...

  10. Proteomic analysis of rice endosperm cells in response to expression of hGM-CSF.

    Science.gov (United States)

    Luo, Junling; Ning, Tingting; Sun, Yunfang; Zhu, Jinghua; Zhu, Yingguo; Lin, Qishan; Yang, Daichang

    2009-02-01

    The accumulation of significant levels of transgenic products in plant cells is required not only for crop improvement, but also for molecular pharming. However, knowledge about the fate of transgenic products and endogenous proteins in grain cells is lacking. Here, we utilized a quantitative mass spectrometry-based proteomic approach for comparative analysis of expression profiles of transgenic rice endosperm cells in response to expression of a recombinant pharmaceutical protein, human granulocyte-macrophage colony stimulation factor (hGM-CSF). This study provided the first available evidence concerning the fate of exogenous and endogenous proteins in grain cells. Among 1883 identified proteins with a false positive rate of 5%, 103 displayed significant changes (p-value < 0.05) between the transgenic and the wild-type endosperm cells. Notably, endogenous storage proteins and most carbohydrate metabolism-related proteins were down-regulated, while 26S proteasome-related proteins and chaperones were up-regulated in the transgenic rice endosperm. Furthermore, it was observed that expression of hGM-CSF induced endoplasmic reticulum stress and activated the ubiquitin/26S-proteasome pathway, which led to ubiquitination of this foreign gene product in the transgenic rice endosperm.

  11. Reorganization of microtubular cytoskeleton and formation of cellular processes during post-telophase in haemanthus endosperm.

    Science.gov (United States)

    Bajer, A S; Smirnova, E A

    1999-10-01

    We followed time-dependent post-telophase reorganization of the microtubule cytoskeleton on immunostained preparations of endosperm of the higher plant Haemanthus. After completion of mitosis, the phragmoplast continued to reorganize for several hours. This prompted the formation of phragmoplast-like derivatives (secondary and accessory phragmoplasts and peripheral microtubular ring). Next, elongated cellular protrusions (processes) appeared at the cell periphery. These processes contained long microtubule bundles and disorderly arranged actin filaments. Microtubule converging centers or accessory phragmoplasts were often present at the tips of the processes. Observation in vivo demonstrated that processes were formed at the cell periphery as extensions of lammelipodia or filopodia-type protrusions that commonly terminated with cytoplasmic blobs. We suggest that processes are derivatives of a peripheral microtubular ring that reorganizes gradually into cellular protrusions. Endosperm processes have several features of neuronal cells, or animal somatic cells with overexpressed MAPs. Since microtubule-containing processes were never detected shortly after extrusion of the cells from the embryo sac, this course of events might be restricted specifically to extruded endosperm and triggered either by removal of cells, their placement in monolayer on agar substrate, or both. Thus, post telophase behavior of endosperm cells offers a novel experimental system for studies of cytoskeleton in higher plants.

  12. Fertilization in Torenia fournieri: actin organization and nuclear behavior in the central cell and primary endosperm

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Studies of the living embryo sacs of Torenia fournieri reveal that the actin cytoskeleton undergoes dramatic changes that correlate with nuclear migration within the central cell and the primary endosperm. Before pollination, actin filaments appear as short bundles randomly distributed in the cortex of the central cell. Two days after anthesis, they become organized into a distinct actin network. At this stage the secondary nucleus, which is located in the central region of the central cell, possesses an associated array of short actin filaments. Soon after pollination, the actin filaments become fragmented in the micropylar end and the secondary nucleus is located next to the egg apparatus. After fertilization, the primary endosperm nucleus moves away from the egg cell and actin filaments reorganize into a prominent network in the cytoplasm of the primary endosperm. Disruption of the actin cytoskeleton with latrunculin A and cytochalasin B indicates that actin is involved in the migration of the nucleus in the central cell. Our data also suggest that the dynamics of actin cytoskeleton may be responsible for the reorganization of the central cell and primary endosperm cytoplasm during fertilization.

  13. Proteomics of Wheat Endosperm: a Tool to Find Genes Involved in Kernel Composition and Quality

    Institute of Scientific and Technical Information of China (English)

    G. Branlard; E. Bancel; I. Nadaud

    2007-01-01

    @@ The composition of the wheat kernel is the result of the expression of thousands of genes translated in enzymes involved in all the biochemical pathways that are needed for endosperm cell functions and also for the accumulation of storage proteins and starch.

  14. EFFECT OF PHYSIOLOGICAL AGE AND GROWTH REGULATORS ON CALLUS BROWNING OF COCONUT ENDOSPERM CULTURE IN VITRO

    Directory of Open Access Journals (Sweden)

    LAZARUS AGUS SUKAMTO

    2011-01-01

    Full Text Available The possibility of physiological age and growth regulators affecting callus browning ofcoconut endosperm was investigated. Solid endosperm explants of four coconut fruits fromsame brunches of two coconut cultivars “Samoan Dwarf ” were grown on modified Murashigeand Skoog (MS formula with addition of 10 mg l putresine, 2.50 g l activated charcoal (AC,1.70 g l phytagel, 0, 10 , 10 , 10 , 10 M 2,4-dichlorophenoxyacetic acid (2,4-D or 4-amino-3,5,6-trichloropicolinic acid (Picloram combined with 10 M 6-benzylaminopurine (BA.Callogenesis occurred on 98.83% of explants. Callus browning between different physiologicalages (antipodal and micropylar tissues of coconut endosperm at 9, 26 and 31 weeks of culture(WOC was significantly different, but not at 16 and 21 WOC. Auxins of 2,4-D and Picloramdid not affect significantly callus browning of endosperm cultures. Auxin doses at 10 , 10 , and10 M decreased significantly callus browning at 9 and 16 WOC, respectively, but at 10 Mbrowning was less significant compared to other doses at 21 WOC. Auxin dose at 10 M causedless significant browning compared to other doses at 31 WOC. The addition of BA decreasedsignificantly callus browning at 9 WOC, but did not affect callus browning thereafter.

  15. Physicochemical and Sensory Properties of Boiled Prosopis africana Seed Endosperm Macerated in Various Ethanol-water Mixtures

    Directory of Open Access Journals (Sweden)

    James E. Obiegbuna

    2013-09-01

    Full Text Available The processing of boiled Prosopis africana endosperm for better utilization using ethanol-water mixtures was explored. Prosopis africana seeds were boiled for 5 h to softness and the endosperm fraction separated from the kernel (cotyledon and the hull. The endosperm was divided into five equal parts which were individually macerated in absolute (Abs ethanol, 80, 60 and 40% ethanol in water prior to sun-drying (32±2°C, 3 days. The fifth sample, which served as control, was left untreated with ethanol. The samples were ground using a hand milling machine and analyzed for the proximate composition, water and oil absorption capacities, foaming capacity and foam stability, bulk density, emulsion activity and stability, colour preference, texture preference and overall acceptability. The results revealed that treatment of the endosperm significantly affected the moisture, protein, fat, ash and carbohydrate contents; water and oil absorption capacities, foaming capacity and foam stability; and the sensory properties. The moisture and protein contents, oil absorption capacity, foam stability, appearance, texture and overall acceptability of endosperm treated with 40% ethanol in water differed significantly (p<0.05 from that treated with absolute ethanol. There was also significant (p<0.05 differences in moisture, protein and carbohydrate contents, oil absorption capacity and foam stability of the 40% ethanol in water treated endosperm and the control. Slightly above 40% ethanol in water (50-60% should be used to macerate Prosopis africana endosperm to reduce the cost of using absolute ethanol.

  16. 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...

  17. Effects of Different Comsteep Liquids on Structure of Starch and Protein Binding in Corn Endosperm

    Directory of Open Access Journals (Sweden)

    Rong Yan

    2015-07-01

    Full Text Available The changes in the structures of corn endosperms immersed in different reagents were studied, in order to preliminarily determine the mode of binding of starch and protein in the endosperm. The corn endosperm was immersed in Sulfurous acid, NaOH, ethanol, L-cysteine and morel berry, respectively. The dissociated starch content was analyzed using water as the reference. The results of the treatment were analyzed by the dissociated starch content, optical microscopy and scanning electron microscopy. The test results showed that by immersing in NaOH solution, the protein matrix was effectively decomposed, the protein in the endosperm was dissolved and the highest amount of starch granules were released. Sulfurous acid and L-cysteine solution could make the protein matrix into pieces by destroying intermolecular disulfide bridge and the starch granules would be released. There were some holes on the surface of starch granules after L-cysteine steeping. Immersed in the morel berry, there were holes on the protein matrix surface and the water absorption of corn endosperm was improved. In case of the sample immersed in ethanol, the zein was dissolved, whereas the matrix protein was still intact with the starch grains that still bound to it. By analysis of the microscopic structure and morphological change of starch and protein in corn endospers in different comsteep liquor, it is demonstrated that the main reason which the starch grains were tightly wrapped by Glutenin is the intermolecular disulfide bonds in glutelin. In addition, a part of the protein combined with starch granules directly. The study provided a reference for exploring the structure of binding of starch and protein and their interaction in corn.

  18. Effects of Zn fertilization on hordein transcripts at early developmental stage of barley grain and correlation with increased Zn concentration in the mature grain.

    Directory of Open Access Journals (Sweden)

    Mohammad Nasir Uddin

    Full Text Available Zinc deficiency is causing malnutrition for nearly one third of world populations. It is especially relevant in cereal-based diets in which low amounts of mineral and protein are present. In biological systems, Zn is mainly associated with protein. Cereal grains contain the highest Zn concentration during early developmental stage. Although hordeins are the major storage proteins in the mature barley grain and suggested to be involved in Zn binding, very little information is available regarding the Zn fertilization effects of hordein transcripts at early developmental stage and possible incorporation of Zn with hordein protein of matured grain. Zinc fertilization experiments were conducted in a greenhouse with barley cv. Golden Promise. Zn concentration of the matured grain was measured and the results showed that the increasing Zn fertilization increased grain Zn concentration. Quantitative real time PCR showed increased level of total hordein transcripts upon increasing level of Zn fertilization at 10 days after pollination. Among the hordein transcripts the amount of B-hordeins was highly correlated with the Zn concentration of matured grain. In addition, protein content of the matured grain was analysed and a positive linear relationship was found between the percentage of B-hordein and total grain Zn concentration while C-hordein level decreased. Zn sensing dithizone assay was applied to localize Zn in the matured grain. The Zn distribution was not limited to the embryo and aleurone layer but was also present in the outer part of the endosperm (sub-aleurone layers which known to be rich in proteins including B-hordeins. Increased Zn fertilization enriched Zn even in the endosperm. Therefore, the increased amount of B-hordein and decreased C-hordein content suggested that B-hordein upregulation or difference between B and C hordein could be one of the key factors for Zn biofortification of cereal grains due to the Zn fertilization.

  19. 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

  20. 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...

  1. 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 ...

  2. Programmed cell death: The life ambition of the barley aleurone layer

    DEFF Research Database (Denmark)

    Mark, Christina; Zor, Kinga; Heiskanen, Arto

    We have developed a 24-well multiplate tissue culture system with electrochemical and optical detection techniques for cultivation of immobilised barley aleurone layers. We have applied the system for the purpose of studying the underlying mechanisms of programmed cell death (PCD) in plants. We h...

  3. High diversity of genes for nonhost resistance of barley to heterologous rust fungi

    NARCIS (Netherlands)

    Jafary, H.; Albertazzi, G.; Marcel, T.C.; Niks, R.E.

    2008-01-01

    Inheritance studies on the nonhost resistance of plants would normally require interspecific crosses that suffer from sterility and abnormal segregation. Therefore, we developed the barley¿Puccinia rust model system to study, using forward genetics, the specificity, number, and diversity of genes in

  4. Genetic mapping of the barley lodging resistance locus Erectoides-k

    DEFF Research Database (Denmark)

    Kristensen, Peter Skov; Dockter, Christoph; Lundqvist, Udda;

    2016-01-01

    The barley (Hordeum vulgare L.) mutant erectoides-k.32 (ert-k.32) was isolated in 1947 from an X-ray-mutant population of cultivar ‘Bonus’. The mutant was released as a cultivar in 1958 with the name ‘Pallas’ – one of the first cereal crop cultivars developed from induced mutants. ‘Pallas’ is a s...

  5. 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.

  6. Recombinant human proinsulin from transgenic corn endosperm: solvent screening and extraction studies

    Directory of Open Access Journals (Sweden)

    C. S. Farinas

    2007-09-01

    Full Text Available Recombinant pharmaceutical proteins are being produced in different systems such as bacteria and mammalian cell cultures. The use of transgenic plants as bioreactors has recently arisen as an alternative system offering many practical and economic advantages. However, finding an optimum strategy for the downstream processing (DSP of recombinant proteins from plants still remains a challenge. In this work, we studied the extraction of recombinant human proinsulin (rhProinsulin produced in the endosperm of transgenic corn seeds. An efficient extraction solvent was selected and the effects of temperature, solvent-to-solid ratio, time, and impeller rotational speed on the extraction were evaluated using an experimental design. After an extraction kinetics study, temperature was further evaluated to maximize rhProinsulin concentration in the extracts and to minimize the native corn components carbohydrates, phenolic compounds, and proteins. A high efficiency condition for extracting rhProinsulin with the selected solvent - 50 mM sodium bicarbonate buffer pH 10.0 and 5 mM DTT - was an extraction time of 2 h at a solvent-to-solid ratio of 10:1 and 25º C. The maximum rhProinsulin concentration in the extracts at that condition was 18.87 mg l-1 or 0.42% of the total soluble protein. These values are within the range in which the production of pharmaceutical proteins in plants can be competitive with other expression systems. The results presented provide information for the development of an additional production platform for the hormone insulin.

  7. Early stages of spindle formation and independence of chromosome and microtubule cycles in Haemanthus endosperm.

    Science.gov (United States)

    Smirnova, E A; Bajer, A S

    1998-01-01

    We analyzed transformation of the interphase microtubular cytoskeleton into the prophase spindle and followed the pattern of spindle axis determination. Microtubules in endosperm of the higher plant Haemanthus (Scadoxus) were stained by the immunogold and immunogold silver-enhanced methods. Basic structural units involved in spindle morphogenesis were "microtubule converging centers." We emphasized the importance of relative independence of chromosomal and microtubular cycles, and the influence of these cycles on the progress of mitosis. Cells with moderately desynchronized cycles were functional, but extreme desynchronization led to aberrant mitosis. There were three distinct phases of spindle development. The first one comprised interphase and early to mid-prophase. During this phase, the interphase microtubule meshwork radiating from the nuclear surface into the cytoplasm rearranged and formed a dense microtubule cage around the nucleus. The second phase comprised mid to late prophase, and resulted in the formation of normal (bipolar) or transitory aberrant (apolar or multipolar) prophase spindles. The third phase comprised late prophase with prometaphase. The onset of prometaphase was accompanied by a rapid association of microtubule converging centers with kinetochores. In this stage aberrant spindles transformed invariably into bipolar ones. Lateral association of a few bipolar kinetochore fibers at early prometaphase established the core of the bipolar spindle and its alignment. We concluded that (1) spindle formation is a largely independent microtubular process modified by the chromosomal/kinetochore cycle; and (2) the initial polarity of the spindle is established by microtubule converging centers, which are a functional substitute of the centrosome/MTOC. We believe that the dynamics of microtubule converging centers is an expression of microtubule self-organization driven by motor proteins as proposed by Mitchison [1992: Philos. Trans. R. Soc. Lond. B

  8. Proteomic and genetic analysis of wheat endosperm albumins and globulins using deletion lines of cultivar Chinese Spring

    DEFF Research Database (Denmark)

    Merlino, Marielle; Bousbata, Sabrina; Svensson, Birte;

    2012-01-01

    Albumins and globulins from the endosperm of Triticum aestivum L. cv Chinese Spring (CS) were analysed to establish a proteome reference map for this standard wheat cultivar. Approximately, 1,145 Coomassie-stained spots were detected by two-dimensional gel electrophoresis (2DE), 410 of which were...... the composition and genetics of a complex tissue, such as the wheat endosperm.......Albumins and globulins from the endosperm of Triticum aestivum L. cv Chinese Spring (CS) were analysed to establish a proteome reference map for this standard wheat cultivar. Approximately, 1,145 Coomassie-stained spots were detected by two-dimensional gel electrophoresis (2DE), 410 of which were...... in endosperm proteins due to chromosomal deletions. This differential analysis of spots allowed structural or regulatory genes, encoding 211 proteins, to be located on segments of the 21 wheat chromosomes. In addition, variance analysis of quantitative variations in spot volume showed that the expression...

  9. Starch granule formation and protein deposition in wheat (Triticum aestivum L.) starchy endosperm cells is altered by high temperature during grain fill

    Science.gov (United States)

    Hurkman, William J.; Wood, Delilah F.

    2010-06-01

    High temperatures during wheat grain fill decrease starch and protein levels, adversely affecting wheat yield and flour quality. To determine the effect of high temperature on starchy endosperm cell development, grain (Triticum aestivum L. 'Butte 86') was produced under a 24/17°C or 37/28°C day/night regimen imposed from flowering to maturity and starch and protein deposition examined using scanning electron microscopy. The high temperature regimen shortened the duration of grain fill from 40 to 18 days. Under the 37/28°C regimen, A- and B-type starch granules decreased in size. A-type starch granules also exhibited pitting, suggesting enhanced action of starch degradative enzymes. Under both temperature regimens, protein bodies originated early in development and coalesced during mid to late development to form a continuous protein matrix surrounding the starch granules. Under the 37/28°C regimen, the proportion of protein matrix increased in endosperm cells of mature grain. Taken together, the changes in starch granule number and size and in protein matrix amount provide clues for understanding how high temperature during grain fill can affect end use properties of wheat flour.

  10. Mobilization of lipid reserves during germination of oat (Avena sativa L.), a cereal rich in endosperm oil.

    Science.gov (United States)

    Leonova, Svetlana; Grimberg, Asa; Marttila, Salla; Stymne, Sten; Carlsson, Anders S

    2010-06-01

    Since the cereal endosperm is a dead tissue in the mature grain, beta-oxidation is not possible there. This raises the question about the use of the endosperm oil in cereal grains during germination. In this study, mobilization of lipids in different tissues of germinating oat grains was analysed using thin-layer and gas chromatography. The data imply that the oat endosperm oil [triacylglycerol (TAG)] is not a dead-end product as it was absorbed by the scutellum, either as free fatty acids (FFAs) released from TAG or as intact TAG immediately degraded to FFAs. These data were supported by light and transmission electron microscopy (LM and TEM) studies where close contact between endosperm lipid droplets and the scutellum was observed. The appearance of the fused oil in the oat endosperm changed into oil droplets during germination in areas close to the aleurone and the scutellar epithelium. However, according to the data obtained by TEM these oil droplets are unlikely to be oil bodies surrounded by oleosins. Accumulation of FFA pools in the embryo suggested further transport of FFAs from the scutellum. Noticeably high levels of TAG were also accumulated in the embryo but were not synthesized by re-esterification from imported FFAs. Comparison between two oat cultivars with different amounts of oil and starch in the endosperm suggests that an increased oil to starch ratio in oat grains does not significantly impact the germination process.

  11. 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...

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 454 sequencing of pooled BAC clones on chromosome 3H of barley

    Directory of Open Access Journals (Sweden)

    Yamaji Nami

    2011-05-01

    Full Text Available Abstract Background Genome sequencing of barley has been delayed due to its large genome size (ca. 5,000Mbp. Among the fast sequencing systems, 454 liquid phase pyrosequencing provides the longest reads and is the most promising method for BAC clones. Here we report the results of pooled sequencing of BAC clones selected with ESTs genetically mapped to chromosome 3H. Results We sequenced pooled barley BAC clones using a 454 parallel genome sequencer. A PCR screening system based on primer sets derived from genetically mapped ESTs on chromosome 3H was used for clone selection in a BAC library developed from cultivar "Haruna Nijo". The DNA samples of 10 or 20 BAC clones were pooled and used for shotgun library development. The homology between contig sequences generated in each pooled library and mapped EST sequences was studied. The number of contigs assigned on chromosome 3H was 372. Their lengths ranged from 1,230 bp to 58,322 bp with an average 14,891 bp. Of these contigs, 240 showed homology and colinearity with the genome sequence of rice chromosome 1. A contig annotation browser supplemented with query search by unique sequence or genetic map position was developed. The identified contigs can be annotated with barley cDNAs and reference sequences on the browser. Homology analysis of these contigs with rice genes indicated that 1,239 rice genes can be assigned to barley contigs by the simple comparison of sequence lengths in both species. Of these genes, 492 are assigned to rice chromosome 1. Conclusions We demonstrate the efficiency of sequencing gene rich regions from barley chromosome 3H, with special reference to syntenic relationships with rice chromosome 1.

  17. Nitrogen and phosphorus compounds in the aleurone grains of Iris pseudoacorus endosperm and Pisum sativum cotyledons

    OpenAIRE

    Ligia Konopska

    2015-01-01

    Aleurone grains from Iris pseudoacorus endosperm and Pisum sativum cotyledons were isolated partly according to Tombs's method (1967). Nitrogen compounds content was determined in them by Kjeldahl's micromethod, and in the particular fractions after Thiman and Laloraya (1960). Mainly protein N was detected in the aleurone grains, constituting 14.8 and 15.2 per cent of the dry mass of pea and Iris seeds, respectively. Moreover, phosphorus compounds were fractionated according to Holden and Pir...

  18. 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...

  19. 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.

  20. Identification of Mild Freezing Shock Response Pathways in Barley Based on Transcriptome Profiling.

    Science.gov (United States)

    Wang, Xiaolei; Wu, Dezhi; Yang, Qian; Zeng, Jianbin; Jin, Gulei; Chen, Zhong-Hua; Zhang, Guoping; Dai, Fei

    2016-01-01

    Low temperature is a major abiotic stress affecting crop growth and productivity. A better understanding of low temperature tolerance mechanisms is imperative for developing the crop cultivars with improved tolerance. We herein performed an Illumina RNA-sequencing experiment using two barley genotypes differing in freezing tolerance (Nure, tolerant and Tremois, sensitive), to determine the transcriptome profiling and genotypic difference under mild freezing shock treatment after a very short acclimation for gene induction. A total of 6474 differentially expressed genes, almost evenly distributed on the seven chromosomes, were identified. The key DEGs could be classified into six signaling pathways, i.e., Ca(2+) signaling, PtdOH signaling, CBFs pathway, ABA pathway, jasmonate pathway, and amylohydrolysis pathway. Expression values of DEGs in multiple signaling pathways were analyzed and a hypothetical model of mild freezing shock tolerance mechanism was proposed. Expression and sequence profile of HvCBFs cluster within Frost resistance-H2, a major quantitative trait locus on 5H being closely related to low temperature tolerance in barley, were further illustrated, considering the crucial role of HvCBFs on freezing tolerance. It may be concluded that multiple signaling pathways are activated in concert when barley is exposed to mild freezing shock. The pathway network we presented may provide a platform for further exploring the functions of genes involved in low temperature tolerance in barley.

  1. 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.

  2. 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...

  3. 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...

  4. 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...

  5. 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.

  6. Evaluation and characterization of an endosperm-specific sbeⅡa promoter in wheat

    Institute of Scientific and Technical Information of China (English)

    MIAO Hongmei; Fleming Joy E.; L(U) Debing; HAN Jinfeng

    2004-01-01

    Starch, the main component of the wheat grain,is the product of a complex biochemical pathway. The sbeⅡa gene plays a key role in controlling the synthesis of starch, in particular, the biosynthesis of amylopectin, in maturing wheat grain. To investigate its regulatory mechanisms and endosperm-specific expression pattern, the sbeⅡa promoter (3094 bp in length) was cloned using APCR and sequenced.The effect of a series of deletions was studied using a GUS transient assay system. Results showed that the 3094 bp sequence (sbe.g construct) exhibited full stable promoting activity and that the activities of 5′ or 3′ deletions reduced levels of GUS expression. Some constructs with internal deletions showed only weak activity, however, sbe.e, with a deletion from -1579-1210 bp resulted in higher levels of expression than the full-length promoter sequence, sbe.g. This indicates that motifs such as the -300 bp element, G-box and/or P-box act as positive elements and are necessary in determining the promoter's endosperm-specific pattern and that negative repressor elements or motifs may also be present within the -1579-1210 bp sequence. The age of wheat endosperm tissue used in the GUS-transient assay system is shown to be of significant importance.

  7. DNA content in embryo and endosperm of maize kernel (Zea mays L.): impact on GMO quantification.

    Science.gov (United States)

    Trifa, Youssef; Zhang, David

    2004-03-10

    PCR-based techniques are the most widely used methods for the quantification of genetically modified organisms (GMOs) through the determination of the ratio of transgenic DNA to total DNA. It is shown that the DNA content per mass unit is significantly different among 10 maize cultivars. The DNA contents of endosperms, embryos, and teguments of individual kernels from 10 maize cultivars were determined. According to our results, the tegument's DNA ratio reaches at maximum 3.5% of the total kernel's DNA, whereas the endosperm's and the embryo's DNA ratios are nearly equal to 50%. The embryo cells are diploid and made of one paternal and one maternal haploid genome, whereas the endosperm is constituted of triploid cells made of two maternal haploid genomes and one paternal haploid genome. Therefore, it is shown, in this study, that the accuracy of the GMO quantification depends on the reference material used as well as on the category of the transgenic kernels present in the mixture.

  8. 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.

  9. 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.

  10. 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....

  11. From field barley to malt: detection and specification of microbial activity for quality aspects.

    Science.gov (United States)

    Noots, I; Delcour, J A; Michiels, C W

    1999-01-01

    Barley grain carries a numerous, variable, and complex microbial population that mainly consists of bacteria, yeasts, and filamentous fungi and that can partly be detected and quantified using plating methods and microscopic and molecular techniques. The extent and the activity of this microflora are determined by the altering state of the grain and the environmental conditions in the malt production chain. Three ecological systems can be distinguished: the growing cereal in the field, the dry barley grain under storage, and the germinating barley kernel during actual malting. Microorganisms interact with the malting process both by their presence and by their metabolic activity. In this respect, interference with the oxygen uptake by the barley grain and secretion of enzymes, hormones, toxins, and acids that may affect the plant physiological processes have been studied. As a result of the interaction, microorganisms can cause important losses and influence malt quality as measured by brewhouse performance and beer quality. Of particular concern is the occurrence of mycotoxins that may affect the safety of malt. The development of the microflora during malt production can to a certain extent be controlled by the selection of appropriate process conditions. Physical and chemical treatments to inactivate the microbial population on the barley grain are suggested. Recent developments, however, aim to control the microbial activity during malt production by promoting the growth of desirable microbial cultures, selected either as biocontrol agents inhibiting mycotoxin-producing molds or as starter cultures actively contributing to malt modification. Such techniques may offer natural opportunities to improve the quality and safety of malt.

  12. CONTRIBUTION OF PRIMARY SPIKES VS TILLERS TO TOTAL DEOXYNIVALENOL IN HARVESTED GRAIN OF WHEAT AND BARLEY

    Directory of Open Access Journals (Sweden)

    Pravin Gautam

    2012-01-01

    Full Text Available Formation of higher number of tillers is desirable for higher grain yield in both wheat and barley; tillers have delayed physiological development. Studies addressing the contribution of secondary tillers in final DON concentration for wheat and barley are limited. This project aims to improve our understanding of the relationship between FHB symptoms and DON concentration by examining the contribution of primary spikes and secondary tillers as it relates to the final DON concentration. Experiments, established as split-split plot design with five replications, was be conducted in South Dakota with moderately resistant and susceptible cultivars of each spring wheat, winter wheat and barley. Two inoculation time treatments was applied; at the anthesis of primary spikes (Feekes 10.5 and at anthesis of tillers (Feekes 11.2. Primary spikes (inoculated at Feekes 10.5 and tillers (inoculated at Feekes 11.2 were harvested separately, analyzed for visually scabby kernels and deoxynivalenol following standard protocols. Levels of DON were statistically higher in main heads of barley and spring wheat in each cultivar compared to its tillers. In winter wheat, though there was higher DON level in tillers than main heads in each cultivar, it was not statistically different. Based upon our result, it is suggested that cultivars with high tillers number should be selected in breeding programs to develop cultivars with high yield and low DON. Similarly, planting density of wheat and barley can be adjusted such that it will result in higher numbers of tillers, increasing tillers proportion that might lower DON concentration in final harvest.

  13. Evolutionary history of barley cultivation in Europe revealed by genetic analysis of extant landraces

    Directory of Open Access Journals (Sweden)

    Jones Huw

    2011-11-01

    Full Text Available Abstract Background Understanding the evolution of cultivated barley is important for two reasons. First, the evolutionary relationships between different landraces might provide information on the spread and subsequent development of barley cultivation, including the adaptation of the crop to new environments and its response to human selection. Second, evolutionary information would enable landraces with similar traits but different genetic backgrounds to be identified, providing alternative strategies for the introduction of these traits into modern germplasm. Results The evolutionary relationships between 651 barley landraces were inferred from the genotypes for 24 microsatellites. The landraces could be divided into nine populations, each with a different geographical distribution. Comparisons with ear row number, caryopsis structure, seasonal growth habit and flowering time revealed a degree of association between population structure and phenotype, and analysis of climate variables indicated that the landraces are adapted, at least to some extent, to their environment. Human selection and/or environmental adaptation may therefore have played a role in the origin and/or maintenance of one or more of the barley landrace populations. There was also evidence that at least some of the population structure derived from geographical partitioning set up during the initial spread of barley cultivation into Europe, or reflected the later introduction of novel varieties. In particular, three closely-related populations were made up almost entirely of plants with the daylength nonresponsive version of the photoperiod response gene PPD-H1, conferring adaptation to the long annual growth season of northern Europe. These three populations probably originated in the eastern Fertile Crescent and entered Europe after the initial spread of agriculture. Conclusions The discovery of population structure, combined with knowledge of associated phenotypes and

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

    Science.gov (United States)

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

    2015-05-01

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

  15. 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.

  16. Comparison of the potential rate of population increase of brown and green color morphs of Sitobion avenae (Homoptera: Aphididae) on barley infected and uninfected with Barley yellow dwarf virus.

    Science.gov (United States)

    Hu, Zu-Qing; Zhao, Hui-Yan; Thieme, Thomas

    2014-06-01

    Life tables of brown and green color morphs of the English grain aphid, Sitobion avenae (Fabricius) reared on barley under laboratory conditions at 20 ± 1°C, 65% ± 5% relative humidity and a photoperiod of 16 : 8 h (L : D) were compared. The plants were either: (i) infected with the Barley yellow dwarf virus (BYDV); (ii) not infected with virus but previously infested with aphids; or (iii) healthy barley plants, which were not previously infested with aphids. Generally, both color morphs of S. avenae performed significantly better when fed on BYDV-infected plants than on plants that were virus free but had either not been or had been previously infested with aphids. Furthermore, when fed on BYDV-infected plants, green S. avenae developed significantly faster and had a significantly shorter reproductive period than the brown color morph. There were no significant differences in this respect between the two color morphs of S. avenae when they were reared on virus-free plants that either had been or not been previously infested with aphids. These results indicate that barley infected with BYDV is a more favorable host plant than uninfected barley for both the color morphs of S. avenae tested, particularly the green color morph.

  17. 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

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. Starch Branching Enzyme 3 Occurs Within Starch Granules in Rice Endosperms%淀粉分支酶3存在于水稻胚乳的淀粉粒之中

    Institute of Scientific and Technical Information of China (English)

    佃蔚敏; 姜华武; 吴平

    2003-01-01

    对水稻胚乳淀粉颗粒结合的淀粉分支酶进行了研究.酶活性分析表明水稻胚乳中存在着与淀粉颗粒结合的淀粉分支酶.氨基酸测序分析结果表明结合于水稻胚乳淀粉粒的淀粉分支酶是分子量为84 kD的淀粉分支酶3(rice starch branching enzyme 3; RBE3).从开花后5 d到种子成熟,淀粉颗粒结合的RBE3蛋白都保持较为稳定的含量.Northern 分析表明水稻胚乳发育过程中RBE4最先表达而RBE3和RBE1的表达滞后.综合以上研究结果说明RBE3存在于水稻胚乳的淀粉之中是由于RBE3与淀粉葡聚糖链具有较高亲和性而难以和葡聚糖链解离,进而随着淀粉粒的增长而被其包裹.%The starch branching enzyme bound to starch granule in rice endosperm was investigated in the present paper. The starch branching enzyme activity assay indicated that some branching enzymes were bound to the starch granule of endosperm in rice. The result of N-terminal protein sequencing suggests that RBE3 (rice starch branching enzyme 3, RBE3), an 84 kD protein, bound to the starch granule of endosperm in rice. The level of starch granule-bound RBE3 protein remained approximately constant during the rice seed development. Northern blot analysis revealed that RBE4 was the earliest expressed starch branching enzyme during the development of rice endosperm and, subsequently RBE3 and RBE1 were expressed. These results suggest that RBE3 is bound to starch granule due to its high affinity to starch in rice endosperms.

  4. 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

  5. Improving nitrogen use efficiency in barley (Hordeum vulgare L.) through the cisgenic approach

    DEFF Research Database (Denmark)

    Kichey, Thomas; Holme, Inger; Møller, Inge Skrumsager

    2009-01-01

    Barley is one of the major crops cultivated worldwide and constitutes an important basis for animal feed. However, the production is facing a number of challenges that will be accentuated in the years to come, in particular restrictions on the use of nitrogen (N) fertilizer. In order to improve t...... the N use efficiency in barley, we are developing a new generation of genetically modified plants based on the concept of cisgenesis. In this approach, plants are transformed only with their own genetic material. The genes encoding the cytosolic isoform of the glutamine synthetase (GS1...... into the pGreenII binary vector. The genes have been inserted into barley by Agrobacterium-mediated transformation using the hygromycin phosphotransferase gene for selection of transformed lines on hygromycin. In this system, the resistance gene is placed on the helper plasmid pSoup allowing for separate...

  6. 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...

  7. 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

  8. 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....

  9. 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.

  10. Specific patterns of gene space organisation revealed in wheat by using the combination of barley and wheat genomic resources

    Directory of Open Access Journals (Sweden)

    Waugh Robbie

    2010-12-01

    Full Text Available Abstract Background Because of its size, allohexaploid nature and high repeat content, the wheat genome has always been perceived as too complex for efficient molecular studies. We recently constructed the first physical map of a wheat chromosome (3B. However gene mapping is still laborious in wheat because of high redundancy between the three homoeologous genomes. In contrast, in the closely related diploid species, barley, numerous gene-based markers have been developed. This study aims at combining the unique genomic resources developed in wheat and barley to decipher the organisation of gene space on wheat chromosome 3B. Results Three dimensional pools of the minimal tiling path of wheat chromosome 3B physical map were hybridised to a barley Agilent 15K expression microarray. This led to the fine mapping of 738 barley orthologous genes on wheat chromosome 3B. In addition, comparative analyses revealed that 68% of the genes identified were syntenic between the wheat chromosome 3B and barley chromosome 3 H and 59% between wheat chromosome 3B and rice chromosome 1, together with some wheat-specific rearrangements. Finally, it indicated an increasing gradient of gene density from the centromere to the telomeres positively correlated with the number of genes clustered in islands on wheat chromosome 3B. Conclusion Our study shows that novel structural genomics resources now available in wheat and barley can be combined efficiently to overcome specific problems of genetic anchoring of physical contigs in wheat and to perform high-resolution comparative analyses with rice for deciphering the organisation of the wheat gene space.

  11. Additional nitrogen fertilization at heading time of rice down-regulates cellulose synthesis in seed endosperm.

    Science.gov (United States)

    Midorikawa, Keiko; Kuroda, Masaharu; Terauchi, Kaede; Hoshi, Masako; Ikenaga, Sachiko; Ishimaru, Yoshiro; Abe, Keiko; Asakura, Tomiko

    2014-01-01

    The balance between carbon and nitrogen is a key determinant of seed storage components, and thus, is of great importance to rice and other seed-based food crops. To clarify the influence of the rhizosphere carbon/nitrogen balance during the maturation stage of several seed components, transcriptome analysis was performed on the seeds from rice plants that were provided additional nitrogen fertilization at heading time. As a result, it was assessed that genes associated with molecular processes such as photosynthesis, trehalose metabolism, carbon fixation, amino acid metabolism, and cell wall metabolism were differentially expressed. Moreover, cellulose and sucrose synthases, which are involved in cellulose synthesis, were down-regulated. Therefore, we compared cellulose content of mature seeds that were treated with additional nitrogen fertilization with those from control plants using calcofluor staining. In these experiments, cellulose content in endosperm from plants receiving additional nitrogen fertilization was less than that in control endosperm. Other starch synthesis-related genes such as starch synthase 1, starch phosphorylase 2, and branching enzyme 3 were also down-regulated, whereas some α-amylase and β-amylase genes were up-regulated. On the other hand, mRNA expression of amino acid biosynthesis-related molecules was up-regulated. Moreover, additional nitrogen fertilization caused accumulation of storage proteins and up-regulated Cys-poor prolamin mRNA expression. These data suggest that additional nitrogen fertilization at heading time changes the expression of some storage substance-related genes and reduces cellulose levels in endosperm.

  12. Bipolaris sorokiniana (teleomorph Cochliobolus Sativus: Causer of barley leaf lesions and root rot in Macedonia

    Directory of Open Access Journals (Sweden)

    Karov Ilija K.

    2009-01-01

    Full Text Available Diseased barley plants (Hordeum vulgare, were noticed in the area of Kumanovo, Bitola, Probistip, Skopje and Kocani, at the beginning of March, 2006. Our investigations were carried out in the period from 2006 to 2009. The plants were highly diseased, probably in the stage of germination, dwarfed with necrotic leaves and with poorly developed root. A rotten root collar was noticed notice in some plants, which could be easily pulled out from the soil. Plants infected in a later developing stage became yellow from the top of the leaf, and many brown-olive, oval shape lesions were noticed. Conidia of Bipolaris sorokiniana (Sacc. Shoen., were isolated from symptomatic lesions. Pseudothecia with asci and ascospores from teleomorph Cochliobolus sativus, were found on the barley straw in the same field the previous year.

  13. SSR ANALYSIS IN THE STUDY OF GENETIC DIVERSITY AND SIMILARITY OF BARLEY CULTIVARS

    Directory of Open Access Journals (Sweden)

    O. R. Lakhneko

    2016-06-01

    Full Text Available The aim of research was to develop an evaluation system of the genetic polymorphism for barley cultivars of Ukrainian and foreign origin based on the analysis of simple sequence repeats and valuable agricultural trait loci as well as to compose the molecular genetic passports for those cultivars. PCRs with the following separation of amplification products by agarose and polyacrylamide electrophoresis were performed to find out genetic polymorphism. Unweighted Pair Group Method with Arithmetic Mean was used for phylogenetic relationship detection. The dandrogram of phylogenetic relationships of 55 barley cultivars was constructed and molecular genetic passports were developed. Molecular genetic passports can be involved in verification for the compliance with standards cultivars, stability and seed purity.

  14. Nitrogen and phosphorus compounds in the aleurone grains of Iris pseudoacorus endosperm and Pisum sativum cotyledons

    Directory of Open Access Journals (Sweden)

    Ligia Konopska

    2015-05-01

    Full Text Available Aleurone grains from Iris pseudoacorus endosperm and Pisum sativum cotyledons were isolated partly according to Tombs's method (1967. Nitrogen compounds content was determined in them by Kjeldahl's micromethod, and in the particular fractions after Thiman and Laloraya (1960. Mainly protein N was detected in the aleurone grains, constituting 14.8 and 15.2 per cent of the dry mass of pea and Iris seeds, respectively. Moreover, phosphorus compounds were fractionated according to Holden and Pirie (1955. Analyses demonstrated the presence in aleurone grains of inorganic P, acid-soluble organophosphorus compounds, phospholipids and RNA.

  15. Combinatorial pooling enables selective sequencing of the barley gene space.

    Directory of Open Access Journals (Sweden)

    Stefano Lonardi

    2013-04-01

    Full Text Available For the vast majority of species - including many economically or ecologically important organisms, progress in biological research is hampered due to the lack of a reference genome sequence. Despite recent advances in sequencing technologies, several factors still limit the availability of such a critical resource. At the same time, many research groups and international consortia have already produced BAC libraries and physical maps and now are in a position to proceed with the development of whole-genome sequences organized around a physical map anchored to a genetic map. We propose a BAC-by-BAC sequencing protocol that combines combinatorial pooling design and second-generation sequencing technology to efficiently approach denovo selective genome sequencing. We show that combinatorial pooling is a cost-effective and practical alternative to exhaustive DNA barcoding when preparing sequencing libraries for hundreds or thousands of DNA samples, such as in this case gene-bearing minimum-tiling-path BAC clones. The novelty of the protocol hinges on the computational ability to efficiently compare hundred millions of short reads and assign them to the correct BAC clones (deconvolution so that the assembly can be carried out clone-by-clone. Experimental results on simulated data for the rice genome show that the deconvolution is very accurate, and the resulting BAC assemblies have high quality. Results on real data for a gene-rich subset of the barley genome confirm that the deconvolution is accurate and the BAC assemblies have good quality. While our method cannot provide the level of completeness that one would achieve with a comprehensive whole-genome sequencing project, we show that it is quite successful in reconstructing the gene sequences within BACs. In the case of plants such as barley, this level of sequence knowledge is sufficient to support critical end-point objectives such as map-based cloning and marker-assisted breeding.

  16. 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.

  17. 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....

  18. 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.

  19. Isolation and characterization of esters of indole-3-acetic acid from the liquid endosperm of the horse chestnut (Aesculus species)

    Science.gov (United States)

    Domagalski, W.; Schulze, A.; Bandurski, R. S.

    1987-01-01

    Esters of indole-3-acetic acid were extracted and purified from the liquid endosperm of immature fruits of various species of the horse chestnut (Aesculus parviflora, A. baumanni, A. pavia rubra, and A. pavia humulis). The liquid endosperm contained, at least 12 chromatographically distinct esters. One of these compounds was purified and characterized as an ester of indole-3-acetic acid and myo-inositol. A second compound was found to be an ester of indole-3-acetic acid and the disaccharide rutinose (glucosyl-rhamnose). A third compound was partially characterized as an ester of indole-3-acetic acid and a desoxyaminohexose.

  20. 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.

  1. 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.

  2. QTLs for barley yield adaptation to Mediterranean environments in the ‘Nure’ × ‘Tremois’ biparental population

    NARCIS (Netherlands)

    Tondelli, A.; Francia, E.; Visioni, A.; Comadran, J.; Mastrangelo, A.M.; Akar, T.; Al-Yassina, A.; Ceccarelli, S.; Grando, S.; Eeuwijk, van F.A.; Thomas, W.T.B.; Stanca, A.M.; Romagosa, I.; Pecchioni, N.

    2014-01-01

    Multi-environment trials represent a highly valuable tool for the identification of the genetic bases of crop yield potential and stress adaptation. A Diversity Array Technology®-based barley map has been developed in the ‘Nure’ × ‘Tremois’ biparental Doubled Haploid population, harbouring the genom

  3. A Simple Method for Assessing Severity of Common Root Rot on Barley

    Directory of Open Access Journals (Sweden)

    Mohammad Imad Eddin Arabi

    2013-12-01

    Full Text Available Common root rot caused by Cochliobolus sativus is a serious disease of barley. A simple and reliable method for assessing this disease would enhance our capacity in identifying resistance sources and developing resistant barley cultivars. In searching for such a method, a conidial suspension of C. sativus was dropped onto sterilized elongated subcrown internodes and incubated in sandwich filter paper using polyethylene transparent envelopes. Initial disease symptoms were easily detected after 48h of inoculation. Highly significant correlation coefficients were found in each experiment (A, B and C between sandwich filter paper and seedling assays, indicating that this testing procedure was reliable. The method presented facilitates a rapid pre-selection under uniform conditions which is of importance from a breeder’s point of view.

  4. Effect of induced lodging on grain yield and quality of brewing barley

    Directory of Open Access Journals (Sweden)

    Eduardo Caierão

    2006-01-01

    Full Text Available Lodging is one of the main factors of constraint to grain yield stability in barley. The objective of this study wasto evaluate the effects of lodging on agronomic and qualitative traits, when induced at different stages of the crop development.The trial was carried out in Victor Graeff, RS, using a randomized complete block design with four replications and 3 factors:year, lodging date and lodging intensity. The analyzed parameters were grain yield (GY, kernel plumpness (KP, germination(G, and score of lodging at harvest (SLH. No significant interaction was observed for GY and G. The effects of inducedlodging at the booting and physiologic maturity stages were distinct for GY, KP and G. Unlike G, the variables GY and KPwere not significantly affected by lodging intensity. Quantitative and qualitative losses in barley can be predicted based onlodging.

  5. Cytosolic glutamine synthetase in barley

    DEFF Research Database (Denmark)

    Thomsen, Hanne Cecilie

    Improving crop nitrogen (N) utilization efficiency (NUE) is of major importance in modern agriculture in order to reduce the amount of N fertilizer used for crop production. There is a high demand for development of crops which are able to produce high yields but with a concomitantly lower N...... fertilizer requirement. The enzyme glutamine synthetase (GS) has been a major topic in plant nitrogen research for decades due to its central role in plant N metabolism. The cytosolic version of this enzyme (GS1) plays an important role in relation to primary N assimilation as well as in relation to N...... and wildtype control. However, when grown to maturity the differences between transgenic lines and wildtype were highly dependent on the growth conditions applied. The transgenic lines had a higher N utilization efficiency (NUtE) than wildtype control, but only when exposed to a mild N stress following...

  6. A cor verde do endosperma do café On the green color of coffee endosperm

    Directory of Open Access Journals (Sweden)

    Paulo Mazzafera

    1988-01-01

    Full Text Available Realizaram-se comparações entre as sementes dos cultivares Mundo Novo de Coffea arabica, cujo endosperma é verde, com as do cultivar Cera, dessa espécie, de endosperma amarelo, com o objetivo de determinar os componentes responsáveis por aquela cor. Nas análises de clorofilas, flavonóides, diterpenos totais, ácido clorogênico e íons Mg, Ca, K, Fe e B, nenhuma diferença foi verificada entre os dois cultivares, sugerindo que a coloração verde se deva à presença de outros componentes ou que o 'Cera' apresente um componente que não ocorre no 'Mundo Novo' e que inibe o desenvolvimento da cor verde no seu endosperma.A comparison was made between the green seeded Mundo Novo and the yellow seeded Cera cultivars of Coffea arabica, in order to determine the components responsible for the green color of the endosperm. Both cultivars produce beverage of good quality. Chlorophylls, flavonoids, total diterpens, chlorogenic acid, and K, Ca, Mg, Fe and B ions were determined in beans of both cultivars. No differences between cultivars were found on these component contents, what suggests that the green color may be due to other compounds or that the Cera endosperm may have an inhibitor component of the green color of the coffee beans.

  7. Evaluation of Caesalpinia pulcherrima endospermic gum as affinity matrices for galactose-binding lectins interaction

    Directory of Open Access Journals (Sweden)

    Renata Chastinet Braga

    2011-04-01

    Full Text Available Lectins are proteins or glycoproteins able to bind, specifically and reversibly carbohydrates and glycoconjugates. Considering this ability, the utilization of Caesalpinia pulcherrima seeds polysaccharides as an affinity matrix was tested. The endospermic gum were solubilized in distinct concentrations of NaOH and treated with different amounts of epichlorohydrin (ECH forming affinity gels with variable capacity for interaction with galactose- binding lectins. The gel with an ECH/gum ration of 6.0mmol/g was selected as the best affinity matrix. The matrix presented different efficiencies in terms of isolating galactose-binding lectins. C. pulcherrima endospermic galactomannans were purified by ethanol precipitation and the purified galactomannan was crosslinked with the best formulation of gel. The Artocarpus incisa, Ricinus communis and Abrus precatorius lectins showed interactions of 11.5, 17.7 and 47.2mg of retained protein in 1g of gel, respectively; the Artocarpus integrifolia lectin showed the highest affinities (79.7mg/g. The heamaglutination assays confirmed the activity and SDS-PAGE electrophoresis confirmed the isolation of the lectins in a single-step procedure.

  8. Pullulanase and Starch Synthase III Are Associated with Formation of Vitreous Endosperm in Quality Protein Maize.

    Directory of Open Access Journals (Sweden)

    Hao Wu

    Full Text Available The opaque-2 (o2 mutation of maize increases lysine content, but the low seed density and soft texture of this type of mutant are undesirable. Lines with modifiers of the soft kernel phenotype (mo2 called "Quality Protein Maize" (QPM have high lysine and kernel phenotypes similar to normal maize. Prior research indicated that the formation of vitreous endosperm in QPM might involve changes in starch granule structure. In this study, we focused on analysis of two starch biosynthetic enzymes that may influence kernel vitreousness. Analysis of recombinant inbred lines derived from a cross of W64Ao2 and K0326Y revealed that pullulanase activity had significant positive correlation with kernel vitreousness. We also found that decreased Starch Synthase III abundance may decrease the pullulanase activity and average glucan chain length given the same Zpu1 genotype. Therefore, Starch Synthase III could indirectly influence the kernel vitreousness by affecting pullulanase activity and coordinating with pullulanase to alter the glucan chain length distribution of amylopectin, resulting in different starch structural properties. The glucan chain length distribution had strong positive correlation with the polydispersity index of glucan chains, which was positively associated with the kernel vitreousness based on nonlinear regression analysis. Therefore, we propose that pullulanase and Starch Synthase III are two important factors responsible for the formation of the vitreous phenotype of QPM endosperms.

  9. In vivo gibberellin biosynthesis in endosperm of Sechium edule Sw. seeds

    Energy Technology Data Exchange (ETDEWEB)

    Ceccarelli, N.; Lorenzi, R. (Universita degli Studi, Pisa (Italy))

    1990-07-01

    Biosynthesis of gibberellins (GAs) was studied in vivo in endosperms of Sechium edule Sw. Exogenous ent-({sup 14}C)kaurene was metabolized into four major products: GA{sub 12}, GA{sub 4}, GA{sub 7} and 16,17-dihydro-16-hydroxy-GA{sub 15} alcohol glucoside. Other minor metabolites were also observed including ent-kaurenol and ent-kaurenal. Conversion of ent-({sup 14}C)kaurene to ent-karuenol glucoside by endosperm cell-free preparations in the presence of UDPG was observed. However, the finding was not confirmed in in vivo studies and is probably artifactual. Overall evidence coming from the analysis of endogenous GAs and in vitro and in vivo biosynthetic studies are discussed in relation to the possible existence in the Sechium seeds of a different route, along with the known pathway, branching from ent-kaurene or ent-7-{alpha}-hydroxykaurenoic acid and this also leading to biologically active GAs.

  10. Metabolic pathways of the wheat (Triticum aestivum endosperm amyloplast revealed by proteomics

    Directory of Open Access Journals (Sweden)

    Dupont Frances M

    2008-04-01

    Full Text Available Abstract Background By definition, amyloplasts are plastids specialized for starch production. However, a proteomic study of amyloplasts isolated from wheat (Triticum aestivum Butte 86 endosperm at 10 days after anthesis (DPA detected enzymes from many other metabolic and biosynthetic pathways. To better understand the role of amyloplasts in food production, the data from that study were evaluated in detail and an amyloplast metabolic map was outlined. Results Analysis of 288 proteins detected in an amyloplast preparation predicted that 178 were amyloplast proteins. Criteria included homology with known plastid proteins, prediction of a plastid transit peptide for the wheat gene product or a close homolog, known plastid location of the pathway, and predicted plastid location for other members of the same pathway. Of these, 135 enzymes were arranged into 18 pathways for carbohydrate, lipid, amino acid, nucleic acid and other biosynthetic processes that are critical for grain-fill. Functions of the other proteins are also discussed. Conclusion The pathways outlined in this paper suggest that amyloplasts play a central role in endosperm metabolism. The interacting effects of genetics and environment on starch and protein production may be mediated in part by regulatory mechanisms within this organelle.

  11. 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

  12. 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.

  13. 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...

  14. 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...

  15. 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....

  16. 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’) ...

  17. 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....

  18. 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.

  19. Imidiazolium based ionic liquids: effects of different anions and alkyl chains lengths on the barley seedlings.

    Science.gov (United States)

    Cvjetko Bubalo, Marina; Hanousek, Karla; Radošević, Kristina; Gaurina Srček, Višnja; Jakovljević, Tamara; Radojčić Redovniković, Ivana

    2014-03-01

    We studied the effects of five imidiazolium based ionic liquids with different anions and length of alkyl chains linked to imidazolium ring on the early development of barley (Hordeum vulgare). The inhibitory effect depends on the ionic liquids concentration and chemical structure, whereby the most toxic one was [C10mim][Br], followed by [C7mim][Br], [C4mim][Br], [C4mim][CH3CO2] and [C4mim][BF4]. Both anion and cation structures affected the toxicity of ionic liquid indicating that selection of more biocompatible anions such as [CH3CO2] does not necessarily indicate lower toxicity. Alternation in the extent of oxidative stress and antioxidant enzymes activities were found in barley plants due to ionic liquid treatments. When seedlings were exposed to higher concentrations of ionic liquids, antioxidant system could not effectively remove reactive oxidative species, leading to lipid peroxidation and damage of the photosynthetic system. However, overall data indicated that the performance of barley seedling was improved when all measured enzymes involved in scavenging of reactive oxygen species (ROS) were increased with special emphasis on GPX activities. Since there are no studies about ionic liquid (IL) toxicity in plants, that simultaneously evaluates the antioxidative enzyme system in response to different ILs, this work is valuable for gaining knowledge about the protection mechanism of plants from oxidative stress caused by IL exposure.

  20. 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).

  1. Water balance and soil moisture dynamics of field plots with barley and grass ley

    Science.gov (United States)

    Johnsson, Holger; Jansson, Per-Erik

    1991-12-01

    A physically based soilwater and heat model was used to estimate the water balance of an arable field in central Sweden for each of three different crop covers (barley with and without N fertilization and grass ley). Annual water balances were calculated for each year from 1981 to 1985. On-site measurements of soil physical properties, meteorological variables and plant development were used as input to the model. Simulated soil forst, snow cover, soilwater contents, soilwater tensions and relative differences in simulated drainage between treatments were in agreement with the corresponding measured values. In the simulation, surface runoff (70 mm year -1 in all treatments) mainly occurred during snowmelt periods and accounted for much of the variation in the total runoff estimate. Annual mean precipitation amounted to 610 mm year -1, whereas average evapotranspiration was calculated to be 320, 360 and 435 mm year -1 in barley without N fertilization, barley with N fertilization and grass ley, respectively. Soil evaporation accounted for 60, 43 and 23% whereas evaporation of intercepted water accounted for 5, 12 and 19% of the total evapotranspiration, respectively. Drainage estimates amounted to 205, 170 and 110 mm year -1.

  2. Stable gene replacement in barley by targeted double-strand break induction.

    Science.gov (United States)

    Watanabe, Koichi; Breier, Ulrike; Hensel, Götz; Kumlehn, Jochen; Schubert, Ingo; Reiss, Bernd

    2016-03-01

    Gene targeting is becoming an important tool for precision genome engineering in plants. During gene replacement, a variant of gene targeting, transformed DNA integrates into the genome by homologous recombination (HR) to replace resident sequences. We have analysed gene targeting in barley (Hordeum vulgare) using a model system based on double-strand break (DSB) induction by the meganuclease I-SceI and a transgenic, artificial target locus. In the plants we obtained, the donor construct was inserted at the target locus by homology-directed DNA integration in at least two transformants obtained in a single experiment and was stably inherited as a single Mendelian trait. Both events were produced by one-sided integration. Our data suggest that gene replacement can be achieved in barley with a frequency suitable for routine application. The use of a codon-optimized nuclease and co-transfer of the nuclease gene together with the donor construct are probably the components important for efficient gene targeting. Such an approach, employing the recently developed synthetic nucleases/nickases that allow DSB induction at almost any sequence of a genome of interest, sets the stage for precision genome engineering as a routine tool even for important crops such as barley.

  3. 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...

  4. 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

  5. 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.

  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. 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.

  8. Characterization of SSIIIa-deficient mutants of rice: the function of SSIIIa and pleiotropic effects by SSIIIa deficiency in the rice endosperm.

    Science.gov (United States)

    Fujita, Naoko; Yoshida, Mayumi; Kondo, Tomonori; Saito, Kaori; Utsumi, Yoshinori; Tokunaga, Takashi; Nishi, Aiko; Satoh, Hikaru; Park, Jin-Hee; Jane, Jay-Lin; Miyao, Akio; Hirochika, Hirohiko; Nakamura, Yasunori

    2007-08-01

    Starch synthase IIIa (SSIIIa)-deficient rice (Oryza sativa) mutants were generated using retrotransposon insertion and chemical mutagenesis. The lowest migrating SS activity bands on glycogen-containing native polyacrylamide gel, which were identified to be those for SSIIIa, were completely absent in these mutants, indicating that they are SSIIIa null mutants. The amylopectin B(2) to B(4) chains with degree of polymerization (DP) >/= 30 and the M(r) of amylopectin in the mutant were reduced to about 60% and 70% of the wild-type values, respectively, suggesting that SSIIIa plays an important part in the elongation of amylopectin B(2) to B(4) chains. Chains with DP 6 to 9 and DP 16 to 19 decreased while chains with DP 10 to 15 and DP 20 to 25 increased in the mutants amylopectin. These changes in the SSIIIa mutants are almost opposite images of those of SSI-deficient rice mutant and were caused by 1.3- to 1.7-fold increase of the amount of SSI in the mutants endosperm. Furthermore, the amylose content and the extralong chains (DP >/= 500) of amylopectin were increased by 1.3- and 12-fold, respectively. These changes in the composition in the mutants starch were caused by 1.4- to 1.7-fold increase in amounts of granules-bound starch synthase (GBSSI). The starch granules of the mutants were smaller with round shape, and were less crystalline. Thus, deficiency in SSIIIa, the second major SS isozyme in developing rice endosperm affected the structure of amylopectin, amylase content, and physicochemical properties of starch granules in two ways: directly by the SSIIIa deficiency itself and indirectly by the enhancement of both SSI and GBSSI gene transcripts.

  9. Genome-wide screen of genes imprinted in sorghum endosperm, and the roles of allelic differential cytosine methylation.

    Science.gov (United States)

    Zhang, Meishan; Li, Ning; He, Wenan; Zhang, Huakun; Yang, Wei; Liu, Bao

    2016-02-01

    Imprinting is an epigenetic phenomenon referring to allele-biased expression of certain genes depending on their parent of origin. Accumulated evidence suggests that, while imprinting is a conserved mechanism across kingdoms, the identities of the imprinted genes are largely species-specific. Using deep RNA sequencing of endosperm 14 days after pollination in sorghum, 5683 genes (29.27% of the total 19 418 expressed genes) were found to harbor diagnostic single nucleotide polymorphisms between two parental lines. The analysis of parent-of-origin expression patterns in the endosperm of a pair of reciprocal F1 hybrids between the two sorghum lines led to identification of 101 genes with ≥ fivefold allelic expression difference in both hybrids, including 85 maternal expressed genes (MEGs) and 16 paternal expressed genes (PEGs). Thirty of these genes were previously identified as imprinted in endosperm of maize (Zea mays), rice (Oryza sativa) or Arabidopsis, while the remaining 71 genes are sorghum-specific imprinted genes relative to these three plant species. Allele-biased expression of virtually all of the 14 tested imprinted genes (nine MEGs and five PEGs) was validated by pyrosequencing using independent sources of RNA from various developmental stages and dissected parts of endosperm. Forty-six imprinted genes (30 MEGs and 16 PEGs) were assayed by quantitative RT-PCR, and the majority of them showed endosperm-specific or preferential expression relative to embryo and other tissues. DNA methylation analysis of the 5' upstream region and gene body for seven imprinted genes indicated that, while three of the four PEGs were associated with hypomethylation of maternal alleles, no MEG was associated with allele-differential methylation.

  10. 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.

  11. The effects of reduced gluten barley diet on humoral and cell-mediated systemic immune responses of gluten-sensitive rhesus macaques.

    Science.gov (United States)

    Sestak, Karol; Thwin, Hazel; Dufour, Jason; Aye, Pyone P; Liu, David X; Moehs, Charles P

    2015-03-06

    Celiac disease (CD) affects approximately 1% of the general population while an estimated additional 6% suffers from a recently characterized, rapidly emerging, similar disease, referred to as non-celiac gluten sensitivity (NCGS). The only effective treatment of CD and NCGS requires removal of gluten sources from the diet. Since required adherence to a gluten-free diet (GFD) is difficult to accomplish, efforts to develop alternative treatments have been intensifying in recent years. In this study, the non-human primate model of CD/NCGS, e.g., gluten-sensitive rhesus macaque, was utilized with the objective to evaluate the treatment potential of reduced gluten cereals using a reduced gluten (RG; 1% of normal gluten) barley mutant as a model. Conventional and RG barleys were used for the formulation of experimental chows and fed to gluten-sensitive (GS) and control macaques to determine if RG barley causes a remission of dietary gluten-induced clinical and immune responses in GS macaques. The impacts of the RG barley diet were compared with the impacts of the conventional barley-containing chow and the GFD. Although remission of the anti-gliadin antibody (AGA) serum responses and an improvement of clinical diarrhea were noted after switching the conventional to the RG barley diet, production of inflammatory cytokines, e.g., interferon-gamma (IFN-γ), tumor necrosis factor (TNF) and interleukin-8 (IL-8) by peripheral CD4+ T helper lymphocytes, persisted during the RG chow treatment and were partially abolished only upon re-administration of the GFD. It was concluded that the RG barley diet might be used for the partial improvement of gluten-induced disease but its therapeutic value still requires upgrading-by co-administration of additional treatments.

  12. The Effects of Reduced Gluten Barley Diet on Humoral and Cell-Mediated Systemic Immune Responses of Gluten-Sensitive Rhesus Macaques

    Directory of Open Access Journals (Sweden)

    Karol Sestak

    2015-03-01

    Full Text Available Celiac disease (CD affects approximately 1% of the general population while an estimated additional 6% suffers from a recently characterized, rapidly emerging, similar disease, referred to as non-celiac gluten sensitivity (NCGS. The only effective treatment of CD and NCGS requires removal of gluten sources from the diet. Since required adherence to a gluten-free diet (GFD is difficult to accomplish, efforts to develop alternative treatments have been intensifying in recent years. In this study, the non-human primate model of CD/NCGS, e.g., gluten-sensitive rhesus macaque, was utilized with the objective to evaluate the treatment potential of reduced gluten cereals using a reduced gluten (RG; 1% of normal gluten barley mutant as a model. Conventional and RG barleys were used for the formulation of experimental chows and fed to gluten-sensitive (GS and control macaques to determine if RG barley causes a remission of dietary gluten-induced clinical and immune responses in GS macaques. The impacts of the RG barley diet were compared with the impacts of the conventional barley-containing chow and the GFD. Although remission of the anti-gliadin antibody (AGA serum responses and an improvement of clinical diarrhea were noted after switching the conventional to the RG barley diet, production of inflammatory cytokines, e.g., interferon-gamma (IFN-γ, tumor necrosis factor (TNF and interleukin-8 (IL-8 by peripheral CD4+ T helper lymphocytes, persisted during the RG chow treatment and were partially abolished only upon re-administration of the GFD. It was concluded that the RG barley diet might be used for the partial improvement of gluten-induced disease but its therapeutic value still requires upgrading—by co-administration of additional treatments.

  13. 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

  14. 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

    -regulated by both GA and seed germination. Those genes and metabolic pathways are likely to be important parts of transcriptional regulatory networks underlying GA and ABA regulation of seed germination and seedling growth. Conclusions The studies developed a model depicting transcriptional regulatory programs underlying barley germination and GA and ABA regulation of germination at gene, pathway and systems levels, and established a standard transcriptome reference for further integration with various -omics and biological data to illustrate biological networks underlying seed germination. The studies also generated a great amount of systems biological evidence for previously proposed hypotheses, and developed a number of new hypotheses on transcriptional regulation of seed germination for further experimental validation.

  15. Rice endosperm protein slows progression of fatty liver and diabetic nephropathy in Zucker diabetic fatty rats.

    Science.gov (United States)

    Kubota, Masatoshi; Watanabe, Reiko; Yamaguchi, Miki; Hosojima, Michihiro; Saito, Akihiko; Fujii, Mikio; Fujimura, Shinobu; Kadowaki, Motoni

    2016-10-01

    We previously reported that rice endosperm protein (REP) has renoprotective effects in Goto-Kakizaki rats, a non-obese diabetic model. However, whether these effects occur in obese diabetes remains unclear. This study aimed to clarify the effects of REP on obese diabetes, especially on fatty liver and diabetic nephropathy, using the obese diabetic model Zucker diabetic fatty (ZDF) rats. In total, 7-week-old male ZDF rats were fed diets containing 20 % REP or casein (C) for 8 weeks. Changes in fasting blood glucose levels and urinary markers were monitored during the experimental period. Hepatic lipids and metabolites were measured and renal glomeruli were observed morphologically. HbA1c levels were significantly lower in rats fed REP, compared with C (Pdiabetes, fatty liver and diabetic nephropathy.

  16. Ways of Developing Plants in Interspecific Hybridization of Cotton

    Institute of Scientific and Technical Information of China (English)

    RAKHMANKULOV; S; DAMINOVA; D; RAKHMANKULOV; M

    2008-01-01

    It is known,that there are various barriers to fertilization,development of embryos,and endosperm because of different number of chromosomes in parents in the interspecific hybridization of cotton.Thus the factors providing normal cell fission of a germ and endosperm are necessary.It is necessary to culture embryos in vitro on the artificial environments containing various phytohormones,or to

  17. SSH Analysis of Endosperm Transcripts and Characterization of Heat Stress Regulated Expressed Sequence Tags in Bread Wheat

    Science.gov (United States)

    Goswami, Suneha; Kumar, Ranjeet R.; Dubey, Kavita; Singh, Jyoti P.; Tiwari, Sachidanand; Kumar, Ashok; Smita, Shuchi; Mishra, Dwijesh C.; Kumar, Sanjeev; Grover, Monendra; Padaria, Jasdeep C.; Kala, Yugal K.; Singh, Gyanendra P.; Pathak, Himanshu; Chinnusamy, Viswanathan; Rai, Anil; Praveen, Shelly; Rai, Raj D.

    2016-01-01

    Heat stress is one of the major problems in agriculturally important cereal crops, especially wheat. Here, we have constructed a subtracted cDNA library from the endosperm of HS-treated (42°C for 2 h) wheat cv. HD2985 by suppression subtractive hybridization (SSH). We identified ~550 recombinant clones ranging from 200 to 500 bp with an average size of 300 bp. Sanger's sequencing was performed with 205 positive clones to generate the differentially expressed sequence tags (ESTs). Most of the ESTs were observed to be localized on the long arm of chromosome 2A and associated with heat stress tolerance and metabolic pathways. Identified ESTs were BLAST search using Ensemble, TriFLD, and TIGR databases and the predicted CDS were translated and aligned with the protein sequences available in pfam and InterProScan 5 databases to predict the differentially expressed proteins (DEPs). We observed eight different types of post-translational modifications (PTMs) in the DEPs corresponds to the cloned ESTs-147 sites with phosphorylation, 21 sites with sumoylation, 237 with palmitoylation, 96 sites with S-nitrosylation, 3066 calpain cleavage sites, and 103 tyrosine nitration sites, predicted to sense the heat stress and regulate the expression of stress genes. Twelve DEPs were observed to have transmembrane helixes (TMH) in their structure, predicted to play the role of sensors of HS. Quantitative Real-Time PCR of randomly selected ESTs showed very high relative expression of HSP17 under HS; up-regulation was observed more in wheat cv. HD2985 (thermotolerant), as compared to HD2329 (thermosusceptible) during grain-filling. The abundance of transcripts was further validated through northern blot analysis. The ESTs and their corresponding DEPs can be used as molecular marker for screening or targeted precision breeding program. PTMs identified in the DEPs can be used to elucidate the thermotolerance mechanism of wheat—a novel step toward the development of

  18. SSH Analysis of Endosperm Transcripts and Characterization of Heat Stress Regulated Expressed Sequence Tags in Bread Wheat.

    Science.gov (United States)

    Goswami, Suneha; Kumar, Ranjeet R; Dubey, Kavita; Singh, Jyoti P; Tiwari, Sachidanand; Kumar, Ashok; Smita, Shuchi; Mishra, Dwijesh C; Kumar, Sanjeev; Grover, Monendra; Padaria, Jasdeep C; Kala, Yugal K; Singh, Gyanendra P; Pathak, Himanshu; Chinnusamy, Viswanathan; Rai, Anil; Praveen, Shelly; Rai, Raj D

    2016-01-01

    Heat stress is one of the major problems in agriculturally important cereal crops, especially wheat. Here, we have constructed a subtracted cDNA library from the endosperm of HS-treated (42°C for 2 h) wheat cv. HD2985 by suppression subtractive hybridization (SSH). We identified ~550 recombinant clones ranging from 200 to 500 bp with an average size of 300 bp. Sanger's sequencing was performed with 205 positive clones to generate the differentially expressed sequence tags (ESTs). Most of the ESTs were observed to be localized on the long arm of chromosome 2A and associated with heat stress tolerance and metabolic pathways. Identified ESTs were BLAST search using Ensemble, TriFLD, and TIGR databases and the predicted CDS were translated and aligned with the protein sequences available in pfam and InterProScan 5 databases to predict the differentially expressed proteins (DEPs). We observed eight different types of post-translational modifications (PTMs) in the DEPs corresponds to the cloned ESTs-147 sites with phosphorylation, 21 sites with sumoylation, 237 with palmitoylation, 96 sites with S-nitrosylation, 3066 calpain cleavage sites, and 103 tyrosine nitration sites, predicted to sense the heat stress and regulate the expression of stress genes. Twelve DEPs were observed to have transmembrane helixes (TMH) in their structure, predicted to play the role of sensors of HS. Quantitative Real-Time PCR of randomly selected ESTs showed very high relative expression of HSP17 under HS; up-regulation was observed more in wheat cv. HD2985 (thermotolerant), as compared to HD2329 (thermosusceptible) during grain-filling. The abundance of transcripts was further validated through northern blot analysis. The ESTs and their corresponding DEPs can be used as molecular marker for screening or targeted precision breeding program. PTMs identified in the DEPs can be used to elucidate the thermotolerance mechanism of wheat-a novel step toward the development of "climate-smart" wheat.

  19. SSH analysis of endosperm transcripts and characterization of heat stress regulated expressed sequence tags in bread wheat

    Directory of Open Access Journals (Sweden)

    Suneha Goswami

    2016-08-01

    Full Text Available Heat stress is one of the major problems in agriculturally important cereal crops, especially wheat. Here, we have constructed a subtracted cDNA library from the endosperm of HS-treated (42°C for 2 h wheat cv. HD2985 by suppression subtractive hybridization (SSH. We identified ~550 recombinant clones ranging from 200 to 500 bp with an average size of 300 bp. Sanger’s sequencing was performed with 205 positive clones to generate the differentially expressed sequence tags (ESTs. Most of the ESTs were observed to be localized on the long arm of chromosome 2A and associated with heat stress tolerance and metabolic pathways. Identified ESTs were BLAST search using Ensemble, TriFLD and TIGR databases and the predicted CDS were translated and aligned with the protein sequences available in pfam and InterProScan 5 databases to predict the differentially expressed proteins (DEPs. We observed eight different types of post-translational modifications (PTMs in the DEPs corresponds to the cloned ESTs—147 sites with phosphorylation, 21 sites with sumoylation, 237 with palmitoylation, 96 sites with S-nitrosylation, 3066 calpain cleavage sites, and 103 tyrosine nitration sites, predicted to sense the heat stress and regulate the expression of stress genes. Twelve DEPs were observed to have transmembrane helixes (TMH in their structure, predicted to play the role of sensors of HS. Quantitative Real-Time PCR of randomly selected ESTs showed very high relative expression of HSP17 under HS; up-regulation was observed more in wheat cv. HD2985 (thermotolerant, as compared to HD2329 (thermosusceptible during grain-filling. The abundance of transcripts was further validated through northern blot analysis. The ESTs and their corresponding DEPs can be used as molecular marker for screening or targeted precision breeding program. PTMs identified in the DEPs can be used to elucidate the thermotolerance mechanism of wheat – a novel step towards the development of

  20. Improved Evidence-Based Genome-scale Metabolic Models for Maize Leaf, Embryo, and Endosperm.

    Directory of Open Access Journals (Sweden)

    Samuel eSeaver

    2015-03-01

    Full Text Available There is a growing demand for genome-scale metabolic reconstructions for plants, fueled by the need to understand the metabolic basis of crop yield and by progress in genome and transcriptome sequencing. Methods are also required to enable the interpretation of plant transcriptome data to study how cellular metabolic activity varies under different growth conditions or even within different organs, tissues, and developmental stages. Such methods depend extensively on the accuracy with which genes have been mapped to the biochemical reactions in the plant metabolic pathways. Errors in these mappings lead to metabolic reconstructions with an inflated number of reactions and possible generation of unreliable metabolic phenotype predictions. Here we introduce a new evidence-based genome-scale metabolic reconstruction of maize, with significant improvements in the quality of the gene-reaction associations included within our model. We also present a new approach for applying our model to predict active metabolic genes based on transcriptome data. This method includes a minimal set of reactions associated with low expression genes to enable activity of a maximum number of reactions associated with high expression genes. We apply this method to construct an organ-specific model for the maize leaf, and tissue specific models for maize embryo and endosperm cells. We validate our models using fluxomics data for the endosperm and embryo, demonstrating an improved capacity of our models to fit the available fluxomics data. All models are publicly available via the DOE Systems Biology Knowledgebase and PlantSEED, and our new method is generally applicable for analysis transcript profiles from any plant, paving the way for further in silico studies with a wide variety of plant genomes.

  1. Introgression of opaque2 into Waxy Maize Causes Extensive Biochemical and Proteomic Changes in Endosperm.

    Directory of Open Access Journals (Sweden)

    Zhiqiang Zhou

    Full Text Available Waxy maize is prevalently grown in China and other countries due to the excellent characters and economic value. However, its low content of lysine can't meet the nutritional requirements of humans and livestock. In the present study, we introgressed the opaque2 (o2 allele into waxy maize line Zhao OP-6/O2O2 by using marker-assisted selection (MAS technique and successfully improved the lysine content and quality of waxy maize. Transcript abundance analysis indicated that the wx1 expression levels had no difference between Zhao OP-6/o2o2 and Zhao OP-6/O2O2. However, Zhao OP-6/o2o2 was characterized by a phenotype of hard and vitreous kernels and accumulation of protein bodies at smaller size (one third of that of parents but in larger numbers. Biochemical analyses showed that Zhao OP-6/o2o2 had 16.7% less free amino acids than Zhao OP-6/O2O2, especially those derived from glycolytic intermediates, but its content of lysine was increased by 51.6% (0.47% vs. 0.31%. The content of amylopectin was 98.5% in Zhao OP-6/o2o2, significantly higher than that in Zhao OP-6/O2O2 (97.7%. Proteomic analyses indicated that o2 introgression not only decreased the accumulation of various zein proteins except for 27-kDa γ-zein, but also affected other endosperm proteins related to amino acid biosynthesis, starch-protein balance, stress response and signal transduction. This study gives us an intriguing insight into the metabolism changes in endosperm of waxy maize introgressed with opaque2.

  2. Characterization and Ectopic Expression of CoWRI1, an AP2/EREBP Domain-Containing Transcription Factor from Coconut (Cocos nucifera L.) Endosperm, Changes the Seeds Oil Content in Transgenic Arabidopsis thaliana and Rice (Oryza sativa L.)

    Science.gov (United States)

    Sun, RuHao; Ye, Rongjian; Gao, Lingchao; Zhang, Lin; Wang, Rui; Mao, Ting; Zheng, Yusheng; Li, Dongdong; Lin, Yongjun

    2017-01-01

    Coconut (Cocos nucifera L.) is a key tropical crop and a member of the monocotyledonous family Arecaceae (Palmaceae). Few genes and related metabolic processes involved in coconut endosperm development have been investigated. In this study, a new member of the WRI1 gene family was isolated from coconut endosperm and was named CoWRI1. Its transcriptional activities and interactions with the acetyl-CoA carboxylase (BCCP2) promoter of CoWRI1 were confirmed by the yeast two-hybrid and yeast one-hybrid approaches, respectively. Functional characterization was carried out through seed-specific expression in Arabidopsis and endosperm-specific expression in rice. In transgenic Arabidopsis, high over-expressions of CoWRI1 in seven independent T2 lines were detected by quantitative real-time PCR. The relative mRNA accumulation of genes encoding enzymes involved in either fatty acid biosynthesis or triacylglycerols assembly (BCCP2, KASI, MAT, ENR, FATA, and GPDH) were also assayed in mature seeds. Furthermore, lipid and fatty acids C16:0 and C18:0 significantly increased. In two homozygous T2 transgenic rice lines (G5 and G2), different CoWRI1 expression levels were detected, but no CoWRI1 transcripts were detected in the wild type. Analyses of the seed oil content, starch content, and total protein content indicated that the two T2 transgenic lines showed a significant increase (P < 0.05) in seed oil content. The transgenic lines also showed a significant increase in starch content, whereas total protein content decreased significantly. Further analysis of the fatty acid composition revealed that palmitic acid (C16:0) and linolenic acid (C18:3) increased significantly in the seeds of the transgenic rice lines, but oleic acid (C18:1) levels significantly declined. PMID:28179911

  3. Characterization and Ectopic Expression of CoWRI1, an AP2/EREBP Domain-Containing Transcription Factor from Coconut (Cocos nucifera L.) Endosperm, Changes the Seeds Oil Content in Transgenic Arabidopsis thaliana and Rice (Oryza sativa L.).

    Science.gov (United States)

    Sun, RuHao; Ye, Rongjian; Gao, Lingchao; Zhang, Lin; Wang, Rui; Mao, Ting; Zheng, Yusheng; Li, Dongdong; Lin, Yongjun

    2017-01-01

    Coconut (Cocos nucifera L.) is a key tropical crop and a member of the monocotyledonous family Arecaceae (Palmaceae). Few genes and related metabolic processes involved in coconut endosperm development have been investigated. In this study, a new member of the WRI1 gene family was isolated from coconut endosperm and was named CoWRI1. Its transcriptional activities and interactions with the acetyl-CoA carboxylase (BCCP2) promoter of CoWRI1 were confirmed by the yeast two-hybrid and yeast one-hybrid approaches, respectively. Functional characterization was carried out through seed-specific expression in Arabidopsis and endosperm-specific expression in rice. In transgenic Arabidopsis, high over-expressions of CoWRI1 in seven independent T2 lines were detected by quantitative real-time PCR. The relative mRNA accumulation of genes encoding enzymes involved in either fatty acid biosynthesis or triacylglycerols assembly (BCCP2, KASI, MAT, ENR, FATA, and GPDH) were also assayed in mature seeds. Furthermore, lipid and fatty acids C16:0 and C18:0 significantly increased. In two homozygous T2 transgenic rice lines (G5 and G2), different CoWRI1 expression levels were detected, but no CoWRI1 transcripts were detected in the wild type. Analyses of the seed oil content, starch content, and total protein content indicated that the two T2 transgenic lines showed a significant increase (P < 0.05) in seed oil content. The transgenic lines also showed a significant increase in starch content, whereas total protein content decreased significantly. Further analysis of the fatty acid composition revealed that palmitic acid (C16:0) and linolenic acid (C18:3) increased significantly in the seeds of the transgenic rice lines, but oleic acid (C18:1) levels significantly declined.

  4. Sanitary state and yielding of spring barley as dependent on soil tillage method

    Directory of Open Access Journals (Sweden)

    Tomasz P. Kurowski

    2012-12-01

    Full Text Available The effects of traditional tillage cultivation (control treatment, no tillage (instead of tillage the soil was loosened with scruff, and direct sowing (with a special drill into unploughed soil on the health of spring barley cultivar. Klimek were compared in three-field crop rotation (field bean, winter wheat, spring barley in an experiment performed in the years 1997-1999 on the soil of a good wheat complex. The results of phytopathological observations carried out over the vegetation season are presented in the form of an injury index. The following diseases were recorded on spring barley: net blotch (Drechslera teres - net type and spot type, powdery mildew (Blumeria graminis, leaf blotch (Rhynchosporium secalis, eyespot (Tapesia yallundae and foot rot (fungal complex. Tillage system had no a significant influence on the occurrence of both types of net blotch. The intensity of powdery mildew and leaf blotch was the highest in the case of traditional tillage cultivation, and the lowest - in that of no tillage. Direct sowing was conductive to the development of eyespot, and no tillage - to foot rot. Fungi of the genus Fusarium, mainly F. culmorum, and the species Bipolaris sorokiniana, were isolated most frequently from infested stem bases. The weather conditions differed during spring barley grown in the three years analyzed. Mean air temperature in 1997 and 1998 was similar to the many-year average for the city of Olsztyn and its surroundings (13.8°C. In the vegetation season 1999 mean air temperature reached 14.6°C, and was considerably higher than the many-year average. Taking into account total precipitation and distribution in the three-year experimental cycle, 1997 and 1998 can be considered average, and 1999 - wet.The weather conditions had a significant effect on the intensity of all diseases observed on spring barley. The highest yield grain was obtained in the case of traditional tillage cultivation (on average 3.06 t·ha-1 for the

  5. Structural and Histochemical Characterization of Developing Rice Caryopsis

    Institute of Scientific and Technical Information of China (English)

    YU Xu-run; ZHOU Liang; XIONG Fei; WANG Zhong

    2014-01-01

    The development of pericarp, seed coat, starchy endosperm and aleurone of the rice caryopsis was investigated, histochemically and structurally, from the time of flowering to maturity. The results showed that during its growth, the maximum length of the caryopsis was attained first, followed by width and then thickness. Histochemical examination of the caryopsis showed that starch was mainly accumulated in the endosperm, but the endosperm showed no metabolic activity, while embryo and pericarp contained a few starch grains, and embryo and aleurone were strongly active. Aleuronic cells contained many aleurone grains and spherosomes, and aleurone in the dorsal region developed earlier and contained more layers of cells. Amyloplasts in endosperm contained many starch granules and were spherical at early stages but polyhedric at late stages. The protein bodies appeared later than amyloplasts, and the number of protein bodies in subaleurone was greater than those in the starchy endosperm. The white-belly portion of endosperm might be relative to the status of amyloplast development.

  6. 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.

  7. Breaking through the Endosperm during Seed Germination%种子萌发过程中胚乳的突破性研究

    Institute of Scientific and Technical Information of China (English)

    徐振江; 陈兵先; 赵晟楠; 袁红霞; 郜珊珊; 张瑜; 陈靖; 王晓峰

    2012-01-01

    Endosperm completely encloses the embryo of many seeds and is a mechanical restraint for the completion of germination. Thus, rupture of the endosperm is the last key step controlling the process of germination. Rupture of the endosperm requires the growth potential of embryo and weakening of the endosperm after imbibition. Weakening of the endosperm involves enzymatic and non-enzymatic mechanisms. This review summarizes research progress in growth potential of embryo, location and ways of endosperm rupture, structure of the endosperm and chemical composition of endosperm cell walls, roles of cell-wall-degrading enzymes, ex-pansin and reactive oxygen species.%胚乳将许多种子的胚完全包裹,是这些种子萌发的物理屏障,其破裂与否是决定种子萌发与否的最后开关.胚乳破裂是胚生长产生由内向外“顶”的机械力量以及胚乳组织本身机械强度下降(胚乳弱化)的共同结果,而胚乳弱化则包括细胞壁的酶促和非酶促松弛机制.本文综述胚生长产生的机械力量、胚乳破裂的部位和方式、胚乳的组织结构及其细胞壁的化学组成、各种细胞壁降解酶及非酶的扩展蛋白(expansin)和活性氧在胚乳弱化中的作用等方面的研究进展.

  8. Concerted suppression of all starch branching enzyme genes in barley produces amylose-only starch granules

    Directory of Open Access Journals (Sweden)

    Carciofi Massimiliano

    2012-11-01

    Full Text Available Abstract Background Starch is stored in higher plants as granules composed of semi-crystalline amylopectin and amorphous amylose. Starch granules provide energy for the plant during dark periods and for germination of seeds and tubers. Dietary starch is also a highly glycemic carbohydrate being degraded to glucose and rapidly absorbed in the small intestine. But a portion of dietary starch, termed “resistant starch” (RS escapes digestion and reaches the large intestine, where it is fermented by colonic bacteria producing short chain fatty acids (SCFA which are linked to several health benefits. The RS is preferentially derived from amylose, which can be increased by suppressing amylopectin synthesis by silencing of starch branching enzymes (SBEs. However all the previous works attempting the production of high RS crops resulted in only partly increased amylose-content and/or significant yield loss. Results In this study we invented a new method for silencing of multiple genes. Using a chimeric RNAi hairpin we simultaneously suppressed all genes coding for starch branching enzymes (SBE I, SBE IIa, SBE IIb in barley (Hordeum vulgare L., resulting in production of amylose-only starch granules in the endosperm. This trait was segregating 3:1. Amylose-only starch granules were irregularly shaped and showed peculiar thermal properties and crystallinity. Transgenic lines retained high-yield possibly due to a pleiotropic upregualtion of other starch biosynthetic genes compensating the SBEs loss. For gelatinized starch, a very high content of RS (65 % was observed, which is 2.2-fold higher than control (29%. The amylose-only grains germinated with same frequency as control grains. However, initial growth was delayed in young plants. Conclusions This is the first time that pure amylose has been generated with high yield in a living organism. This was achieved by a new method of simultaneous suppression of the entire complement of genes encoding starch

  9. 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

  10. 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...

  11. 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...

  12. Rhizopus oligosporus and yeast co-cultivation during barley tempeh fermentation--nutritional impact and real-time PCR quantification of fungal growth dynamics.

    Science.gov (United States)

    Feng, Xin Mei; Passoth, Volkmar; Eklund-Jonsson, Charlotte; Alminger, Marie Larsson; Schnürer, Johan

    2007-06-01

    Barley tempeh was produced by fermenting barley kernels with Rhizopus oligosporus. The potential of the yeasts Saccharomyces cerevisiae (three strains), S. boulardii (one strain), Pichia anomala (one strain) and Kluyveromyces lactis (one strain) to grow together with R. oligosporus during barley tempeh fermentation was evaluated. All yeast strains grew during the fermentation and even during cold storage of tempeh (P<0.01). The growth of yeasts slightly increased the ergosterol contents, but did not influence amino acid contents and compositions, and did not reduce phytate contents. Slight increases of vitamins B(6) and niacinamide, and slight decreases of B(1) and biotin were observed. Quantification of fungal growth is difficult during mixed species fermentations because ergosterol is found in all fungal species, and colony-forming-unit (cfu) estimations are not reliable for R. oligosporus and other sporulating fungi. Therefore, we developed a quantitative real-time PCR method for individually quantifying S. cerevisiae and R. oligosporus growth in barley tempeh. The PCR results were highly correlated with the ergosterol content of R. oligosporus and with the number of cfu of S. cerevisiae. Thus, real-time PCR is a rapid and selective method to quantify yeasts and R. oligosporus during mixed species fermentation of inhomogenous substrate such as barley tempeh.

  13. 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.

  14. 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.

  15. 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.

  16. 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...

  17. Purification and characterization of recombinant high pI Barley α-Glucosidase

    DEFF Research Database (Denmark)

    Næsted, Henrik; Bojsen, Kirsten; Svensson, Birte

    α-glucosidase activity in barley seeds plays a crucial role in embryonic development. The concerted action of α-glucosidase, α-amylase, α-amylase, and limit dextrinase serves as the machinery responsible for the supply of glucose as energy source for the developing embryo in the germinating seed......-glucosidase containing culture using a four step purification strategy yielding 42 mg of pure enzyme. remarkably, the purified enzyme exhibits a higher molecular mass in sds-page than expected from the primary structure. the apparent molecular mass of 100 kda compared to 92 kda calculated from the amino acid sequence...

  18. Mechanisms of Induced Resistance in Barley Against Drechslera teres.

    Science.gov (United States)

    Lyngs Jørgensen, H J; Lübeck, P S; Thordal-Christensen, H; de Neergaard, E; Smedegaard-Petersen, V

    1998-07-01

    ABSTRACT Quantitative and qualitative histopathological methods and molecular analyses were used to study the mechanisms by which preinoculation with either of the nonbarley pathogens, Bipolaris maydis and Septoria nodorum, inhibited growth of Drechslera teres. Collectively, our data suggest that induced resistance is the principal mechanism responsible for impeding the pathogen. The enhancement of resistance in the host was primarily manifested during penetration by D. teres, and after penetration, where growth of D. teres ceased soon after development of infection vesicles. Thus, 24 h after pretreatment with B. maydis or S. nodorum, the penetration frequency from D. teres appressoria was reduced from 42.7% in the controls to 9.5 and 14.8%, respectively. The reductions were associated with increased formation of fluorescent papillae in induced cells (early defense reaction). The postpenetrational inhibition of D. teres completely stopped fungal growth and was apparently linked to an enhancement of multicellular hypersensitive responses in inducer-treated leaves (late defense reaction). Papillae formation and multicellular hypersensitive reactions were also observed in fully susceptible, noninduced control leaves, but they were inadequate to stop fungal progress. Northern blots from leaves treated with either inducer alone support the conclusion that induced resistance is involved in suppression of D. teres by increased formation of papillae and hypersensitive reactions. Thus, the blots showed strong expression of several defense response genes that are involved in these reactions in barley attacked by Erysiphe graminis f. sp. hordei.

  19. Reduction of 13 kD prolamins increases recombinant protein yield and recovery rate in rice endosperm.

    Science.gov (United States)

    Kawakatsu, Taiji; Takaiwa, Fumio

    2012-11-01

    RNA silencing inducible sequence (RSIS) causes post-transcriptional gene silencing (PTGS) of 5' or 3' flanking sequence-containing genes by inhibiting proper transcriptional termination. Exploiting this nature, 13 kD Pro-less, in which major seed storage protein (SSP) 13 kD prolamins are reduced, has been generated. In 13 kD Pro-less, other SSPs, such as glutelins, are increased as a compensation effect to maintain amino acid pool. 7Crp is the seven-linked epitope peptide derived from major cedar pollen allergens Cry j 1 and Cry j 2. When 7Crp is expressed in 13 kD Pro-less endosperm, accumulation level of 7Crp increased. Furthermore, recovery rate of 7Crp without reducing reagent increased. These findings indicate that 13 kD Pro-less endosperm provides a good production platform for recombinant proteins.

  20. 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

  1. 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...

  2. 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...

  3. 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...

  4. 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.

  5. An Endosperm-Associated Cuticle Is Required for Arabidopsis Seed Viability, Dormancy and Early Control of Germination.

    Directory of Open Access Journals (Sweden)

    Julien De Giorgi

    2015-12-01

    Full Text Available Cuticular layers and seeds are prominent plant adaptations to terrestrial life that appeared early and late during plant evolution, respectively. The cuticle is a waterproof film covering plant aerial organs preventing excessive water loss and protecting against biotic and abiotic stresses. Cutin, consisting of crosslinked fatty acid monomers, is the most abundant and studied cuticular component. Seeds are dry, metabolically inert structures promoting plant dispersal by keeping the plant embryo in an arrested protected state. In Arabidopsis thaliana seeds, the embryo is surrounded by a single cell endosperm layer itself surrounded by a seed coat layer, the testa. Whole genome analyses lead us to identify cutin biosynthesis genes as regulatory targets of the phytohormones gibberellins (GA and abscisic acid (ABA signaling pathways that control seed germination. Cutin-containing layers are present in seed coats of numerous species, including Arabidopsis, where they regulate permeability to outer compounds. However, the role of cutin in mature seed physiology and germination remains poorly understood. Here we identify in mature seeds a thick cuticular film covering the entire outer surface of the endosperm. This seed cuticle is defective in cutin-deficient bodyguard1 seeds, which is associated with alterations in endospermic permeability. Furthermore, mutants affected in cutin biosynthesis display low seed dormancy and viability levels, which correlates with higher levels of seed lipid oxidative stress. Upon seed imbibition cutin biosynthesis genes are essential to prevent endosperm cellular expansion and testa rupture in response to low GA synthesis. Taken together, our findings suggest that in the course of land plant evolution cuticular structures were co-opted to achieve key physiological seed properties.

  6. An Endosperm-Associated Cuticle Is Required for Arabidopsis Seed Viability, Dormancy and Early Control of Germination.

    Science.gov (United States)

    De Giorgi, Julien; Piskurewicz, Urszula; Loubery, Sylvain; Utz-Pugin, Anne; Bailly, Christophe; Mène-Saffrané, Laurent; Lopez-Molina, Luis

    2015-12-01

    Cuticular layers and seeds are prominent plant adaptations to terrestrial life that appeared early and late during plant evolution, respectively. The cuticle is a waterproof film covering plant aerial organs preventing excessive water loss and protecting against biotic and abiotic stresses. Cutin, consisting of crosslinked fatty acid monomers, is the most abundant and studied cuticular component. Seeds are dry, metabolically inert structures promoting plant dispersal by keeping the plant embryo in an arrested protected state. In Arabidopsis thaliana seeds, the embryo is surrounded by a single cell endosperm layer itself surrounded by a seed coat layer, the testa. Whole genome analyses lead us to identify cutin biosynthesis genes as regulatory targets of the phytohormones gibberellins (GA) and abscisic acid (ABA) signaling pathways that control seed germination. Cutin-containing layers are present in seed coats of numerous species, including Arabidopsis, where they regulate permeability to outer compounds. However, the role of cutin in mature seed physiology and germination remains poorly understood. Here we identify in mature seeds a thick cuticular film covering the entire outer surface of the endosperm. This seed cuticle is defective in cutin-deficient bodyguard1 seeds, which is associated with alterations in endospermic permeability. Furthermore, mutants affected in cutin biosynthesis display low seed dormancy and viability levels, which correlates with higher levels of seed lipid oxidative stress. Upon seed imbibition cutin biosynthesis genes are essential to prevent endosperm cellular expansion and testa rupture in response to low GA synthesis. Taken together, our findings suggest that in the course of land plant evolution cuticular structures were co-opted to achieve key physiological seed properties.

  7. 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.

  8. 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.

  9. Effect of gamma-radiation and microwave heating on endosperm microstructure in relation to some technological properties of wheat grain.

    Science.gov (United States)

    Błaszczak, W; Gralik, J; Klockiewicz-Kamińska, E; Fornal, J; Warchalewski, J R

    2002-04-01

    Grains of the Polish winter wheat variety Begra were subject to gamma-radiation (60Co) within the dose range of 0.05-10 kGy and microwave treatment from 15 to 180 s, the latter resulting in a bulk grain temperature ranging from 28 to 98 degrees C. Both processes were responsible for marked structural changes of wheat kernel endosperm and were pronounced with increasing dose of ionizing radiation or with prolonged time of microwave treatment. Endosperm microstructures of wheat kernels treated with gamma-radiation ranging from 0.05 to 0.5 kGy did not differ from that of untreated ones. Only some changes in the structure of starch granules and proteins were observed at doses of 1, 5 and 10 kGy. Simultaneously at the doses of 5 and 10 kGy a statistically significant decrease of falling number, sodium dodecyl sulfate (SDS) sedimentation values, dough stability and energy were observed, while dough weakening progressed. Microwave treatment longer than 90 s caused marked changes in kernel endosperm structure. Some changes of proteins caused by denaturation created visible fibrils as well as high swelling and deformation of starch granules. This was followed by an increase in the falling number value, decrease in the estimated results of SDS sedimentation test, lowering of the percent of gluten washed out and of the dough energy, respectively. Also a significant decrease in bread quality expressed by volume and score was observed with increasing time of exposure to microwave.

  10. Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L.

    Institute of Scientific and Technical Information of China (English)

    Nurul Izzati Osman; Norrizah Jaafar Sidik; Asmah Awal

    2016-01-01

    Objective: To induce callus from the medicinally valuable species, Barringtonia racemosa L.(B. racemosa) whereby the formation of callus is essential for micropropagation studies and in vitro plant secondary metabolites production.Methods: The callus induction potential in B. racemosa was assessed from endosperm explant cultured on different culture media and plant hormonal treatments. Lloyd and Mc Cown’s woody plant medium and Murashige and Skoog’s medium were used in the study as culture media. On the other hand, various concentrations and combinations of2,4-dichlorophenoxyacetic acid(1.0–2.0 mg/L) and kinetin(0.5–2.5 mg/L) had been incorporated in the culture media to exert the effects of auxin and cytokinin on callus induction.Results: From the present study, it was found that the profuse [(1.681 ± 0.770) g fresh weight,(0.239 ± 0.239) g dry weight] and friable callus formation was optimally produced with desirable morphology and considerable percentage of callus induction(56.70%) in endosperm explants cultured on 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.5 mg/L kinetin in Murashige and Skoog’s medium.Conclusions: A reliable protocol for inducing callus formation of profuse and friable morphology in endosperm explant of B. racemosa had therefore been successfully established.

  11. 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.

  12. 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...

  13. 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...

  14. Supplementation of Reduced Gluten Barley Diet with Oral Prolyl Endopeptidase Effectively Abrogates Enteropathy-Associated Changes in Gluten-Sensitive Macaques

    Directory of Open Access Journals (Sweden)

    Karol Sestak

    2016-06-01

    Full Text Available Celiac disease (CD is an autoimmune disorder that affects approximately three million people in the United States. Furthermore, non-celiac gluten sensitivity (NCGS affects an estimated additional 6% of the population, e.g., 20 million in the U.S. The only effective treatment of CD and NCGS requires complete removal of gluten sources from the diet. While required adherence to a gluten-free diet (GFD is extremely difficult to accomplish, efforts to develop additional supportive treatments are needed. To facilitate these efforts, we developed a gluten-sensitive (GS rhesus macaque model to study the effects of novel therapies. Recently reported results from phase one of this project suggest that partial improvement—but not remission—of gluten-induced disease can be accomplished by 100-fold reduction of dietary gluten, i.e., 200 ppm—by replacement of conventional dietary sources of gluten with a mutant, reduced gluten (RG barley (lys3a-derived source. The main focus of this (phase two study was to determine if the inflammatory effects of the residual gluten in lys3a mutant barley grain could be further reduced by oral supplementation with a prolylendopeptidase (PE. Results reveal that PE supplementation of RG barley diet induces more complete immunological, histopathological and clinical remission than RG barley diet alone. The combined effects of RG barley diet and PE supplementation resulted in a further decrease of inflammatory mediators IFN-γ and TNF secretion by peripheral lymphocytes, as well as decreased plasma anti-gliadin and anti-intestinal tissue transglutaminase (TG2 antibodies, diminished active caspase production in small intestinal mucosa, and eliminated clinical diarrhea—all comparable with a gluten-free diet induced remission. In summary, the beneficial results of a combined RG barley and PE administration in GS macaques may warrant the investigation of similar synergistic approaches.

  15. Supplementation of Reduced Gluten Barley Diet with Oral Prolyl Endopeptidase Effectively Abrogates Enteropathy-Associated Changes in Gluten-Sensitive Macaques.

    Science.gov (United States)

    Sestak, Karol; Thwin, Hazel; Dufour, Jason; Liu, David X; Alvarez, Xavier; Laine, David; Clarke, Adam; Doyle, Anthony; Aye, Pyone P; Blanchard, James; Moehs, Charles P

    2016-06-28

    Celiac disease (CD) is an autoimmune disorder that affects approximately three million people in the United States. Furthermore, non-celiac gluten sensitivity (NCGS) affects an estimated additional 6% of the population, e.g., 20 million in the U.S. The only effective treatment of CD and NCGS requires complete removal of gluten sources from the diet. While required adherence to a gluten-free diet (GFD) is extremely difficult to accomplish, efforts to develop additional supportive treatments are needed. To facilitate these efforts, we developed a gluten-sensitive (GS) rhesus macaque model to study the effects of novel therapies. Recently reported results from phase one of this project suggest that partial improvement-but not remission-of gluten-induced disease can be accomplished by 100-fold reduction of dietary gluten, i.e., 200 ppm-by replacement of conventional dietary sources of gluten with a mutant, reduced gluten (RG) barley (lys3a)-derived source. The main focus of this (phase two) study was to determine if the inflammatory effects of the residual gluten in lys3a mutant barley grain could be further reduced by oral supplementation with a prolylendopeptidase (PE). Results reveal that PE supplementation of RG barley diet induces more complete immunological, histopathological and clinical remission than RG barley diet alone. The combined effects of RG barley diet and PE supplementation resulted in a further decrease of inflammatory mediators IFN-γ and TNF secretion by peripheral lymphocytes, as well as decreased plasma anti-gliadin and anti-intestinal tissue transglutaminase (TG2) antibodies, diminished active caspase production in small intestinal mucosa, and eliminated clinical diarrhea-all comparable with a gluten-free diet induced remission. In summary, the beneficial results of a combined RG barley and PE administration in GS macaques may warrant the investigation of similar synergistic approaches.

  16. Supplementation of Reduced Gluten Barley Diet with Oral Prolyl Endopeptidase Effectively Abrogates Enteropathy-Associated Changes in Gluten-Sensitive Macaques

    Science.gov (United States)

    Sestak, Karol; Thwin, Hazel; Dufour, Jason; Liu, David X.; Alvarez, Xavier; Laine, David; Clarke, Adam; Doyle, Anthony; Aye, Pyone P.; Blanchard, James; Moehs, Charles P.

    2016-01-01

    Celiac disease (CD) is an autoimmune disorder that affects approximately three million people in the United States. Furthermore, non-celiac gluten sensitivity (NCGS) affects an estimated additional 6% of the population, e.g., 20 million in the U.S. The only effective treatment of CD and NCGS requires complete removal of gluten sources from the diet. While required adherence to a gluten-free diet (GFD) is extremely difficult to accomplish, efforts to develop additional supportive treatments are needed. To facilitate these efforts, we developed a gluten-sensitive (GS) rhesus macaque model to study the effects of novel therapies. Recently reported results from phase one of this project suggest that partial improvement—but not remission—of gluten-induced disease can be accomplished by 100-fold reduction of dietary gluten, i.e., 200 ppm—by replacement of conventional dietary sources of gluten with a mutant, reduced gluten (RG) barley (lys3a)-derived source. The main focus of this (phase two) study was to determine if the inflammatory effects of the residual gluten in lys3a mutant barley grain could be further reduced by oral supplementation with a prolylendopeptidase (PE). Results reveal that PE supplementation of RG barley diet induces more complete immunological, histopathological and clinical remission than RG barley diet alone. The combined effects of RG barley diet and PE supplementation resulted in a further decrease of inflammatory mediators IFN-γ and TNF secretion by peripheral lymphocytes, as well as decreased plasma anti-gliadin and anti-intestinal tissue transglutaminase (TG2) antibodies, diminished active caspase production in small intestinal mucosa, and eliminated clinical diarrhea—all comparable with a gluten-free diet induced remission. In summary, the beneficial results of a combined RG barley and PE administration in GS macaques may warrant the investigation of similar synergistic approaches. PMID:27367722

  17. The effect of various long-term tillage systems on soil properties and spring barley yield

    OpenAIRE

    MALECKA, Irena; Blecharczyk, Andrzej; SAWINSKA, Zuzanna; DOBRZENIECKI, Tomasz

    2012-01-01

    This study, performed on a soil that is classified as Albic Luvisols that developed on loamy sands overlying loamy material (1.4% organic matter and pH 6.5), concerns the impact of tillage systems on soil properties and the yield of spring barley. The experiment design included 3 tillage systems: conventional tillage, reduced tillage, and no-tillage. Continuous cultivation for 7 consecutive years by reduced tillage and no-tillage led to changes in the physical properties of the surface soil l...

  18. PRODUCTION OF RECOMBINANT HIGH pI-BARLEY α-GLUCOSIDASE

    DEFF Research Database (Denmark)

    Næsted, Henrik; Svensson, Birte

    α-glucosidase of glycoside hydrolase family 31 plays in barley seeds a crucial role in embryonic development. The concerted action of α-glucosidase, β-amylase, α-amylase, and limit dextrinase is responsible for supply of glucose in the germinating seed as energy source for the embryo and growing......-glucosidase was purified in a yield of 42 mg from the culture (3.5 liter) by a four step purification procedure. Remarkably, the purified enzyme exhibited slightly higher molecular mass of 100 kDa in SDS-PAGE than 92 kDa calculated from the primary structure. This difference in apparent molecular mass suggested...

  19. Extraction of light filth from barley, oatmeal, and mixed dry infant cereals: collaborative study.

    Science.gov (United States)

    Dent, R G

    1980-03-01

    A new method has been developed for the extraction of light filth from oatmeal, barley, and mixed dry infant cereals which involves HCl digestion followed by wet sieving. The residue is defatted in a paper cup, using isopropanol, and transferred to a 2 L Wildman trap flask with 40% isopropanol. Filth elements are extracted with light mineral oil. Interlaboratory studies resulted in 92 and 86% average recoveries of rodent hairs and insect fragments, respectively. The proposed method has been adopted as official first action.

  20. Desenvolvimento de populações duplo-haplóides de cevada cervejeira associadas à atividade das enzimas (1-3, 1-4-β-glucanases Development of doubled-haploids populations in malting barley associated to activity of enzymes (1-3, 1-4-β-glucanases

    Directory of Open Access Journals (Sweden)

    Janaína Endres Georg-Kraemer

    2011-05-01

    Full Text Available Populações duplo-haplóides apresentam especial vantagem para análises genéticas, uma vez que a informação que elas oferecem pode ser maximizada, devido ao fato que todos os locos encontram-se em homozigose. O objetivo deste trabalho foi o desenvolvimento de duas populações duplo-haplóides (DHs de cevada (Hordeum vulgare ssp. vulgare L. segregantes para a atividade das enzimas (1-3, 1-4-β-glucanases, através da técnica de cultura de anteras. Foram realizados dois cruzamentos com cultivares contrastantes para esta característica. As cultivares parentais selecionadas foram 'MN 698' e 'CEV 97047', para o desenvolvimento da população "malte verde" (MV, e 'Embrapa 127' e 'CEV 96025' para o desenvolvimento da população "malte seco" (MS. Foram cultivadas 10.734 anteras da população MS e 4.139 anteras da população MV. A população MV produziu 50% mais plântulas verdes quando comparada à população MS, refletindo a importância do genótipo na resposta à cultura de anteras e na regeneração. A maioria das plantas adultas duplo-haplóides foi obtida através da duplicação espontânea in vitro do genoma haplóide, ocorrendo em 66% das plantas da população MS e 76% das plantas da população MV. Também foram observadas, em menor frequência, plantas haplóides, triplóides e tetraplóides. Através da técnica de cultura de anteras, foram desenvolvidas 204 linhagens duplo-haplóides, das quais 72 linhagens são da população "malte seco" e 132 linhagens são da população "malte verde". Este material constitui um importante germoplasma para o melhoramento genético da cevada.Doubled haploid populations offer special advantages in genetic analyses, since the information they provide may be maximized due to the fact that all loci are homozygous. The aim of this study was to develop two barley (Hordeum vulgare ssp.vulgare L. doubled-haploid (DHs populations segregating to (1-3, 1-4-β-glucanases activity, utilizing the anther

  1. Determination of Four Major Saponins in Skin and Endosperm of Seeds of Horse Chestnut (Aesculus Hippocastanum L.) Using High Performance Liquid Chromatography with Positive Confirmation by Thin Layer Chromatography

    Science.gov (United States)

    Abudayeh, Zead Helmi Mahmoud; Al Azzam, Khaldun Mohammad; Naddaf, Ahmad; Karpiuk, Uliana Vladimirovna; Kislichenko, Viktoria Sergeevna

    2015-01-01

    Purpose: To separate and quantify four major saponins in the extracts of the skin and the endosperm of seeds of horse chestnut (Aesculus hippocastanum L.) using ultrasonic solvent extraction followed by a high performance liquid chromatography-diode array detector (HPLC-DAD) with positive confirmation by thin layer chromatography (TLC). Methods: The saponins: escin Ia, escin Ib, isoescin Ia and isoescin Ib were extracted using ultrasonic extraction method. The optimized extraction conditions were: 70% methanol as extraction solvent, 80 °C as extraction temperature, and the extraction time was achieved in 4 hours. The HPLC conditions used: Zorbax SB-ODS-(150 mm × 2.1 mm, 3 μm) column, acetonitrile and 0.10% phosphoric acid solution (39:61 v/v) as mobile phase, flow rate was 0.5 mL min−1 at 210 nm and 230 nm detection. The injection volume was 10 μL, and the separation was carried out isothermally at 30 °C in a heated chamber. Results: The results indicated that the developed HPLC method is simple, sensitive and reliable. Moreover, the content of escins in seeds decreased by more than 30% in endosperm and by more than 40% in skin upon storage for two years. Conclusion: This assay can be readily utilized as a quality control method for horse chestnut and other related medicinal plants. PMID:26819933

  2. High night temperatures during grain number determination reduce wheat and barley grain yield: a field study.

    Science.gov (United States)

    García, Guillermo A; Dreccer, M Fernanda; Miralles, Daniel J; Serrago, Román A

    2015-11-01

    Warm nights are a widespread predicted feature of climate change. This study investigated the impact of high night temperatures during the critical period for grain yield determination in wheat and barley crops under field conditions, assessing the effects on development, growth and partitioning crop-level processes driving grain number per unit area (GN). Experiments combined: (i) two contrasting radiation and temperature environments: late sowing in 2011 and early sowing in 2013, (ii) two well-adapted crops with similar phenology: bread wheat and two-row malting barley and (iii) two temperature regimes: ambient and high night temperatures. The night temperature increase (ca. 3.9 °C in both crops and growing seasons) was achieved using purpose-built heating chambers placed on the crop at 19:000 hours and removed at 7:00 hours every day from the third detectable stem node to 10 days post-flowering. Across growing seasons and crops, the average minimum temperature during the critical period ranged from 11.2 to 17.2 °C. Wheat and barley grain yield were similarly reduced under warm nights (ca. 7% °C(-1) ), due to GN reductions (ca. 6% °C(-1) ) linked to a lower number of spikes per m(2) . An accelerated development under high night temperatures led to a shorter critical period duration, reducing solar radiation capture with negative consequences for biomass production, GN and therefore, grain yield. The information generated could be used as a starting point to design management and/or breeding strategies to improve crop adaptation facing climate change.

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

    Directory of Open Access Journals (Sweden)

    Corinna Brit Liller

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

  4. Expression and genomic integration of transgenes after Agrobacterium-mediated transformation of mature barley embryos.

    Science.gov (United States)

    Uçarlı, C; Tufan, F; Gürel, F

    2015-02-06

    Mature embryos in tissue cultures are advantageous because of their abundance and rapid germination, which reduces genomic instability problems. In this study, 2-day-old isolated mature barley embryos were infected with 2 Agrobacterium hypervirulent strains (AGL1 and EHA105), followed by a 3-day period of co-cultivation in the presence of L-cystein amino acid. Chimeric expression of the b-glucuronidase gene (gusA) directed by a viral promoter of strawberry vein banding virus was observed in coleoptile epidermal cells and seminal roots in 5-day-old germinated seedlings. In addition to varying infectivity patterns in different strains, there was a higher ratio of transient b-glucuronidase expression in developing coleoptiles than in embryonic roots, indicating the high competency of shoot apical meristem cells in the mature embryo. A total of 548 explants were transformed and 156 plants developed to maturity on G418 media after 18-25 days. We detected transgenes in 74% of the screened plant leaves by polymerase chain reaction, and 49% of these expressed neomycin phosphotransferase II gene following AGL1 transformation. Ten randomly selected T0 transformants were analyzed using thermal asymmetric interlaced polymerase chain reaction and 24 fragments ranged between 200-600 base pairs were sequenced. Three of the sequences flanked with transferred-DNA showed high similarity to coding regions of the barley genome, including alpha tubulin5, homeobox 1, and mitochondrial 16S genes. We observed 70-200-base pair filler sequences only in the coding regions of barley in this study.

  5. A chemometric approach to determine the phenolic compounds in different barley samples by two different stationary phases: a comparison between C18 and pentafluorophenyl core shell columns.

    Science.gov (United States)

    Gómez-Caravaca, Ana María; Verardo, Vito; Berardinelli, Annachiara; Marconi, Emanuele; Caboni, Maria Fiorenza

    2014-08-15

    Barley (Hordeum vulgare L.) is a cereal crop that has been cultivated since ancient times. However, its interest as nutritional food and as food ingredient is relatively new. Thus, in this study, the phenolic compounds of eighteen different varieties of barley (4 waxy and 14 non-waxy) grown under the same agronomic conditions in the same experimental field have been determined by HPLC-DAD-MS. Two new methodologies were developed using new generation superficially porous HPLC columns with different stationary phases: C18 and pentafluorophenyl (PFP). Twelve free phenolic compounds and eight bound phenolic compounds could be identified in barley samples in less than 22min. The study of different method parameters showed that C18 column was more suitable for the analysis of phenolic compounds of barley. Hierarchical cluster analysis (HCA) was conducted in order to assess the different ability of the two different core shell HPLC columns in the discrimination between "waxy" and "non-waxy" varieties, and only HCA of C18 column could separate waxy and non-waxy genotypes. Significant differences in the content of phenolic compounds between waxy and non-waxy samples were found, being waxy barley samples the ones which presented higher content of free and bound phenolic compounds. Once the best discriminant HPLC column was established, principal component analysis (PCA) was applied and it was able to discriminate between "waxy" and "non-waxy" varieties; however it discriminated the barley samples based only in free phenolic compounds. Because of that, partial least squares discriminant analysis (PLS-DA) and Artificial Neural Networks (ANN) were carried out. PLS-DA and ANN permitted the classification of waxy and non-waxy genotypes from both free and bound phenolic compounds.

  6. 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

  7. 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...

  8. 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

  9. 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

  10. 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...

  11. 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...

  12. 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.

  13. 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....

  14. 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...

  15. 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

  16. 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

  17. 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.

  18. 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...

  19. 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,

  20. 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...

  1. 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.

  2. 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...

  3. 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

  4. 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.

  5. Detection of nitrogen deficiency QTL in juvenile wild barley introgression lines growing in a hydroponic system

    Directory of Open Access Journals (Sweden)

    Hoffmann Astrid

    2012-10-01

    Full Text Available Abstract Background In this report we studied the genetic regulation of juvenile development of wild barley introgression lines (S42ILs under two contrasting hydroponic nitrogen (N supplies. Ten shoot and root related traits were examined among 42 S42ILs and the recurrent parent ‘Scarlett’. The traits included tiller number, leaf number, plant height, leaf and root length, leaf to root length ratio, shoots and root dry weight, shoot to root weight ratio, and chlorophyll content. Our aims were (1 to test the suitability of a hydroponic system for early detection of favourable S42ILs, (2 to locate quantitative trait loci (QTL that control the examined traits, (3 to identify favourable wild barley alleles that improve trait performances in regard to N treatment and, finally, (4 to validate the identified QTL through comparison with previously reported QTL originating from the same parental cross. Results The phenotypic data were analysed in a mixed model association study to detect QTL. The post-hoc Dunnett test identified 28 S42ILs that revealed significant (P Hsp effects for tiller number, leaf number, leaf length, plant height and leaf to root ratio on the long arm of chromosome 7H. These QTL correspond to QTL for ears per plant and plant height that were previously detected in field trials conducted with the same S42ILs or with the S42 population. Conclusion Our results suggest that the QTL we identified under hydroponic N cultivation partly correspond to QTL detected in field experiments. Due to this finding, screening of plants in early developmental stages grown in a hydroponic system may be a fast and cost effective method for early QTL detection and marker-assisted allelic selection, potentially speeding up elite barley breeding programs.

  6. Comparative transcriptome profiling of two Tibetan wild barley genotypes in responses to low potassium.

    Directory of Open Access Journals (Sweden)

    Jianbin Zeng

    Full Text Available Potassium (K deficiency is one of the major factors affecting crop growth and productivity. Development of low-K tolerant crops is an effective approach to solve the nutritional deficiency in agricultural production. Tibetan annual wild barley is rich in genetic diversity and can grow normally under poor soils, including low-K supply. However, the molecular mechanism about low K tolerance is still poorly understood. In this study, Illumina RNA-Sequencing was performed using two Tibetan wild barley genotypes differing in low K tolerance (XZ153, tolerant and XZ141, sensitive, to determine the genotypic difference in transcriptome profiling. We identified a total of 692 differentially expressed genes (DEGs in two genotypes at 6 h and 48 h after low-K treatment, including transcription factors, transporters and kinases, oxidative stress and hormone signaling related genes. Meanwhile, 294 low-K tolerant associated DEGs were assigned to transporter and antioxidant activities, stimulus response, and other gene ontology (GO, which were mainly involved in starch and sucrose metabolism, lipid metabolism and ethylene biosynthesis. Finally, a hypothetical model of low-K tolerance mechanism in XZ153 was presented. It may be concluded that wild barley accession XZ153 has a higher capability of K absorption and use efficiency than XZ141 under low K stress. A rapid response to low K stress in XZ153 is attributed to its more K uptake and accumulation in plants, resulting in higher low K tolerance. The ethylene response pathway may account for the genotypic difference in low-K tolerance.

  7. Genotype by environment interaction of quantitative traits: a case study in barley.

    Science.gov (United States)

    Zhao, Fuping; Xu, Shizhong

    2012-07-01

    Genotype by environment interaction is a phenomenon that a better genotype in one environment may perform poorly in another environment. When the genotype refers to a quantitative trait locus (QTL), this phenomenon is called QTL by environment interaction, denoted by Q×E. Using a recently developed new Bayesian method and genome-wide marker information, we estimated and tested QTL main effects and Q×E interactions for a well-known barley dataset produced by the North American Barley Genome Mapping Project. This dataset contained seven quantitative traits collected from 145 doubled-haploid (DH) lines evaluated in multiple environments, which derived from a cross between two Canadian two-row barley lines, Harrington and TR306. Numerous main effects and Q×E interaction effects have been detected for all seven quantitative traits. However, main effects seem to be more important than the Q×E interaction effects for all seven traits examined. The number of main effects detected varied from 26 for the maturity trait to 75 for the heading trait, with an average of 61.86. The heading trait has the most detected effects, with a total of 98 (75 main, 29 Q×E). Among the 98 effects, 6 loci had both the main and Q×E effects. Among the total number of detected loci, on average, 78.5% of the loci show the main effects whereas 34.9% of the loci show Q×E interactions. Overall, we detected many loci with either the main or the Q×E effects, and the main effects appear to be more important than the Q×E interaction effects for all the seven traits. This means that most detected loci have a constant effect across environments. Another discovery from this analysis is that Q×E interaction occurs independently, regardless whether the locus has main effects.

  8. Mapping regulatory genes as candidates for cold and drought stress tolerance in barley.

    Science.gov (United States)

    Tondelli, A; Francia, E; Barabaschi, D; Aprile, A; Skinner, J S; Stockinger, E J; Stanca, A M; Pecchioni, N

    2006-02-01

    Cereal crop yield is greatly affected in many growing areas by abiotic stresses, mainly low temperature and drought. In order to find candidates for the tolerance genes for these stresses, 13 genes encoding for transcription factors and upstream regulators were screened by amplification and SSCP on six parental genotypes of three barley mapping populations ('Nure' x 'Tremois', 'Proctor' x 'Nudinka', and 'Steptoe' x 'Morex'), and mapped as newly developed STS, SNP, and SSCP markers. A new consensus function map was then drawn using the three maps above, including 16 regulatory candidate genes (CGs). The positions of barley cold and drought tolerance quantitative trait loci (QTLs) presently described in the literature were added to the consensus map to find positional candidates from among the mapped genes. A cluster of six HvCBF genes co-mapped with the Fr-H2 cold tolerance QTL, while no QTLs for the same trait were positioned on chromosome 7H, where two putative barley regulators of CBF expression, ICE1 and FRY1, found by homology search, were mapped in this work. These observations suggest that CBF gene(s) themselves, rather than their two regulators, are at present the best candidates for cold tolerance. Four out of 12 drought tolerance QTLs of the consensus map are associated with regulatory CGs, on chromosomes 2H, 5H, and 7H, and two QTLs with effector genes, on chromosomes 5H and 6H. The results obtained could be used to guide MAS applications, allowing introduction into an ideal genotype of favourable alleles of tolerance QTLs.

  9. High-throughput phenotyping to detect drought tolerance QTL in wild barley introgression lines

    KAUST Repository

    Honsdorf, Nora

    2014-05-13

    Drought is one of the most severe stresses, endangering crop yields worldwide. In order to select drought tolerant genotypes, access to exotic germplasm and efficient phenotyping protocols are needed. In this study the high-throughput phenotyping platform "The Plant Accelerator", Adelaide, Australia, was used to screen a set of 47 juvenile (six week old) wild barley introgression lines (S42ILs) for drought stress responses. The kinetics of growth development was evaluated under early drought stress and well watered treatments. High correlation (r = 0.98) between image based biomass estimates and actual biomass was demonstrated, and the suitability of the system to accurately and non-destructively estimate biomass was validated. Subsequently, quantitative trait loci (QTL) were located, which contributed to the genetic control of growth under drought stress. In total, 44 QTL for eleven out of 14 investigated traits were mapped, which for example controlled growth rate and water use efficiency. The correspondence of those QTL with QTL previously identified in field trials is shown. For instance, six out of eight QTL controlling plant height were also found in previous field and glasshouse studies with the same introgression lines. This indicates that phenotyping juvenile plants may assist in predicting adult plant performance. In addition, favorable wild barley alleles for growth and biomass parameters were detected, for instance, a QTL that increased biomass by approximately 36%. In particular, introgression line S42IL-121 revealed improved growth under drought stress compared to the control Scarlett. The introgression line showed a similar behavior in previous field experiments, indicating that S42IL-121 may be an attractive donor for breeding of drought tolerant barley cultivars. © 2014 Honsdorf et al.

  10. 麦绿素的生理功能及开发研究进展%Research Advancement on Applications and Function of Barley Green

    Institute of Scientific and Technical Information of China (English)

    李一卓

    2011-01-01

    This paper mainly introduced research advances on the physiological function of barley green,health food production status and its development prospect.%主要介绍麦绿素的生理功能的研究进展、保健食品开发现状及其发展前景等.

  11. Registration of LP1-2581, LP1-2163H, LP3-1159, and LP640-1304 low phytate spring barley germplasm lines

    Science.gov (United States)

    The Agricultural Research Service, U.S. Dept. of Agriculture (USDA-ARS) has developed and released four low-phytate spring barley (Hordeum vulgare L.) germplasm lines: LP1-2581 (Reg. No._____, PI 658245), LP1-2163H (Reg. No._____, PI 658248 ), LP3-1159 (Reg. No._____, PI 658247), and LP640-1304 (...

  12. A Transient Expression System for the Functional Assessment of Early Response Genes on the Powdery Mildew Infected Barley or Wheat Leaves

    Institute of Scientific and Technical Information of China (English)

    LI Ai-li

    2003-01-01

    The principle and the basic steps of the transient assay system for the functional assessment ofearly response genes on the powdery mildew infected barley or wheat leaves were summarized in brief. The development of this technology and its extensive application were reviewed. Future studies on this approach wererecommended in this paper.

  13. Scale-up of ethanol production from winter barley by the EDGE (enhanced dry grind enzymatic) process in fermentors up to 300 liters

    Science.gov (United States)

    A fermentation process, which was designated the EDGE (enhanced dry grind enzymatic) process, has recently been developed for barley ethanol production. In the EDGE process, in addition to the enzymes normally required for starch hydrolysis, commercial Beta-glucanases were used to hydrolyze (1,3)(1,...

  14. 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.

  15. 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...

  16. 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...

  17. Improving the efficiency of isolated microspore culture in six-row spring barley: I-optimization of key physical factors.

    Science.gov (United States)

    Esteves, Patricio; Belzile, François

    2014-06-01

    An improved isolated microspore culture protocol alleviating the recalcitrance typically observed in six-row spring barley was developed by optimizing four key physical factors to increase embryogenesis and reduce albinism. Doubled haploid (DH) plants are completely homozygous individuals that can be generated in just a few months via androgenesis in vitro. DHs are useful tools in genetic research and in plant breeding. Isolated microspore culture (IMC) is the most efficient way to produce DHs, but a strong genotype dependency imposes limitations to its wide application. Six-row, spring barley genotypes are considered as particularly recalcitrant due to a low frequency of embryogenesis and a high rate of albinism. Seeking to develop an efficient IMC protocol for this type of barley, we explored four important factors: (1) the harvest stage of immature spikes, (2) the type of pretreatment applied, (3) the osmotic potential in the induction medium, and (4) the plating density of microspores. This work was first performed using four barley genotypes: two typical six-row spring cultivars (ACCA and Léger), a two-row spring (Gobernadora) and a two-row winter (Igri) cultivar. First, by optimizing the harvest stage for each genotype we obtained a twofold to fourfold increase in the yield of embryogenic microspores. Second, two pretreatments (0.3 M mannitol for 2 days, or a combination of cold and heat over 15 days) both performed significantly better than the commonly used cold pretreatment (28 days at 4 °C). Third, an induction medium-containing mannitol (32 g/l) doubled green plant regeneration. Fourth, a plating density of 10(6) microspores/ml yielded the highest number of green regenerated plants. Our most important findings were then confirmed using sets of F1s from a six-row, spring-type breeding program.

  18. PEMANFAATAN FRAKSI KAYA ASAM LAURAT HASIL HIDROLISIS DARI ENDOSPERM KELAPA MENGGUNAKAN LIPASE ENDOGENEUS SEBAGAI PENGAWET SUSU KEDELAI KEMASAN (Utilization of High Lauric Fraction that Produced from Coconut Endosperm Using Lipase Endogenous as Preservation of Soybean Milk Packaging

    Directory of Open Access Journals (Sweden)

    Moh. Su'i

    2016-10-01

    Full Text Available Results of previous studies show that the high lauric fraction isolated from coconut endosperm is able to inhibit pathogenic and non-pathogenic bacteria. This research aims to study the addition of high lauric fraction that hydrolysed of coconut endosperm of the storability of soy milk packaging. High lauric fraction isolated from coconut milk, then the fraction analized of the fatty acid composition with gas chromatography (GC and then used as a preservative soy milk. The fraction is added to the soy milk with concentrations of 0, 10, 15 and 20%, then stored for 3 days. Every day is observed until soy milk damaged. The results showed that the fraction isolated from coconut milk contains 50.45% lauric acid, 17.52% myristic acid, 7.02% palmitic acid, 6.46% capric acid, 5.52% caprylic acid, 5.12% linoleic acid, 1.89% oleic acid, and 0.11% caproic acid. The addition of lauric acid-rich fraction of 20% were able to preserve soy milk for 2 days with a total microbe 1.00 x 104 cfu/ml, free fatty acids 0.12 m mol/ml, pH 5.05 and a balanced aroma 4 (nice. Keywords: Coconut, lauric acid, soy milk, storage ABSTRAK Hasil penelitian sebelumnya menunjukkan bahwa fraksi kaya asam laurat hasil isolasi dari endosperm kelapa mampu menghambat bakteri patogen dan non patogen. Penelitian ini bertujuan mempelajari penambahan fraksi kaya asam laurat hasil hidrolisis dari endosperm kelapa terhadap daya simpan susu kedelai kemasan. Fraksi yang kaya asam laurat diisolasi dari santan kelapa kemudian fraksi tersebut diuji komposisi asam lemaknya menggunakan chromatografi gas (GC dan selanjutnya digunakan sebagai bahan pengawet susu kedelai. Fraksi kaya asam laurat ditambahkan ke dalam susu kedelai dengan konsentrasi 0, 10, 15 dan 20%, kemudian disimpan selama 3 hari. Setiap hari dilakukan pengamatan hingga susu mengalami kerusakan. Hasil penelitian menunjukkan bahwa fraksi hasil isolasi dari santan kelapa mengandung asam laurat 50,45%, asam miristat 17,52%, asam palmitat

  19. Microscopic Structures of Endosperms Before and After Gelatinization in Rice Varieties with Varied Grain Quality

    Institute of Scientific and Technical Information of China (English)

    YANG Ze-min; WANG Wei-jin; LAN Sheng-yin; XU Zhen-xiu; ZHOU Zhu-qing; WANG Meng

    2003-01-01

    The microscopic structures of the endosperm of indica rice varieties with different quality be-fore and after gelatinization were observed using scanning electron microscope. The results showed that the de-gree of gelatinization varied in different parts of the grain and in different varieties under the same experimen-tal conditions. The gelatinization of dorsal side was the most complete. Its cells were decomposed totally intopuff-like or flocculent materials. The ventral side gelatinized less thoroughly, appearing agglomerate and somecell frames were still visible. The middle part gelatinized most incompletely and the cells were still integrated.Evident differences in gelatinization were observed among different varieties, the dorsal, ventral and middleparts of high quality varieties gelatinized more thoroughly than those of the corresponding parts of low qualityvarieties respectively. An obvious concavity often appeared in the middle of the cross-section of the low qualitygrains while the cross-section of high quality grains was normally flat. The same phenomenon was noted whencomparing the early maturing indica rice and the late maturing indica rice. Varietal difference of gelatiniza-tion in dorsal sides was not as distinct as in middle parts and ventral sides. The difference among dorsal side,middle part and ventral side in gelatinization was greater in low quality grains than that of high quality grains.In addition, a lot of ruptured cells were observed in the cross-section of high quality rice, while few of themcould be found in the low quality rice. Apparently, the number of ruptured cells is positively correlated withrice quality. Quality of rice grain also has positive correlation with the rate of water absorption and extension. Highrates of water absorption and extension lead to better gelatinization of rice grain, and hence indicate good quality.

  20. High-resolution analysis of parent-of-origin allelic expression in the Arabidopsis Endosperm.

    Directory of Open Access Journals (Sweden)

    Philip Wolff

    2011-06-01

    Full Text Available Genomic imprinting is an epigenetic phenomenon leading to parent-of-origin specific differential expression of maternally and paternally inherited alleles. In plants, genomic imprinting has mainly been observed in the endosperm, an ephemeral triploid tissue derived after fertilization of the diploid central cell with a haploid sperm cell. In an effort to identify novel imprinted genes in Arabidopsis thaliana, we generated deep sequencing RNA profiles of F1 hybrid seeds derived after reciprocal crosses of Arabidopsis Col-0 and Bur-0 accessions. Using polymorphic sites to quantify allele-specific expression levels, we could identify more than 60 genes with potential parent-of-origin specific expression. By analyzing the distribution of DNA methylation and epigenetic marks established by Polycomb group (PcG proteins using publicly available datasets, we suggest that for maternally expressed genes (MEGs repression of the paternally inherited alleles largely depends on DNA methylation or PcG-mediated repression, whereas repression of the maternal alleles of paternally expressed genes (PEGs predominantly depends on PcG proteins. While maternal alleles of MEGs are also targeted by PcG proteins, such targeting does not cause complete repression. Candidate MEGs and PEGs are enriched for cis-proximal transposons, suggesting that transposons might be a driving force for the evolution of imprinted genes in Arabidopsis. In addition, we find that MEGs and PEGs are significantly faster evolving when compared to other genes in the genome. In contrast to the predominant location of mammalian imprinted genes in clusters, cluster formation was only detected for few MEGs and PEGs, suggesting that clustering is not a major requirement for imprinted gene regulation in Arabidopsis.

  1. A Quantitative Profiling Method of Phytohormones and Other Metabolites Applied to Barley Roots Subjected to Salinity Stress

    Science.gov (United States)

    Cao, Da; Lutz, Adrian; Hill, Camilla B.; Callahan, Damien L.; Roessner, Ute

    2017-01-01

    As integral parts of plant signaling networks, phytohormones are involved in the regulation of plant metabolism and growth under adverse environmental conditions, including salinity. Globally, salinity is one of the most severe abiotic stressors with an estimated 800 million hectares of arable land affected. Roots are the first plant organ to sense salinity in the soil, and are the initial site of sodium (Na+) exposure. However, the quantification of phytohormones in roots is challenging, as they are often present at extremely low levels compared to other plant tissues. To overcome this challenge, we developed a high-throughput LC-MS method to quantify ten endogenous phytohormones and their metabolites of diverse chemical classes in roots of barley. This method was validated in a salinity stress experiment with six barley varieties grown hydroponically with and without salinity. In addition to phytohormones, we quantified 52 polar primary metabolites, including some phytohormone precursors, using established GC-MS and LC-MS methods. Phytohormone and metabolite data were correlated with physiological measurements including biomass, plant size and chlorophyll content. Root and leaf elemental analysis was performed to determine Na+ exclusion and K+ retention ability in the studied barley varieties. We identified distinct phytohormone and metabolite signatures as a response to salinity stress in different barley varieties. Abscisic acid increased in the roots of all varieties under salinity stress, and elevated root salicylic acid levels were associated with an increase in leaf chlorophyll content. Furthermore, the landrace Sahara maintained better growth, had lower Na+ levels and maintained high levels of the salinity stress linked metabolite putrescine as well as the phytohormone metabolite cinnamic acid, which has been shown to increase putrescine concentrations in previous studies. This study highlights the importance of root phytohormones under salinity stress and

  2. Construction of the Seed-Coat cDNA Microarray and Screening of Differentially Expressed Genes in Barley

    Institute of Scientific and Technical Information of China (English)

    Jin-Song PANG; Meng-Yuan HE; Bao LIU

    2004-01-01

    Some barley mutants can synthesize neither anthocyanins nor proanthocyanidins in the seed coat, which is related to several genes in locus Ant13, but the exact model of action remains unknown. We used the cDNA microarray technology with barley transcription-deficient mutant (ant13-152) that does not synthesize proanthocyanidins as the tester, and its wild type genotype (Triumph) as the driver, to study this question. Six-thousand and forty-eight clones from the wild type Morex testa+pericarp cDNA library were amplified using PCR, and the DNA fragments were spotted on commercial amino-modified glass slide as microarray. The mRNAs from the developing seed coat (8-15 days) of both the mutant and the wild-type barley plants were isolated, and labeled respectively with Cy3-dUTP and Cy5-dUTP when reversely transcribed to cDNAs. The labeled cDNAs were used as probes, mixed at the same molar concentration, and hybridized with the DNA fragments on the slide. Seventy clones exhibiting marked differential expression (ratio>4) were identified from the microarray. All the 25 cDNA clones that showed an over-expression in wild type in comparison to the mutant ant13-152 were sequenced. It was found that most of these overexpressing clones were transcription/translation and hordein-associated genes. These results have laid a solid material basis for further elucidation of the metabolic pathway in proanthocyanidin synthesis in barley and likely other plants.

  3. BAC library resources for map-based cloning and physical map construction in barley (Hordeum vulgare L.

    Directory of Open Access Journals (Sweden)

    Wu Cheng-Cang

    2011-05-01

    Full Text Available Background Although second generation sequencing (2GS technologies allow re-sequencing of previously gold-standard-sequenced genomes, whole genome shotgun sequencing and de novo assembly of large and complex eukaryotic genomes is still difficult. Availability of a genome-wide physical map is therefore still a prerequisite for whole genome sequencing for genomes like barley. To start such an endeavor, large insert genomic libraries, i.e. Bacterial Artificial Chromosome (BAC libraries, which are unbiased and representing deep haploid genome coverage, need to be ready in place. Result Five new BAC libraries were constructed for barley (Hordeum vulgare L. cultivar Morex. These libraries were constructed in different cloning sites (HindIII, EcoRI, MboI and BstXI of the respective vectors. In order to enhance unbiased genome representation and to minimize the number of gaps between BAC contigs, which are often due to uneven distribution of restriction sites, a mechanically sheared library was also generated. The new BAC libraries were fully characterized in depth by scrutinizing the major quality parameters such as average insert size, degree of contamination (plate wide, neighboring, and chloroplast, empty wells and off-scale clones (clones with 250 fragments. Additionally a set of gene-based probes were hybridized to high density BAC filters and showed that genome coverage of each library is between 2.4 and 6.6 X. Conclusion BAC libraries representing >20 haploid genomes are available as a new resource to the barley research community. Systematic utilization of these libraries in high-throughput BAC fingerprinting should allow developing a genome-wide physical map for the barley genome, which will be instrumental for map-based gene isolation and genome sequencing.

  4. Isolation and characterization of wheat bran starch and endosperm starch of selected soft wheats grown in Michigan and comparison of their physicochemical properties.

    Science.gov (United States)

    Liu, Ya; Ng, Perry K W

    2015-06-01

    Three soft wheat varieties with relatively high crop yields and different levels of milling softness equivalence were studied to characterize bran starch properties compared with those of endosperm starch from the same wheat sample. Bran starch had more short chains than had endosperm starch, and was found to have a higher percentage of B-type granules, higher amylose content, higher crystallinity, broader gelatinization temperature range, higher enthalpy of gelatinization, lower retrogradation degree, and lower pasting peak and setback viscosities than had the counterpart endosperm starch. Bran starch of variety Aubrey had the highest crystallinity (21.75%) and gelatinization temperature (62.9°C), while bran starch of variety D8006 had the highest percentage of B-type granules and lowest retrogradation degree (21.7%). Results of this study provide a foundation for better utilization of bran starch during whole grain food processing.

  5. Antioxidant activity in barley (Hordeum Vulgare L.) grains roasted in a microwave oven under conditions optimized using response surface methodology.

    Science.gov (United States)

    Omwamba, Mary; Hu, Qiuhui

    2010-01-01

    Microwave processing and cooking of foods is a recent development that is gaining momentum in household as well as large-scale food applications. Barley contains phenol compounds which possess antioxidant activity. In this study the microwave oven roasting condition was optimized to obtain grains with high antioxidant activity measured as the ability to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. Antioxidant activity of grains roasted under optimum conditions was assessed based on DPPH radical scavenging activity, reducing power and inhibition of oxidation in linoleic acid system. The optimum condition for obtaining roasted barley with high antioxidant activity (90.5% DPPH inhibition) was found to be at 600 W microwave power, 8.5 min roasting time, and 61.5 g or 2 layers of grains. The roasting condition influenced antioxidant activity both individually and interactively. Statistical analysis showed that the model was significant (P < 0.0001). The acetone extract had significantly high inhibition of lipid peroxidation and DPPH radical scavenging activity compared to the aqueous extract and alpha-tocopherol. The reducing power of acetone extracts was not significantly different from alpha-tocopherol. The acetone extract had twice the amount of phenol content compared to the aqueous extract indicating its high extraction efficiency. GC-MS analysis revealed the presence of phenol acids, amino phenols, and quinones. The aqueous extract did not contain 3,4-dihydroxybenzaldehyde and 4-hydroxycinnamic acid which are phenol compounds reported to contribute to antioxidant activity in barley grain.

  6. Creation of the first ultra-low gluten barley (Hordeum vulgare L.) for coeliac and gluten-intolerant populations.

    Science.gov (United States)

    Tanner, Gregory J; Blundell, Malcolm J; Colgrave, Michelle L; Howitt, Crispin A

    2016-04-01

    Coeliac disease is a well-defined condition that is estimated to affect approximately 1% of the population worldwide. Noncoeliac gluten sensitivity is a condition that is less well defined, but is estimated to affect up to 10% of the population, and is often self-diagnosed. At present, the only remedy for both conditions is a lifelong gluten-free diet. A gluten-free diet is often expensive, high in fat and low in fibre, which in themselves can lead to adverse health outcomes. Thus, there is an opportunity to use novel plant breeding strategies to develop alternative gluten-free grains. In this work, we describe the breeding and characterization of a novel ultra-low gluten (ULG) barley variety in which the hordein (gluten) content was reduced to below 5 ppm. This was achieved using traditional breeding strategies to combine three recessive alleles, which act independently of each other to lower the hordein content in the parental varieties. The grain of the initial variety was shrunken compared to wild-type barleys. We implemented a breeding strategy to improve the grain size to near wild-type levels and demonstrated that the grains can be malted and brewed successfully. The ULG barley has the potential to provide novel healthy foods and beverages for those who require a gluten-free diet.

  7. 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.

  8. Nitrogen availability to Pseudomonas fluorescens DF57 is limited during decomposition of barley straw in bulk soil and in the barley rhizosphere.

    Science.gov (United States)

    Jensen, L E; Nybroe, O

    1999-10-01

    The availability of nitrogen to Pseudomonas fluorescens DF57 during straw degradation in bulk soil and in barley rhizosphere was studied by introducing a bioluminescent reporter strain (DF57-N3), responding to nitrogen limitation, to model systems of varying complexity. DF57-N3 was apparently not nitrogen limited in the natural and sterilized bulk soil used for these experiments. The soil was subsequently amended with barley straw, representing a plant residue with a high carbon-to-nitrogen ratio (between 60 and 100). In these systems the DF57-N3 population gradually developed a nitrogen limitation response during the first week of straw decomposition, but exclusively in the presence of the indigenous microbial population. This probably reflects the restricted ability of DF57 to degrade plant polymers by hydrolytic enzymes. The impact of the indigenous population on nitrogen availability to DF57-N3 was mimicked by the cellulolytic organism Trichoderma harzianum Rifai strain T3 when coinoculated with DF57-N3 in sterilized, straw-amended soil. Limitation occurred concomitantly with fungal cellulase production, pointing to the significance of hydrolytic activity for the mobilization of straw carbon sources, thereby increasing the nitrogen demand. Enhanced survival of DF57-N3 in natural soil after straw amendment further indicated that DF57 was cross-fed with carbon/energy sources. The natural barley rhizosphere was experienced by DF57-N3 as an environment with restricted nitrogen availability regardless of straw amendment. In the rhizosphere of plants grown in sterilized soil, nitrogen limitation was less severe, pointing to competition with indigenous microorganisms as an important determinant of the nitrogen status for DF57-N3 in this environment. Hence, these studies have demonstrated that nitrogen availability and gene expression in Pseudomonas is intimately linked to the structure and function of the microbial community. Further, it was demonstrated that the

  9. Genevestigator Transcriptome Meta-Analysis and Biomarker Search Using Rice and Barley Gene Expression Databases

    Institute of Scientific and Technical Information of China (English)

    Philip Zimmermann; Oliver Laule; Josy Schmitz; Tomas Hruz; Stefan Bleuler; Wilhelm Gruissem

    2008-01-01

    The wide-spread use of microarray technologies to study plant transcriptomes has Ied to important discoveries and to an accumulation of profiling data covering a wide range of different tissues,developmental stages,perturbations,and genotypes.Querying a large number of microarray experiments can provide insights that cannot be gained by analyzing single experiments.However,such a meta-analysis poses.significant challenges with respect to data comparability and normalization,systematic sample annotation,and analysis tools.Genevestigator addresses these issues using a large curated expression database and a set of specifically developed analysis tools that are accessible over the internet.This combination has already proven to be usefuIin the area of plant research based on a Iarge set of Arabidopsis data (Grennan,2006).Here,we present the release of the Genevestigator rice and barley gene expression databases that contain quailty-controlled and welI annotated microarray experiments using ontologies.The databases currently comprise experiments from pathology,plant nutrition.abiotic stress,hormone treatment,genotype,and spatial or temporal analysis,but are expected to cover a broad variety of research areas as more experimentaI data become available.The transcriptome meta-analysis of the modeI species rice and barley iS expected to deliver results that can be used for functional genomics and biotechnoIogical applications in cereals.

  10. 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.

  11. Rice alcohol dehydrogenase 1 promotes survival and has a major impact on carbohydrate metabolism in the embryo and endosperm when seeds are germinated in partially oxygenated water

    Science.gov (United States)

    Takahashi, Hirokazu; Greenway, Hank; Matsumura, Hideo; Tsutsumi, Nobuhiro; Nakazono, Mikio

    2014-01-01

    Background and Aims Rice (Oryza sativa) has the rare ability to germinate and elongate a coleoptile under oxygen-deficient conditions, which include both hypoxia and anoxia. It has previously been shown that ALCOHOL DEHYDROGENASE 1 (ADH1) is required for cell division and cell elongation in the coleoptile of submerged rice seedlings by means of studies using a rice ADH1-deficient mutant, reduced adh activity (rad). The aim of this study was to understand how low ADH1 in rice affects carbohydrate metabolism in the embryo and endosperm, and lactate and alanine synthesis in the embryo during germination and subsequent coleoptile growth in submerged seedlings. Methods Wild-type and rad mutant rice seeds were germinated and grown under complete submergence. At 1, 3, 5 and 7 d after imbibition, the embryo and endosperm were separated and several of their metabolites were measured and compared. Key results In the rad embryo, the rate of ethanol fermentation was halved, while lactate and alanine concentrations were 2·4- and 5·7- fold higher in the mutant than in the wild type. Glucose and fructose concentrations in the embryos increased with time in the wild type, but not in the rad mutant. The rad mutant endosperm had lower amounts of the α-amylases RAMY1A and RAMY3D, resulting in less starch degradation and lower glucose concentrations. Conclusions These results suggest that ADH1 is essential for sugar metabolism via glycolysis to ethanol fermentation in both the embryo and endosperm. In the endosperm, energy is presumably needed for synthesis of the amylases and for sucrose synthesis in the endosperm, as well as for sugar transport to the embryo. PMID:24431339

  12. 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...

  13. 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...

  14. 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.

  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. Linking stomatal traits and expression of slow anion channel genes HvSLAH1 and HvSLAC1 with grain yield for increasing salinity tolerance in barley.

    Science.gov (United States)

    Liu, Xiaohui; Mak, Michelle; Babla, Mohammad; Wang, Feifei; Chen, Guang; Veljanoski, Filip; Wang, Gang; Shabala, Sergey; Zhou, Meixue; Chen, Zhong-Hua

    2014-01-01

    Soil salinity is an environmental and agricultural problem in many parts of the world. One of the keys to breeding barley for adaptation to salinity lies in a better understanding of the genetic control of stomatal regulation. We have employed a range of physiological (stomata assay, gas exchange, phylogenetic analysis, QTL analysis), and molecular techniques (RT-PCR and qPCR) to investigate stomatal behavior and genotypic variation in barley cultivars and a genetic population in four experimental trials. A set of relatively efficient and reliable methods were developed for the characterization of stomatal behavior of a large number of varieties and genetic lines. Furthermore, we found a large genetic variation of gas exchange and stomatal traits in barley in response to salinity stress. Salt-tolerant cultivar CM72 showed significantly larger stomatal aperture under 200 mM NaCl treatment than that of salt-sensitive cultivar Gairdner. Stomatal traits such as aperture width/length were found to significantly correlate with grain yield under salt treatment. Phenotypic characterization and QTL analysis of a segregating double haploid population of the CM72/Gairdner resulted in the identification of significant stomatal traits-related QTLs for salt tolerance. Moreover, expression analysis of the slow anion channel genes HvSLAH1 and HvSLAC1 demonstrated that their up-regulation is linked to higher barley grain yield in the field.

  18. Linking stomatal traits and expression of slow anion channel genes HvSLAH1 2 HvSLAC1 with grain yield for increasing salinity tolerance in barley

    Directory of Open Access Journals (Sweden)

    Xiaohui eLiu

    2014-11-01

    Full Text Available Soil salinity is an environmental and agricultural problem in many parts of the world. One of the keys to breeding barley for adaptation to salinity lies in a better understanding of the genetic control of stomatal regulation. We have employed a range of physiological and molecular techniques (stomata assay, gas exchange, phylogenetic analysis, QTL analysis, and gene expression to investigate stomatal behaviour and genotypic variation in barley cultivars and a genetic population in four experimental trials. A set of relatively efficient and reliable methods were developed for the characterisation of stomatal behaviour of large numbers of varieties and genetic lines. Furthermore, we have found a large genetic variation of gas exchange and stomatal traits in barley in response to salinity stress. Salt-tolerant CM72 showed significantly larger stomatal aperture in 200 mM NaCl treatment than that of salt-sensitive Gairdner. Stomatal traits such as aperture width/length were found to significantly correlate with grain yield in salt treatment. Phenotypic characterisation and QTL analysis of a segregating double haploid population of the CM72/Gairdner resulted in the identification of significant stomatal traits-related QTLs for salt tolerance. Moreover, expression analysis of the slow anion channel genes HvSLAH1 and HvSLAC1 demonstrated that their up-regulation is linked to high barley grain yield in the field.

  19. 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...

  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. 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.

  2. 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.

  3. Barley grain enrichement with essential elements by agronomic biofortification

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    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

  4. 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)

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. Transcriptome Comparative Profiling of Barley eibi1 Mutant Reveals Pleiotropic Effects of HvABCG31 Gene on Cuticle Biogenesis and Stress Responsive Pathways

    Directory of Open Access Journals (Sweden)

    Eviatar Nevo

    2013-10-01

    Full Text Available Wild barley eibi1 mutant with HvABCG31 gene mutation has low capacity to retain leaf water, a phenotype associated with reduced cutin deposition and a thin cuticle. To better understand how such a mutant plant survives, we performed a genome-wide gene expression analysis. The leaf transcriptomes between the near-isogenic lines eibi1 and the wild type were compared using the 22-k Barley1 Affymetrix microarray. We found that the pleiotropic effect of the single gene HvABCG31 mutation was linked to the co-regulation of metabolic processes and stress-related system. The cuticle development involved cytochrome P450 family members and fatty acid metabolism pathways were significantly up-regulated by the HvABCG31 mutation, which might be anticipated to reduce the levels of cutin monomers or wax and display conspicuous cuticle defects. The candidate genes for responses to stress were induced by eibi1 mutant through activating the jasmonate pathway. The down-regulation of co-expressed enzyme genes responsible for DNA methylation and histone deacetylation also suggested that HvABCG31 mutation may affect the epigenetic regulation for barley development. Comparison of transcriptomic profiling of barley under biotic and abiotic stresses revealed that the functions of HvABCG31 gene to high-water loss rate might be different from other osmotic stresses of gene mutations in barley. The transcriptional profiling of the HvABCG31 mutation provided candidate genes for further investigation of the physiological and developmental changes caused by the mutant.

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. Effect of gibberellic acid on α-amylase and ribonuclease activities in rye endosperm after various periods of grain storage

    Directory of Open Access Journals (Sweden)

    T. Mierzwińska

    2015-01-01

    Full Text Available α-Amylase and ribonuclease activities were determined in embryoless rye half-seeds exhibiting different viaibility after various times of storage (2, 5 and 8 years. The half-seeds were incubated for 24, 48, 72 and 96 h with and without GA3. α-Amylase activity in the endosperm of rye grain stored for 2 and 5 years and incubated without the hormone increased considerably, whereas in 8-year grain, practically incapable of germination, it remained almost unchanged. GA3caused a considerable increase of α-amylase activity in all the tested samples. In grain stored for 2 and 5 years the enzyme activity reached its peak under the influence of the hormone earlier than in the control samples. The differences in ribonuclease activity between the particular samples were less pronounced, and the reaction to the hormone much weaker. The results indicate that the reaction of rye endosperm to GA3depends on the physiological properties of the grain.

  16. Minus end-directed kinesin-like motor protein, Kcbp, localizes to anaphase spindle poles in Haemanthus endosperm.

    Science.gov (United States)

    Smirnova, E A; Reddy, A S; Bowser, J; Bajer, A S

    1998-01-01

    Microtubule-based motor proteins assemble and reorganize acentrosomal mitotic and meiotic spindles in animal cells. The functions of motor proteins in acentrosomal plant spindles are unknown. The cellulosic cell wall and relative small size of most plant cells precludes accurate detection of the spatial distribution of motors in mitosis. Large cell size and absence of a cellulosic cell wall in Haemanthus endosperm make these cells ideally suited for studies of the spatial distribution of motor proteins during cell division. Immunolocalization of a kinesin-like calmodulin-binding protein (KCBP) in Haemanthus endosperm revealed its mitotic distribution. KCBP appears first in association with the prophase spindle. Highly concentrated within the cores of individual kinetochore fibers, KCBP decorates microtubules of kinetochore-fibers through metaphase. By mid-anaphase (when a barrel-shaped spindle becomes convergent), the protein redistributes and accumulates at the spindle polar regions. In telophase, KCBP relocates toward the phragmoplast and cell plate. These data suggest a role for KCBP in anaphase spindle microtubule convergence, which assures coherence of kinetochore-fibers within each sister chromosome group. Increasing coherence of kinetochore-fibers prevents splitting within each sister chromosome group and formation of multinucleated cells.

  17. Mode of endosperm and wholemeal arabinoxylans solubilisation during rye breadmaking: genotypic diversity in the level, substitution degree and macromolecular characteristics.

    Science.gov (United States)

    Cyran, Malgorzata R; Dynkowska, Wioletta M

    2014-02-15

    Insight into solubilisation mechanisms of rye arabinoxylans during breadmaking is important for understanding the biochemical processes that affect bread attributes. Purified ethanol precipitated water-extractable arabinoxylans (WE-AX) and residual unextractable counterparts (WU-AX) were isolated from rye flours and resulting breads. While the endosperm flours had lower endoxylanase activities and higher arabinose-to-xylose ratios of WU-AX than those of corresponding wholemeals, there were not any significant differences between them in the mean amounts of WU-AX hydrolysed during breadmaking. Nevertheless, they were highly affected by rye cultivar used for breadmaking. On average, 42% and 36% of WU-AX were recovered in bread WE-AX fraction, causing its 11% and 8% increase, respectively for endosperm and wholemeal breads. Bread WE-AX, however, had lower molecular weights than those of starting flours, implying chains depolymerisation. Degree of AX solubilisation depends mainly on rye genotype used, determining combined effect of enzymatic and acid hydrolyses, associations and fine structure of AX.

  18. Construction of chimeric enzymes out of maize endosperm branching enzymes I and II: activity and properties.

    Science.gov (United States)

    Kuriki, T; Stewart, D C; Preiss, J

    1997-11-14

    Branching enzyme I and II isoforms from maize endosperm (mBE I and mBE II, respectively) have quite different properties, and to elucidate the domain(s) that determines the differences, chimeric genes consisting of part mBE I and part mBE II were constructed. When expressed under the control of the T7 promoter in Escherichia coli, several of the chimeric enzymes were inactive. The only fully active chimeric enzyme was mBE II-I BspHI, in which the carboxyl-terminal part of mBE II was exchanged for that of mBE I at a BspHI restriction site and was purified to homogeneity and characterized. Another chimeric enzyme, mBE I-II HindIII, in which the amino-terminal end of mBE II was replaced with that of mBE I, had very little activity and was only partially characterized. The purified mBE II-I BspHI exhibited higher activity than wild-type mBE I and mBE II when assayed by the phosphorylase a stimulation assay. mBE II-I BspHI had substrate specificity (preference for amylose rather than amylopectin) and catalytic capacity similar to mBE I, despite the fact that only the carboxyl terminus was from mBE I, suggesting that the carboxyl terminus may be involved in determining substrate specificity and catalytic capacity. In chain transfer experiments, mBE II-I BspHI transferred more short chains (with a degree of polymerization of around 6) in a fashion similar to mBE II. In contrast, mBE I-II HindIII transferred more long chains (with a degree of polymerization of around 11-12), similar to mBE I, suggesting that the amino terminus of mBEs may play a role in the size of oligosaccharide chain transferred. This study challenges the notion that the catalytic centers for branching enzymes are exclusively located in the central portion of the enzyme; it suggests instead that the amino and carboxyl termini may also be involved in determining substrate preference, catalytic capacity, and chain length transfer.

  19. Protein bodies of castor bean endosperm: isolation, fractionation, and the characterization of protein components.

    Science.gov (United States)

    Tully, R E; Beevers, H

    1976-12-01

    Protein bodies in the endosperm of castor bean seeds (Ricinus communis L.) contain phytin globoids and protein crystalloids embedded in an amorphous proteinaceous matrix. The protein bodies are apparently surrounded by a single membrane. The protein bodies were isolated by grinding and centrifuging in glycerol. Such isolated protein bodies were almost identical (after cytological fixation) to those observed in situ, except that the globoids were lost. However, membrane-like structures appear to have surrounded the globoids. Histochemical analysis of the isolated protein bodies showed that carbohydrates (glycoproteins) are localized only in the matrix region.Addition of water to protein bodies in glycerol caused dissolution of the matrix, and release of the globoids and crystalloids. When the crystalloids were centrifuged on sucrose density gradients, they were recovered at an equilibrium density of 1.29 to 1.30 g/ml. The crystalloids were only slightly soluble in most aqueous buffers but were very soluble in sodium dodecyl sulfate, urea, or NaOH solutions.Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and chromatography on ion exchange celluloses show that the protein bodies are composed of one major and several minor anodic proteins. The major protein, along with a few of the minor proteins, is localized in the crystalloids.The major protein (molecular weight 65,000) was converted by mercaptoethanol into subunits with molecular weights of 32,000 and 15,800. It is proposed that the protein is made up of two of the smaller subunits and one of the larger, linked by disulfide bridges. None of the crystalloid proteins appear to be glycosylated.The water-soluble matrix fraction is composed mainly of two proteins, with molecular weights of 12,500 and 10,300 on the gels. Neither is a glycoprotein, and neither can be reduced with mercaptoethanol to give subunits. The soluble fraction also contains other lesser components among which are

  20. 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....