Biogenesis of auxin in the coleptile of a semi-brachytic barley, uzu

The coleoptile of a semi-brachytic barley, uzu(Hordeum vulgareL. cv. Akashinriki), elongated ca 1/2 as much as the coleoptileof the contrasting normal form which is isogenic excepting theuz gene. This retarded growth of the uzu coleoptile, as comparedto the normal coleoptile, is not due to changes in the rateof basipetal transport of auxin, neither to the destructionof auxin during transport, nor to sensitivity to auxin. Furthermore,less of the extractable and bound auxins were found in the uzu(uzuz)coleoptile than in the normal(UZUZ)coleoptile, suggesting thatthe retarded growth of uzu coleoptile may be due to less auxinproduction. Apical tips of the normal coleoptile grown under sterile conditionsresponded to both tryptophan and tryptamine, but uzu coleoptiletips responded only to tryptamine. Thus, growth retardationof the uzu coleoptile may be due to lower activity of the enzymewhich converts tryptophan to tryptamine in the uzu coleoptile. (Received August 20, 1973; )     

Uzu Mutation in Barley (Hordeum vulgare L.) Reduces the Leaf Unrolling Response to Brassinolide

A sensitive method to examine the brassinolide (BL) response of barley (Hordeum vulgare L.) using dark-grown leaf segments was established based on the known method for wheat. BL responses of 53 dwarf isogenic lines of barley were examined, and two lines were found having a uzu gene that doesn't respond significantly. These results indicate that uzu dwarfism may be caused by the non-responding character to BL.     

普通小麦×大麦杂交后代中间材料的GISH及PAGE鉴定

利用基因组荧光原位杂交 (GISH)及种子贮藏蛋白聚丙烯酰胺凝胶电泳 (PAGE)对普通小麦×大麦杂交后代中间材料进行了鉴定分析。GISH结果表明 ,WBA984和WBA9812为二体小大麦异附加系 ,WBS0 2 15和WBS0 2 6 4为小大麦二体异代换系 ,WBT0 2 12 5和WBT0 2 183为端部易位系 ;种子贮藏蛋白PAGE分析表明 ,WBA9812和WBS0 2 6 4含有大麦特有的高分子量麦谷蛋白亚基和在γ区含有大麦特有的醇溶蛋白带型 ,WBA9812为大麦 5H附加系 ,WBS0 2 6 4为 1B/ 5H代换系 ,WBT0 2 12 5为 1BL/ 5HL端部易位系。     

Growth Regulation of Dark-grown Dwarf Barley Coleoptile by the Endogenous IAA Content

Growth curves of dark-grown coleoptiles of 11 isogenic coleoptilardwarf strains of barley (Hordeum vulagare L. cv. Akashinriki:uzu, 5, 77, 97, 105, 125, 131, 133, 136, 145 and 148) were simulatedwith a logistic equation and the endogenous IAA contents ofthe barley strains were determined. Growth analysis of the dwarfbarley coleoptiles revealed that the final coleoptile lengthwas correlated with the growth rate on the 2nd day after germination(r=0.897), when the growth rate was about maximum. The endogenousIAA Content of the barley strains, measured fluorometrically,indicated that on the 2nd day, the dwarf strains contained lessendogenous IAA than the normal Strain. The IAA content on the2nd day was correlated to the growth rate on the 2nd day (r=0.907,except for Strain 145) and the final coleoptile length (r=0.933,except for strains 77 and 145). The correlation, however, wasnot significant on the 3rd day. These results suggested thatthe dwarfism of the dark-grown coleoptiles of the barley Strainsexamined is primarily controlled by the endogenous IAA content. ¹ Present address: Department of Biology, Faculty of Science,Osaka City University, Osaka 558, Japan. (Received February 1, 1982; Accepted April 13, 1982)     

Genotypic variation in response of barley to boron deficiency

Responses of a range of barley (Hordeum vulgare L.) genotypes to boron (B) deficiency were studied in two experiments carried out in sand culture and in the field at Chiang Mai, Thailand. In experiment 1, two barley genotypes, Stirling (two-row) and BRB 2 (six-row) and one wheat (Triticum aestivum L.) genotype, SW 41, were evaluated in sand culture with three levels of applied B (0, 0.1 and 1.0 μM B) to the nutrient solution. It was found that B deficiency depressed flag leaf B concentration at booting, grain number and grain yield of all genotypes. In barley Stirling, B deficiency also depressed number of spikes plant^-1, spikelets spike^-1 and straw yield. However, no significant difference between genotypes in flag leaf B concentration was found under low B treatments. Flag leaf B concentration below 4 mg kg^-1 was associated with grain set reduction and could, therefore, be used as a general indicator for B status in barley. In experiment 2, nine barley and two wheat genotypes were evaluated in the field on a low B soil with three levels of B. Boron levels were varied by applying either 2 t of lime ha^-1 (BL), no B (B0) or 10 kg Borax ha^-1 (B+) to the soil prior to sowing. Genotypes differed in their B response for grain spike^-1, grain spikelet^-1 and grain set index (GSI). The GSI of the B efficient wheat, Fang 60, exceeded 90% in all B treatments. The B inefficient wheat SW 41 and most of the barley genotypes set grain normally (GSI >80%) only at the B+. In B0 GSI of the barley genotypes ranged from 23% to 84%, and in BL from 19% to 65%. Three of the barley with severely depressed GSI in B0 and BL also had a decreased number of spikelets spike^-1. In experiment 3, 21 advanced barley lines from the Barley Thailand Yield Nursery 1997/98 (BTYN 1997/98) were screened for B response in sand culture with no added B. Grain Set Index of the Fang 60 and SW 41 checks were 98 and 65%, respectively, and GSI of barley lines ranged between 5 and 90%. One advanced line was identified as B efficient and two as moderately B efficient. The remaining lines ranked between moderately inefficient to inefficient. These experiments have established that there is a range of responses to B in barley genotypes. This variation in the B response was observed in vegetative as well as reproductive growth. Boron efficiency should be considered in breeding and selection of barley in low B soils. This revised version was published online in June 2006 with corrections to the Cover Date.     

Identification of barley genome segments introgressed into wheat using PCR markers.

Barley has several important traits that might be used in the genetic improvement of wheat. For this report, we have produced wheat-barley recombinants involving barley chromosomes 4 (4H) and 7 (5H). Wheat-barley disomic addition lines were crossed with 'Chinese Spring' wheat carrying the phlb mutation to promote homoeologous pairing. Selection was performed using polymerase chain reaction (PCR) markers to identify lines with the barley chromosome in the ph1b background. These lines were self pollinated, and recombinants were identified using sequence-tagged-site (STS) primer sets that allowed differentiation between barley and wheat chromosomes. Several recombinant lines were isolated that involved different STS-PCR markers. Recombination was confirmed by allowing the lines to self pollinate and rescreening the progeny via STS-PCR. Progeny testing confirmed 9 recombinants involving barley chromosome 4 (4H) and 11 recombinants involving barley chromosome 7 (5H). Some recombinants were observed cytologically to eliminate the possibility of broken chromosomes. Since transmission of the recombinant chromosomes was lower than expected and since seed set was reduced in recombinant lines, the utility of producing recombinants with this method is uncertain.     

Long-term effects of blue or red light on ATP and ADP contents in primary barley leaves

Levels of ATP and ADP were studied in primary leaves of barley (Hordeum vulgare L. cv. Viner) seedlings grown under blue (BL) or red light (RL) of various irradiances. In mature leaf segments, BL stimulated a greater accumulation of adenylates than RL. Transfer of barley seedlings from RL to BL for 48 h caused about a twofold increase in the content of adenylates, probably due to de-novo synthesis of adenine nucleotides. Weak BL was found to stimulate an increase in the adenylate content and a higher irradiance enhanced the stimulatory effect. The adenylate content increased markedly from the base towards the tip of barley leaves grown under BL but not in those grown under RL. However, the adenylate content was higher in the basalmost segment of barley leaves grown under RL, indicating that the action of RL on adenylate content proceeded more rapidly than that of BL. The same conclusion could be drawn from the results of experiments with de-etiolated leaves. A linear relationship was established between the maximum rate of CO₂ fixation and the ATP or ADP content in mature segments of primary barley leaves. The possible involvement of two photoreceptors, phytochrome and cryptochrome, in the long-term light regulation of the total content of adenylates in primary barley leaves is discussed.Abbreviations BL blue light - Chl chlorophyll - RL red light     

Identification of wheat-barley translocations by sequential GISH and two-colour FISH in combination with the use of genetically mapped barley SSR markers.

Five wheat-barley translocations in a wheat background were characterized through the combination of cytogenetic and molecular genetic approaches. The wheat chromosome segments involved in the translocations were identified using sequential GISH and two-colour FISH with the probes pSc119.2 and pAs1. The barley chromatin in these lines was identified using SSR markers. A total of 45 markers distributed over the total barley genome were selected from a recently published linkage map of barley and tested on the translocation lines. The following translocations were identified: 2DS.2DL-1HS, 3HS.3BL, 6BS.6BL-4HL, 4D-5HS, and 7DL.7DS-5HS. Wheat-barley disomic and ditelosomic addition lines for the chromosomes 3HS, 4H, 4HL, 5H, 5HL, and 6HS were used to determine the correct location of 21 markers and the position of the centromere. An intragenomic translocation breakpoint was detected on the short arm of the barley chromosome 5H with the help of SSR marker analysis. Physical mapping of the SSR markers on chromosomes 1H and 5H was carried out using the intragenomic and the interspecific translocation breakpoints, as well as the centromere, as physical landmarks.     

Genetic studies of Triticeae dehydrins: assignment of seed proteins and a regulatory factor to map positions

 A collection of 200 wheat (Triticum aestivum L. cv ‘Chinese Spring’) cytogenetic stocks (nullisomic, tetrasomic, nulli-tetrasomic, ditelosomic and deletion lines, addition and substitution stocks from intra- and inter-specific crosses) was utilized to determine the proteins encoded by some of the wheat and barley dehydrin genes, using a western blot procedure. Proteins extracted from seeds were reacted with antibodies that recognize dehydrins in a wide range of plants, including wheat and barley. Proteins encoded by dehydrin loci in chromosome arms 4DS, 5BL and 6AL of ‘Chinese Spring’ wheat were assigned by this method. There was also evidence of a regulatory factor on 5B in the vicinity of the dhn genes, and on 5H in wheat-barley addition lines, that is required for a normal level of expression of seed dehydrins in hexaploid wheat. Further understanding of this putative regulatory factor would be helpful for the interpretation of linkage studies that may relate dehydrin gene expression to phenotypes such as dehydration, salinity or low-temperature tolerance. Received: 27 August 1997 / Accepted: 4 February 1998     

The aspartic proteinase of barley is a vacuolar enzyme that processes probarley lectin in vitro.

Previous work suggested that the aspartic proteinase from Hordeum vulgare (HvAP) would be a vacuolar protein in plant cells. Based on N-terminal sequencing we show that the in vitro-translated protein was translocated into the lumen of microsomal membranes, causing a concomitant removal of 25 amino acid residues from the protein. Vacuoles were purified from barley leaf protoplasts and were shown to contain all of the aspartic proteinase activity found in the protoplasts. This vacuolar localization of HvAP was confirmed with immunocytochemical electron microscopy using antibodies to HvAP in both barley leaf and root cells. In an attempt to discern a function for this protease, we investigated the ability of HvAP to process the C-terminal proregion of barley lectin (BL) in vitro. Prolectin (proBL), expressed in bacteria, was processed rapidly when HvAP was added. Using several means, we were able to determine that 13 amino acid residues at the C terminus of proBL were cleaved off, whereas the N terminus stayed intact during this incubation. Immunohistochemical electron microscopy showed that HvAP and BL are co-localized in the root cells of developing embryos and germinating seedlings. Thus, we propose that the vacuolar HvAP participates in processing the C terminus of BL.     

Screening of certain barley lines for resistance to root rot disease caused by Fusarium graminearum

This investigation was conducted in 2005/2006 and 2006/2007 to test 235 barley lines plus two varieties Giza 127 and Giza 128 for resistance and susceptibility to Fusarium graminearum. All screened barley lines showed varied significant degrees of infestation to root rot pathogen. A screening system is described for identifying barley lines which are effective in controlling resistant or susceptible lines. By detecting small but consistent differences in root rot severity, the bioassay proved effective in large-scale screening for partial resistance: already 335 barley lines and two varieties have been screened. We found five groups (7.12%), 22 barley lines and both varieties are resistant (R) (8.31%); 28 barley lines are moderately resistant (MR) (19.29%); 65 barley lines are moderately susceptible (MS) (27.89%); 94 barley lines are susceptible (S) and (37.39%) 126 barley lines are highly susceptible (HS). The high degree of precision makes this an invaluable tool in the understanding of pathogen aggressiveness, host specialisation and parasitic fitness. Disease scale was strongly negative and had moderate correlation with germination (?0.309?? and ?0.649??) under normal and disease treatment. The correlation between yield and normal and disease treatment during two seasons was strong and negative (?0.834?? and ?0.847??, respectively were detected).     

Brassinolide alleviates the drought-induced adverse effects in barley by modulation of enzymatic antioxidants and ultrastructure

In the present study, possible role of exogenously applied brassinolide (BL) in alleviating the detrimental effects of polyethylene glycol (PEG)-inducted drought stress in barley was evaluated. Barley seedlings were subjected to two PEG levels (0 and 8% w/v) for 2 weeks followed by a foliar spray of BL (0, 0.01, 0.1 and 1 µM) to assess the changes in growth, gas exchange, chlorophyll contents, antioxidant enzyme activities and ultrastructural changes. PEG-induced drought considerably inhibited the barley growth in terms of shoot and root length, shoot, root fresh and dry weights, and chlorophyll contents. Moreover, extensive decrease by PEG-induced drought in gas exchange attributes i.e. net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO₂ were also recorded. However, exogenous application of BL remarkably improved the gas exchange attributes, chlorophyll contents and plant growth under either drought or control condition. Furthermore, foliar spray of BL decreased the production of malondialdehyde (MDA) and reactive oxygen species (ROS) in the leaves and roots by increasing enzymatic antioxidant activities under drought stress. The microscopic examination indicated that exogenous application of BL improved the cell structure and enabled a clean mesophyll cell having a well-developed chloroplast with thylakoid membranes of the drought stressed plants. On the basis of these findings, it can be concluded that exogenous application of BL could alleviate drought stress to barley by ameliorating photosynthetic parameters, improving antioxidants enzyme activities, reducing ultrastructural changes in leaf mesophyll and finally maintaining the plant growth.     

The distribution of transgene insertion sites in barley determined by physical and genetic mapping

The exact site of transgene insertion into a plant host genome is one feature of the genetic transformation process that cannot, at present, be controlled and is often poorly understood. The site of transgene insertion may have implications for transgene stability and for potential unintended effects of the transgene on plant metabolism. To increase our understanding of transgene insertion sites in barley, a detailed analysis of transgene integration in independently derived transgenic barley lines was carried out. Fluorescence in situ hybridization (FISH) was used to physically map 23 transgene integration sites from 19 independent barley lines. Genetic mapping further confirmed the location of the transgenes in 11 of these lines. Transgene integration sites were present only on five of the seven barley chromosomes. The pattern of transgene integration appeared to be nonrandom and there was evidence of clustering of independent transgene insertion events within the barley genome. In addition, barley genomic regions flanking the transgene insertion site were isolated for seven independent lines. The data from the transgene flanking regions indicated that transgene insertions were preferentially located in gene-rich areas of the genome. These results are discussed in relation to the structure of the barley genome.     

An alternative to radiation hybrid mapping for large-scale genome analysis in barley

The presence of a monosomic gametocidal chromosome (GC) in a barley chromosome addition line of common wheat generates structural aberrations in the barley chromosome as well as in the wheat chromosomes of gametes lacking the GC. A collection of structurally aberrant barley chromosomes is analogous to a panel of radiation hybrid (RH) mapping and is valuable for high-throughput physical mapping. We developed 90 common wheat lines (GC lines) containing aberrant barley 7H chromosomes induced by a gametocidal chromosome, 2C. DNAs isolated from these GC lines provided a panel of 7H chromosomal fragments in a wheat genetic background, comparable with RH mapping panels in mammals. We used this 7H GC panel and the methodology for RH mapping to physically map PCR-based barley markers, SSRs and AFLPs, onto chromosome 7H, relying on polymorphism between the 7H chromosome and the wheat genome. We call this method GC mapping. This study describes a novel adaptation and combination of methods of inducing chromosomal rearrangements to produce physical maps of markers. The advantages of the presented method are similar to RH mapping in that non-polymorphic markers can be used and the mapping panels can be relatively easily obtained. In addition, mapping results are cumulative when using the same mapping set with new markers. The GC lines will be available from the National Bioresources Project-KOMUGI (). Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

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

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

Evaluation of regeneration potential of mature embryo derived callus in Indian cultivars of barley (Hordeum vulgare L.)

A protocol for plant regeneration in Indian cultivars of barley (Hordeum vulgare L.) has been developed using mature embryo culture. The influence of various auxins 2,4-D (2,4-dichlorophenoxyacetic acid), Dicamba (3,6-dichloro-o-anisic acid) and Picloram (4-amino-3,5,6-trichloropicolinic acid) on the callus induction and subsequent plant regeneration revealed highest percent of callus induction form cultivar (cv) BL 2 on MSB₅ medium (MS salts + B₅ vitamins) supplemented with 6 mg l^?1 Picloram, but maximum number of shoot buds (6–13) were regenerated on MSB₅ medium containing 0.5 mg l^?1 Picloram. Regenerated shoots were rooted on half-strength MSB₅ medium. Plantlets were successfully transferred to soil and grown to maturity in greenhouse. The effect of copper sulphate revealed significant improvement in callus induction and plant regeneration when the concentration of CuSO₄ was increased to 3 μM (30 times higher than normal MS medium) for cv BL 2. Regeneration potential differed for different cultivars of barley used, with highest for cv BL 2 and lowest for cv BH 924. We conclude that the Indian barley genotypes exhibit plant regeneration from mature embryo cultures. The protocol has potential application in barley improvement through genetic engineering.     

Effect of Growth Stage on Resistance to Drechslera teres f. teres in Barley

Three experiments were conducted to evaluate seedling and adult plant resistance to Drechslera teres f. teres in barley lines with a potential use as resistance sources in barley breeding. Disease resistance of barley seedlings, inoculated in the glasshouse and growth chamber at the one- and two-leaf stages, was significantly (P 0.05) correlated with disease reactions on the fourth and flag leaves of barley plants grown in growth chambers ( r = 0.84- 0.94) as well as with disease levels recorded on the three uppermost leaves in a field experiment ( r = 0.64). Several of the barley lines showed no correlation, demonstrating either improved or decreased disease resistance at later growth stages. Six barley lines (Alexis, Heartland, Kinnan, CI4922, CI9776 and SWl114-93) had the same ranking in disease resistance to D. teres when damaged (punctured) leaves were inoculated with a drop of spore suspension as when undamaged leaves were spray-inoculated with a spore suspension.     

Chromosomal assignment and deletion mapping of barley EST markers

From about 10000 PCR-based EST markers of barley we chose 1421 EST markers that were demonstrated to be amplified differently by PCR between wheat (Triticum aestivum cv. Chinese Spring) and barley (Hordeum vulgare cv. Betzes). We assigned them to the seven barley chromosomes (1H to 7H) by PCR analysis using a set of wheat-barley chromosome addition lines. We successfully assigned 701 (49.3%) EST markers to the barley chromosomes: 75 to 1H, 127 to 2H, 119 to 3H, 94 to 4H, 108 to 5H, 81 to 6H and 97 to 7H. By using a set of Betzes barley telosomic addition lines of Chinese Spring, we could successfully determine the chromosome-arm (S or L) location of at least 90% of the EST markers assigned to each barley chromosome. We conducted a trial mapping using 90 EST markers assigned to 7HS (49) or 7HL (41) and 19 wheat lines carrying 7H structural changes. More EST markers were found in the distal region than in the proximal region.