Marker-assisted characterization of frost tolerance in barley (Hordeum vulgare L.)

Five molecular markers associated to two frost tolerance QTLs ( Fr-H1 and Fr-H2 ) were tested both on nine Turkish accessions, classified by breeders as highly frost-tolerant, and on a previously described sample of 26 barleys, winter, facultative and spring. Accessions were characterized in terms of frost tolerance under both field conditions and artificial freezing test at −12°C. The Turkish lines resulted to be equal or superior to the most tolerant European genotypes tested, showing that they can be used to improve the frost tolerance of the EU barley germplasm. The marker Hv BM5A ( Vrn-H1 and Fr-H1 ) resulted to be the best predictor for assisted selection within this germplasm, because of its high correlation between allelic variation and phenotypic traits. Only Hv CBF4 of the three Hv CBF markers tested at Fr-H2 was associated to the trait, but at lower significance than HvBM5A . The PCR-based molecular marker of Vrn-H1 can thus be used in barley breeding not only for selection of facultative and winter types, but also for fast routine selection of frost tolerant genotypes.     

SSR analysis of introgression of drought tolerance from the genome of Hordeum spontaneum into cultivated barley (Hordeum vulgare ssp vulgare)

Wild barley (Hordeum vulgare ssp. spontaneum) and landraces are important sources of resistance to biotic and abiotic stresses since they possess wide genetic diversity that may be missing in current elite varieties. In this study, we evaluated a set of 57 barley introgression lines divided in groups depending on the expected levels of introgression (50, 25, and 12.5 %) from one Hordeum spontaneum accessions (Hsp 41-1) and on those (50 and 25 %) from a second (Hsp 41-5); in both cases the 25 % level was represented by two groups depending on the other parent. The two H. spontaneum accessions have been used as the best sources of drought tolerance in the ICARDA barley-breeding program. Graphical genotyping and genetic diversity analysis were used to examine the relative contribution of H. spontaneum and the extent of genetic differences among the 57 lines using 74 microsatellite markers that cover 941 cM of the barley genome. The average proportion of the genome containing H. spontaneum alleles in each group was of 44.5 %, group 1; 24.6 %, group 2; 21.6 %, group 3; 45.4 %, group 4; 19 %, group 5; 15.5 %, group 6 and 11.4 %, group 7. Introgression lines in group 1 and 4, with the highest observed introgression with Hsp 41-1 and Hsp 41-5, showed higher grain yield and better agronomic performance under field conditions in Breda and Khanaser, i.e., the two most stressed environments in which the groups were phenotyped, indicating the usefulness of using H. spontaneum as a source of chromosomal linkage blocks important for improved drought tolerance. However, more extensive genome coverage will be needed to identify the specific chromosomal regions associated with superior performance under extreme drought.     

Inheritance of resistance to Rhynchosporium secalis in spring barley (Hordeum vulgare L.)

The inheritance of durable resistance of selected spring barley varieties to Rhynchosporium secalis was investigated. Data from the F₂ generation of a 4 × 4 diallel, without reciprocals and the F₄ generation of three crosses selected out of this diallel, suggest that resistance in this sample of varieties tested is complex in inheritance. Significant additive effects were detected indicating that the resistance level of barley cultivars may be improved by the hybridisation of suitable varieties. However, the genes conferring resistance seem to be concealed by the expression of one completely dominant resistance gene in our set of varieties. These results are partly in conflict with previous results on the inheritance of resistance to R. secalis in the breeder's line ‘11258/2286₁₃A’ indicating that the effectiveness of this resistance gene may be greatly influenced by the genetic background of the current population of R. secalis.     

Salinity tolerance of wild barley Hordeum vulgare ssp. spontaneum

Salinity tolerance of 47 wild barley genotypes and six barley cultivars was evaluated under control and salinity stress (300 mM NaCl) conditions. Shoot and root dry weight (DW), plant height, membrane stability index (MSI), relative water content, survival rate, leaf malondialdehyde (MDA) and proline contents, root and leaf Na, K, Ca and K/Na ratio, and chlorophyll a fluorescence were measured. Salinity stress caused significant increase in the MDA, proline content, Na and Ca concentrations of the roots and leaves, but resulted in a decrease in the other traits. H. spontaneum genotypes were considerably less affected by the salinity than the genotypes of H. vulgare. Plant survivability was negatively correlated with the Na concentration (r =−.66) but positively correlated with the leaf K/Na ratio (r = .67) and MSI (r = .68). Tolerance mechanisms such as ion exclusion (Na) were likely to be present in the wild barley causing K/Na homeostasis as well as the much lower root and shoot Na, resulting in the higher survival rate.     

RAPD characterization of Brazilian barley (Hordeum vulgare ssp. vulgare) varieties

Thirteen varieties of Brazilian barley selected for malting qualities were analysed by RAPD (Random Amplified Polymorphic DNA). Amplification with 18 random primers generated 221 reproducible bands, of which 206 bands were polymorphic (93%):of this number, 137 fragments (62%) detected diversity among varieties and 56 bands (25.34%) allowed the distinction of varieties or groups of them. Variation was detected in all Brazilian varieties studied. Within-variety similarities estimated by Jaccard Similarity Coefficient ranged from 0.28 to 0.94, with averages ranging from 0.57 to 0.83, and an overall average of 0.72. Nevertheless, in the cluster analysis representatives of the same variety always fell into the same group and only later joined the other varieties. The average intervarietal similarities estimated by Jaccard Similarity Coefficient ranged from 0.45 to 0.62, with an overall average of 0.52. Many bands or combinations of bands which were responsible for the differentiation of all varieties were detected. Nevertheless, the majority of these bands cannot be considered as diagnostic markers because a great number of them did not occur in many representatives of the variety or had low intensity or even because they were not easily identified in the total pattern of bands.     

Mapping quantitative trait loci for salt tolerance at germination and the seedling stage in barley (Hordeum vulgare L.)

Quantitative trait loci (QTLs) controlling salt tolerance at germination and the seedling stage in barley (Hordeum vulgare L.) were identified by interval mapping analysis using marker information from two doubled haploid (DH) populations derived from the crosses, Steptoe/Morex and Harrington/TR306. Interval mapping analysis revealed that the QTLs for salt tolerance at germination in the DH lines of Steptoe/Morex were located on chromosomes 4 (4H), 6(6H), and 7(5H), and in the DH lines of Harrington/TR306 on chromosomes 5(1H) and 7(5H). In both DH populations, the most effective QTLs were found at different loci on chromosome 7(5H). Genetic linkage between salt tolerance at germination and abscisic acid (ABA) response was found from QTL mapping. The QTLs for the most effective ABA response at germination were located very close to those for salt tolerance on chromosome 7 (5H) in both crosses. The QTLs for salt tolerance at the seedling stage were located on chromosomes 2(2H), 5(1H), 6(6H), and 7(5H) in the DH lines of Steptoe/Morex, and on chromosome 7(5H) in the DH lines of Harrington/TR 306. Their positions were different from those of QTLs controlling salt tolerance at germination, indicating that salt tolerance at germination and at the seedling stage were controlled by different loci. This revised version was published online in July 2006 with corrections to the Cover Date.     

Rapid phenotyping of adult plant resistance in barley (Hordeum vulgare) to leaf rust under controlled conditions

Breeding for adult plant resistance (APR) is currently impeded by the low frequency of annual field‐based testing and variable environmental conditions. We developed and implemented a greenhouse‐based methodology for the rapid phenotyping of APR to leaf rust in barley to improve the efficacy of gene discovery and cloning. We assessed the effects of temperature (18 and 23°C) and growth stage (1–5 weeks) on the expression of APR in the greenhouse using 28 barley genotypes with both known and uncharacterized APR. All lines were susceptible in week 1, while lines carrying Rph20 and several with uncharacterized resistance expressed resistance as early as week 2. In contrast, lines lacking Rph20 and carrying either Rph23 and/or Rph24 expressed resistance from week 4. Resistant phenotypes were clearest at 18°C. A subset of 16 of the 28 lines were assessed for leaf rust across multiple national and international field sites. The greenhouse screening data reported in this study were highly correlated to most of the field sites, indicating that they provide comparable data on APR phenotypes for screening purposes.     

Identification of isolate specific sources of scald resistance in Turkish barley (Hordeum vulgare) accessions

Disease reactions to specific Rhynchosporium secalis isolates from western Canada were characterized on forty Turkish Hordeum vulgare accessions. One accession, MEH151-1, exhibited resistance against isolate WRS1860 and was also resistant to isolates WRS1493 and WRS1824 which were avirulent to the Rh gene. A related line, MEH151-2 exhibited resistance to WRS1493 but not to WRS1860. Crosses between both MEH151-1 and MEH151-2, and the susceptible cultivar, Harrington, were used to demonstrate a linkage between resistance to WRS1493 and an allele specific amplicon, Falc666. Resistance to WRS1860 imparted by MEH151-1 was mapped to the other side of the Falc666 locus. Falc666 was previously shown to be located near the centromere on the long arm of chromosome 3. Characterization of the genetic basis for the scald resistance phenotype exhibited by these Turkish accessions, coupled with the identification of marker linkages, provides evidence for genetic variation in scald resistance in this chromosomal region. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mapping genes for deep-seeding tolerance in barley

Deep-seeding tolerance, the emergence of seedlings from deep seeded conditions, is involved in stand establishment in semi-arid regions, where the soil surface is too dry for seed germination. Genes determining deep-seeding tolerance in barley were mapped using two doubled haploid populations derived from the following crosses: Harrington × TR306 (H/T)and Step toe × Morex (S/M). Significant quantitative trait loci (QTLs) for deep-seeding tolerance were found in each population. Two QTL sex plained 40% of the phenotypic variation in the H/T population and one QTL (S/M) 8% of the total phenotypic variance. Multiple QTLs accounting for coleoptile length and first internode length were detected in both populations. In the H/T population, there were coincident QTLs for deep-seeding tolerance, coleoptile length and first internode length on the long arm of chromosome 5H. These QTLs correspond with previously reported QTLs for abscisic acid and gibberellic acid response. QTL coincidence may be due to the pleiotropic effects of alleles at a single locus. This information may be useful for breeding programs manipulating morphological and physiological traits in order to develop varieties for semi-arid regions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic control of Russian wheat aphid (Diuraphis noxia) resistance in wheat accession PI 47545

The Russian wheat aphid, Diuraphis noxia (Mordvilko), is a major pest of cereal crops in many areas of the world, causing serious reduction in grain yield in wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.). Incorporating genetic resistance to D. noxia into wheat cultivars is paramount to effectively reduce damage inflicted by this pest. Genetic resistance to D. noxia has been identified in wheat, barley and rye germplasm, and several resistance genes are available for use for cultivar improvement. In the United States of America, only a few Russian wheat aphid (RWA) resistant winter wheat cultivars are currently available, and these cultivars contain only one of the six known RWA resistance genes. The objective of this study was to determine the inheritance of RWA resistance in wheat accession PI 47545, using a screening method based on differences in the leaf morphology of resistant and susceptible types following insect challenge. PI 47545 was selected for study, since it displayed high levels of resistance in a white-grained wheat background, the predominant wheat class produced in the Pacific Northwest of the USA. Segregation analysis was conducted on an F₂ population developed by cross-hybridizing the susceptible soft white winter wheat cultivar ‘Daws’ to the resistant accession PI 47545. Russian wheat aphid screening data from this population indicated that the resistance in PI 47545 is controlled by a single, dominant gene (χ² = 1.72; p ≤ 0.189). This revised version was published online in August 2006 with corrections to the Cover Date.     

大麦蛋白质二硫键异构酶基因家族的鉴定与表达分析

蛋白质二硫键异构酶(protein disulfide isomerases,PDIs)是定位于真核生物内质网中的一类蛋白,具有催化蛋白质二硫键氧化还原反应与协助蛋白质发生构型变化、类似分子伴侣的功能,参与生长发育、生物或非生物胁迫应答等方面的调控。真核生物中PDI基因家族的分子鉴定已有报道,但是对大麦PDI基因家族的鉴定分析鲜有报道。本研究利用生物信息学分析手段鉴定了10个大麦PDI-Like基因(HvPDILs),并分析了相应编码蛋白的结构特征。与其他物种PDILs的系统发育分析发现,10个大麦PDILs分处在8个不同的系统分支,与小麦PDILs高度同源。对公共数据库中的组织和时空表达数据分析发现, HvPDILs基因均表达,且具组织和时空表达特异性。对接种大麦温性花叶病毒(BaMMV)之后的叶片样品进行基因表达水平分析,发现其中5个HvPDILs基因出现显著的表达差异,说明其参与病毒感染过程,但参与方式及作用机制还有待后续研究。     

Genetic dissection of endosperm hydration in malting barley (Hordeum vulgare)

Hydration of the endosperm is a critical part of the malting process that ensures proper modification of the grain. However, little is known about the genetic controls of endosperm hydration and its relationship to agronomic and malt quality traits. The extent of endosperm hydration is estimated through hydration index (HYI). We measured HYI, agronomic, and malt quality traits on a 169-line subset of the NSGC Barley Core Panel, which includes global malt lines, some dating from the inception of European breeding programmes. Utilizing GWAS, 61 QTLs were identified for HYI, dormancy, agronomic, and malt quality traits. Of these, six were found to be related to HYI and were located on 1H, 2H, 3H, 6H, and 7H. We found HYI QTLs cosegregating with kernel size and hardness (1H and 3H), malting quality (2H and 6H), and dormancy (2H and 6H). These results indicate that endosperm hydration after steeping can be improved by selecting high HYI alleles on 2H, 6H, and 7H, positively impacting malting quality without negatively impacting kernel size or dormancy.     

Genes for scald resistance from wild barley (Hordeum vulgare ssp spontaneum) and their linkage to isozyme markers

Summary Accessions of Hordeum vulgare ssp. spontaneum, the wild progenitor of barley, collected in Israel (70), Iran (15) and Turkey (6) were screened for seedling response to four isolates of Rhynchosporium secalis, the pathogen causing leaf scald in barley. Resistance was very common in the collection (77%) particularly among accessions from the more mesic sites (90%). The genetics of this resistance were investigated in fifteen backcross (BC₃) lines that contained an isozyme variant from H.v. ssp. spontaneum in a H.v. ssp. vulgare (cv. Clipper) background and were resistant to scald. Segregation in the BC₃F₂ families conformed with a single dominant resistance gene in 9 of the 15 lines. Scald resistance and the isozyme marker were closely linked in three of the BC₃-lines, loosely linked in four and unlinked in the remaining eight. Scald resistance genes were identified on barley chromosomes 1, 3, 4 and 6. Crosses between several of the scald resistant BC-lines together with the linkage data indicated that at least five genetically independent resistances are available for combining together for deployment in barley. The linkage of scald resistance in several BC₃-lines to the isozyme locus Acp2 is of special interest as this locus is highly polymorphic in wild barley.     

Complementary competition in cultivated barley (Hordeum vulgare L.)

Summary Grain yield and its endpoint components were studied for three barley varieties, Dayton, Hudson, and Tenn 60-34, as pure stands and two-way and three-way mixtures at three widely separated sites for two years. The populations, including the pure stands, produced similar grain yields. Complementary competition was observed in all mixed populations in which Tenn 60-34 occurred. Its yield was suppressed in mixtures about 20% while Dayton and Hudson increased a like amount so that total mixed population yield was equal to that expected from pure stand performance. Tenn 60-34 occurred at 33, 50, and 80% frequencies in mixed populations with no difference in competition, indicating a lack of frequency dependency for the effect.Yield component data indicated that the reduced yield of Tenn 60-34 in mixed stands was due entirely to a decreased number of spikes produced per unit area. It was inferred that this was due to competition at time of tillering although this could not be determined since the varieties were indistinguishable during vegetative growth.An examination of population-environment interaction provided some evidence for the idea that heterogeneous populations in which there is intense competition show greater micro-environment (within-site) instability. Heterogeneous populations generally showed lower than expected contribution to the population-environment variance although the differences were quite small.These results have important implications for the utilization of heterogeneous populations in agricultural production and for the management of genetically diverse populations in plant breeding practice since genotypic frequencies are expected to change rapidly with complementary competition.     

Monosomic analysis of Russian wheat aphid (Diuraphis noxia) resistance in Triticum aestivum line PI137739

Summary The Russian wheat aphid, Diuraphis noxia (Mordvilko) (Homoptera: Aphididae), has become an important pest of wheat (Triticum aestivum L.) in the United States. The aphid causes a phytotoxemic reaction in wheat evidenced by local and systemic chlorosis and rolling of infested leaves. Developing resistance in wheat cultivars to D. noxia is an essential factor in controlling the damage caused by this pest. Several sources of genetic resistance to D. noxia have been identified in wheat germplasm. Monosomic analysis of the monogenic resistant T. aestivum accession PI137739 has shown that the gene (Dn1) for resistance is carried on chromosome 7D. It appears that chromosome 7B may carry a second resistance gene for D. noxia that might be a source of minor or complementary gene action for resistance.     

Mapping and candidate gene identification of loci induced by phytohormones in barley (Hordeum vulgare L.)

Phytohormones, a group of structurally unrelated small molecules are plant-signalling compounds that trigger induced resistance against certain pathogens and herbivores. The hormones jasmonic acid (JA), ABA, salicilic acid (SA) and ethylene (ET) are known to play major roles in regulating plant defence responses. In order to determine the changes in growth and in the chlorophyll content induced by the exogenous application of these elicitors, a set of DH lines of the Oregon-Wolfe Barley mapping population, previously screened to locate aphid resistant genes, was investigated. The aim of the current research was to map the induced defence genes and to reveal the relationship with aphid resistance. There were highly significant differences between controls and hormone treated plants in the aerial fresh and dry weights (AFW, ADW), the foliar area (FA) and the root dry weight (RDW). More than 15 JA and ET-induced lines exceeded the chlorophyll (Ch) values of their controls. Most of the plant traits were associated with the same genetic windows on chromosomes 3H, 5H and 7H in the controls and hormone treated plants. QTL(s) identified on chromosome 3H and 5H explained most of the variation of AFW, ADW, FA and RDW of controls and treated plants. QTL(s) located on chromosome 5H were associated with the variation of chlorophyll contents on JA-treated plants. The Ch in ET and ABA-treated plants was associated with two different regions on chromosome 7H. One of the latter genetic windows also explained the variation of RDW of ET- and ABA-treated plants. A sequence homology search was performed to derive the putative function of the genes linked to the QTLs. Several QTLs were identified located close to aphid resistance genes previously mapped. This is the first report of genes associated with hormone response in barley that could be involved with insect resistance. Those recombinant lines carrying the appropriate alleles could be useful for breeding barley to enlarge the genetic base of defence against stress.     

Adaptation of barley (Hordeum vulgare L.) to harsh Mediterranean environments

Summary Differences in development of the apex may be the reason for cultivar differences in adaptation of barley to terminal drought in Mediterranean environments. The present study was conducted to identify apical development patterns of barley adapted to terminal drought stressed Mediterranean environments and to determine plant characteristics which can be used as criteria to select for an adapted development. Thirty-five two-rowed barley (Hordeum vulgare L.) entries were grown at two sites in northern Syria (Tel Hadya and Breda) in 1988/89. Four apical development patterns were observed: a slow or fast vegetative development, depending on the vernalization requirement, combined with a slow or fast generative development, depending on the daylength response of the crop. Early heading was related to fast generative development. Leaf appearance rates on the main shoot were constant during a major part of the pre-anthesis period, but significant differences were observed among development patterns. Genotypic differences in main shoot tiller number were associated with differences in the onset of tiller appearance and not with differences in tiller appearance rate or final leaf number on the main shoot. Since vernalization requirements and daylength responses are largely independent of terminal drought stress, selection for an adapted phenology can be done in favourable environments. Morphological traits related to these responses (winter growth habit, cold tolerance, plant colour, growth vigour, heading date) can be used as criteria for selection for adaptation to low-rainfall Mediterranean environments.     

Greenhouse evaluation of red winter wheats for resistance to the Russian wheat aphid (Diuraphis noxia,Mordvilko)

Summary The Russian wheat aphid (Diuraphis noxia, Mordvilko) (RWA) is responsible for significant economic damage to cereal crops in arid and semi-arid environments. In this research 20 red winter wheats originating from Iran were evaluated for resistance to RWA. Leaf rolling, leaf folding, and leaf chlorosis were measured using 0 to 3 scales. An overall mean damage score was calculated as the average of the three measured damage symptoms. Plants from seven central Asian accessions (PI222666, PI222668, PI225226, PI225267, PI225271, PI243630, and PI243642) had mean damage scores significantly lower (p < 0.001) than Stephens wheat (RWA susceptible) and not significantly different from Border oat (RWA resistant). These results are consistent with previous studies which found a high frequency of resistant wheats collected from the central Asian region.     

Host plant resistance in some wild wheats to the Russian wheat aphid, Diuraphis noxia (Mordvilko) (Homoptera: Aphididae)

A total of 259 accessions of wild Triticum species originating from different countries, along with 91 triticale (6×)× bread wheat true-breeding derivatives, two bread wheat, and three triticale cultivars were screened for resistance to the Russian wheat aphid, a serious insect pest of the wheat crop. Twenty-four entries with low damage ratings on the basis of amount of leaf rolling and leaf chlorosis were retested along with resistant and susceptible controls. On the basis of leaf roll damage ratings, eight entries including four Triticum monococcum var. boeoticum (T. boeoticum), one T. monococcum var. monococcum (T. monococcum), two T. timopheevii var. araraticum (T. araraticum), and one triticale cultivar were significantly superior to ‘Karl’ (susceptible control) wheat. Among these, four accessions — three T. boeoticum and one T. araraticum— were significantly superior to all other entries and were equal to the resistant control (PI 372129) in resistance rating based on leaf rolling and leaf chlorosis (except T. boeoticum TA 202). The leaf chlorosis damage rating of all accessions were significantly lower than that of the susceptible check.