Molecular mapping of major genes and quantitative trait loci determining flowering time in response to photoperiod in barley

Two major genes (eam8 and eam10) and two quantitative trait loci (QTL) determining flowering time in barley were associated with restriction fragment length polymorphism markers. The loci eam8 and eam10 were found to map in regions of chromosomes 1HL and 3HL, respectively, already estimated from previous classical linkage analyses. While investigating doubled haploid lines of a spring habit barley mapping population, two QTL for flowering time were detected on chromosomes 1HL and 7HS, respectively, when the material was grown under long photoperiod conditions. When growing the same lines under short photoperiod, no QTL were discernible. Allelic and homoeologous relationships with flowering time loci described earlier in barley and other Triticeae species are discussed.     

RFLP mapping of a new cereal cyst nematode resistance locus in barley

Cereal cyst nematode (CCN) ( Heterodera avenae Woll.) is an economically damaging pest of barley in many of the worlds cereal growing areas. The development of CCN-resistant cultivars may be accelerated with the application of molecular markers. Three resistance genes against the pest have been mapped previously to chromosome 2 ( Ha 1, Ha 2 and Ha 3). In this study, a third gene present in the Australian barley variety 'Galleon' derived from the landrace 'CI3576' was located. Segregation analysis of CCN resistance data derived from doubled haploid populations of the cross 'Haruna Nijo'×'Galleon' identified a single major locus controlling CCN resistance in the variety 'Galleon'. This locus mapped to the long arm of chromosome 5H estimated to be 6.2 cM from the known function restriction fragment length polymorphism marker XYL (xylanase). While five genes for CCN resistance, including Ha2, have been mapped to group 2 chromosomes in the Triticeae, no gene other than Ha4 has been identified on group 5 chromosomes.     

Resistance to powdery mildew in a doubled haploid barley population and its association with marker loci

The genetic basis of resistance to powdery mildew (Erysiphe graminis DC. f.sp. hordei Marchal) was analyzed using doubled haploid barley (Hordeum vulgare L.) lines from the cross Harrington/TR306. Based on infection types observed after inoculation with defined single-conidium isolates, the lines were classified into four groups. The observed phenotypic ratio fit a two-locus model. The two putative loci were mapped relative to molecular markers. One coincided with the previously mapped dMlg locus on chromosome 4. Based on the observed infection types, Harrington carries the Mlg resistance allele, and TR306 carries a second locus on chromosome 7 (5H); this was tentatively designated Ml(TR). It is the first reported race-specific powdery mildew resistance gene located on that chromosome. These two loci were also detected by simple interval mapping of disease severity data from naturally infected field plots. Composite interval mapping with the first two resistance loci as co-factors detected an additional locus on chromosome 6, with a minor effect on resistance. Finally, superimposing the race-specific classification onto the field data provided evidence for a minor-effect locus on chromosome 7 (5H). The Mlg locus had the largest effect, the Ml(TR) locus had an intermediate effect and the other two loci had very small effects. This study demonstrates the effectiveness of an integrated approach to identifying and mapping resistance loci using classification data from inoculated experiments and quantitative data from field experiments. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cloning, genetic and physical mapping of resistance gene analogs in barley (Hordeum vulgare L.)

The majority of verified plant disease resistance genes isolated to date belong to the NBS‐LRR class, encoding proteins with a predicted nucleotide binding site (NBS) and a leucine‐rich repeat (LRR) region. Using degenerate primers, designed from the conserved motifs of the NBS region in tobacco N and Arabidopsis RPS2 genes, we isolated 190 resistance gene analogs (RGA) clones from barley genomic DNA. A total of 13 single‐ and low‐copy RGAs were genetically mapped onto chromosomes 1H–7H (except 5H) using three barley double haploid (DH) mapping populations: Steptoe × Morex, Harrington × TR306 and LUGC × Bowman. Sequence analysis of the RGAs showed that they are members of a diverse group. As a result of BLAST searches, one RGA proved unique as it did not detect any significant hit. Another RGA is putatively functional, because it detected several barley expressed sequence tag (EST) matches. To physically map the RGAs, 13 sequences were used to screen a 6.3 × cv. ‘Morex’ bacterial artificial chromosome (BAC) library. After fingerprint analysis, eight contigs were constructed incorporating 62 BAC clones. These BAC contigs are of great value for positional cloning of disease resistance genes, because they span the regions where various barley R genes have been genetically mapped.     

Use of component analysis in QTL mapping of complex crop traits: a case study on yield in barley

Genes contributing to the quantitative variation of a complex crop trait can be numerous. However, using existing approaches, the number of quantitative trait loci (QTL) detected for a trait is limited. Therefore, rather than looking for QTL for a complex trait itself, determining QTL for underlying component traits might give more information. In this study the potential of component analysis in QTL mapping of complex traits was examined using grain yield in spring barley as an example. Grain yield was divided into three components: number of spikes/m², number of kernels/spike, and 1000‐kernel weight. These traits were measured for individuals of a recombinant inbred‐line population in field trials conducted over 2 years. By the use of an approximate multiple QTL model, one to eight QTL were detected for each trait in a year. Some QTL were mapped to similar positions in both years. Almost all QTL for yield were found at the position of or in close proximity to QTL for its component traits. A number of QTL for component traits were not detected when yield itself was subjected to QTL analysis. However, relative to the QTL for yield itself, all component‐trait QTL did not explain the variation in yield better. The results in relation to the potential of using component analysis in studying complex crop traits are discussed.     

Identification of QTLs associated with salinity tolerance at late growth stage in barley

Salinity is a major abiotic stress to barley (Hordum vulgare L.) growth and yield. In the current study, quantitative trait loci (QTL) for yield and physiological components at the late growth stage under salt stress and non-stress environments were determined in barley using a double haploid population derived from a cross between CM72 (salt-tolerant) and Gairdner (salt-sensitive). A total of 30 QTLs for 10 traits, including tiller numbers (TN), plant height, spikes per line (SPL), spikes per plant (SPP), dry weight per plant, grains per plant, grain yield, shoot Na⁺ (NA) and K⁺ concentraitions (K) in shoot, and Na⁺/K⁺ ratio (NAK), were detected, with 17 and 13 QTLs under non-stress and salt stress, respectively. The phenotypic variation explained by individual QTL ranged from 3.25 to 29.81%. QTL flanked by markers bPb-1278 and bPb-8437 on chromosomes 4H was associated with TN, SPL, and SPP under salt stress. This locus may be useful in the breeding program of marker-assisted selection for improving salt tolerance of barley. However, QTLs associated with NA, K, and NAK differed greatly between non-stress and salt stress environments. It may be suggested that only the QTLs detected under salt stress are really associated with salt tolerance in barley. D. Xue and Y. Huang contributed equally to the article.     

Screening for resistance in the primary and secondary gene pool of barley against the root-lesion nematode Pratylenchus neglectus

Root-lesion nematodes of the genus Pratylenchus are significant pests in crop cultivation throughout many parts of the world. A study was initiated to determine the resistance of Hordeum vulgare and H. vulgare ssp. spontaneum (wild barley) against one major representative of the genus Pratylenchus , P. neglectus . A glasshouse test was first established. Barley seedlings were grown in 20 cm³ tubes filled with sand. Each plant was inoculated with 400 P. neglectus juveniles. After 12 weeks of cultivation nematodes were isolated from roots and sand using a misting chamber. The nematodes were counted under a microscope. A representative collection of 565 barley and wild barley accessions was tested in this way. The average number of nematodes per accession ranged from 350 to 12 000. In a verification experiment, 35 accessions with low and high infection rates were tested. This experiment identified a number of accessions with low infection rates. The perspectives for future breeding of barley cultivars resistant to root-lesion nematodes are discussed.     

Mapping of QTL controlling NIR predicted hot water extract and grain nitrogen content in a spring barley cross using marker-regression

A restriction fragment length polymorphism (RFLP) map constructed from 99 doubled haploid lines of a cross between two spring barley varieties (‘;Blenheim’בKym’) was used to map QTL controlling hot water extract and grain nitrogen content (predicted by analysis with near-infrared reflectance spectroscopy). Eight QTL affecting predicted hot water extract were identified by a marker-regression approach. The largest effects were found on chromosomes 3HL, associated with the denso dwarfing gene which is present in‘Blenheim’and conferred poorer predicted hot water extract quality, and 4HL. Other QTL were detected on chromosomes IHS. IHL. 2HS, 2HL. 5HL and 6HS. Analysis of single markers by analysis of variance detected an additional effect on chromosome 1H. Eight QTL affecting predicted grain nitrogen content were identified by marker-regression, on chromosomes 1HS, 1HL. 2HL. 5HS, 6H, 7HS and 7HL. There was also evidence for an additional QTL on chromosome 5HL. The positions of the grain nitrogen content QTL on 5HS and 5HL are comparable to QTL on wheat chromosomes 5A and 5D that affect grain protein content. The denso gene had no detectable effect on grain nitrogen content.     

Locating supplementary RFLP markers on barley chromosome 7 and synteny with homoeologous wheat group 5

In order to develop QTL applications, eight new loci were mapped on barley chromosome 7 using 124 doubled haploid lines of the North American Barley Genome Mapping Project (NABGMP) progeny (‘Steptoe’בMorex’)- These loci involve six genomic DNA restriction fragment length polymorphisms (RFLPs) and two cDNA-RFLPs including a puroindoline gene. The distribution of these markers on barley chromosome 7 was compared with that of homoeologous wheat counterparts, i.e. wheat group 5. One locus on chromosome 7 was associated with a QTL for β-glucanase activity measured in green and finished barley malt.     

Ribosomal DNA spacer-length polymorphisms in three samples of wild and cultivated barleys and their relevance to the origin of cultivated barley

This study was conducted to provide additional data for evaluating two important issues surrounding the origin of cultivated barley: (i) the level of genetic diversity of the two‐rowed wild barley from Tibet, and (ii) the distribution of rDNA allele 104 in wild and cultivated barleys in the Occidental region. A total of 198 accessions consisting of three distinct samples were used: 82 entries of two‐rowed wild barley from Tibet, 57 accessions of two‐rowed wild barley from 8 countries with a broad range of representation of two‐rowed wild barley in the world, and 59 landrace accessions from four countries representing a part of the barley‐growing areas in the Middle East. These were assayed for rDNA spacer‐length variants (slvs). In all, 27 rDNA space length pheno types were detected, from which 10 slvs were identified as alleles at the two rDNA loci. The two‐rowed wild barley samples from Tibet had the lowest level of genetic variation as evaluated by rDNA polymorphism. Together with results of previous studies, the two wild forms (two‐rowed and six‐rowed) from Tibet could not account for the large genetic diversity observed in the cultivated barley of this region, suggesting that Tibet is unlikely a centre of origin for cultivated barley. In samples from the Occidental region, allele 104 of Rm2 was very rare in wild barley, but occurred at the highest frequency in cultivated barley, while the reverse is the case for allele 107, which is consistent with previous results. The implications of such a contrasting distribution of these rDNA alleles between wild and cultivated barleys in the origin and evolution of cultivated barley were discussed.     

Inheritance and chromosome location of Alp, a gene controlling aluminum tolerance in 'Dayton' barley

Aluminum (Al) toxicity is a major limiting factor in acid soils and more adequate genetic tolerance is needed to improve barley adaptation and production in affected regions. To study the inheritance and chromosome location of the Alp gene controlling Al tolerance in ‘Dayton’ barley the primary trisomics of sensitive ‘Shin Ebisu 16’ were crossed to ‘Dayton’. Parental, F₁ and F₂ seedlings were grown in nutrient solution containing 0.03, 0.06 and 0.09mM Al. and classified for tolerance by haematoxylin staining of the roots. In diploid F₂ progeny, Alp was inherited as a single gene, dominant at 0.06mM and recessive at 0.09 mM concentrations, as indicated by the 3:1 and 1:3 (tolerant: sensitive) segregation ratios, respectively. Segregation of the trisomic Frderived F₂ seedlings at 0.06mM Al deviated significantly from the 3:1 only for the triplo 4/‘Dayton’ cross. Data for this cross fit the expected trisomic ratios, indicating that the Alp gene is distally located from the centromere on chromosome 4. These results confirm that tolerance is simply inherited, but expression of tolerance is dependent on Al concentration and allele dose.     

Waterlogging Affects Leaf and Tillering Dynamics in Wheat and Barley

The aim of this study was to analyse (i) the crop attributes that determine flowering time (i.e. final leaf number, FLN; and phyllochron, Phy), (ii) the dynamics of tiller appearance and (iii) the synchrony between leaf and tiller appearance in wheat and barley plants exposed to waterlogging. Two experiments were carried out in pots, in which wheat and barley cultivars were exposed to five waterlogging treatments, during different periods throughout the crop cycle, from emergence to maturity. The appearance of leaves and tillers on the main stem was measured twice a week in labelled plants. Waterlogging from emergence to flag leaf appearance significantly delayed time to flowering. The delay was greater when waterlogging occurred at the beginning of tillering, lengthening the period from emergence to flowering 24 % (13–15 days) in barley and 10–15 % (6–10 days) in wheat, as compared to control. Phy was the main attribute explaining the delay in flowering, as FLN was not altered. Waterlogging during the early stages of development reduced tiller appearance rate (TAR) in both species, but this effect was partially counterbalanced by a lengthening of the tillering phase, so the effect on final tiller number at maturity was limited. In conclusion, the exposure of wheat and barley to waterlogging during early stages of development delayed time to flowering and reduce TAR in both species. Waterlogging during more advanced crop stages produced slight effects on tillering dynamics, which would indicate that waterlogging affected structure generation more than mortality.     

Genetic analysis of heading date and other agronomic characters in barley (Hordeum vulgare L.)

Genetic analyses of heading date, tiller number, plant height, grain yield, kernel weight, and plump and thin kernels were made in three six-rowed barley crosses (Hordeum vulgare L.) involving four cultivars. Six populations, P₁ , P₂ , F₁ , F₂ , BC₁ , and BC₂ , from each cross were grown and evaluated at Fargo and Prosper, North Dakota, 1982. Parental means within crosses generally were different except for tiller number. Comparison of generation means suggested that late heading was dominant to early, high kernel weight was dominant to low, and kernel plumpness was influenced by additive gene action. The relationship between yield and heading date was not consistent among crosses and positive r values were quite low. It should be possible to select early maturing, high yielding segregates with plump kernels. Heterosis over the mid-parent was quite similar among crosses for heading date, but there was no heterosis over the high parent. Inbreeding depression was fairly constant for heading date, but was less consistent for yield. The lack of uniformity for estimates of inbreeding depression can be related to environmental variation and to its influence on type of gene action. The ratio of additive to dominance variance was inconsistent among crosses for heading date and yield. These data suggest selection for these characters should be delayed past the F 2 generation. Broad sense heritabilities for heading date ranged from 42 to 86%. Values obtained for grain yield were more consistent among broad sense than narrow sense estimates. Genetic advance estimates were low due to lack of additive variance. This revised version was published online in July 2006 with corrections to the Cover Date.     

Quantitative trait loci for scald resistance in barley localized by a non-interval mapping procedure

Three quantitative trait loci (QTL) for scald resistance in barley were identified and mapped in relation to molecular markers using a population of chromosome doubled‐haploid lines produced from the F₁ generation of a cross between the spring barley varieties ‘Alexis’ and ‘Regatta’. Two field experiments were conducted in Denmark and two in Norway to assess disease resistance. The percentage leaf area covered with scald (Rhynchosporium secalis) ranged from 0 to 40% in the 189 doubled‐haploid (DH) lines analysed. One quantitative trait locus was localized in the centromeric region of chromosome 3H, Qryn3, using the MAPQTL program. MAPQTL was unable to provide proper localization of the other two resistance genes and so a non‐interval QTL mapping method was used. One was found to be located distally to markers on chromosome 4H (Qryn4) and the other, Qryn6, was located distally to markers on chromosome 6H. The effects of differences between the Qryn3, Qryn4 and Qryn6 alleles in two barley genotypes for the QTL were estimated to be 8.8%, 7.3% and 7.0%, respectively, of leaf covered by scald. No interactions between the QTLs were found.     

RFLP diversity within and between major groups of barley in Europe

Restriction fragment length polymorphism (RFLP) diversity has been determined and analyzed as expressed by 33 single‐ or low‐copy clone/ enzyme combinations at 32 loci distributed over all chromosomes of the barley genome within a sample of 223 European barley accessions comprised of pure line (single‐head progenies) genotypes. The accessions have been selected to include landraces and widely grown cultivars derived from crossbreeding during the 20th century in North‐, West‐ and Central European countries. Genetic diversity obtained from 83 alleles across all accessions is characterized by the diversity index H = 0.385. The diversity indices determined for landraces and cultivars were almost equal, with the difference between spring (H = 0.260) and winter (H = 0.415) barley approaching statistical significance, while comparisons of other groupings only revealed statistically insignificant trends. A more detailed analysis based on differences in allele frequency distributions at each locus (clone/enzyme combinations resp.) revealed very clear differences related to the existence, continuity and dynamics of changes in group‐specific RFLP profiles. With the majority (69%) of RFLP alleles at 23 out of 32 loci on all barley chromosomes involved, contributions from chromosomes 1H, 3H, 4H and 5H seem to be of special importance. Differences in the overall average of abundance indicate higher levels of genetic diversity within both groups of winter barley compared with both groups of spring barley, from which the most frequent alleles at 15 (2‐rowed spring barley) and 17 (6‐rowed spring barley) RFLP loci approach fixation. The results of this study are discussed in relation to the history of barley cultivation and barley breeding in Europe, and possible explanations for group‐specific differences in the RFLP profiles of landraces and cultivars as well as for the high levels of (nearly) fixed alleles of both subsets of spring barley, and with respect to progress in barley breeding that it has been possible to obtain within the rather narrow RFLP profiles.     

Genetic variation for dry matter and nitrogen accumulation and translocation in two-rowed spring barley II. Nitrogen translocation

An understanding of the partition between pre-anthesis and post-anthesis N uptake and their contribution to total grain N and protein content in spring two-rowed barley (Hordeum vulgare spp. distichum L.) is important to achieve additional breeding progress for both fodder and malting barley. N translocation from the vegetative tissues at anthesis to the kernel, N translocation efficiency, and N harvest index (NHI) were studied in field experiments during 4 years (1995–1998). Plants were harvested at anthesis and maturity and divided into leaf+culm, chaff and grain. Significant cultivar differences in N translocation, N translocation efficiency and NHI were determined. Across cultivars, the highest N translocation was in a favorable year (93 kg ha⁻¹) and the lowest in a year with poor growing conditions (40 kg ha⁻¹). Cultivar differences in N translocation were related to dry matter and pre-anthesis N accumulation (R²>0.70). N translocation efficiency varied more among the cultivars (0.27–0.66) than years (0.47–0.52). Post-anthesis N uptake was negatively correlated (P<0.01) with N translocation. NHI ranged among the cultivars from 0.49 to 0.73 and among the years from 0.57 to 0.74. The cultivars Arapiles, Schooner, Cantala, Kaskade and Pek stored in the grain more than 70% and Hiproly less than 50% of above-ground N at maturity. Translocated N participated with 85, 56, 42, and 61% in grain N in 1995, 1996, 1997, and 1998, respectively. The ratio of translocated N to grain N could be an indicator of growing conditions; a higher ratio indicates good growing conditions over the entire growth period, a lower ratio indicates poor conditions during pre-anthesis, and a medium ratio indicates some temperature and water deviations from the long-term average. Straw N concentration was in significant positive (P<0.01) correlation with N translocation and translocation efficiency. Straw N concentration adequately represents N efficiency utilization for synthesis of grain protein, and because it saves time and money compared to N harvest index determination, it can be used for the testing of breeding materials for the development of new barley cultivars.     

优良水稻染色体片段代换系Z746的鉴定及重要农艺性状QTL定位及其验证

粒型、株高及穗部组成性状与产量形成密切相关,是水稻重要农艺性状,但遗传基础复杂。染色体片段代换系是用于复杂性状遗传研究的良好材料。本研究鉴定了一个以日本晴为受体、西恢18为供体亲本的水稻优良染色体片段代换系Z746。Z746携带来自西恢18的7个代换片段,平均代换长度为3.99 Mb,其株高、粒长和穗部性状均与受体存在显著差异。进一步通过日本晴与Z746杂交构建的次级F2群体共检测到36个相关QTL,分布于2号、3号、4号、6号和11号染色体。其中5个可能与已克隆基因等位,如qPH3-1等, 8个可被多次检出,表明这些是遗传稳定的主效QTL。Z746的粒长主要由4个QTL(qGL3、qGL4、qGL2、qGL6)控制,其中qGL3和qGL4对粒长变异的贡献率分别为60.28%和27.47%。株高由5个QTL控制,穗长由4个QTL控制,每穗粒数由2个QTL控制,千粒重由2个QTL控制。然后以MAS在F3共选出8个单片段代换系,并以此在F4进行了相关QTL验证,共有24个QTL可被8个单片段代换系(SSSL)检出,重复检出率为66.7%,表明这些QTL遗传稳定。本研究为目的 QTL的进一步...     

Diversity and geographical distribution of seed lipoxygenase‐1 thermostability types in barley

To understand the diversity in the thermostability of the seed lipoxygenase‐1 (LOX‐1), 1040 cultivars of worldwide barley (Hordeum vulgare ssp. vulgare) genetic resources were investigated. The relative thermostability of LOX‐1 (LOX‐RTS) in these lines showed a bimodal frequency distribution and these lines were categorized into the high and low thermostability types (H‐type and L‐type, respectively). The H‐type lines predominated in the wild progenitor, ssp. spontaneum. The geographical distribution of these types in the cultivars was surveyed. The frequencies of the H‐ and L‐types were almost equal to one another in southwestern Asia. The occurrence of the H‐type predominated in eastern Asia and Africa, whereas in Europe and Turkey, the L‐type did. The predominance of the L‐type in Europe and Turkey can be understood through the hypothesis that the Fertile Crescent domestication contributed the majority of diversity in Europe. The uneven geographical distribution of the LOX‐1 thermostability types in the cultivars may reflect a polyphyletic origin of barley.