Application of quantitative trait locus mapping to the development of winter-habit malting barley*

Malting quality and winter-hardiness in barley are ‘ultimate’ phenotypes composed of constituent quantitatively inherited traits. A synthesis of molecular-marker linkage data and field phenotyping to reveal the location of quantitative trait loci (QTL) may assist in the development of winter-habit malting barley varieties. One-hundred doubled haploid progeny from a winter x spring cross were evaluated under fall and spring-planted conditions. Malting quality phenotypes and a 76-point map were used to identify QTL and to assess the effect of spring-and autumn-sowing on QTL expression. Many QTL effects were common to both environments and corresponded to QTL detected in other barley germplasm. While there were significant differences in the magnitude of effects across environments, there were no changes in the favourable allele phase. QTL effects for grain protein and diastatic power level coincided with the locations of known function genes. Coincident QTL for a number of mailing-quality traits on chromosome 7 suggests either the presence of a multi-locus cluster of genes controlling components of malting quality or a regulatory gene or genes controlling the cascade of enzymatic processes that function during the malting process. Based on these results, simultaneous selection for malting quality and cold tolerance should be possible in this genetic background.     

QTL alleles from a winter feed type can improve malting quality in barley

Mapping quantitative trait loci (QTLs) responsible for malting quality traits in barley populations has been the main genetic approach to malting quality breeding. A ‘winter × spring’ doubled haploid barley population ‘Nure’ × ‘Tremois’, where such traits were segregating, has been recently developed. Our objective was to map QTLs for malting quality from 2 years of trials in two contrasting locations. QTLs were found on six chromosomes, with a main cluster on chromosome 1H. For wort viscosity and malt extract, favourable alleles at two loci on chromosome 5H were carried by the winter feeding parent ‘Nure’. Doubled‐haploids with higher quality than the spring malting cultivar ‘Tremois’ showed either a facultative or a winter growth habit and a level of frost tolerance comparable with that of the winter tolerant parent ‘Nure’. Markers and QTLs of quality traits were further validated on a separate set of DH lines, coming from the same cross, by means of marker‐assisted selection. This showed that, at least in the present cross combination, positive contributions to malting quality can be found in winter feed barley.     

Evaluation of a barley (Hordeum vulgare) chromosome 2 drought tolerance quantitative trait locus in genomic backgrounds of two cultivars

A barley drought tolerance Quantitatif Trait Locus (QTL) on chromosome 2 was transferred from tolerant cultivar ‘Tadmor’ to susceptible ‘Baronesse’ and ‘Aydanhanım’. Effects of this QTL on drought tolerance and other traits were studied using near-isogenic lines under controlled environments and field trials for two years. This QTL resulted in 5.0% and 9.1% improvement in leaf relative water content of ‘Baronesse’ and ‘Aydanhanım’ cultivars, respectively, under controlled environments. The QTL accelerated heading and maturity by 2.5 days in ‘Baronesse’ and by 5–6 days in ‘Aydanhanım’. It was associated with shorter stature and more ears. This QTL region increased grain yields by 1.1 and 0.6 t/ha in ‘Baronesse’ and ‘Aydanhanım’, respectively, mainly by increasing the number of tillers. There were previous reports related to yield promoting effects of this region harbouring flowering locus eps2 (barley HvCEN gene). However, sequencing of 1025 bp fragment encompassing HvCEN coding region revealed that our parents and near-isogenic lines had no Single Nucleotide Polymorphism (SNP) variation, ruling out direct involvement of eps2. These findings pointed to the possible effect of another flowering locus in the QTL region.     

Grain dormancy QTL identified in a doubled haploid barley population derived from two non-dormant parents

Grain dormancy provides protection against pre-harvest sprouting (PHS) in cereals. Composite interval mapping and association analyses were performed to identify quantitative trait loci (QTL) contributing grain dormancy in a doubled haploid (DH) barley population (ND24260?×?Flagship) consisting of 321 lines genotyped with DArT markers. Harvest-ripe grain collected from three field experiments was germinated over a 7-day period to determine a weighted germination index for each line. DH lines displaying moderate to high levels of grain dormancy were identified; however, both parental lines were non-dormant and displayed rapid germination within the first two?days of testing. Genetic analysis identified two QTL on chromosome 5H that were expressed consistently in each of the three environments. One QTL (donated by Flagship) was located close to the centromeric region of chromosome 5H (qSDFlag), accounting for up to 15% of the phenotypic variation. A second QTL with a larger effect (from ND24260) was detected on chromosome 5HL (qSDND), accounting for up to 35% of the phenotypic variation. qSDFlag and qSDND displayed an epistatic interaction and DH lines that had the highest levels of grain dormancy carried both genes. We demonstrate that qSDND in the ND24260?×?Flagship DH population is positioned proximal and independent to the well-characterised SD2 region that is associated with both high levels of dormancy and inferior malt quality. This indicates that it should be possible to develop cultivars that combine acceptable malting quality and adequate levels of grain dormancy for protection against PHS by utilizing these alternate QTL.     

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

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 genomic position of a gene set with a putative role in the regulation of flowering time and abiotic stress response in barley. The population has been evaluated in eighteen location-by-year combinations across the Mediterranean basin. QTL mapping identified several genomic regions responsible for barley adaptation to Mediterranean conditions in terms of phenology, grain yield and yield component traits. The most frequently detected yield QTL had the early flowering HvCEN_EPS2 locus (chromosome 2H) as peak marker, showing a positive effect from the early winter parent ‘Nure’ in eight field trials, and explaining up to 45.8 % of the observed variance for grain yield. The HvBM5A_VRN-H1 locus on chromosome 5H and the genomic region possibly corresponding to PPD-H2 on chromosome 1H were significantly associated to grain yield in five and three locations, respectively. Environment-specific QTLs for grain yield, and clusters of yield component QTLs not related to phenology and or developmental genes (e.g. on chromosome 4H, BIN_09) were observed as well. The results of this work provide a valuable source of knowledge and tools for both explaining the genetic bases of barley yield adaptation across the Mediterranean basin, and using QTL-associated markers for MAS pre-breeding and breeding programmes.     

Quantitative trait loci for grain yield and yield components in a cross between a six-rowed and a two-rowed barley

Summary The positions of quantitative trait loci (QTL) for yield and yield components were estimated using a 85-point linkage map and phenotype data from a F₁-derived doubled haploid (DH) population of barley. Yield and its components were recorded in two growing seasons. Highly significant QTL effects were found for all traits at several sites in the genome. A major portion of the QTL was found on chromosome 2. The effect of the alleles in locus v on thousand grain weight and kernels per ear explained 70–80% of the genetic variation in the traits. QTL × year interaction was found for grain yield. Several different QTL were found within the two-rowed DH lines compared to those found in the six-rowed DH lines. Epistasis between locus v and several loci for yield and yield components indicates that genes are expressed differently in the two ear types. This may explain the difficulties of selecting high yielding lines from crosses between two-rowed and six-rowed barley.Abbreviations DH doubled haploid - QTL quantitative trait locus/loci - RAPD random amplified polymorphic DNA - RFLP restriction fragment length polymorphism - T. Prentice Tystofte Prentice - V. Gold Vogelsanger Gold     

Possibility of exploiting the Yd2 resistance to BYDV in spring barley breeding

The effects of the Yd2 gene on tolerance to barley yellow dwarf virus (BYDV) and other agronomically important characters in spring barley were evaluated in a set of randomly selected doubled haploid (DH) lines of an‘Igri’/‘Atlas 68’ cross and three crosses between CIMMYT Yd2 materials and the Czech malting barley ‘Akcent’. The cleaved amplified polymorphic site (CAPS) diagnostic marker Yd2 was used for identification of the Yd2 gene and this analysis showed high agreement with the results of field infection tests. Yd2 lines exhibited significantly lower symptom scores and lower reductions of some grain yield characters, but their resistance level was not consistent over the years. The presence of secondary stresses (high temperature/drought) in 2000 led to relatively higher sensitivity to BYDV infection, strengthened by the long life cycle of genotypes. In cases where secondary stresses were mild (in 2002), the longer life cycle significantly increased sensitivity to BYDV infection only in the absence of the Yd2 gene (in susceptible genotypes). The examination of different vegetative, grain yield and malting quality characters separately for groups of Yd2 and non‐ Yd2 lines did not show any evidence of adverse effect of the Yd2 gene on any character.     

Quantitative trait loci for dry matter digestibility and particle size traits in two-rowed × six-rowed barley population

More than half of the barley grown in the USA is used for livestock feed, with the remaining stocks diverted for human food and malting purposes. The use of barley grain as a major source of cattle feed has been criticized because of its rapid digestion in the rumen, which can result in digestive disorders in cattle. In sacco dry matter digestibility (ISDMD) and particle size (PS) after dry rolling have been found to play a role in the feedlot performance of barley as a feed grain. Reducing the rate of ISDMD is predicted to result in significantly improved animal health and average daily gain. A recombinant inbred line population derived from a cross between a high ISDMD, two-rowed barley cultivar (Valier) and a six-rowed Swiss landrace line (PI370970) exhibiting far slower ISDMD has been developed for studying the underlying genetic locations and mechanisms of these traits. To detect associated quantitative trait loci (QTLs), we collected and analyzed data from irrigated and rain-fed environments. A significant negative correlation was observed between ISDMD and PS. High heritability estimates for ISDMD and PS suggest that early selection for these traits during breeding would be achievable. Four QTLs were identified on chromosomes 2H, 6H, and 7H, explaining 73–85% of ISDMD phenotypic variation, while three QTLs on 2H and 7H were associated with variation in PS and explained 58–77% of its variation. A major QTL on chromosome 2H tightly linked to the morphology-modifying gene vrs1 was found to dramatically control 35–62% of the phenotypic variation of ISDMD and 26–53% of that of PS. The impact of the vrs1 locus on ISDMD was validated in two populations representing different genetic backgrounds. Our results suggest that it may also be advantageous to simultaneously overlap these QTLs around the vrs1 locus.     

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.     

Comparative analysis of Australian and Canadian barleys for seed dormancy and malting quality

The association between high malting quality and pre-harvest sprouting (PHS) susceptibility is a key challenge when developing new malting barley varieties. A new malting barley variety Baudin has successfully combined high malting quality and PHS tolerance. A doubled haploid population was developed for mapping PHS tolerance and seed dormancy from a cross of Baudin?×?AC Metcalfe using 233 molecular markers. Three QTLs were mapped for seed dormancy based on the standard germination test at 24, 48 and 72?h. One major QTL was mapped to the long arm of chromosome 5H controlling seed dormancy and PHS tolerance from Baudin. Two other minor QTLs were identified from Baudin on chromosomes 3 and 7H. QTL/QTL interaction was detected for seed dormancy between chromosomes 3 and 5H. The PHS tolerance allele of the 5H QTL from Baudin contributes to higher malt yield without significant impact on diastatic power, beta-glucan content and wort viscosity. QTL from Baudin provide new sources to integrate PHS tolerance and high malting quality.     

Mapping a factor controlling the thermostability of seed lipoxygenase-1 in barley

Barley lipoxygenase (LOX)‐1 is believed to affect the stability of flavour and the foam of beer. The purpose of this study was to investigate the genetic variation of the LOX‐1 thermostability, and to analyse the mode of inheritance of this trait. A simple method was established to evaluate the LOX‐1 relative thermostability (LOX‐RTS). With this method, 153 barley cultivars were screened for LOX‐RTS. The frequency of the LOX‐RTS values was distributed in a bimodal manner. Based on these values, the barley lines were categorized into two groups: an H‐type with relatively thermostable LOX‐1 and an L‐type with relatively thermolabile LOX‐1. Using a ‘Steptoe’/‘Morex’ doubled haploid population, a major quantitative trait locus (QTL) associated with LOX‐RTS was identified on chromosome 4H of barley, explaining 82% of the variance. Mapping of a CAPS marker specific for the LoxA locus revealed co‐segregation with this QTL. In this study, the existence of the thermostability types of barley seed LOX‐1 and the locus controlling the thermostability were made clearer.     

Identification and mapping of a QTL for rachis internode length associated with cleistogamy in barley

General knowledge of the closed flowering trait, or cleistogamy, of barley is still limited. The relationship between cleistogamy and spike morphology characters was studied and linkage of cleistogamy genes with a highly significant quantitative trait locus (QTL) for rachis internode length on the long arm of chromosome 2H was detected. The mapping populations consisted of 129 doubled haploid lines of ‘Mikamo Golden’ × ‘Harrington’ and 150 F₂ plants of ‘Misato Golden’ × ‘Satsuki Nijo’. The phenotypic variance explained by this QTL accounted for 77.5% and 82.6% of the variance in rachis internode lengt, respectively, in these two populations. The peaks of the QTL coincided with the positions of the cleistogamy gene loci.     

Use of barley seed vigour to discriminate drought and cold tolerance in crop years with high seed vigour and low trait variation

Seed vigour is a precondition for early and homogenous field emergence of barley, in addition to effective malting. This study aimed to assess the selection of barley varieties by using seed vigour as the indicator. Seed vigour of barley (quantified as the germination percentage) was evaluated under drought (?0.2 MPa) and temperature stress (10°C). At two locations over a 3‐year period, 1 population of 133 Derkado × B83‐12/21/5 doubled haploid (DH) lines (and parents) was evaluated for seed vigour, of which 108 DH lines were assessed for three malting parameters. The relatively high values of vigour during the 3‐year period (overall average 94–95%) probably impeded high variations in genetic potential. A total of 27 DH lines of the 133 evaluated showed transgression for vigour (up to 98%) in comparison with the parents (Derkado: 96%; B83: 92%). In conclusion, caution should be applied when selecting for seed vigour, even in good crop years with high levels of seed vigour and low trait variations. Such selection might improve vigour, particularly in crop years with unsuitable weather conditions.     

Employment of hordein subunit polymorphisms in establishing selection criteria for high quality malting barley (Hordeum vulgare L.)

Cereal seed storage proteins are encoded by complex multigene families and their subunit profiles are highly related to end-use qualities. Each fraction of albumin and hordein was extracted and its subunit profile was evaluated in related to malt and grain quality parameters. The purpose of this study was to provide selection criteria for high quality malting barley using grain and malt quality parameters and biochemical-genetic information. Grain and malt quality of 13 local adaptability test (LAT) lines were evaluated for malting process. A total of 16 germplasm accessions of high or low seed storage protein content were also evaluated for biochemicalgenetic analysis. The correlation coefficients among quality parameters were analyzed. Several important quality parameters in brewing process showed significant positive or negative correlations. Seed storage protein subunits of albumin and hordein of all tested lines and accessions were evaluated using 12% 1D SDS-PAGE. Scored data of protein subunit’s presence or absence was applied to Agglomerative Hierarchical Clustering (AHC) for statistical analysis and showed specific grouping patterns among tested lines. Clustered lines with subunit information were highly related with agricultural performance and grain and malt qualities. Based on the profiles of seed storage protein subunits, association of hordein subunit of 38, 43, and 65 kDa with high malt scored lines was found. The obtained results would provide improved selection criteria for high quality malting barley in the malting barley breeding program.     

Construction of chromosomal segment substitution lines and genetic dissection of introgressed segments associated with yield determination in the parents of a super‐hybrid rice

Heterosis is a phenomenon whereby hybrids of inbred lines produce favourable phenotypes that exceed those of their parents. Traits of interest are higher yield and stronger stress tolerance. The two‐line super‐hybrid rice ‘Liangyoupei9’ (LYP9) shows superiority to both its elite inbred line ‘93‐11’ and ‘Pei'ai64s’ (‘PA64s’) parents and conventional hybrids. However, the genetic basis of its hybrid vigour, especially yield determination, remains elusive. In the present study, a set of 156 chromosome segment substitution lines (CSSLs) carrying overlapping segments from ‘PA64s’ in a genetic background of ‘93‐11’ were constructed and planted in six environments. Three major agronomic traits, viz. panicle length (PL), heading date (HD) and plant height (PH), and five yield‐related traits, viz. grain weight per panicle (GWP), number of grains per panicle (GPP), 1000‐grain weight (TGW), seed set (SS) and number of panicles of per plant (PPP), were evaluated over 3 years. Quantitative trait loci (QTL) analysis was conducted using a likelihood ratio test based on stepwise regression. Forty‐six putative QTL distributed on 11 chromosomes were detected in more than one year. Remarkably, GWP of four CSSLs carrying positive yield QTL outperformed the recurrent parent ‘93‐11’ by more than 15%, in at least two environments. These results indicate that CSSLs are effective in identifying yield‐associated traits, and lines harbouring such QTL will be rich in resources for future molecular breeding programmes.     

Genetic variation in barley of crossability with wheat and its quantitative trait loci analysis

To study genetic variation in crossability, 80 barley accessions of diverse geographic origin consisting of 50 wild barleys (H. vulgare ssp. spontaneum or ssp. agriocrithon) and 30 cultivated barleys (H. vulgare ssp. vulgare) were crossed as the male parent with a highly crossable wheat variety, Shinchunaga. Crossabilities, expressed as the percentage of pollinated florets giving embryo-containing caryopses, ranged from 0% to 68.6%. Barley accessions from East Asia had generally a low crossability, while barley accessions from other regions exhibited a wider range of crossability including highly crossable genotypes. No significant difference in mean crossability was found between wild and cultivated barleys. To estimate the number and location of barley genes controlling the crossability, doubled haploid lines derived from the cross between the barley varieties Steptoe and Morex were crossed as the male parent with wheat. Quantitative trait loci (QTL) analysis using molecular markers identified four QTL. These were mapped to the centromeric regions of chromosomes 2H, 3H and 5H and the short arm of chromosome 7H. The QTL on chromosomes 3H and 5H had larger effects than those on chromosomes 2H and 7H. The four QTL collectively explained 35.4% of the total variance under a multiple QTL model. Relationships of the QTL identified in the present study with previously reported crossability genes of barley and wheat are discussed. This revised version was published online in August 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.     

Quantitative trait loci for stay-greenness and agronomic traits provide new insights into chlorophyll homeostasis and nitrogen use in rice

Functional stay-green (FSG) is characterized by delayed senescence during the reproductive stage in rice. A recombinant inbred line (RIL) population derived from ‘Pusa 677’/‘PSG16’ was used to study the genetics of FSG in a rice mutant, ‘PSG16’. The RILs exhibited significant variation under two N regimes for agro-morphological traits, leaf chlorophyll content, flowering time, yield components and nitrogen (N) use. Using a genome wide linkage map spanning 1910.8 cM with 104 polymorphic markers, we have mapped six major quantitative trait loci (QTLs). One robust QTL on chromosome 1 was associated consistently across different N regimes with chlorophyll content and flowering time. The QTL on chromosome 7 was associated with grain number, whereas the QTL on chromosome 6 was found related to N harvest index and spikelet fertility. Although ‘PSG16’ showed a clear advantage in grain yield as well as having better N assimilation, we could not establish a direct genetic relationship SG trait and N use. Based on the QTL information, FSG trait of ‘PSG16’ could be useful in deciphering multiple stress responses in rice.     

A major QTL effect controlling resistance to powdery mildew in winter wheat at the adult plant stage

Powdery mildew is one of the major diseases of wheat in regions with a maritime or semi‐continental climate which can strongly affect grain yield. The objective of the study was to identify and compare quantitative resistance to powdery mildew of line RE9001 at the adult plant and vernalized seedling stages. RE9001 has no known Pm gene and shows a high level of adult plant resistance in the field. Using 104 recombinant inbred lines (RILs) of an RE9001 × ‘Courtot’ F8 population, a genetic map was developed with 363 markers distributed over 26 linkage groups and covering 3825 cM. The global map density was 1 locus/10.3 cM. RILs were assessed under field and tunnel greenhouse conditions for 2 years in two locations. Eleven quantitative trait loci (QTL) were detected at the adult stage and they explained 63% of the variation, depending on the environment. Three QTLs were found, at least, in the two environments. One QTL from RE9001, mapped on chromosome 2B, was stable in each environment. This QTL, QPm.inra.2B, explained 10.3–36.6% of the variation and could be mapped in the vicinity of the Pm6 gene. At the vernalized seedling stage, one QTL detected by the isolate 93‐27 could be an allele of the Pm3g gene present in ‘Courtot’. No residual effect of the Pm3g gene was detected at either stage. Markers flanking the QTL 2B could be useful tools to combine resistance to powdery mildew in wheat cultivars.     

Mapping quantitative trait loci for yield-related traits in Chinese cabbage (Brassica rapa L. ssp. pekinensis)

Chinese cabbage (Brassica rapa L. ssp. pekinensis) is one of the most important vegetables in China. However, the inheritance of yield-related traits in Chinese cabbage is poorly understood to date. To map quantitative trait loci (QTL) for yield-related traits in Chinese cabbage, a genetic linkage map was constructed with 192 doubled haploid (DH) lines. The genetic map was constructed based on 190 sequence-related amplified polymorphisms and 43 simple sequence repeats. QTL mapping was conducted for 11 yield-related traits in 170 DH lines derived from a cross between two diverse Chinese cabbage lines, ‘WZ’ and ‘FT’, under different environmental conditions. A total of 46 main QTL (M-QTL) and 7 epistatic QTL (E-QTL) were identified. The phenotypic variation explained by each M-QTL and E-QTL ranged from 4.85 to 25.06 % and 1.85 to 13.29 %, respectively. The QTL-by-environment interactions were detected using the QTLNetwork 2.0 program in joint analyses of multi-environment phenotypic values. The phenotypic variation explained by each QTL and by QTL × environment interaction was 1.14–4.24 % and 0.00–1.26 %, respectively. Our results provide a better understanding of the genetic factors controlling leaf and head-related traits in Chinese cabbage.