A genome-wide association study of malting quality across eight US barley breeding programs

We report malt quality QTLs relevant to breeding with greater precision than previous mapping studies. The distribution of favorable alleles suggests strategies for marker-assisted breeding and germplasm exchange. This study leverages the breeding data of 1,862 barley breeding lines evaluated in 97 field trials for genome-wide association study of malting quality traits in barley. The mapping panel consisted of six-row and two-row advanced breeding lines from eight breeding populations established at six public breeding programs across the United States. A total of 4,976 grain samples were subjected to micro-malting analysis and mapping of nine quality traits was conducted with 3,072 SNP markers distributed throughout the genome. Association mapping was performed for individual breeding populations and for combined six-row and two-row populations. Only 16 % of the QTL we report here had been detected in prior bi-parental mapping studies. Comparison of the analyses of the combined two-row and six-row panels identified only two QTL regions that were common to both. In total, 108 and 107 significant marker-trait associations were identified in all six-row and all two-row breeding programs, respectively. A total of 102 and 65 marker-trait associations were specific to individual six-row and two-row breeding programs, respectively indicating that most marker-trait associations were breeding population specific. Combining datasets from different breeding program resulted in both the loss of some QTL that were apparent in the analyses of individual programs and the discovery of new QTL not identified in individual programs. This suggests that simply increasing sample size by pooling samples with different breeding history does not necessarily increase the power to detect associations. The genetic architecture of malting quality and the distribution of favorable alleles suggest strategies for marker-assisted selection and germplasm exchangeThis is the publisher’s final pdf. The published article is copyrighted by Springer and can be found at: http://link.springer.com/journal/122. Supporting information located at: http://link.springer.com/article/10.1007%2Fs00122-015-2465-

SNP Discovery and Chromosome Anchoring Provide the First Physically-Anchored Hexaploid Oat Map and Reveal Synteny with Model Species

A physically anchored consensus map is foundational to modern genomics research; however, construction of such a map in oat (Avena sativa L., 2n = 6x = 42) has been hindered by the size and complexity of the genome, the scarcity of robust molecular markers, and the lack of aneuploid stocks. Resources developed in this study include a modified SNP discovery method for complex genomes, a diverse set of oat SNP markers, and a novel chromosome-deficient SNP anchoring strategy. These resources were applied to build the first complete, physically-anchored consensus map of hexaploid oat. Approximately 11,000 high-confidence in silico SNPs were discovered based on nine million inter-varietal sequence reads of genomic and cDNA origin. GoldenGate genotyping of 3,072 SNP assays yielded 1,311 robust markers, of which 985 were mapped in 390 recombinant-inbred lines from six bi-parental mapping populations ranging in size from 49 to 97 progeny. The consensus map included 985 SNPs and 68 previously-published markers, resolving 21 linkage groups with a total map distance of 1,838.8 cM. Consensus linkage groups were assigned to 21 chromosomes using SNP deletion analysis of chromosome-deficient monosomic hybrid stocks. Alignments with sequenced genomes of rice and Brachypodium provide evidence for extensive conservation of genomic regions, and renewed encouragement for orthology-based genomic discovery in this important hexaploid species. These results also provide a framework for high-resolution genetic analysis in oat, and a model for marker development and map construction in other species with complex genomes and limited resources

Model SNP development for complex genomes based on hexaploid oat using high-throughput 454 sequencing technology

<p>Abstract</p> <p>Background</p> <p>Genetic markers are pivotal to modern genomics research; however, discovery and genotyping of molecular markers in oat has been hindered by the size and complexity of the genome, and by a scarcity of sequence data. The purpose of this study was to generate oat expressed sequence tag (EST) information, develop a bioinformatics pipeline for SNP discovery, and establish a method for rapid, cost-effective, and straightforward genotyping of SNP markers in complex polyploid genomes such as oat.</p> <p>Results</p> <p>Based on cDNA libraries of four cultivated oat genotypes, approximately 127,000 contigs were assembled from approximately one million Roche 454 sequence reads. Contigs were filtered through a novel bioinformatics pipeline to eliminate ambiguous polymorphism caused by subgenome homology, and 96 <it>in silico </it>SNPs were selected from 9,448 candidate loci for validation using high-resolution melting (HRM) analysis. Of these, 52 (54%) were polymorphic between parents of the Ogle1040 × TAM O-301 (OT) mapping population, with 48 segregating as single Mendelian loci, and 44 being placed on the existing OT linkage map. Ogle and TAM amplicons from 12 primers were sequenced for SNP validation, revealing complex polymorphism in seven amplicons but general sequence conservation within SNP loci. Whole-amplicon interrogation with HRM revealed insertions, deletions, and heterozygotes in secondary oat germplasm pools, generating multiple alleles at some primer targets. To validate marker utility, 36 SNP assays were used to evaluate the genetic diversity of 34 diverse oat genotypes. Dendrogram clusters corresponded generally to known genome composition and genetic ancestry.</p> <p>Conclusions</p> <p>The high-throughput SNP discovery pipeline presented here is a rapid and effective method for identification of polymorphic SNP alleles in the oat genome. The current-generation HRM system is a simple and highly-informative platform for SNP genotyping. These techniques provide a model for SNP discovery and genotyping in other species with complex and poorly-characterized genomes.</p

A new genetic linkage map of barley (\u3ci\u3eHordeum vulgare\u3c/i\u3e L.) facilitates genetic dissection of height and spike length and angle

Plant height and spike length and angle are important agronomic traits in the production of barley (Hordeum vulgare L.) due to strong correlations with lodging and disease. The objective of this study was to use QTL analysis to identify genetic regions associated with each trait in a recombinant inbred line (RIL) mapping population derived from a cross of Falcon by Azhul. Falcon is a spring six-row hulless feed barley with long spikes displaying obtuse angles, while Azhul is a spring dwarf, six-row hulless food barley with short spikes displaying acute angles. The population was genotyped using SNP, DArT and SSR markers and quantitative trait loci (QTL) were detected on chromosomes 2H (102.8 cM, spikelength), 3H (89.2 cM, plant height and 38.2, spike angle and length), 4H (19.0 cM, spike length), and 5H(106.7 cM, spike angle). In conclusion, we developed a barley genetic map, which incorporated SNP, DArT and SSR markers, for detection of height and spike length and angle QTL. Three spike angle, one spike length and one plant height QTL were novel and by using comparative genomics we identified possible candidate genes involved in gibberellic acid signaling and auxin- and ethylene-responsive pathways. This knowledge can be used to generate suitable markers for barley breeding improvement

Comparative Systems Biology Reveals Allelic Variation Modulating Tocochromanol Profiles in Barley (Hordeum vulgare L.)

Tocochromanols are recognized for nutritional content, plant stress response, and seed longevity. Here we present a systems biological approach to characterize and develop predictive assays for genes affecting tocochromanol variation in barley. Major QTL, detected in three regions of a SNP linkage map, affected multiple tocochromanol forms. Candidate genes were identified through barley/rice orthology and sequenced in genotypes with disparate tocochromanol profiles. Gene-specific markers, designed based on observed polymorphism, mapped to the originating QTL, increasing R2 values at the respective loci. Polymorphism within promoter regions corresponded to motifs known to influence gene expression. Quantitative PCR analysis revealed a trend of increased expression in tissues grown at cold temperatures. These results demonstrate utility of a novel method for rapid gene identification and characterization, and provide a resource for efficient development of barley lines with improved tocochromanol profiles

Motifs affected by promoter sequence polymorphism between Falcon and Azhul.

<p>Motifs affected by promoter sequence polymorphism between Falcon and Azhul.</p

Tocochromanol means of Falcon and Azhul parents and two barley checks grown over four location years.

<p>Means and heritability estimates were calculated for Falcon x Azhul (FA) RILs using combined years at each location.</p>a<p>Concentrations given in µg/g. Values followed by the same letter or number of asterisks are not significantly different within a column (p<0.05). For means of genotypes within an form, a single asterisk is equivalent to A, two asterisks to B, and three asterisks to C.</p>+<p>indicates an intermediate value.</p>b<p>Irrigated field trial in Aberdeen.</p>c<p>Non-irrigated field trial at Tetonia.</p>d<p>Broad sense heritability calculated as genotype variance divided by cumulative variance including error.</p

Predicted secondary structure (A) and three-dimensional structure (B) of HGGT proteins in barley cultivars Falcon and Azhul.

<p>Sequence polymorphism contributing to folding changes is indicated by the green consensus guide (A). Blue curved arrows represent turns, orange arrows represent beta strands, pink cylinders represent alpha helices, and gray wavy lines represent coils. Pink labels within ribbon diagrams correspond to the predicted active site.</p

Quantitative PCR analysis of <i>VTE</i>4 (top) and <i>HGGT</i> (bottom) in Falcon and Azhul barley cultivars.

<p>Tissue samples used for cDNA preparation were taken from shoots and embryos grown at room temperature and at 4°C.</p