BackgroundHigh density genetic maps of plants have, nearly without exception, made use of marker datasets containing missing or questionable genotype calls derived from a variety of genic and non-genic or anonymous markers, and been presented as a single linear order of genetic loci for each linkage group. The consequences of missing or erroneous data include falsely separated markers, expansion of cM distances and incorrect marker order. These imperfections are amplified in consensus maps and problematic when fine resolution is critical including comparative genome analyses and map-based cloning. Here we provide a new paradigm, a high-density consensus genetic map of barley based only on complete and error-free datasets and genic markers, represented accurately by graphs and approximately by a best-fit linear order, and supported by a readily available SNP genotyping resource.ResultsApproximately 22,000 SNPs were identified from barley ESTs and sequenced amplicons; 4,596 of them were tested for performance in three pilot phase Illumina GoldenGate assays. Data from three barley doubled haploid mapping populations supported the production of an initial consensus map. Over 200 germplasm selections, principally European and US breeding material, were used to estimate minor allele frequency (MAF) for each SNP. We selected 3,072 of these tested SNPs based on technical performance, map location, MAF and biological interest to fill two 1536-SNP "production" assays (BOPA1 and BOPA2), which were made available to the barley genetics community. Data were added using BOPA1 from a fourth mapping population to yield a consensus map containing 2,943 SNP loci in 975 marker bins covering a genetic distance of 1099 cM.ConclusionThe unprecedented density of genic markers and marker bins enabled a high resolution comparison of the genomes of barley and rice. Low recombination in pericentric regions is evident from bins containing many more than the average number of markers, meaning that a large number of genes are recombinationally locked into the genetic centromeric regions of several barley chromosomes. Examination of US breeding germplasm illustrated the usefulness of BOPA1 and BOPA2 in that they provide excellent marker density and sensitivity for detection of minor alleles in this genetically narrow material.

Diversity Arrays Technology (DArT) can detect and type DNA variation at several hundred genomic loci in parallel without relying on sequence information. Here we show that it can be effectively applied to genetic mapping and diversity analyses of barley, a species with a 5,000-Mbp genome. We tested several complexity reduction methods and selected two that generated the most polymorphic genomic representations. Arrays containing individual fragments from these representations generated DArT fingerprints with a genotype call rate of 98.0% and a scoring reproducibility of at least 99. A lthough 50 years have passed since the structure of DNA was deciphered (1), the study of DNA variation emerged as a field of scientific endeavor only in the last 25 years. Two groups of technologies were developing in parallel from the very beginning: DNA sequencing and molecular markers. DNA sequencing technology developed quickly from proof of concept (2, 3) to an automated process (4), enabling the field of genomics. Molecular marker technologies progressed rapidly as well. Based on the Southern blot technique (5), Botstein et al. (6) developed the restriction fragment length polymorphism (RFLP) technique as a method for creating genetic linkage maps.Development of the PCR technique spawned two important molecular marker techniques: amplified fragment length polymorphism (AFLP) (7) and simple sequence repeats (8). Thousands of studies using molecular markers in plants, including hundreds in barley, have been published but are not referenced because of space limitations.DNA sequencing and molecular marker technologies started to merge when the accumulated sequence data began to yield information on sequence variation among different accessions of the same species. It was soon noted that single-nucleotide polymorphism (SNP) is the most abundant marker type, promising nearly unlimited supply of markers (9). Many alternatives were developed for the SNP assay (primer extension, selective ligation) and the platform to type assays in high throughput (DNA chip, printed and self-assembling arrays, matrix-assisted laser desorption ionization͞ time-of-flight mass spectroscopy) (10)(11)(12)(13)(14).For humans and a limited number of model organisms, the throughput of SNP assays has increased impressively, and assay costs have decreased correspondingly. Yet discovering sequence polymorphism in nonmodel species is difficult, which is particularly true for many crops with limited resources and often complex, polyploid genomes. We have developed Diversity Arrays Technology (DArT) to enable whole-genome profiling of such crops without the need of sequence information. DArT is based on microarray hybridizations that detect the presence versus absence of individual fragments in genomic representations as described by Jaccoud et al. (15).For our initial proof-of-concept study, we selected a species with a simple genome (rice) and used AFLP-like complexity reduction methods to generate genomic representations (15). Here we apply a non-AFLP version ...

A map of the barley genome consisting of 295 loci was constructed. These loci include 152 cDNA restriction fragment length polymorphism (RFLP), 114 genomic DNA RFLP, 14 random amplified polymorphic DNA (RAPD), five isozyme, two morphological, one disease resistance and seven specific amplicon polymorphism (SAP) markers. The RFLP-identified loci include 63 that were detected using cloned known function genes as probes. The map covers 1,250 centiMorgans (cM) with a 4.2 cM average distance between markers. The genetic lengths of the chromosomes range from 124 to 223 cM and are in approximate agreement with their physical lengths. The centromeres were localized to within a few markers on all of the barley chromosomes except chromosome 5. Telomeric regions were mapped for the short (plus) arms of chromosomes 1, 2 and 3 and the long (minus) arm of chromosomes 7.

Background: Molecular marker technologies are undergoing a transition from largely serial assays measuring DNA fragment sizes to hybridization-based technologies with high multiplexing levels. Diversity Arrays Technology (DArT) is a hybridization-based technology that is increasingly being adopted by barley researchers. There is a need to integrate the information generated by DArT with previous data produced with gel-based marker technologies. The goal of this study was to build a high-density consensus linkage map from the combined datasets of ten populations, most of which were simultaneously typed with DArT and Simple Sequence Repeat (SSR), Restriction Enzyme Fragment Polymorphism (RFLP) and/or Sequence Tagged Site (STS) markers.

In recent years, access to complete genomic sequences, coupled with rapidly accumulating data related to RNA and protein expression patterns, has made it possible to determine comprehensively how genes contribute to complex phenotypes. However, for major crop plants, publicly available, standard platforms for parallel expression analysis have been limited. We report the conception and design of the new publicly available, 22K Barley1 GeneChip probe array, a model for plants without a fully sequenced genome. Array content was derived from worldwide contribution of 350,000 high-quality ESTs from 84 cDNA libraries, in addition to 1,145 barley (Hordeum vulgare) gene sequences from the National Center for Biotechnology Information nonredundant database. Conserved sequences expressed in seedlings of wheat (Triticum aestivum), oat (Avena strigosa), rice (Oryza sativa), sorghum (Sorghum bicolor), and maize (Zea mays) were identified that will be valuable in the design of arrays across grasses. To enhance the usability of the data, BarleyBase, a MIAME-compliant, MySQL relational database, serves as a public repository for raw and normalized expression data from the Barley1 GeneChip probe array. Interconnecting links with PlantGDB and Gramene allow BarleyBase users to perform gene predictions using the 21,439 non-redundant Barley1 exemplar sequences or cross-species comparison at the genome level, respectively. We expect that this first generation array will accelerate hypothesis generation and gene discovery in disease defense pathways, responses to abiotic stresses, development, and evolutionary diversity in monocot plants.

Quantitative trait locus (QTL) and QTL x environment (E) interaction effects for agronomic and malting quality traits were measured using a 123-point linkage map and multi-environment phenotype data from an F1-derived doubled haploid population of barley (Hordeum vulgare). The QTL × E interactions were due to differences in magnitude of QTL effects. Highly significant QTL effects were found for all traits at multiple sites in the genome. Yield QTL peaks and support intervals often coincided with plant height and lodging QTL peaks and support intervals. QTL were detected in the vicinity of a previously mapped Mendelian maturity locus and known function probes forα- andβ-amylase genes. The average map density (9.6 cM) should be adequate for molecular marker-assisted selection, particularly since there were few cases of alternative favorable alleles for different traits mapping to the same or adjacent intervals.

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