7 research outputs found
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From the Oregon Wolfe Barley to fall-sown food barley : markers, maps, marker-assisted selection and quantitative trait loci
Understanding complex traits is a fundamental challenge in plant genetics and a prerequisite for molecular breeding. Tools for trait dissection are markers, maps, and quantitative trait locus (QTL) analysis. Marker-assisted selection (MAS) is an application that integrates these tools. In this thesis research, a new sequence-based marker was evaluated, maps were constructed and used, and QTLs were detected using two types of populations. Marker-assisted selection was used to develop a novel class of barley. Restriction-site Associated DNA (RAD), a sequence based-marker technology, allows for simultaneous high-density single nucleotide polymorphism (SNP) discovery and genotyping. We assessed the value of RAD markers for linkage map construction using the Oregon Wolfe Barley (OWB) mapping population. We compared a RAD-based map to a map generated using Illumina GoldenGate Assay (EST-based SNPs). The RAD markers generated a high quality map with complete genome coverage. We then used the RAD map to locate QTL for agronomic fitness traits. A paper describing this research was published (Chutimanitsakun et al., 2011). Marker-assisted selection was used to rapidly develop fall-sown barley germplasm for human food uses. The target traits were high grain β-glucan, vernalization sensitivity (VS) and low temperature tolerance (LTT). The target loci were WX and VRN-H2. Marker-assisted selection was effective in fixing target alleles at both loci and waxy starch led to increase in grain β-glucan. Unexpected segregation at VRN-H1 and VRN-H3, revealed by genome-wide association mapping (GW-AM), led to unanticipated phenotypic variation in VS and LTT. We found that GW-AM is an efficient and powerful method for identifying the genome coordinates of genes determining target traits. Precise information is obtained with perfect markers; additional research may be needed when multiple alleles are segregating at target loci and significant associations are with markers in linkage disequilibrium (LD) with the target loci. A paper describing this research will be submitted for publication
Comparative mapping of the Oregon Wolfe Barley using doubled haploid lines derived from female and male gametes
28 Pag., 2 Tabl. 4 Fig. The definitive version is available at: http://www.springerlink.com/content/0040-5752/The Oregon Wolfe Barley mapping population is a resource for genetics research and instruction. Prior reports are based on a population of doubled haploid (DH) lines developed by the Hordeum bulbosum (H.b.) method, which samples female gametes. We developed new DH lines from the same cross using anther culture (A.C.), which samples male gametes. Linkage maps were generated in each of the two subpopulations using the same 1,328 single nucleotide polymorphism markers. The linkage maps based on DH lines derived from the products of megasporogeneis and microsporogenesis revealed minor differences in terms of estimated recombination rates. There were no differences in locus ordering. There was greater segregation distortion in the A.C.-derived subpopulation than in the H.b.-derived subpopulation, but in the region showing the greatest distortion, the cause was more likely allelic variation at the ZEO1 plant height locus rather than to DH production method. The effects of segregation distortion and pleiotropy had greater impacts on estimates of quantitative trait locus effect than population size for reproductive fitness traits assayed under greenhouse conditions. The Oregon Wolfe Barley (OWB) population and data are community resources. Seed is available from three distribution centers located in North America, Europe, and Asia. Details on ordering seed sets, as well as complete genotype and phenotype data files, are available at http://wheat.pw.usda.gov/ggpages/maps/OWB/.L. Cistué was recipient of a Senior Research Fellowship from the Spanish Ministry of Science and Innovation during his time with the Oregon State University Barley Project and his research was financed by the Spanish Ministry of Science and Innovation thought the National Plan Projects AGL2005-07195-C02-01 and AGL2008-05541-C02-01. Alfonso Cuesta-Marcos was supported by a postdoctoral fellowship from the Spanish Ministerio de Ciencia e Innovación (MICINN). The BOPA SNPs were developed under the auspices of USDA-CSREES-NRI Grant No 2006- 55606-16722 “Barley Coordinated Agricultural Project: Leveraging Genomics, Genetics, and Breeding for Gene Discovery and Barley Improvement”.Peer reviewe
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Application of marker-assisted selection and genome-wide association scanning to the development of winter food barley germplasm resources
Barley (Hordeum vulgare) is an important component of heart-healthy whole grain diets because it contains β-glucan. All current US barley varieties with high β-glucan are spring habit and have waxy starch. Winter varieties have agronomic advantages but require low-temperature tolerance (LTT). Vernalization sensitivity (VS) is associated with higher levels of LTT. To rapidly develop fall-sown varieties with LTT and higher grain β-glucan, we therefore used marker-assisted selection (MAS) at the WX and VRN-H2 loci. The MAS-derived lines, together with unrelated non-waxy germplasm developed via phenotypic selection (PS), were used for a genome-wide association scan (GWAS). The panel was phenotyped for grain β-glucan, LTT and VS. It was genotyped with 3072 single-nucleotide polymorphisms (SNPs) and allele-specific primers. Marker-assisted selection fixed target alleles at both loci but only one of the target phenotypes (higher β-glucan percentage) was achieved. Variation for VS and LTT is attributable to (i) incomplete information about VRN-H1 at the outset of the project and (ii) unexpected allelic variation at VRN-H3 with a large effect on VS and LTT.Keywords: food barley, betaglucan, winter barley, marker assisted selection, vernalization, low temperature toleranceKeywords: food barley, betaglucan, winter barley, marker assisted selection, vernalization, low temperature toleranc
Construction and application for QTL analysis of a Restriction Site Associated DNA (RAD) linkage map in barley
<p>Abstract</p> <p>Background</p> <p>Linkage maps are an integral resource for dissection of complex genetic traits in plant and animal species. Canonical map construction follows a well-established workflow: an initial discovery phase where genetic markers are mined from a small pool of individuals, followed by genotyping of selected mapping populations using sets of marker panels. A newly developed sequence-based marker technology, Restriction site Associated DNA (RAD), enables synchronous single nucleotide polymorphism (SNP) marker discovery and genotyping using massively parallel sequencing. The objective of this research was to assess the utility of RAD markers for linkage map construction, employing barley as a model system. Using the published high density EST-based SNP map in the Oregon Wolfe Barley (OWB) mapping population as a reference, we created a RAD map using a limited set of prior markers to establish linakge group identity, integrated the RAD and prior data, and used both maps for detection of quantitative trait loci (QTL).</p> <p>Results</p> <p>Using the RAD protocol in tandem with the Illumina sequence by synthesis platform, a total of 530 SNP markers were identified from initial scans of the OWB parental inbred lines - the "dominant" and "recessive" marker stocks - and scored in a 93 member doubled haploid (DH) mapping population. RAD sequence data from the structured population was converted into allele genotypes from which a genetic map was constructed. The assembled RAD-only map consists of 445 markers with an average interval length of 5 cM, while an integrated map includes 463 RAD loci and 2383 prior markers. Sequenced RAD markers are distributed across all seven chromosomes, with polymorphic loci emanating from both coding and noncoding regions in the <it>Hordeum </it>genome. Total map lengths are comparable and the order of common markers is identical in both maps. The same large-effect QTL for reproductive fitness traits were detected with both maps and the majority of these QTL were coincident with a dwarfing gene (<it>ZEO) </it>and the <it>VRS1 </it>gene, which determines the two-row and six-row germplasm groups of barley.</p> <p>Conclusions</p> <p>We demonstrate how sequenced RAD markers can be leveraged to produce high quality linkage maps for detection of single gene loci and QTLs. By combining SNP discovery and genotyping into parallel sequencing events, RAD markers should be a useful molecular breeding tool for a range of crop species. Expected improvements in cost and throughput of second and third-generation sequencing technologies will enable more powerful applications of the sequenced RAD marker system, including improvements in <it>de novo </it>genome assembly, development of ultra-high density genetic maps and association mapping.</p
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ChutimanitsakunYadaCropSoilScienceApplicationMarker-Assisted.pdf
Barley (Hordeum vulgare) is an important component of heart-healthy
whole grain diets because it contains β-glucan. All current US barley
varieties with high β-glucan are spring habit and have waxy starch. Winter
varieties have agronomic advantages but require low-temperature tolerance
(LTT). Vernalization sensitivity (VS) is associated with higher
levels of LTT. To rapidly develop fall-sown varieties with LTT and
higher grain β-glucan, we therefore used marker-assisted selection
(MAS) at the WX and VRN-H2 loci. The MAS-derived lines, together
with unrelated non-waxy germplasm developed via phenotypic selection
(PS), were used for a genome-wide association scan (GWAS). The panel
was phenotyped for grain β-glucan, LTT and VS. It was genotyped with
3072 single-nucleotide polymorphisms (SNPs) and allele-specific primers.
Marker-assisted selection fixed target alleles at both loci but only
one of the target phenotypes (higher β-glucan percentage) was achieved.
Variation for VS and LTT is attributable to (i) incomplete information
about VRN-H1 at the outset of the project and (ii) unexpected allelic variation
at VRN-H3 with a large effect on VS and LTT.Keywords: vernalization, low temperature tolerance, winter barley, marker assisted selection, betaglucan, food barle
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ChutimanitsakunYadaCropSoilScienceApplicationMarker-Assisted_SupportingInformation.zip
Barley (Hordeum vulgare) is an important component of heart-healthy
whole grain diets because it contains β-glucan. All current US barley
varieties with high β-glucan are spring habit and have waxy starch. Winter
varieties have agronomic advantages but require low-temperature tolerance
(LTT). Vernalization sensitivity (VS) is associated with higher
levels of LTT. To rapidly develop fall-sown varieties with LTT and
higher grain β-glucan, we therefore used marker-assisted selection
(MAS) at the WX and VRN-H2 loci. The MAS-derived lines, together
with unrelated non-waxy germplasm developed via phenotypic selection
(PS), were used for a genome-wide association scan (GWAS). The panel
was phenotyped for grain β-glucan, LTT and VS. It was genotyped with
3072 single-nucleotide polymorphisms (SNPs) and allele-specific primers.
Marker-assisted selection fixed target alleles at both loci but only
one of the target phenotypes (higher β-glucan percentage) was achieved.
Variation for VS and LTT is attributable to (i) incomplete information
about VRN-H1 at the outset of the project and (ii) unexpected allelic variation
at VRN-H3 with a large effect on VS and LTT.Keywords: food barley, low temperature tolerance, winter barley, vernalization, marker assisted selection, betagluca