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High-throughput single nucleotide polymorphism genotyping for breeding applications in rice using the BeadXpress platform |
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| Authors: | Michael J Thomson Keyan Zhao Mark Wright Kenneth L McNally Jessica Rey Chih-Wei Tung Andy Reynolds Brian Scheffler Georgia Eizenga Anna McClung Hyunjung Kim Abdelbagi M Ismail Marjorie de Ocampo Chromewell Mojica Ma Ymber Reveche Christine J Dilla-Ermita Ramil Mauleon Hei Leung Carlos Bustamante Susan R McCouch |
| Institution: | 1. International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines 2. Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, NY, 14853-2601, USA 3. Department of Genetics, Stanford University, Stanford, CA, 94305-5120, USA 4. Department of Plant Breeding and Genetics, Cornell University, 162 Emerson Hall, Ithaca, NY, 14853-1901, USA 5. USDA-ARS Genomics and Bioinformatics Research Unit, 141 Experiment Station Road, P.O. BOX 36, Stoneville, MS, 38776-0350, USA 6. USDA-ARS Dale Bumpers National Rice Research Center, 2890 Hwy 130 East, Stuttgart, AR, 72160, USA |
| Abstract: | Multiplexed single nucleotide polymorphism (SNP) markers have the potential to increase the speed and cost-effectiveness of genotyping, provided that an optimal SNP density is used for each application. To test the efficiency of multiplexed SNP genotyping for diversity, mapping and breeding applications in rice (Oryza sativa L.), we designed seven GoldenGate VeraCode oligo pool assay (OPA) sets for the Illumina BeadXpress Reader. Validated markers from existing 1536 Illumina SNPs and 44?K Affymetrix SNP chips developed at Cornell University were used to select subsets of informative SNPs for different germplasm groups with even distribution across the genome. A 96-plex OPA was developed for quality control purposes and for assigning a sample into one of the five O. sativa population subgroups. Six 384-plex OPAs were designed for genetic diversity analysis, DNA fingerprinting, and to have evenly-spaced polymorphic markers for quantitative trait locus (QTL) mapping and background selection for crosses between different germplasm pools in rice: Indica/Indica, Indica/Japonica, Japonica/Japonica, Indica/O. rufipogon, and Japonica/O. rufipogon. After testing on a diverse set of rice varieties, two of the SNP sets were re-designed by replacing poor-performing SNPs. Pilot studies were successfully performed for diversity analysis, QTL mapping, marker-assisted backcrossing, and developing specialized genetic stocks, demonstrating that 384-plex SNP genotyping on the BeadXpress platform is a robust and efficient method for marker genotyping in rice. |
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