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Heterogeneous inbred populations are useful as sources of near-isogenic lines for RAPD marker localization
Authors:S D Haley  L K Afanador  P N Miklas  J R Stavely  J D Kelly
Institution:(1) Department of Crop and Soil Sciences, Michigan State University, 48824 East Lansing, MI, USA;(2) Tropical Agricultural Research Station, USDA-ARS, P.O. Box 70, 00681 Mayaguez, Puerto Rico;(3) Microbiology and Plant Pathology Laboratory, Plant Sciences Institute, USDA-ARS, 20705 Beltsville, MD, USA;(4) Present address: Department of Plant Sciences, South Dakota State University, 57007 Brookings, SD, USA
Abstract:The development and use of RAPD markers for applications in crop improvement has recently generated considerable interest within the plant breeding community. One potential application of RAPDs is their use for ldquotaggingrdquo simply-inherited (monogenic) pest-resistance genes and enabling more efficient identification and selection of genotypes carrying specific combinations of resistance genes. In this report, we propose and describe the use of heterogeneous inbred populations as sources of near-isogenic lines (NILs) for targeting RAPD markers linked to major pest resistance genes. The development of these NILs for RAPD marker analyses involved a sequence of line and mass selection during successive generations of inbreeding. DNA bulks derived from the NILs were used to identify a RAPD marker (designated OK14620, generated by 5prime-CCCGCTACAC-3prime decamer) that was tightly linked (2.23±1.33 centiMorgans) to an important rust Uromyces appendiculatus (Pers.) Unger var. appendiculatus] resistance gene (Ur-3) in common bean (Phaseolus vulgaris L.). The efficiency of this approach was demonstrated by a low rate of false-positives identified, the tightness of the linkage identified, and the ability to detect polymorphism between genomic regions that are representative of the same gene pool of common bean. This method of deriving NILs should find application by researchers interested in utilizing marker-assisted selection for one or more major pest resistance genes. The identification of OK14620 should help to facilitate continued use of the Ur-3 resistance source and will now enable marker-assisted pyramiding of three different bean rust resistance sources (two previously tagged) to provide effective and stable resistance to this important pathogen.Research supported in part by the grant DAN 1310-G-SS-6008-00 from the USAID Bean/Cowpea Collaborative Research Support Program, the Michigan Agricultural Experiment Station, and the USDA-ARS. Mention of a trademark or a proprietary product does not constitute a guarantee or warranty of the product by the USDA and does not imply its approval to the exclusion of other products that may also be suitable
Keywords:Phaseolus vulgaris L    Bean rust resistance  Polymerase chain reaction (PCR)  DNA markers  Near-isogenic lines (NILs)
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