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BAC end sequences corresponding to the B4 resistance gene cluster in common bean: a resource for markers and synteny analyses
Authors:Perrine David  Mireille Sévignac  Vincent Thareau  Yann Catillon  Jim Kami  Paul Gepts  Thierry Langin  Valérie Geffroy
Institution:Institut de Biotechnologie des Plantes, Université Paris-Sud, 91405, Orsay, France.
Abstract:In common bean, a complex disease resistance (R) gene cluster, harboring many specific R genes against various pathogens, is located at the end of the linkage group B4. A BAC library of the Meso-american bean genotype BAT93 was screened with PRLJ1, a probe previously shown to be specific to the B4 R gene cluster, leading to the identification of 73 positive BAC clones. BAC-end sequencing (BES) of the 73 positive BACs generated 75 kb of sequence. These BACs were organized into 6 contigs, all mapped at the B4 R gene cluster. To evaluate the potential of BES for marker development, BES-derived specific primers were used to check for linkage with two allelic anthracnose R specificities Co-3 and Co-3 ( 2 ), through the analysis of pairs of Near Isogenic Lines (NILs). Out of 32 primer pairs tested, two revealed polymorphisms between the NILs, confirming the suspected location of Co-3 and Co-3 ( 2 ) at the B4 cluster. In order to identify the orthologous region of the B4 R gene cluster in the two model legume genomes, bean BESs were used as queries in TBLASTX searches of Medicago truncatula and Lotus japonicus BAC clones. Putative orthologous regions were identified on chromosome Mt6 and Lj2, in agreement with the colinearity observed between Mt and Lj for these regions.
Keywords:Disease resistance gene            Phaseolus vulgaris            BAC-end sequence  Near isogenic lines  Synteny
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