野败型水稻细胞质雄性不育恢复基因Rf-4的分子标记定位

Molecular Mapping of the Fertility Restorer Gene Rf-4 for WA Cytoplasmic Male Sterility in Rice

为了用分子标记准确定位野败型水稻细胞质雄性不育恢复基因Rf 4 ,将日本水稻基因组项目 (RiceGenomeProgram ,RGP)构建的水稻遗传连锁图谱第 10染色体分子遗传图上的分子标记R1877和G2 15 5之间对应区域YAC物理图上的 6个YAC克隆进行了亚克隆 ,获得 119个片段。对这些探针进行多态性探查 ,获得了 2个多态分子标记。用珍汕 97A和恢复基因近等基因系的杂种F2 分离群体中的 117完全不育株进行连锁分析表明 ,从YAC4 892获得的亚克隆Y3 8与Rf 4座位的连锁距离为 0 .9cM ,从YAC4 6 30获得的亚克隆Y1 10与Rf 4座位的连锁距离为3.2cM。根据以上结果把Rf 4座位定位于第 10染色体的特定位置 ,为该基因的分子标记辅助选择和定位克隆打下了基础。

The cytoplasmic male sterility for wild abortive (CMS WA) has been wildly used for hybrid rice breeding in China. The fertility restoration of CMS WA is controlled mainly by two independent and dominant nuclear fertility restoring genes, Rf 3 and Rf 4 . To map the Rf 4 gene with molecular markers, rice YAC clones of RGP, Japan were used to create new molecular marker. YAC contigs located between RFLP markers R1877 and G2155 on chromosome 10 were confirmed by hybridization with 12 RFLP probes. Six YAC clones, Y4630, Y2670, Y4892, Y2111, Y3821 and Y5528 were identified. Chromosome DNAs of the YAC clones were prepared and separated by CHEF. A total of 119 probes were created by sub cloning of the YAC DNAs. RFLPs were screened between Zhenshan97A and its near isogenic lines with Rf 4Rf 4 genotype. Two probes, Y3 8 from Y4892 and Y1 10 from Y4630, were found to be polymorphic. Using F 2 populations from crosses between Zhenshan97A and its near isogenic lines ZSR11, Y3 8 and Y1 10 were mapped to Rf 4 locus with genetic distances of 0.9cM and 3.2cM, respectively.