滇1型水稻质核雄性不育恢复基因的PCR分子标记

PCR Tagging of A Fertility Restoring Gene of Rice CMS Dian 1

用混合品集分析方法 (Bulked L ine Analysis简称 BL A) ,在保持系和测交父本中的 DNA各取 2 0个样等量混合 ,对PMRF、OSR- 33、RM2 94、RG30 4、RG2 5 7五对引物进行筛选 ,结果发现 PMRF和 OSR- 33俩对 PCR引物在保持系和恢复系之间有遗传差异。用这俩对引物分别对 4 0个保持系和 6 0个测交父本 DNA样品进行扩增。另外 ,用滇 1型杂交粳稻的保持系 4 0份和测交父本 6 0份与不育系进行测交 ,杂种 F1 花粉的育性经 1 %的 I- KI染色。用扩增出的带型与 F1 花粉的育性进行相关分析 ,结果表明扩增出带型与花粉的育性呈显著正相关 ,两种带型对应的花粉可育率均值经 t测验差异极显著 ,表明 PMRF和 OSR- 33标记与滇 1型细胞质雄性不育育性恢复基因连锁 ,由于 PMRF和 OSR- 33PCR引物是设计在第 1 0染色体长臂的中部 ,所以 ,滇 1型细胞质雄性不育育性恢复的一个基因可初步定位于第 1

To make clear fertility restoring inheritance of Rice CWS dian 1, a method,bulked line analysis was applied. Each bulk,contained 20 DNA samples parents of test cross and maintaining lines respectively, and were amplified by selecting primers of PMRF,OSR 33,RM294,RG304 and RG257. Differences were found in the primer PMRF and OSR 33. Then using two primers, 40 DNA samples of maintaining lines and 60 restoring lines were amplified. Test cross between CWS Dian 1 lines with maintaining lines and restoring lines were made. Pollens of F1 progenies were stained by 1% I+KI,the pollen fertilities of F1 progenies were different. According to statistical analysis, based on variance analysis and T test,striking correlation were found between the pollen fertility and the three markers. It demonstrated the two molecular markers linked with fertility restoring gene of CWS dian 1. The three molecular markers were located in the middle of the long arm of chromosome 10, so a fertility restoring gene of CWS dian1 located in the middle of the long arm of chromosome 10.