甘蓝型油菜细胞质雄性不育系L04-05A恢复基因的SSR标记

SSR Markers of B. napus L. Cytoplasmic Male Sterile Lines L04-05A Restoring Genes

以甘肃农业大学农学院育成的甘蓝型油菜细胞质雄性不育系L04-05A与恢复材料L04-05C1,L04-05C2配制的F1,F2分离群体为材料,采用分离群体分组分析法(BSA),进行了油菜细胞质雄性不育育性恢复基因Rfp分子标记的筛选及连锁分析。结果表明,F2可育株与不育株的分离比例为4.30∶1和2.73∶1,χ2C值分别为2.538 4和0.389 8,均小于x20.05,1,符合3∶1的分离比,呈孟德尔式遗传,表明PLCMS(L04-05A)育性恢复基因Rfp是受一对显性基因控制的简单遗传。再分别利用L04-05A×L04-05C1的F2群体和L04-05A×L04-05C2的F2群体构建的可育DNA混合池和不育DNA混合池对60对引物进行了筛选,在2个DNA池之间显示有差异条带的2对引物,再用2个群体单株进行了进一步验证,发现引物Ol13-E08扩增的分子量为150 bp的条带为可育株特异标记,在不育单株中未扩增出该条带,重复性较好。进一步用引物Ol13-E08分别对2个F2群体进行扩增,确定Ol13-E08-150 bp为与PLCMS(L04-05A)育性恢复基因连锁的SSR标记,Ol13-E08-150 bp与育性恢复基因的遗传距离为5.13 c M。

This paper based on the cytoplasmic male sterile line L04-05 A of brassica napus which bred from College of Agronomy,Gansu Agricultural University and F1, F2 segregating population which prepared from restoring material L04-05C1 and L04-05C2, and the Bulked Sergeant Analysis（BSA）was adopted to screen molecular marker and to linkage analysis of the restorer gene of cytoplasmic male sterile. Based on the F2 segregating population, it proved that the fertility restoring of L04-05C1 and L04-05C2 to L04-05 A was controlled by one dominant gene through fertility segregation analysis in florescence. To screen 60 primers by using fertile DNA mixing pool and sterile DNA mixing pool which established from number 06-06 F2 population and number 06-09 F2 population respectively; and it showed that there were two primers of differential band between the two DNA mixing pool, then to further prove by the single plant that used for establishing mixing pool, and it was found that the molecular weight of 150 bp band which amplified by primer Ol13-E08 was the differential marker for fertile plant, and it couldn＇t be amplified this band in sterile plant, with good reproducibility. Then further amplified the F2 population of number 06-06 and 06-09 respectively by primer Ol13-E08 made sure that Ol13-E08-150 bp was the SSR marker linked to the fertility restorer gene of PLCMS（L04-05A）, and the genetic distance between Ol13-E08-150 bp and fertility restorer gene was 5.13 c M.