杂交粳稻BT型雄性不育恢复基因功能标记优化研究

A Study of Optimization of Functional Markers for Restorer Gene of BT-type Cytoplasmic Male Sterility

根据杂交粳稻BT型细胞质雄性不育保持系与恢复系在Rf1a位点上存在574 bp插入或缺失变异,在缺失区域上游设计10条引物,下游设计9条引物,利用保持系和恢复系分别进行PCR扩增。结果显示,引物Rfa-7F/Rfa-7R在恢复系中扩增出957 bp的片段,保持系扩增出383 bp的片段,并且条带特异性强、稳定性高。同时对引物Rfa-7F/Rfa-7R的PCR扩增体系最低试剂用量进行筛选,为分子辅助选育BT型杂交粳稻恢复系研发出简单稳定低成本的功能标记奠定基础。

According to 574-bp insertion/deletions in the restorer and BT-type cytoplasmic male sterility line at Rf1 a locus,10 forward primers and 9 reverse primers were designed to screen for the efficient marker using restorer and maintain lines.The results showed that a specific 957 bp/383 bp fragments were amplified from restorer line or maintain line with Rfa-7F/Rfa-7R primer, which indicate Rfa-7F/Rfa-7R is an efficient marker. The Rfa-7F/Rfa-7R PCR reaction system was further optimized to improve the selection efficiency, which lay a foundation on molecular assistant selection for the BT-type cytoplasmic male sterility lines.