粳稻BT型细胞质雄性不育恢复基因功能标记的开发与应用

Development and Application of a Functional Marker Associated with Fertility-restoring Gene for BT-type Cytoplasmic Male Sterility(CMS) in japonica Rice

为提高粳稻BT型细胞质雄性不育恢复基因的选择效率,在前人克隆的基础上,根据其恢复系与不育系在Rf1a位点574bp的碱基插入/缺失,设计出功能标记InDel-Rf1a。利用该标记对不同地区来源的72份籼、粳稻材料进行检测,结果表明所有的常规籼稻品种、恢复系及保持系在Rf1a位点并不存在缺失,其基因型为Rf1aRf1a。这些材料对粳稻BT型细胞质雄性不育系具有恢复育性的作用;而绝大多数常规粳稻品种(爱知106和伊粳12号除外)在该位点存在缺失,其基因型为rf1arf1a,它们对BT型细胞质雄性不育系具有保持不育的能力。同时,为进一步验证该标记对不同基因型的检测效果,利用其对粳稻恢复系、不育系、杂交组合以及863A/宁恢8号F2分离群体的DNA进行扩增,根据其电泳带型可准确区分出Rf1a位点的3种基因型。

To improve the selection efficiency of restoring gene for BTtype cytoplasmic male sterility （CMS） in japonica rice, we designed a functional marker InDelRf1a based on a 574 bp insertion/deletion （InDel） in Rf1a locus between japonica restorer line and CMS line reported by previous studies and used it to identify the genotypes of Rf1a locus in 72 indica or japonica rice materials. The results showed that there was no 574 bp deletion in indica conventional rice varieties, restorer and maintain lines and they could restore the fertility of BTtype CMS in japonica rice, with genotypes of Rf1aRf1a; However, the same deletion was detected in most of japonica conventional rice varieties, except in Aichi 106 and Yijing 12, and they had the ability to maintain sterility of BTtype CMS lines, with genotypes of Rf1aRf1a. For further verification of the detection effect with this marker for different genotypes in this locus, we used it to amplify the DNA of different japonica rice restorer lines, CMS lines, hybrid F1 and the F2 population derived from a cross between 863A and Ninghui 8, and three genotypes in Rf1a locus could be distinguished distinctly by gel electrophoresis.