Xu X Y, Zhang Y, Zhang L G, Fang Z Y
State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, Yangling, Shaanxi, China.
State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, Yangling, Shaanxi, China
Genet Mol Res. 2014 Nov 14;13(4):9606-14. doi: 10.4238/2014.November.14.4.
In order to map the restorer gene BrRfp of the polima (pol)-like cytoplasmic male sterility (CMS) 06J45 line in heading Chinese cabbage, an F2 segregating population with 258 individuals of CMS06J45 and the restorer line 01S325 were tested by sequence-related amplified polymorphism (SRAP) and insertion-deletion (InDel) technologies combined with the bulked segregant analysis method. As a result, two SRAP markers, me3em3.366 and pm88bg5.263, that were linked with the BrRfp gene were identified from 463 SRAP primer pairs. By cloning, sequencing, and basic local alignment search tool analysis, the two markers were targeted to the BGIScaffold000053 of Brassica rapa in the Brassica database. Using the BGIScaffold000053 sequence, four InDel primer pairs were designed and identified to be linked with the BrRfp gene in this population. Linkage analysis showed that these markers were distributed on both sides of the BrRfp gene, the linkage distances of two nearest markers InDel878.1125 and InDel920.713 were 0.82 and 0.46 cM, respectively, and the BrRfp gene was restricted to a 243-kb genomic region of B. rapa. These specific markers provided basic information for map-based cloning of the BrRfp gene and will be very valuable for the marker-assisted selection of a new restorer line in heading Chinese cabbage.
为定位结球白菜中polima（pol）型细胞质雄性不育（CMS）06J45系的恢复基因BrRfp，以CMS06J45与恢复系01S325构建的含258个单株的F2分离群体为试材，采用序列相关扩增多态性（SRAP）和插入缺失（InDel）技术并结合集群分离分析法进行研究。结果表明，从463对SRAP引物中筛选出2个与BrRfp基因连锁的SRAP标记，分别为me3em3.366和pm88bg5.263。通过克隆、测序及基本局部比对搜索工具分析，将这2个标记定位到甘蓝型油菜数据库中甘蓝的BGIScaffold000053上。利用BGIScaffold000053序列设计并筛选出4对InDel引物，在该群体中均与BrRfp基因连锁。连锁分析表明，这些标记分布在BrRfp基因两侧，最近的2个标记InDel878.1125和InDel920.713与BrRfp基因的连锁距离分别为0.82和0.46 cM，BrRfp基因被定位在甘蓝243 kb的基因组区域内。这些特异性标记为BrRfp基因的图位克隆提供了基础信息，对结球白菜新恢复系的分子标记辅助选择具有重要价值。
