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甘蓝型油菜隐性三系核不育系因不育性稳定、不育基因易转育、恢复谱广、无胞质负效应、制种产量高等优点在生产上得到广泛应用。其不育性状受两对隐性重叠不育基因(ms1和ms2)与一对隐性上位抑制基因(rf)互作控制。ms1和ms2同时纯合(ms1ms1ms2ms2)表现不育,但隐性纯合rf(rfrf)对ms1ms1ms2ms2的表达起抑制作用,又使其表现可育(临保系,ms1ms1ms2ms2rfrf)。本研究利用BSA群分法,通过AFLP分子标记和SRAP分子标记的筛选,利用Rf基因的BC1分离群体(ms1ms1ms2ms2Rfrf+ms1ms1ms2ms2rfrf)进行遗传连锁分析。结果表明,共筛选到4个与Rf基因均共分离的标记,这些标记在该基因不同遗传背景的BC1群体中均与Rf基因紧密连锁。标记测序序列BLASTn结果表明,其位于大白菜A7染色体Scaffold000017(3.3Mb)上。根据该Scaffold序列信息,开发了一系列SSR引物,多态性SSR引物在Rf基因的一个BC1群体中进行连锁分析,最终将该基因限定在245kb的一个范围内。其中SSR标记A7-10、A7-24与Rf基因共分离,A7-24为共显性标记。
Recessive three-line genic male sterile lines of Brassica napus have been widely used in production because of their stable sterility, easy gene transfer of sterile genes, wide spectrum recovery, no negative effects of cytoplasm and high seed yield. Its sterility trait is controlled by two pairs of recessive overlapping sterility genes (ms1 and ms2) and a pair of recessive upper inhibitory genes (rf). ms1 and ms2 are both homozygous (ms1ms1ms2ms2), but implicit homozygous rf (rfrf) inhibits the expression of ms1ms1ms2ms2, which in turn makes it fertile (ms1ms1ms2ms2rfrf). In this study, genetic linkage analysis (BCI) of Rf gene was carried out by using BCA cluster analysis, AFLP molecular marker and SRAP molecular marker screening, and BC1 isolate of Rf gene (ms1ms1ms2ms2Rfrf + ms1ms1ms2ms2rfrf). The results showed that four co-segregation markers with Rf gene were screened, and these markers were closely linked to Rf gene in the BC1 population of different genetic backgrounds. The BLASTn sequence of the marker sequencing sequence indicated that it was located on the Scaffold000017 (3.3 Mb) of chromosome A7 in Chinese cabbage. Based on the Scaffold sequence information, a series of SSR primers were developed, and a polymorphic SSR primer was used for linkage analysis in a BC1 population of Rf gene, and finally the gene was limited to a range of 245kb. SSR markers A7-10, A7-24 were co-segregated with Rf gene, and A7-24 was co-dominant marker.