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通过从 NBS 保守序列设计简并引物 PCR 的方法,以花椰菜(Brassica oleracea vat.botrytis)抗、感黑腐病的近等基因系为材料,分离得到 NBS-LRR 型抗性基因同源序列,并获得1个克隆,命名为 RGA330-7.Southern 杂交表明,该片段在近等基因系中存在明显的多态性,且该片段在抗黑腐病基因位点至少存在3个以上类似 RGA330-7的同源拷贝.序列分析结果认为该克隆与 NBS-LRR 型抗性基因的部分 CDSs 有很高的同源性,说明该片段属于 NBS-LRR 型.系统进化分析该序列与甘蓝型油菜的2个抗病同源序列归为一类,很可能这3个不同来源的抗性基因同源序列同属于一种抗性基因家族.因此推测该序列与花椰菜抗黑腐病基因紧密相关,为进一步克隆花椰菜抗黑腐病基因提供了可靠的候选基因,对分子标记辅助抗性育种具有重要意义。
The NBS-LRR type resistance gene homologous sequences were isolated from Brassica oleracea vat.botrytis resistant and black-susceptible near-isogenic lines by designing PCR from degenerate primers of NBS conserved sequences One clone was obtained and named as RGA330-7. Southern blotting showed that the fragment has obvious polymorphism in near isogenic line, and there are at least three more similar RGA330-7 Of the homologous copies of the NBS-LRR type resistance gene.The results of sequence analysis showed that the fragment shared high homology with some CDSs of NBS-LRR type resistance gene, indicating that the fragment belongs to NBS-LRR type. It is possible that the homologous sequences of the resistance genes of the three different origins belong to one family of resistance genes.Therefore, it is speculated that the sequence is closely related to the resistance genes of cauliflower to black rot, Cloning cauliflower resistance gene to the black rot provides a reliable candidate gene for molecular marker assisted resistance breeding is of great significance.