中国农业科学 ›› 2006, Vol. 39 ›› Issue (12): 2621-2626 .

• 研究简报 • 上一篇    

不结球白菜抗病基因同源序列的克隆及分析

王彦华,侯喜林,申书兴,陈雪平,王梅   

  1. 南京农业大学作物遗传与种质创新国家重点实验室
  • 收稿日期:2005-08-03 修回日期:1900-01-01 出版日期:2006-12-10 发布日期:2006-12-10
  • 通讯作者: 侯喜林

Isolation and Characterization of Resistant Gene Analogs from Brassica campestris ssp.chinensis

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  1. 南京农业大学作物遗传与种质创新国家重点实验室
  • Received:2005-08-03 Revised:1900-01-01 Online:2006-12-10 Published:2006-12-10

摘要: 【目的】利用同源序列法分离不结球白菜抗病基因同源序列。【方法】根据植物抗病基因TIR-NBS-LRR保守区设计简并引物,对不结球白菜基因组DNA及cDNA进行PCR扩增。【结果】获得了10个具有通读氨基酸序列的片段。同源性比较发现,该10个片段均属于TIR-NBS-LRR类抗病基因同源序列(RGA),与已知R基因相应区段的氨基酸序列一致性为50%~66%。与已知抗病基因聚类分析结果显示,该10个RGA序列可以分为5大类。利用RGA950作为探针进行Southern杂交,结果表明,其在基因组中存在多拷贝。【结论】本研究成功获得了不结球白菜RGA序列,为进一步克隆不结球白菜的R基因奠定了基础。

关键词: 不结球白菜, 抗病基因同源序列, TIR-NBS-LRR, Southern杂交

Abstract: The PCR degenerate primers were designed according to the conservative domain TIR-NBS-LRR of most disease resistant genes in plants. DNA fragments of 240 to 980 bp were amplified by genomic DNA and cDNA PCR from B. campestris ssp.chinensis resistant materials Xuekeqing and Suzhouqing. Ten resistant gene analogs were obtained which contain the TIR-NBS-LRR domain.The deduced amino acid sequences have different homology scores compared with those of resistance genes or resistance gene analogs from other plants in GeneBank. A cluster analysis based on the deduced amino acid sequences with several resistance genes from other plants, these RGAs can be classified into five major groups. Using RGA950 as probe, Southern bloting analysis showed there existed multi-copies in genomic DNA of Xuekeqing. These RGAs can further be used as probes for cloning of resistance gene and construction of genetic map in B. campestris ssp.chinensis.

Key words: B. campestris ssp.chinensis, resistance gene analog, TIR-NBS-LRR, Southern blot