甘薯NBS-LRR类抗病基因同源序列的克隆、分析及数目研究  

Cloning and Copy Number Estimation of NBS-LRR Based Disease Resistance Gene Analogs from Ipomoea batatas

作　　者：屈满义[1,2] 查向东[1,2] 王钰[1,2] 杨金环[1,2] 蒋琳[1,2] 阮龙[3] 

机构地区：[1]安徽大学生命科学学院,合肥230039 [2]安徽省生态工程与生物技术重点实验室,合肥230039 [3]安徽省农业科学院,合肥230031

出　　处：《热带作物学报》2008年第5期610-617,共8页Chinese Journal of Tropical Crops

基　　金：安徽省自然科学基金项目(编号:050410104);安徽省高校省级自然科学研究重点项目(编号:KJ2007A091);安徽大学分子药理学创新团队项目(编号:02203109)资助

摘　　要：根据植物抗病基因保守序列设计特异引物,对甘薯高抗茎线虫病品种AB94078-1、感病品种徐18及其8个后代品系进行PCR扩增。获得15个具有完整氨基酸读码框的片段,均具有NBS类抗病蛋白4个保守基序,其中5个片段含LRR结构。利用从亲本AB94078-1扩增的R510-AB作为探针进行Southern杂交,结果表明,其在基因组中存在多个拷贝,并估计其在基因组中的拷贝数目,克隆新的甘薯RGA序列,为进一步获得甘薯抗病基因打下基础。PCR primers were designed according to the conserved NBS-LRR domains of the disease resistance genes cloned in plants of sweet potato （Ipomoea batatas） , and the genomic DNA extracted from Sweet potatoes AB94078-1, Xushul8 and their 8 progenies were utilized as templates. Fifteen resistance gene analogs （RGA） were obtained, which contain 4 motifs of the NBS domain, and 5 of the analogs also possess LRR domain. Southern blotting analysis, using R510-AB as probe, showed that there were multicopies in the genomic DNA of AB94078-1. The origin and organization of NBS-LRR based disease resistance gene analogs in the Ipomoea batatas genome were studied. The knowledge about these RGAs can further be used for cloning of full-length disease resistance gene in Ipomoea batata genome.

关 键 词：甘薯 抗病基因同源序列 NBS-LRR SOUTHERN杂交 

分 类 号：Q943.2[生物学—植物学]