转菰候选基因克隆获得抗白叶枯病水稻植株  

Transgenic Rice Plants Harboring Genomic DNA from Zizania latifolia Confer Bacterial Blight Resistance

作　　者：沈玮玮[1,2] 宋成丽[1] 陈洁[3] 付亚萍[3] 吴建利[3] 江绍玫[2] 

机构地区：[1]江西师范大学生命科学学院,江西南昌330022 [2]江西财经大学,江西南昌330013 [3]中国水稻研究所,浙江杭州310006

出　　处：《中国水稻科学》2010年第5期447-452,共6页Chinese Journal of Rice Science

基　　金：国家自然科学基金资助项目(30760115);国家转基因重大专项资助项目(2008ZX08001-002)

摘　　要：以菰NBS-LRR类抗病基因同源序列FZ14(GenBank登录号:DQ239432)为模板设计特异性引物FZ14P1/P2,通过克隆池PCR法经分级筛选,从菰基因组TAC文库中获得1个阳性克隆(ZR1),序列分析比对证实该阳性克隆为含有菰抗病基因同源序列FZ14的抗病基因候选克隆。同时,ZR1具有植物NBS-LRR类型抗性基因中的P-loop(kinase1a)、kinase2、ki-nase3a和GLPL(Gly-Leu-Pro-Leu)等保守基序,可能为抗性基因的部分序列。通过农杆菌介导转化水稻品种日本晴,获得36个对白叶枯病菌PXO71具有明显抗性的独立转化子,结果表明,菰ZR1克隆中至少含有1个白叶枯病抗性基因。Based on the sequence of resistance gene analog FZ14 derived from Zizania latifolia（Griseb.）,a pair of specific PCR primers FZ14P1/FZ14P2 was designed to isolate candidate disease resistance gene. The pooled-PCR approach was adapted using the primer pairs to screen a genomic transformation-competent artificial chromosome（TAC） library derived from Zizania latifolia. A positive TAC clone （ZR1） was obtained and confirmed by sequence analysis. The results indicated that ZR1 consisted of conserved motifs similar to P-loop （kinase 1a）,kinase 2,kinase 3a and GLPL（Gly-Leu-Pro-Leu）,indicating that it could be a portion of NBS-LRR type of resistance gene. Using Agrobacterium tumafaciens-mediated transformation of Nipponbare mature embryo,a total of 48 independent transgenic T0 plants were obtained. Among them,36 plants were highly resistant to the virulent bacterial strain PXO71. The results indicated that ZR1 contains at least one bacterial blight resistance gene.

关 键 词：菰 可转化人工染色体文库 抗病基因同源序列 水稻 白叶枯病抗性 转基因 

分 类 号：S511[农业科学—作物学]