番木瓜环斑病毒株系的分子生物学方法鉴定  

IDENTIFICATION OF Papaya ringspot potyvirus STRAINS WITH MOLECULAR BIOLOGICAL METHODS

作　　者：贺国安[1] 肖火根[1] 张曙光[1] 李华平[1] 范怀忠[1] 

机构地区：[1]华南农业大学植物病毒研究室,广州510642

出　　处：《植物病理学报》2002年第1期21-25,共5页Acta Phytopathologica Sinica

基　　金：广东省重点科技攻关项目 (99M0 4 2 0 2 G);国家自然科学基金项目 (39870 4 34)

摘　　要：以 PRSV株系特异性引物对 PRSV的 PRSV12 6 (PRSV日本分离物 )、Ys、Vb和 Sm等株系进行 RT- PCR方法鉴定 ,引物 PR2 1/ PR2 2能把 Ys从 Vb和 Sm中鉴定出来 ,PR30 0 / PR30 1则能把 Vb从 Ys、Sm和 PRSV12 6中鉴定出来 ;用限制性内切酶 Hae II、Sau3A I和 Hinf I对 PRSV的PRSV12 6、Ys、Vb和 Sm等株系进行单酶切 RT- PCR- RFLP分析 ,Hinf I能把 PRSV12 6与 Ys、Vb和Sm鉴别开来 ,Sau3A I能把 Ys与 Vb和 Sm鉴别开来 ,Hae II则能把 Ys与 PRSV12 6、Vb和 Sm鉴别开来 ;以 P1/ P2为引物 ,对 Vb、Ys和 Sm株系进行 RT- PCR- RFLP- SSCP分析 ,结果能一次把三者较好地区别开来。The strains of Papaya ringspot potyvirus (PRSV) such as PRSV126?Ys?Vb?Sm were identified with strain specific primers of PRSV. It was found that the primer PR21/PR22 could distinguish Ys from Vb and Sm, while another pair of primer PR300/PR301 could dif ferentiate Vb from Ys,Sm and PRSV126. The PRSV strains could also be identified by RT PCR restriction fragment length polymorphism(RT PCR RFLP) with one restriction enzyme of Hae II,Sau3A I and Hinf I. The results indicated that Ys could be identified with single digestion of Sau3A I or of Hae II, and PRSV126 with Hinf I. The strains of Vb,Ys and Sm could also be identified respectively by RT PCR RFLP single strand conformation polymorphism(RT PCR RFLP SSCP).

关 键 词：分子生物学方法 番木瓜环斑病毒 株系鉴定 RT-PCR RT-PCR-RFLP RT-PCR-RFLP-SSCP 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治] S432.41[农业科学—植物保护]