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

以 PRSV 株系特异性引物对 PRSV 的 PRSV126(PRSV 日本分离物)、Ys、Vb 和 Sm 等株系进行 RT-PCR 方法鉴定,引物 PR21/PR22 能把Ys 从 Vb 和 Sm 中鉴定出来,PR300/PR301则能把 Vb 从 Ys、Sm 和 PRSV126中鉴定出来;用限制性内切酶 Hae Ⅱ、Sau3A Ⅰ和 Hinf Ⅰ对 PRSV 的PRSV126、Ys、Vb 和 Sm 等株系进行单酶切 RT-PCR-RFLP 分析,Hinf Ⅰ能把 PRSV126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 PRSV126、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 differentiate Vb from Ys、Sm and PRSV126.The PRSV strains could also be identi- fied by RT-PCR restriction fragment length polymorphism(RT-PCR-RFLP)with one re- striction enzyme of Hae Ⅱ,Sau 3A Ⅰ and Hinf Ⅰ.The results indicated that Ys could be identified with single digestion of Sau 3A 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).