豇豆病毒病病原的分子鉴定

Molecular Analysis of the Viral Pathogens of Zhejiang Cowpea Disease

为澄清浙江省豇豆病毒病病原及其分类 ,采用马铃薯Y病毒属通用引物Sprimer扩增了浙江豇豆线状病毒基因组 3′ 末端序列。同时又根据已知CMVRNA3序列设计引物 pCMV 4 4 6 3/ 12 0 0 - 1181,扩增了混合侵染的CMV ZJ运动蛋白基因全序列。外壳蛋白氨基酸序列分析表明 ,病样中仅存在一种线状病毒 (CV ZJ) ,该病毒与一泰国豇豆蚜传花叶病毒 (CABMV)具有最高的同源性 (98 6 %) ,与花生条纹病毒 (PStV)、石斛花叶病毒 (DeMV)、赤豆花叶病毒 (AZMV)、黑眼豇豆花叶病毒 (BlCMV)和菜豆普通花叶病毒 (BCMV)分离物间的同源性为 88 5 %～94 4 %,而与其它CABMV分离物的同源性均低于 70 %。进化树分析表明 ,PStV、AZMV、DeMV、BlCMV和BCMV为同种异名 ,应统称为BCMV ,而CABMV为另一种病毒 ;CV ZJ和泰国CABMV分离物应分类为BCMV豇豆株系。CMV ZJ的运动蛋白序列分析表明 ,该分离物属于CMV亚组Ⅰ ,与其它分离物氨基酸序列同源性为 93 1%～97 8%。

There were two viruses detected in the infected cowpea samples collected from Zhejiang Province.One was a member of potyvirus genus,coded as CV-ZJ,and the other one was CMV,coded as CMV-ZJ.A universal PCR primer was designed to amplify the 3'-terminal sequence of the genome(s) of cowpea potyvirus(es) and two primers pCMV44-63/1200-1181 were used to amplify the movement protein(MP)of CMV which coexisted with cowpea potyvirus(es).Sequence analysis on the coat protein peptide showed that CV-ZJ was most closely related to a Thailand isolate CABMV(98.6% identity),and had a high homology(84.5%～94.4%) to PStV,DeMV,AZMV,BlCMV and BCMV isolates,but had less than 70% homology to other isolates of CABMV.Phylogenetic tree of the coat protein sequences was created and suggested that except CABMV,other 4 viruses were synonymous with BCMV.Therefore,CV-ZJ and the Thailand isolate CABMV should be classified as cowpea strains of BCMV.Analysis on the MP sequence of CMV-ZJ suggested a close relationship with isolates of CMV subgroup I(93.1%～97.8% homology).