一种新发现的侵染绿豆的菜豆普通花叶病毒分子鉴定

摘要

This study was performed to identify one isolate with wizened ,mottle and vein clearing mosaic symp-tom collected from greenhouse mung bean(Vigna radiata (L.) Wilczek)in Jangsu.We used electron micrograph, PCR,determination of nucleotide sequence and ELISA .We can see the characteristics of the virus under transmis-sion electron microscopy .We used Potyvirus universal primers for RT-PCR,found the symptom was caused by Bean common mosaic virus(BCMV).This isolate was called BCWV-JAAS.The nucleotides sequences of the genes from the isolate shared homologies from 95% with reported BCMV-HB.Total RNA was amplified by RT-PCR with the specific primers designed on the basis of the reported coat protein gene of Bean common mosaic virus( BCMV) .An 1 082 bp gene fragment was amplified from total RNA of isolate ,and subsequent sequence analysis indicated that the 862 bp fragment contained complete cp gene which encoded 228 amino acids.By amplification cp genes and construct phylogenetic trees get BCMV-JAAS with other Chinese isolates of BCMV have a close genetic relationship . We performed the virus samples by ELISA ,the results shown positive we ensured that was infecting by Bean com-mon mosaic virus(BCMV) to cause the mung bean plant disease .This is the first time found the mung bean was in-fected by BCMV in China .%为确定从江苏省农科院蔬菜所温室大棚所采集到的表现花叶、皱缩、明脉症状的绿豆植株是由何种病毒引起的，采取了透射电子显微镜观察、RT-PCR、序列测序以及酶联免疫吸附法（ ELISA）等方法进行鉴定。结果表明，在透射电镜下可观察到风轮状、束状病毒内含体，与Potyvirus属病毒粒子形态一致；对所提取的病样总RNA使用Poty-virus通用引物进行RT-PCR扩增，得到由1082个核苷酸组成的序列，包含编码228个氨基酸的完整cp基因，产物通过NCBI比对，发现和已报道的菜豆普通花叶病毒（ Bean common mosaic virus，BCMV） HB分离物同源性高达95％，初步判定该病原为菜豆普通花叶病毒（ BCMV），命名为BCMV-JAAS分离物；通过扩增得到的cp基因，构建系统发育树，表明BCMV-JAAS与BCMV的其他中国分离物亲缘关系较近；通过ELISA检测，进一步确定是由菜豆普通花叶病毒（ BCMV）感染引起的绿豆病毒病，这是我国第一次在绿豆上发现菜豆普通花叶病毒。