西瓜花叶病毒辽宁臭椿分离物的鉴定与全序列分析

Detection and Complete Viral Sequence Analysis of Watermelon mosaic virus on Ailanthus altissima in Liaoning

2015年10月,在辽宁省盖州市采集到表现花叶的臭椿样品。应用电子显微镜负染色技术检测到线状病毒粒子(800 nm×18 nm)。利用Potyvirus通用引物PVY-Legpoty-F/PVY-Legpoty-R对三株代表样品进行RTPCR扩增,得到750 bp阳性片段。经Blast搜索,与西瓜花叶病毒(Watermelon mosaic virus,WMV)高度同源。利用WMV-5NF/WMV-5NR、WMV-MNF/WMV-MNR和WMV-3NF/WMV-3NR三对引物扩增WMV全基因组,确定为10070核苷酸(MF418043)。基于cp基因进行同源性分析和系统进化关系分析,发现盖州臭椿分离物与北京臭椿分离物的关系最近。这是辽宁臭椿上WMV分子特征的首次报道。

Samples from the Ailanthus altissima showing yellow mosaic symptoms were collected from Gaizhou,Liaoning Province in October,2015.Virions measuring about 800 nm×18 nm were observed by transmission electron microscope（TEM）.The universal degenerate primer PVY-Legpoty-F/PVY-Legpoty-R was used for the detection of Potyviruses from three symptomatic leaf samples by RT-PCR.The expected fragments about 750 bp in size were amplified.By blast,it shared high nucleotide sequence identity with that of the published Watermelon mosaic virus（WMV）.Then,WMV-5 NF/WMV-5 NR,WMV-MNF/WMV-MNR and WMV-3 NF/WMV-3 NR primers were used to amplify the whole viral genome.It was determined to be 10070 nucleotide（MF418043）.Based on the cp,it showed most closed to another WMV isolate from Ailanthus altissima.This is the first report of the natural occurrence of WMV on Ailanthus altissima in Liaoning.