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为了解马铃薯Y病毒在贵州烟田的发生和流行规律,对2013—2015年采集到的贵州省多个烟区呈现典型茎脉坏死症状的疑似受马铃薯Y病毒(Potato virus Y,PVY)侵染的烟草样品进行单斑分离和病毒鉴定,并以现有PVY序列信息为参考,设计了4对PVY特异引物,经过RT-PCR扩增、T载体连接、序列测定以及序列拼接,并利用MEGA软件进行系统进化分析。结果发现PVY福泉分离物(PVY Fuquan isolate,PVY-FQ)和PVY大方分离物(PVY Dafang isolate,PVY-DF)为优势株系并获得了PVY-FQ和PVY-DF的全基因组序列。PVY FQ整条基因组由9 699个碱基组成,编码多聚蛋白的开放阅读框位于第188~9 373nt;PVY DF整条基因组由9 706个碱基组成,编码多聚蛋白的开放阅读框位于第190~9 375nt。2条序列的基本特征与已报道PVY基因组一致。对PVY-FQ和PVY-DF全序列与已报道PVY序列进行系统进化分析发现,PVY-FQ与PVY NTN株系具有较高的亲缘关系,而PVY-DF与PVYN具有较高亲缘关系。
In order to understand the occurrence and prevalence of potato virus Y in tobacco fields in Guizhou Province, it is suspected to be suspected of being infected by Potato virus Y (PVY) in a number of tobacco-covered areas in Guizhou Province collected from 2013 to 2015 Tobacco samples were single spotted and virus identified. Based on the existing PVY sequence information, four pairs of PVY specific primers were designed, amplified by RT-PCR, T vector connection, sequence determination and sequence splicing. The MEGA software Perform systematic evolutionary analysis. The results showed that PVY Fuquan isolate (PVY-FQ) and PVY Dafang isolate (PVY-DF) were dominant strains and the whole genome sequences of PVY-FQ and PVY-DF were obtained. The entire genome of PVY FQ consists of 9 699 bases. The open reading frame encoding the polyprotein is located between the 188th and 9373nt. The entire PVY DF genome consists of 9 706 bases. The open reading frame encoding the polyprotein is located at The first 190 ~ 9 375nt. The basic characteristics of the two sequences are consistent with the reported PVY genome. Phylogenetic analysis of PVY-FQ and PVY-DF sequences and reported PVY sequences showed that PVY-FQ had a higher genetic relationship with PVY NTN lines, while PVY-DF had a higher genetic relationship with PVYN lines.