扬州蚕豆病毒病的鉴定及野豌豆潜隐病毒M扬州分离物基因组克隆及其序列分析

 对田间疑似病毒侵染的蚕豆样品,提取总RNA,分离siRNA,建库并通过高通量测序,其结果经velvet拼接组装, 经Blastn与NCBI比对分析,发现该样品感染4种病毒:野豌豆潜隐病毒M (vicia cryptic virus M,VCV-M)、野豌豆潜隐病毒 (vicia cryptic virus, VCV)、紫云英矮宿病毒 (milk vetch dwarf virus, MDV) 和三叶草黄脉病毒 (clover yellow vein virus, ClYVV)。其中,有14条VCV-M相关的contigs。RT-PCR扩增并拼接获得VCV-M-YZ基因组,其长度为3 434 nt, 其5′-UTR和3′-UTR 分别为142 nt和117 nt,各自形成与该属代表病毒南方番茄病毒 (southern tomato virus, STV) 5′-UTR和3′-UTR相似的高级结构,且A+U含量均较高。 VCV-M-YZ与已报道的VCV-M基因组序列一致性在98%以上。系统进化分析表明,VCV-M-YZ与已报道的VCV-M属于一个种,无明显的地理分布差异。本研究结果丰富了VCV-M群体的基因组遗传信息,为VCV-M的监测及深入研究提供了基础。

The identification of virus diseases of faba bean in Yangzhou and cloning and sequence analysis of the full-length genome of vicia cryptic virus M Yangzhou isolate

The field-grown faba bean samples exhibiting virus-like symptoms were collected and used for RNA extraction. Then, the siRNA was separated and used for siRNA library construction. After sequencing, velvet assembly, and blastn analysis, we found that collected faba bean samples were infected by 4 different viruses, which included VCV-M, vicia cryptic virus (VCV), milk vetch dwarf virus (MDV), and clover yellow vein virus (ClYVV). Among them, there are 14 VCV-M related contigs. As we know, the Amalgaviridae is a recently recognized family of dsRNA viruses that includes four species of plant viruses. RT-PCR amplification generated the 3 434 nt genome of VCV-M-YZ, of which the 5′-UTR and 3′-UTR were 142 nt and 117 nt, respectively. The 5′-UTR and 3′-UTR have high A+U content and form higher secondary structures, as well as the 5′-UTR and 3′-UTR of southern tomato virus, which is the representative virus of Amalgavirus. All this implies the specific characteristic of Amalgavirus replication. The sequences identity of VCV-M-YZ and reported VCV-M were 98%, and phylogenetic analyses showed that VCV-M-YZ and VCV-M belong to the same species, without obvious difference related to geographical distribution. Our studies enrich the genome genetic information of the VCV-M population, and provide basics of further research and supervisory control of VCV-M.