甜菜西方黄化病毒两个山东辣椒分离物全基因组扩增及序列分析

 通过RACE和RT-PCR技术相结合的方法,在山东省潍坊市和济宁市辣椒上克隆出2个甜菜西方黄化病毒(beet western yellows virus,BWYV)的全基因组序列,基因组全长均为5 699 nt。与GenBank比对发现,2个分离物的5′UTR(Untranslated region,UTR)变异程度小、相对比较保守,而3′UTR变异程度大、为突变热点区域。潍坊、济宁两地分离物与GenBank中该病毒的全基因组序列相似度平均值分别为89.89%和89.72%。系统进化树分析发现BWYV亚洲地区的分离物聚类在3个不同的组别,本研究克隆的2个山东分离物和3个日本分离物(LC428355、LC428356、LC428357)、2个韩国分离物(LC198684、KM076647)聚在一组,而美国分离物与法国分离物聚在其他分支,表明该病毒的进化存在地理相关性。这是首次对BWYV中国辣椒分离物的全基因组序列克隆,丰富了BWYV的序列信息,有助于进一步了解该病毒种群的遗传进化关系。

Whole genome amplification and sequences analysis of two pepper isolates of beet western yellows virus from Shandong Province,China

The whole genome sequences of two isolates of beet western yellows virus (BWYV) were cloned from peppers in Weifang City and Jining City, Shandong Province by RACE technology and RT-PCR method, both with a total length of 5 699 nt. Comparing with the isolates from other origins deposited in GenBank, it was found that 5′UTR of the two isolates were relatively conservative and did not show much variation, while variation of 3′UTR were large, it was considered a mutation hot-spot. The average similarity of the whole genome sequence between Weifang and Jining isolates and other isolates of BWYV in GenBank was 89.89% and 89.72%, respectively. Phylogenetic tree analysis revealed that BWYV isolates from Asian clustered in three groups, the two Shandong isolates clustered with three Japanese isolates (LC428355, LC428356, LC428357), two Korea isolates (LC198684, KM076647), and US and France isolates clustered in other branches, respectively, indicating that the evolution of the virus was related to geographical distribution. This is the first clone about the whole-genome of BWYV isolates form pepper in China, which enriches the sequence information of BWYV and contributes to further understanding the genetic and evolutionary relationship of the virus.