蚕豆萎蔫病毒2号安徽分离物全基因组序列测定与分析

摘要

利用酶联免疫和RT-PCR技术对采自安徽地区的蚕豆病株进行检测,确定其病原为蚕豆萎蔫病毒2号Broad bean wilt virus 2(BBWV2).为明确BBWV2安徽分离物(BBWV2-AH)的分类地位,克隆了该分离物的全基因组序列,分析了其基因组特征.结果表明,BBWV2-AH RNA1全长为5 944 bp(GenBank登录号:KY606992),含有1个ORF;BBWV2-AH RNA2全长3 587 bp (GenBank登录号:KY606993),含有1个ORF.全序列核苷酸和氨基酸相似性分析显示,BBWV2-AH RNA1与BBWV2其他分离物的核苷酸、氨基酸相似性分别为78.4％～96％和87.1％～99％;BBWV2-AH RNA2与BBWV2其他分离物的核苷酸、氨基酸相似性分别为76.8％～95.5％和88.2％～98.3％.全基因组核苷酸序列系统发育分析显示,BBWV2-AH RNA1与中国的BBWV2-Hu-nan RNA1的亲缘关系最近,而BBWV2-AH RNA2与韩国的多个分离物聚集在一起,再与中国的分离物BBWV2-B935形成一个分支.%BBWV2 was detected from broad bean plants in Anhui Province by DAS-ELISA and RT-PCR.To further characterize the BBWV2 Anhui isolate (BBWV2-AH),complete nucleotide sequence of BBWV2-AH was determined.Sequence analysis suggested that BBWV2-AH RNA1 was comprised of 5 944 nucleotides,encoding one ORF (GenBank accession no.KY606992);BBWV2-AH RNA2 was comprised of 3 587 nucleotides,also encoding one ORF (GenBank accession no.KY606993).The comparison of nucleotide sequences suggested that RNA1 and RNA2 of BBWV2-AH shared 78.4％-96％ and 76.8％-95.5％ nucleotide sequence identity with those of other BBWV2 isolates,respectively.Furthermore,the comparison of amino acid sequences suggested that RNA1 and RNA2 of BBWV2-AH shared 87.1％-99％ and 88.2％-98.3％ amino acid sequence identity with those of other BBWV2 isolates,respectively.Phylogenetic analysis based on nucleotide sequences showed that RNA1 of BBWV2-AH isolate was most closely related to BBWV2-Hunan isolate,while RNA2 of BBWV2-AH formed an independent branch with several Korean isolates,and was then closely related to BBWV2-B935 from China.