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本试验在生物学接种的基础上,利用非序列依赖性PCR扩增(sequence-independent amplification,SIA)对感病青椒(Capsicum annuum)进行了分子鉴定.序列测定及分析发现,侵染青椒的病毒为蚕豆萎蔫病毒2号(Broad bean wilt virus 2,BBWV2).为明确BBWV2青椒分离物(BBWV2-Ca)的分类地位,对BBWV2-Ca的全基因组序列进行了分段克隆、序列测定和分析.BBWV2-Ca RNA1全长5 929 bp,含有1个ORF.BBWV2-Ca RNA2全长3 559 bp,含有1个ORF,编码3种蛋白:VP53/VP37、外壳蛋白大亚基(large coat protein,LCP)和外壳蛋白小亚基(small coat protein,SCP).全序列核苷酸同源性分析显示,BBWV2-Ca RNA1与BBWV2其它分离物核苷酸同源性在77.9%~93.7%之间;RNA2与BBWV2其它分离物核苷酸同源性在80.2%~93.8%之间.全序列系统进化分析显示,BBWV2-Ca RNA1与BBWV2-XJ14-3以及BBWV2-RP3株系聚为一簇,亲缘关系最近;RNA2与BBWV2-Am形成一个独立分支,亲缘关系最近.
In this study, based on the biological inoculation, the molecular identification of the susceptible green pepper (Capsicum annuum) was carried out by sequence-independent amplification (SIA) .Sequencing and analysis showed that the virus Was Broad bean wilt virus 2 (BBWV2) .To clarify the taxonomic status of BBWV2-Ca (BBWV2-Ca), the whole genome sequence of BBWV2-Ca was cloned, sequenced and analyzed. The full length of BBWV2-Ca RNA1 is 5 929 bp, containing 1 ORF. The full length of BBWV2-Ca RNA2 is 3 559 bp, containing 1 ORF and encodes three proteins VP53 / VP37, large coat protein (LCP) ) And small coat protein (SCP). Nucleotide homology analysis of full-length nucleotide sequences showed that nucleotide homology between BBWV2-Ca RNA1 and other BBWV2 isolates ranged from 77.9% to 93.7% Nucleotide homology between RNA2 and other BBWV2 isolates ranged from 80.2% to 93.8% .A total sequence phylogenetic analysis showed that BBWV2-Ca RNA1 and BBWV2-XJ14-3 and BBWV2-RP3 clustered into clusters, The recent relationship; RNA2 and BBWV2-Am form an independent branch, the closest relationship.