摘要
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.
To identify the viruses that induced Capsicum annuum disease, biological inoculation and sequence-independent amplification (SIA)were used. Results of SIA and sequencing showed that there was Broad bean wilt virus 2 (BBWV2) in Capsicum annuum leaves. To further characterize the BBWV2 isolate Capsicum annuum (BBWV2-Ca), the complete sequence of BBWV2-Ca was cloned and determined. BBWV2-Ca RNA1 comprised 5 929 bp nucleotides with one ORF. BBWV2-Ca RNA2 comprised 3 559 nucleotides with one ORF and it encoded 3 kinds of proteins, VP53/VP37, large coat protein and small coat protein. Nucleotide sequence similarity searches showed that BBWV2-Ca RNA1 and RNA2 were from 77.9% to 93.7% and from 80.2% to 93.8 % to that of other BBWV2 strains, respectively. Phylogenetic comparisons based on amino acid sequences showed that BBWV2-Ca RNA1 formed an independent branch with BBWV2-XJ14-3 and BBWV2-RP3. BBWV2-Ca RNA1 was most closely related to BBWV2-XJ14-3 and BBWV2-RP3. BBWV2-Ca RNA2 formed an independent branch with BBWV2-Am. Therefore, BBWV2-Ca RNA2 was most closely related to BBWV2-Am.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2014年第9期927-932,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
公益性行业(农业)科研专项经费(No.201303028)
山西省自然科学基金项目(No.2012011032-5)资助~~
关键词
青椒
蚕豆萎蔫病毒2号
全基因组序列
Capsicum annuum
Broad bean wilt virus 2
complete genome sequences