侵染芹菜的甜椒内源RNA病毒鉴定及序列分析

摘要

本试验利用双链RNA (double-stranded RNA,dsRNA)提取技术和非序列依赖PCR扩增(sequence-independent amplification,SIA)方法对表现皱缩症状的芹菜进行分子鉴定,发现芹菜被甜椒内源RNA病毒Bell pepper alphaendornavirus (BPEV)所侵染.为进一步明确山西芹菜BPEV分离物(BPEV-QC,GenBank登录号为KY659226)的分类地位,根据SIA测序结果设计BPEV特异性引物进行RT-PCR检测,并进行序列相似性比较和系统进化分析.序列同源性分析表明:BPEV-QC与BPEV分离物(IS、Maor、Kyosuzu、BPEV、lj、BPEV-YW)的同源性为80.0％～97.8％,与其他内源RNA病毒科分离物的同源性仅为30.6％～37.6％,表明:BPEV-QC与BPEV分离物(IS、Maor、Kyosuzu、BPEV、lj)形成一个独立分支,亲缘关系最近.%Viral pathogen that caused Apium graveolens wrinkled disease,was identified by double-stranded RNA (dsRNA) extraction and sequence-independent amplification (SIA).The results of SIA and sequencing showed that A.graveolens was infected by Bell pepper alphaendornavirus (BPEV).To further characterize the BPEV strain from A.graveolens (BPEV-QC),specific primer pairs were designed for RT-PCR according to the results of SIA.Sequence alignments showed that BPEV-QC shared high homology (80.0％-97.8％) with BPEV isolates (IS,Maor,Kyosuzu,BPEV,lj,BPEV-YW),but shared low homology (30.6％-37.6％) with other Endornaviridae isolates,Phylogenetic analysis based on nucleotide sequences of BPEV-QC and other Endornaviridae isolates showed that BPEV-QC was most closely related to Endornaviridae isolates (IS,Maor,Kyosuzu,BPEV,lj),and these isolates formed an independent branch.