甜菜黑色焦枯病毒新疆分离物基因组RNA的序列分析及侵染性cDNA克隆构建  

Sequence Analysis and Construction of Infectious cDNA Clone of Beet Black Scorch Virus Xinjiang Isolate

作　　者：席德慧[1] 曹云鹤[1] 郭立华[1] 原雪峰[1] 蔡祝南[1] 韩成贵[1] 李大伟[1] 于嘉林[1] 

机构地区：[1]中国农业大学农业生物技术国家重点实验室,北京100094

出　　处：《病毒学报》2006年第4期309-313,共5页Chinese Journal of Virology

基　　金：国家自然科学基金(30325001和30270063)资助项目

摘　　要：甜菜黑色焦枯病毒(Beet black scorch virus,BBSV)新疆分离物(BBSV-X)和宁夏分离物(BBSV-N)分别来自生态环境不同的新疆和宁夏的甜菜产区,自然条件下,BBSV-X含有卫星RNA,BBSV-N不含有卫星RNA。为明确二者的分子差异,以BBSV-X基因组RNA为模板,通过RT-PCR扩增获得了全长cDNA克隆。序列分析显示,BBSV-X基因组RNA全长为3 644个核苷酸,与以往报道的BBSV-N相同,且核苷酸序列一致性高达99.45%。除5′和3′非翻译区各有一个核苷酸发生变异外,核苷酸序列差异主要存在于ORF1通读区和ORF6,氨基酸序列差异仅存在于ORF1的通读区。构建获得了BBSV-X的侵染性cDNA克隆,用体外转录物定量接种了苋色藜(Chenopodiumamaranticolor)和豇豆(Vigna sinensis),结果显示BBSV-X与BBSV-N之间在致病性上存在一定的差异。Two isolates of Beet black scorch virus, BBSV-X and BBSV-N, were isolated from Xinjiang and Ningxia respectively, of which BBSV-X contains a satellite RNA component. To compare their sequence differentiation, full-length cDNA clone of BBSV-X was obtained by RT-PCR. Sequence analysis showed that the genome RNA of BBSV-X consists of 3644 nucleotides in length, same as that of BBSV-N reported previously. It shares 99.45 % identity in terms of nucleotide sequence with BBSV-N. Nucleotide differences mainly appeared in ORF1-RT and ORF6, while the variation of amino acids only in ORF1-RT. By inoculation with in vitro transcripts of the cDNAs, the results revealed different pathogenicity between BBSV-X and BBSV-N on Chenopodiurn arnaranticolor and Vigna sinensis.

关 键 词：甜菜黑色焦枯病毒 新疆分离物 侵染性cDNA克隆 致病性 

分 类 号：S432.4[农业科学—植物病理学]