芜菁花叶病毒山东分离物外壳蛋白基因的克隆及序列分析  

Cloning and Sequence Analysis of Coat Protein Genes of Turnip Mosaic Virus Isolates Obtained from Shandong

作　　者：宋云枝[1] 李玲玲[2] 朱常香[1] 温孚江[3] 温孚凯[2] 

机构地区：[1]山东农业大学生命科学学院,泰安271018 [2]山东农业大学植物保护学院,泰安271018 [3]山东省作物生物学重点实验室,泰安271018

出　　处：《中国农业科学》2005年第3期504-510,共7页Scientia Agricultura Sinica

基　　金：山东省"三 O"工程资助项目

摘　　要：从山东省 3 地市感病的白菜和萝卜上分离到芜菁花叶病毒 6 个分离物,将其分别命名为 TuMV-SD1、TuMV-SD2、TuMV-SD3　、TuMV-SD4、　TuMV-SD5 和 TuMV-SD6。利用 RT-PCR 克隆了这 6 个分离物的外壳蛋白基因,测定了它们的核苷酸序列,并进行了序列分析。结果表明,6 个分离物的 CP 基因均为 867 个碱基,核苷酸序列同源性较高,　达 97.0%～99.4%;它们与 World-B 组分离物的同源性最高,达 91.8%～100%;与 Brassica-　Raphanus　(BR)　致病型分离物的同源性次之,在 89.1%～91.0%之间;与 basal-B 组分离物的同源性最低,仅 86.0%～90.3%。基于 TuMV 的 CP 基因核苷酸序列的分子进化树显示,TuMV 分离物可分为 4 组,本研究所分离到的 6 个 TuMV 山东分离物属于第四组,即 world-B 组。用分离到的 6 个 TuMV 分离物对已获得的转 TuMV　CP 基因的抗病毒大白菜进行攻毒试验。结果表明,尽管作为转基因的 CP 基因与这 6 个分离物的 CP 基因只有 88.2%～88.9%的同源性,但转基因大白菜对这 6 个分离物均具有明显抗性。Six isolates of turnip mosaic virus named TuMV-SD1, TuMV-SD2, TuMV-SD3, TuMV-SD4, TuMV-SD5, and TuMV-SD6 were isolated from infected Chinese cabbages and radishes in three different regions (Taian, Yantai, and Zaozhuang) in Shandong province. A pair of primers was designed and synthesized based on the nucleotide sequences of coat protein (CP) genes of TuMV reported. The CP genes of the six TuMV isolates were cloned by RT-PCR method. The sequences of all the CP genes from the six isolates were sequenced and compared with the CP gene sequences of TuMV reported in GenBank. The results showed that the sequences of the six CP genes were all 867 nucleotides in length, and the isolates shared high sequence homology from 97.0% to 99.4% among them. They also shared high homologies of 91.8% to 100% with the sequences of world-B group isolates, of 89.1% to 91.0% with the sequences of Brassica-Raphanus (BR) pathotype isolates, and of 86.0% to 90.3% with the sequences of basal-B group isolates. TuMV isolates reported so far were devided into four groups based on phylogenetic analysis of the CP gene sequences. All the six isolates analysized in this study belonged to the fourth group, i.e. world-B group. Resistance assay revealed that the transgenic Chinese cabbage containing a CP gene of TuMV was resistant to infections of all the six isolates, although the CP transgene only shared a homology of less than 90%with the CP genes of the six TuMV isolates.

关 键 词：分离物 外壳蛋白基因 同源性 芜菁花叶病毒 大白菜 转基因 核苷酸序列 MV SD CP 

分 类 号：S432.4[农业科学—植物病理学]