新疆加工番茄上番茄花叶病毒的分子鉴定  

Identification on Molecular of Tomato mosaic virus of Processing Tomato in Xinjiang

作　　者：姜玉霞[1] 向本春[1] 安仙丽[1] 黄家风[1] 汪铖华 

机构地区：[1]石河子大学农学院,新疆石河子832000 [2]新疆康正农业科技开发有限责任公司,乌鲁木齐830000

出　　处：《新疆农业科学》2008年第3期484-489,共6页Xinjiang Agricultural Sciences

基　　金：国家教育部“春晖计划”项目(Z2005-2-83004);石河子大学高层次人才科研启动资金专项(RCZX200516)

摘　　要：从表现花叶、畸形的加工番茄病株上获得分离物xj124，用1对ToMVCP基因引物进行RT-PCR，扩增得到了689bp的特异性核苷酸片段。将PCR产物插入到克隆载体pMD18-T中再转化到大肠杆菌TOP10。经限制酶酶切鉴定，重组克隆中含有与PCR产物大小相同的689bp的插入片段。cDNA全序列分析表明，所扩增出的689bpToMVCP基因，含1个480bp开放阅读框架（ORV），编码160氨基酸组成的蛋白，所克隆的基因包含完整的ToMVCP基因。所测得的xj124分离物与国内外已报道的部分ToMV CP基因序列比较。核苷酸同源性高达84．4％-99．8％，氨基酸同源性高达98．7％-100％。因此将该分离物鉴定为番茄病毒病花叶病毒。Xj124 isolate was obtained from naturally infected processing tomatoes with symptoms of mosaic and malformation in Xinjiang. A pair of primers was designed based on ToMV CP gene, and a fragment of about 689 bp was amplified by lit - PCR. The products of PCR were cloned into PMD18 - T and transformed into TOP10. The combined clone was identified by restriction enzyme digesting and the nucleotide sequencing. The results showed that the cloned segment was 689 bp and contained one open reading frames （ORF） that was composed of 480 bp nucleotides encoding a coat protein gene of 160 amino acids. Comparison of the nucleotide and amino acid sequences of the full length of CP gene of ToMV xjl24 with the corresponding sequences of the part of the ToMV isolates reported showed that the homology is 83.1% - 99.1% at nucleotide level , and 99.4% - 100% at amino acid level, respectively. From the above results, the xjl24 isolate was identified as ToMV.

关 键 词：番茄花叶病毒 分子鉴定 克隆外壳蛋白基因 序列分析 

分 类 号：S436.412.19[农业科学—农业昆虫与害虫防治]