新疆加工番茄上一种类似南方番茄病毒的分子检测  

Molecular Detection of a Virus Similar to Southern Tomato Virus(STV) of Processing Tomato in Xinjiang

作　　者：苏海娣[1] 梁学超[1] 崔百明[1] 郑银英[1] 王建江 向本春[1] 

机构地区：[1]石河子大学生命科学学院,农业生物技术重点实验室,石河子832003 [2]新疆石河子蔬菜花卉研究所,石河子832003

出　　处：《石河子大学学报（自然科学版）》2013年第3期271-275,共5页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(31260420)

摘　　要：在对加工番茄里格87-5进行RNA测序时,我们发现存在南方番茄病毒的基因组序列,序列包含STV除5′末端12bp和3′末端35bp的其它全部3390bp的RNA序列,其中只有5个碱基存在差异(与EF442780相比),说明里格87-5感染了STV。为了快速检测加工番茄品种的病毒感染情况,我们设计了STV特异性的PCR引物STV168F/STV745R,用一步法RT-PCR方法,从里格87-5的总RNA中扩增得到了预期的577bp的扩增产物,回收、克隆到pGEM-T上,提取质粒经EcoRI酶切鉴定后进行序列测定,与已报道的STV的序列同源性为99%～100%,表明该片段是STV特异性的。运用这项技术调查了20份加工番茄材料,结果显示,有3份材料的全部植株均带毒,13份材料均不带毒,另外4份材料部分带毒。同时,在新疆石河子蔬菜花卉研究所随机采集了加工番茄叶片样品,统计大田检出率约为28%。We found the presence of the genomic sequence of southern tomato virus （STV） when sequencing processing tomato Lige 87-5. The sequence contains all STV 3390bp RNA sequence except 5′ end of 12bp and a 3′ end 35bp other all 3390bp. Compared with EF442780,the sequence displays differences in only five bases,which means Lige 87-5 has infected STV. A specific primer STV168F/STV745R was designed and one-step RT-PCR detection system was used for rapid detection of viral infection of the processing tomato varieties, specific of 577bp fragment amplified by PCR cloned into pGEM-T plasmid, and the plasmid extraction were sequenced after EcoRI restriction enzyme digestion, with 99%-100% homology compared with the reported STV sequence, indicating that the fragment is STV specificity. 3 out of 20 processing tomato material surveyed by this technology are total infected, 13 non-infected and 4 partial infected. Processing tomato infected with STV was preliminary confirmed, 28% in filed samples.

关 键 词：新疆 加工番茄 南方番茄病毒 一步法RT-PCR检测 

分 类 号：S641.2[农业科学—蔬菜学] S436.412[农业科学—园艺学]