进境大豆种子中烟草环斑病毒的快速检测  

RAPID DETECTION OF TOBACCO RINGSPOT VIRUS IN IMPORTED SOYBEAN SEEDS

作　　者：闻伟刚[1] 崔俊霞[1] 盛蕾[1] 

机构地区：[1]宁波出入境检验检疫局,宁波315012

出　　处：《大豆科学》2007年第5期748-751,共4页Soybean Science

基　　金：宁波出入境检验检疫局科技项目(甬K34-2006)

摘　　要：烟草环斑病毒(Tobacco ringspot virus,TRSV)是进境大豆种子必须检疫的项目,属于我国禁止入境的二类检疫性有害生物,本文建立了大豆种子中TRSV的快速检测方法。采用Trizol试剂直接提取大豆种子总RNA,通过RT-PCR扩增得到359 bp特异性产物。该产物经半巢式PCR进一步验证得到206 bp特异性片段,经测序及同源性比较表明,片段序列与GenBank中登录的TRSV外壳蛋白基因部分序列存在92%～96%的同源性,而未见与其它基因序列存在同源性。同时,大豆种子样品经DAS-ELISA复核检测结果显示TRSV阳性反应。因此,判定该批进境大豆种子携带TRSV。Tobacco tested in imported ringspot virus（TRSV） is the second level quarantine pest which should be soybean. In this paper, a rapid detection method was established which used Trizol for extracting total RNA of soybean seeds directly and adopted RT-PCR for amplification, a 359 bp special band was obtained. Through further verification of semi-nested PCR and sequence homologous comparation,a 206 bp special product was obtained and had 920% ～96% identities with TRSV sequence of coat protein genes published in GeneBank. The homology to other genetic sequences of plant viruses were not found. Meanwhile, the soybean seeds sample showed TRSV positive reaction when checked by DAS-ELISA. Thus,the soybean seeds sample was determined to contain TRSV.

关 键 词：大豆 烟草环斑病毒 快速检测 RT—PCR 半巢式PCR 

分 类 号：S41-31[农业科学—植物保护]