TC-RT-PCR检测菜豆荚斑驳病毒的研究  

Detection of Bean pod mottle virus by TC-RT-PCR

作　　者：沈建国[1,2] 高芳銮[2] 廖富荣[3] 王念武[1] 郭琼霞[1] 吴祖建[2] 

机构地区：[1]福建出入境检验检疫局技术中心福建省检验检疫技术研究重点实验室,福建福州350001 [2]福建农林大学植物病毒研究所,福建福州350002 [3]厦门出入境检验检疫局,福建厦门361012

出　　处：《激光生物学报》2009年第1期108-111,共4页Acta Laser Biology Sinica

基　　金：福建省自然科学基金计划资助项目(B0610001;2006J0047);福建科技重大专项专题项目(2006NZ0002-1;2006NZ0002-2);福建出入境检验检疫局科技项目(FK2007-25)

摘　　要：根据已报道的菜豆荚斑驳病毒(Bean pod mottle virus,BPMV)外壳蛋白(Coat protein,CP)基因序列设计特异性引物,应用试管捕捉RT-PCR(Tube capture RT-PCR,TC-RT-PCR)技术对大豆种皮上的BPMV进行检测。结果表明,TC-RT-PCR方法能从携带BPMV的大豆种皮上扩增到预期大小的基因片段。将TC-RT-PCR扩增产物克隆测序后进行序列分析,结果显示扩增到的片段序列与BPMV的CP基因序列具有高度同源性,进一步证实了该方法的准确性。应用TC-RT-PCR方法,成功检测了一批进境大豆样品。本文建立的TC-RT-PCR方法,为大豆种子上BPMV的检测和诊断提供了一种新的参考方法。A pair of specific primers were designed and synthesized based on the conserved nucleotide sequence of coat protein （CP） gene of Bean pod mottle virus （BPMV）. Tube capture RT-PCR （TC-RT-PCR） was established to detect BPMV in soybean seed coat. The results showed that the target fragment could be amplified from the BPMV-infected soybean seed coats. Cloning and sequencing analysis indicated that nucleotide sequence of the target fragment was highly identical to the reported BPMV genomic sequence, which verified the accuracy of the method. The presence of BPMV in imported soybean from America was successfully detected using TC-RT-PCR method. It could be concluded that TC-RTPCR method provided a new reference for detection and diagnosis of BPMV in soybean seed.

关 键 词：菜豆荚斑驳病毒 检测 TC—RT—PCR 

分 类 号：S431.12[农业科学—农业昆虫与害虫防治]