3种粒线虫多重PCR检测方法  

Multiple-PCR detection on three species of Anguina genus

作　　者：马以桂[1] 王金成[2] 谢辉[1] 周春娜[1] 杜宇[3] 黄国明[2] 李芳荣[4] 

机构地区：[1]华南农业大学,广州510642 [2]天津出入境检验检疫局,天津300456 [3]云南出入境检验检疫局,昆明650228 [4]深圳出入境检验检疫局,深圳518010

出　　处：《植物病理学报》2006年第6期508-511,共4页Acta Phytopathologica Sinica

基　　金：国家食品安全关键技术专项课题(2001BA804A22)

摘　　要：剪股颖粒线虫[Anguina ag rostis(S teinbuch,1799)F ilipjev,1936]、小麦粒线虫[Anguina tritici(S teinbuch,1799)F ilipjev,1936]和维氏粒线虫[Anguina w evelli(Van den Berg,1985)S idd iq i,2000]都是重要的植物病原线虫,它们在成熟种子和虫瘿中的虫态通常是幼虫,而这3种线虫的幼虫形态非常相似,难以根据其特征对它们进行快速准确的种类鉴定。本研究根据这3种线虫的rDNA-ITS区域序列,分别设计筛选了特异性引物AgrF1/AgrR1、TriF1/TriR1、W evF1/W evR1,构建了这3种线虫单条幼虫多重PCR检测体系,获得3个大小差异明显的片段,表明这些引物设计合理,适合这3种线虫的快速准确检测。Anguina agrostis, Anguina tritici and Anguina wevelli are important pests of agriculture. However, only juveniles are generally found in seed galls and similar in morphology. Three pairs of primers, named as AgrF1/AgrR1, TriF1/TriR1 and WevF1/WevR1, were designed based on rDNA-ITS nucleotide sequences of A. agrostis, A. tritici and A. wevelli. PCR techniques were developed and improved for detection of the three species with DNA extrated from single juvenile as templates. The amplified products for A. agrostis, A. tritici and A. wevelli were 499,617 and 377 bp in size and showed a strong unique band in electrophoresis respectively. This study indicated that the multiple-PCR technology could satisfy the need of detection of A. agrostis. A. tritici and A. wevelli.

关 键 词：剪股颖粒线虫 小麦粒线虫 维氏粒线虫 DNA 多重PCR 

分 类 号：S544.2[农业科学—作物学]