PCR技术在印度腥黑穗病菌检测和鉴定中的应用  

APPLICATION OF POLYMERASE CHAIN REACTION (PCR) METHOD TO DETECTION AND IDENTIFICATION OFTilletia indica Mitra

作　　者：易建平[1] 沈禹飞[1] 陶庭典[1] 戚龙君[1] 印丽萍[1] 郑建中[1] 

机构地区：[1]上海出入境检验检疫局,上海200135

出　　处：《上海农业学报》2002年第2期57-61,共5页Acta Agriculturae Shanghai

基　　金：国家检验检疫局课题"小麦印度腥黑穗病菌检测鉴定技术"的一部分 (编号 :K0 39- 1999)

摘　　要：利用 2对小麦印度腥黑穗病菌 (Tilletiaindica Mitra)特异性引物 (Tin3/Tin4 ,F3/R1)和 2对黑麦草腥黑穗病菌T .walkeri特异性引物 (Tin7/Tin8,Tin11/Tin4 )建立快速鉴定T .indica及其近似种T .walkeri的PCR程序。引物Tin3/Tin4 ,F3/R1可特异性扩增T .indica分别得到 4 15bp和 4 98bp产物 ,扩增 T .walkeri无产物 ;引物Tin7/Tin8,Tin11/Tin4可特异性扩增T .walkeri 得到 391bp和 4 15bp产物 ,扩增 T .indica 无产物。 4对引物扩增 T .horrida、T .controversa和T .laevis无产物。产物片段的测序验证了PCR反应的可靠性。为进口粮检疫中T .indica的鉴定提供了快速、简便。A PCR program for quick identification of Tille tia indica and T.walkeri was set up by use of two pairs of T.indica s pecific primers (Tin3/Tin4 and F3/R1) and two pairs of  T.walkeri  speci fic primers (Tin7/Tin8 and Tin11/Tin4). Primers Tin3/Tin4 and F3/R1 could specif ically amplify T.indica and 415bp and 498bp products were gained, respective ly, but they amplified T.walkeri and no PCR products were gained. Primers Ti n7/Tin8 and Tin11/Tin4 could specifically amplify T.walkeri and 391bp and 41 5bp products were obtained, respectively, but they amplified T.indica and no PCR products were obtained. The four pairs of primers amplified T.horrida, T. controversa and T.laevis and no products were gained. The specificity and reliability of the PCR were confirmed by sequencing the PCR products fragment. T his provided a rapid, simple and practical PCR diagnostic method for identifyin g T.indica in imported grain inspection.

关 键 词：PCR技术 印度腥黑穗病菌 检测 鉴定 小麦 黑麦草 

分 类 号：S432.44[农业科学—植物病理学] S435.121.4[农业科学—农业昆虫与害虫防治]