杜鹃芽枯病菌PCR及实时荧光PCR检测  

Detection of Seifertia azaleae by PCR and real-time PCR

作　　者：龚静如 段维军 龚婕 王寅鹏 崔金璐 易建平 Gong Jingru;Duan Weijun;Gong Jie;Wang Yinpeng;Cui Jinlu;Yi Jianping(The Technical Center for Animal&Plant&Food Inspection and Quarantine of Shanghai Customs District,Shanghai 200135,China;Ningbo Customs District)

机构地区：[1]上海海关动植物与食品检验检疫技术中心,上海200135 [2]宁波海关

出　　处：《植物检疫》2024年第4期49-54,共6页Plant Quarantine

摘　　要：根据GenBank中杜鹃芽枯病菌(Seifertia azaleae)及其相关种ITS序列设计特异引物SA1/SA2和探针SA-P,建立杜鹃芽枯病菌常规PCR和实时荧光PCR检测方法。引物SA1/SA2能扩增5株S. azaleae菌株DNA,得到351 bp的预期扩增产物,其他供试菌株均不能扩增出预期产物,检测灵敏度达60 pg菌体DNA。探针SA-P对5株S. azaleae均表现为阳性扩增,其他供试菌株均为阴性扩增,检测灵敏度可达60 fg菌体DNA,比常规PCR高1000倍。对杜鹃花苞样品和叶片样品的检测结果表明常规PCR和实时荧光PCR检测方法均可用于进境杜鹃种苗中杜鹃芽枯病菌的快速检测。The primer SA1/SA2 and probe SA-P based on the ITS sequence of S. azalea and related species in GenBank was developed. A PCR and real-time PCR method for the detection of Seifertia azaleae were established. The continental PCR with primer SA1/SA2 and the real time PCR with probe SA-P could got the positive result for 5 strains of S. azaleae,but no positive amplification was obtained for other strains. The detection sensitivity of continental PCR can reach 60 pg of mycelium DNA,Probe SA-P can reach 60 fg of mycelium DNA,which is 1 000 times higher than conventional PCR. The detection assay for the flower bud and leaf samples of rhododendron showed potential role for rapid detection of S. azaleae in imported rhododendron seedlings.

关 键 词：杜鹃芽枯病菌 检测 常规PCR 实时荧光PCR 

分 类 号：S41[农业科学—植物保护]