新疆4种林木腐烂病菌PCR快速检测技术研究  

Rapid PCR Detection Technology for Four Species of Valsa Canker Pathogens of Trees in Xinjiang

作　　者：郭开发[1] 姚兆群[1] 吴彩兰[1] 向本春[1] 赵思峰[1] 

机构地区：[1]新疆绿洲农业病虫害治理与植保资源利用自治区高校重点实验室/石河子大学农学院,新疆石河子832003

出　　处：《新疆农业科学》2016年第10期1843-1849,共7页Xinjiang Agricultural Sciences

基　　金：新疆维吾尔自治区科技援疆项目(201491150);新疆维吾尔自治区高层次人才引进项目(2015-2017);The science and technology aid project of the Xinjiang Uygur Autonomous Region(201491150);The project of the introduction of the higher-grade talented persons of the Xinjiang Uygur Autonomous Region(2015-2017)

摘　　要：【目的】建立新疆林木腐烂病菌的快速PCR检测方法,为新疆林木腐烂病的早期测报和防治提供技术依据。【方法】针对新疆林木腐烂病菌主要种苹果黑腐皮壳(Valsa mali),污黑腐皮壳(Valsa.sordida),Leucostoma niveum和Valsa.malicola的r DNA-ITS特异性区段设计属专化型引物和种特异性引物。【结果】属专化型引物VF/VR可以从腐烂病菌中扩增一条424 bp的条带,检测灵敏度为10 pg/m L;设计的4对种特异性引物分别可对4种腐烂病菌V.mali,V.sordida,L.niveum,V.malicola检测到263 bp、423 bp、307 bp、308 bp的条带,检测灵敏度均为10 pg/m L。【结论】采用腐烂病菌Valsa的r DNA-ITS特异性区段设计的引物及PCR方法,可用于新疆林木腐烂病的快速分子检测。【Objective】To develop a rapid PCR assay for detection of trees Valsa canker in Xinjiang and provide the technical support for forecast and prevention of trees Valsa canker.【Method】Based on r DNA-ITS conservative sequence of the Valsa genus,a pair of genus-specific primers and four pairs of species-specific primers were designed for Valsa mali,Valsa sordida,Leucostoma niveum and Valsa malicola. 【Result】Using genus-specific primers VF/VR to amplify a unique 424 pb band from Valsa and using four pairs of species-specific primers to amplify 263 bp,423 bp,307 bp and 308 bp band from V. mali,V. sordida,L.niveum and V. malicola respectively. And the detection sensitivity was 10 pg/m L. 【Conclusion】The results suggested that developed rapid molecular detection method in this study based on r DNA-ITS conservative sequence of the Valsa genus can be used in the Xinjiang trees Valsa canker.

关 键 词：林木 腐烂病菌 r DNA-ITS PCR检测 特异性引物 

分 类 号：S436.6[农业科学—农业昆虫与害虫防治]