引起玉米穗腐病的两种镰孢菌双重PCR快速检测体系的建立  

Duplex PCR detection of two Fusariums causing maize ear rot

作　　者：史亚娟[1] 孙新艳[1] 袁虹霞[1] 邢小萍[1] 王振跃[1] 张晓婷[1] 施艳[1] 李洪连[1] SHI Ya-juan SUN Xin-yan YUAN Hong-xia XING Xiao-ping WANG Zhen-yue ZHANG Xiao-ting SHI Yan LI Hong-lian(College of Plant Protection, Henan Agricultural University/Collaborative Innovation Centre of Henan Grain Crops/State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 450002, Chin)

机构地区：[1]河南农业大学/河南省粮食作物协同创新中心/小麦玉米作物学国家重点实验室,郑州450002

出　　处：《植物病理学报》2017年第1期35-39,共5页Acta Phytopathologica Sinica

基　　金：河南省玉米产业技术体系植保岗位科学家专项(S2015-02-G05);国家科技支撑计划粮食丰产科技工程河南课题(2013BAD07B07)

摘　　要：禾谷镰孢(Fusarium graminearum)和拟轮枝镰孢(Fusarium verticillioides)是引起玉米穗腐病的两种主要病原菌。为了建立这两种病原菌的快速和定量检测体系,本研究分别设计了针对F.graminearum和F.verticillioides的基因特异性引物,建立了双重PCR反应体系,并从DNA浓度、引物浓度和退火温度3个方面对反应体系进行了优化。结果表明,退火温度为54℃和58℃时扩增效果较好,DNA模版的检测灵敏度可以达到6.25 ng·μL^(-1);F.graminearum和F.verticillioides引物最佳终浓度配比为0.2μmol·L^(-1)/0.4μmol·L^(-1)。本研究建立的双重PCR对同时鉴别引起玉米穗腐病的两种主要镰孢菌提供了准确、快速、灵敏的检验检疫技术。Fusarium graminearum and F.verticillioides,the two main pathogens causing maize ear rot,were set as targets for the quick and quantitative analyses.In this study the duplex PCR was established using the specific primer pairs for F.graminearum and F.verticillioides.The reaction was optimized from the aspects of DNA concentration,primer ratio and annealing temperature.The results showed that the annealing temperature of54℃ and 58℃ displayed efficient amplification,the optimum primer ratio of F.graminearum to F.verticillioides was 0.2 μmol·L^（-1）/0.4μmol·L^（-1）,and the sensitivity for the reaction reached as low as 6.25 ng·μL^（-1）.The duplex PCR developed in the study provides an accurate and sensitive approach to simultaneously detect F.graminearum and F.verticillioides.

关 键 词：玉米穗腐病 双重PCR 禾谷镰孢 拟轮枝镰孢 

分 类 号：S432.44[农业科学—植物病理学]