草莓灰霉病菌对啶酰菌胺抗性及其SdhB基因序列差异的分子检测  

Resistance Mechanism to Boscalid and Molecular Mutation-detection Based on SdhB Gene of Botrytis cinerea from Strawberry

作　　者：刘欣 曾蓉[1] 徐丽慧[1] 高士刚[1] 戴富明[1] LIU Xin;ZENG Rong;XU Lihui;GAO Shigang;DAI Fuming(Institute of Eco-Environment and Plant Protection,Shanghai Key Laboratory of Protection Horticultural Technology,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China)

机构地区：[1]上海市农业科学院生态环境保护研究所设施园艺重点实验室

出　　处：《园艺学报》2019年第11期2155-2163,共9页Acta Horticulturae Sinica

基　　金：上海市瓜果产业技术体系建设项目[沪农科产字(2017)第1号];上海市自然科学基金项目(16ZR1424100);上海市农业科学院卓越团队项目[农科创2018(B-01)]

摘　　要：啶酰菌胺被广泛用于草莓灰霉病的防治,然而灰葡萄孢菌(Botrytis cinerea)对啶酰菌胺存在较大的抗性风险,抗性出现主要与SdhB基因突变有关。AS-PCR(Allele-specificPCR)方法常用于检测基因点突变。为明确上海地区草莓灰霉病菌对啶酰菌胺的抗性机制,2018年测定了126株草莓灰霉病菌中的61株抗性菌株与5株随机选取敏感菌株的SdhB基因序列。结果表明,上海地区草莓灰霉病菌目前已存在4种抗性突变类型:P225F、N230I、H272R和H272Y,其中H272R和P225F为优势突变类型。基于AS-PCR原理,设计及筛选了包含2条特异性引物的三引物组,优化了反应体系,建立了可通过1轮PCR反应即可区别啶酰菌胺敏感菌株与4种抗性突变菌株的快速检测方法。Boscalid is widely used in control of strawberry gray mold disease.However,it is at a high risk of resistance development for Botrytis cinerea to boscalid.The resistance was mainly caused by mutations of SdhB gene.The method of AS-PCR(Allele-specific PCR)is always used to detect point mutations in DNA.In order to evaluate the resistance mechanism to boscalid of Botrytis cinerea from strawberry in Shanghai,resistance strains in 126 isolates of 2018 were SdhB-sequenced.The results showed 4 types of mutations associated to boscalid resistance in Shanghai,P225 F,N230 I,H272 R and H272 Y.Among then,H272 R and P225 F were the dominant type.Therefore,for rapid detection,the tri-primer including 2 specific primers was designed and selected based on AS-PCR principle,and the PCR system was optimized to develop the method,by which the 4 types of mutations could be distinguished from the wild sensitive type in one PCR reaction.

关 键 词：草莓 灰葡萄孢 AS-PCR 啶酰菌胺 杀菌剂抗性检测 

分 类 号：S668.4[农业科学—果树学]