Molecular and Biochemical Characterization of Laboratory and Field Mutants of Botrytis cinerea Resistant to Fludioxonil.
Molecular and Biochemical Characterization of Laboratory and Field Mutants of Botrytis cinerea Resistant to Fludioxonil.
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DOI：
10.1094/pdis-11-15-1290-re
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发表时间：
2016-04
期刊：
影响因子：
4.5
通讯作者：
Weichao Ren;W. Shao;Xu Han;Ming-guo Zhou;Changjun Chen
中科院分区：
文献类型：
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作者：
Weichao Ren;W. Shao;Xu Han;Ming-guo Zhou;Changjun Chen
灰葡萄孢是一种丝状植物病原体，具有对杀菌剂产生抗性的高风险。据报道，苯基吡咯杀真菌剂咯菌腈对B具有优异的活性。灰霉病在我国已得到越来越多的应用。本研究对B的实验室和田间突变体的分子生物学特性进行了研究。已经研究了对咯菌腈具有抗性的灰霉病菌。2012年至2014年期间，B。采用离体培养法测定了采自江苏、山东两省的灰霉病菌对防治黄瓜、番茄灰霉病常用药剂的敏感性。在2013年从黄瓜收集的75个菌株中，有两个对咯菌腈具有高抗性（HR）。在2014年从番茄上采集的308个分离物中，有4个是咯菌腈-HR。田间检测到对咯菌腈有抗药性的灰霉病菌。通过室内筛选，获得了6株咯菌腈抗性突变株。这些突变体对咯菌腈表现出稳定的抗性，如抗性因子值从34.38到> 10，000所示。与咯菌腈敏感的B.灰霉病，所有的田间和实验室突变体表现出降低的健身，定义为菌丝生长，孢子形成，毒力，和渗透胁迫的敏感性。当用1 μg/ml咯菌腈处理时，敏感菌株表现出菌丝体中甘油含量和Bchog 1表达水平的增加，而田间和实验室HR突变体中的水平仅略有增加。对田间和实验室的咯菌腈-HR突变体的Bos 1基因序列分析表明，田间突变体的突变位于N-末端区域的组氨酸激酶、腺苷酸环化酶、甲基接受趋化蛋白和磷酸酶（HAMP）结构域，而实验室突变体的突变则分布在C-末端区域的HAMP结构域或HATP_c结构域。这些结果将加深我们对B抗性机制的理解。灰霉病菌对咯菌腈。
Botrytis cinerea is a filamentous phytopathogen with a high risk of developing resistance to fungicides. The phenylpyrrole fungicide fludioxonil has been reported to have excellent activity against B. cinerea and increasingly has been applied to control gray mold in China. In this study, molecular and biochemical characteristics of laboratory and field mutants of B. cinerea resistant to fludioxonil has been investigated. During 2012 to 2014, B. cinerea isolates collected from Jiangsu and Shandong Provinces in China were tested in vitro for sensitivity to fungicides commonly used to suppress gray mold of cucumber and tomato. Among the 75 isolates collected from cucumber in 2013, two were highly resistant (HR) to fludioxonil. Of the 308 isolates collected from tomato in 2014, four were fludioxonil-HR. This was the first time that B. cinerea isolates HR to fludioxonil had been detected in the field. Six fludioxonil-resistant mutants were obtained in the laboratory by selection on fungicide-amended media. These mutants exhibited stable resistance to fludioxonil, as indicated by resistance factor values that ranged from 34.38 to >10,000. In comparison with fludioxonil-sensitive isolates of B. cinerea, all field and laboratory mutants showed reduced fitness, as defined by mycelial growth, sporulation, virulence, and sensitivity to osmotic stress. When treated with fludioxonil at 1 μg/ml, sensitive isolates showed increased glycerol contents in mycelium and expression levels of Bchog1, while levels in field and laboratory HR mutants increased only slightly. Sequences of the Bos1 gene of field and laboratory fludioxonil-HR mutants showed that mutations in field mutants were located in the histidine kinase, adenylyl cyclase, methyl-accepting chemotaxis protein, and phosphatase (HAMP) domains of the N-terminal region, whereas mutations in the laboratory mutants were distributed in HAMP domains or in the HATPase_c domain of the C-terminal region. These results will enhance our understanding of the resistance mechanism of B. cinerea to fludioxonil.