数字农科院2.0

Lamp Detection Of The Genetic E.lement 'Mona' Associated With D mi Resistance In Monilinia Fructicola

文献类型： 外文期刊

作者： Chen, Shuning;Yuan, Huizhu;Schnabel, Guido;Luo, Chaoxi;Luo, Chaoxi

作者机构：

关键词： DMI fungicides; fungicide resistance; LAMP method; Monilinia fructicola; peach brown rot

期刊名称： PEST MANAGEMENT SCIENCE

ISSN： 1526-498X

年卷期： 2019 年 75 卷 3.0 期

页码：

收录情况： JCR（2021版）

摘要： BACKGROUND The increasing use of demethylation inhibitor (DMI) fungicides for the control of peach brown rot has resulted in resistance in Monilinia fructicola. Resistance in the southeastern USA is caused by overexpression of the MfCYP51 gene due to the presence of a 65-bp inserted element 'Mona' located in the upstream regulatory region of MfCYP51. A rapid diagnostic assay would be useful to detect the presence and monitor further spread of this resistance mechanism. RESULTS A loop-mediated isothermal amplification (LAMP) method was developed for rapid detection of 'Mona'-based DMI resistance. The assay was optimized for specificity and sensitivity, and was shown to detect the presence of 10 fg of purified target DNA per reaction within 85 min. Only DNA isolated from DMI-resistant isolates containing 'Mona' resulted in a fluorescent signal after LAMP assay amplification. DNA from sensitive isolates from China and the USA and six other common fungal species of peach did not yield a signal. The method also positively identified 'Mona' from crude DNA extracts (using Lyse and Go reagents heated to 100 degrees C for 10 min) obtained from the mycelium and conidia of symptomatic fruit. CONCLUSION Considering its specificity, stability and repeatability, the LAMP assay could be a valuable tool for rapid on-site diagnosis of M. fructicola isolates resistant to DMI fungicides in the southeastern USA. (c) 2018 Society of Chemical Industry

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