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通过扩增阻滞突变系统聚合酶链反应(ARMS-PCR)快速检测对啶酰菌胺耐药菌株中的 和 突变

Rapid Detection of and Mutations in Boscalid Resistant Strains by ARMS-PCR.

作者信息

Liu Xin, Zeng Rong, Gao Shigang, Xu Lihui, Dai Fuming

机构信息

Institute of Eco-Environment and Plant Protection, Shanghai Key Laboratory of Protection Horticultural Technology, Shanghai Academy of Agricultural Sciences, 1000 Jinqi Road, Shanghai 201403, China.

出版信息

Plant Pathol J. 2019 Feb;35(1):71-76. doi: 10.5423/PPJ.NT.08.2018.0151. Epub 2019 Feb 1.

Abstract

and mutations have been found associated with boscalid resistance in from strawberry in Shanghai, China. For rapid detection of two mutations, tetra-primers were designed and optimized to gain the relatively high accuracy and specificity based on the ARMS-PCR technique, by which resistance can be identified with different lengths of products on agarose gels. The tetra-primer ARMS-PCR systems for and were validated by 9 -squenced strains repeatedly. Then, sensitivity of 30 more strains were also tested by the methods, which were accordant with genotypes by sequencing and the sensitivity of conidial germination to boscalid by 100%. Thus, the methods developed in this study are proved to be rapid, inexpensive, accurate and practical for resistance detection of caused by and mutations.

摘要

在中国上海的草莓中发现了与啶酰菌胺抗性相关的突变。为了快速检测这两种突变,基于扩增受阻突变系统聚合酶链反应(ARMS-PCR)技术设计并优化了四引物,以获得相对较高的准确性和特异性,通过该技术可在琼脂糖凝胶上根据产物长度不同鉴定抗性。用于[具体基因1]和[具体基因2]的四引物ARMS-PCR系统经9株测序菌株反复验证。然后,还用这些方法检测了另外30株菌株的敏感性,其结果与测序基因型一致,且分生孢子萌发对啶酰菌胺的敏感性为100%。因此,本研究开发的方法被证明对于由[具体基因1]和[具体基因2]突变引起的[病原菌名称]抗性检测是快速、廉价、准确且实用的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2f5/6385654/976fe221a745/ppj-35-071f1.jpg

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