A Rapid and Simple Method for DNA Preparation of Magnaporthe oryzae from Single Rice Blast Lesions for PCR-Based Molecular Analysis
文献类型： 外文期刊
作者： Dong, Liying 1 ; Liu, Shufang 1 ; Li, Jing 2 ; Tharreau, Didier 3 ; Liu, Pei 1 ; Tao, Dayun 2 ; Yang, Qinzhong 1 ;
作者机构： 1.Yunnan Acad Agr Sci, Agr Environm & Resources Inst, Key Lab Green Prevent & Control Agr Transboundary, Kunming 650205, Yunnan, Peoples R China
2.Yunnan Acad Agr Sci, Food Crops Res Inst, Kunming 650205, Yunnan, Peoples R China
3.Ctr Cooperat Int Rech Agron Dev CIRAD, TA 54K, UMR BGPI, F-34398 Montpellier, France
关键词： DNA extraction; infected rice tissue; Magnaporthe oryzae
期刊名称：PLANT PATHOLOGY JOURNAL （ 影响因子：2.321； 五年影响因子：2.541 ）
ISSN： 1598-2254
年卷期： 2022 年 38 卷 6 期
页码：
收录情况： SCI
摘要： Rice blast is one of the most destructive diseases of rice worldwide, and the causative agent is the filamentous ascomycete Magnaporthe oryzae. With the success-ful cloning of more and more avirulence genes from M. oryzae, the direct extraction of M. oryzae genomic DNA from infected rice tissue would be useful alter-native for rapid monitoring of changes of avirulence genes without isolation and cultivation of the pathogen. In this study, a fast, low-cost and reliable method for DNA preparation of M. oryzae from a small piece of infected single rice leaf or neck lesion was established. This single step method only required 10 min for DNA preparation and conventional chemical reagents com-monly found in the laboratory. The AvrPik and AvrPi9 genes were successfully amplified with the prepared DNA. The expected DNA fragments from 570 bp to 1,139 bp could be amplified even three months after DNA preparation. This method was also suitable for DNA preparation from M. oryzae strains stored on the filter paper. All together these results indicate that the DNA preparation method established in this study is reliable, and could meet the basic needs for polymerase chain reaction-based analysis of M. oryzae.
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