The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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A Rapid and Simple Method for DNA Preparation of Magnaporthe oryzae from Single Rice Blast Lesions for PCR-Based Molecular Analysis

  • Liying, Dong (Agricultural Environment and Resources Institute/Key Laboratory of Green Prevention and Control of Agricultural Transboundary Pests of Yunnan Province, Yunnan Academy of Agricultural Sciences) ;
  • Shufang, Liu (Agricultural Environment and Resources Institute/Key Laboratory of Green Prevention and Control of Agricultural Transboundary Pests of Yunnan Province, Yunnan Academy of Agricultural Sciences) ;
  • Jing, Li (Food Crops Research Institute, Yunnan Academy of Agricultural Sciences) ;
  • Didier, Tharreau (Centre de Cooperation Internationale en Recherche Agronomique pour le Developpement (CIRAD)) ;
  • Pei, Liu (Agricultural Environment and Resources Institute/Key Laboratory of Green Prevention and Control of Agricultural Transboundary Pests of Yunnan Province, Yunnan Academy of Agricultural Sciences) ;
  • Dayun, Tao (Food Crops Research Institute, Yunnan Academy of Agricultural Sciences) ;
  • Qinzhong, Yang (Agricultural Environment and Resources Institute/Key Laboratory of Green Prevention and Control of Agricultural Transboundary Pests of Yunnan Province, Yunnan Academy of Agricultural Sciences)
  • Received : 2022.02.10
  • Accepted : 2022.10.09
  • Published : 2022.12.01
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Abstract

Rice blast is one of the most destructive diseases of rice worldwide, and the causative agent is the filamentous ascomycete Magnaporthe oryzae. With the successful 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 alternative 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 commonly 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.

Keywords

Acknowledgement

This work was supported by the Applied Basic Research Key project of Yunnan (202001AS070006), the National Natural Science Foundation of China (31560493, 31860524), and the Major Special Project of Yunnan Province (202102AE090003).

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