A rapid and simple method for DNA preparation of Magnaporthe oryzae from single rice blast lesions for PCR-based molecular analysis
2022
Dong, Liying | Liu, Shuguang | Li, Jing | Tharreau, Didier | Liu, Pei | Tao, Dayun | Yang, Qinzhong | Yunnan Academy of Agricultural Sciences | Plant Health Institute of Montpellier (UMR PHIM) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Montpellier ; Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université de Montpellier (UM) | Applied Basic Research Key Project of Yunnan;;CHN; | National Natural Science Foundation of China;NNSF;CHN;http://dx.doi.org/10.13039/501100001809 | Major Special Project of Yunnan;;CHN;
Source Agritrop Cirad (https://agritrop.cirad.fr/603215/)
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Mostrar más [+] Menos [-]Inglés. 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.
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