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Quick and Accurate Detection and Quantification of Magnaporthe oryzae in Rice Using Real-Time Quantitative Polymerase Chain Reaction

February 2015 , Volume 99 , Number  2
Pages  219 - 224

Geng Sun, College of Plant Sciences, Jilin University, Jilin, Changchun 130062, China, and State Key Laboratory of the Discovery and Development of Novel Pesticide, Shenyang Research Institute of Chemical Industry Co., Ltd., Shenyang 110021, China; Jinliang Liu, Guihua Li, Xianghui Zhang, Tingting Chen, and Jingyuan Chen, College of Plant Sciences, Jilin University; Hao Zhang, College of Resource and Environment, Jilin Agricultural University; Dongping Wang, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87544; Fengjie Sun, School of Science and Technology, Georgia Gwinnett College, Lawrenceville, GA 30043; and Hongyu Pan, College of Plant Sciences, Jilin University

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Accepted for publication 28 July 2014.

Rice blast, caused by Magnaporthe oryzae, is one of the most severe fungal diseases in rice worldwide. In this study, we developed methods to quickly and accurately detect and quantify M. oryzae in the pure cultures of the fungus, rice plants, and rice seed by using SYBR Green I of the real-time quantitative polymerase chain reaction (qPCR). Results of absolute qPCR show that Magnaporthe oryzae can be detected at as low as 6.9 × 10−5 ng of genomic DNA. Results also show that all 10 varieties of rice seed examined in this study contain this fungus, indicating that M. oryzae is generally widespread in rice seed. We report the quantification of DNA of M. oryzae in rice leaves collected in the field, instead of in the lab, using relative qPCR by using rice actin gene as a housekeeping gene. Our results show great practical significance because we would know the potential fungal infection even before planting.

Copyright © 2015 The American Phytopathological Society