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국화에 발생하는 반쪽시들음병균 Verticillium dahliae 검출용 등온 증폭법 개발

Development of a Loop-mediated Isothermal Amplification Detection Assay for Verticillium dahliae Infection in Chrysanthemum

한국균학회지 2019년 47권 4호 p.437 ~ 441
백창기 ( Back Chang-Gi ) - 국립원예특작과학원 원예특작환경과

박미정 ( Park Mi-Jeong ) - 국립원예특작과학원 원예특작환경과
한경숙 ( Han Kyung-Sook ) - 국립원예특작과학원 기획조정과
박종한 ( Park Jong-Han ) - 국립원예특작과학원 원예특작환경과

Abstract


Verticillium wilt disease is caused by a fungal plant pathogen Verticillium dahliae, which attacks commercial crops such as chrysanthemum. The conventional methods so far used to identify this fungal pathogen require high expertise and are time-consuming. Therefore, in this study, we developed an assay for the rapid and specific detection of V. dahliae infection using loop-mediated isothermal amplification (LAMP) method. For this assay, four primers for LAMP were designed for targeting cellulose-growth-specific protein partial mRNA gene in Verticillium dahliae . Under standard condition, the optimum reaction temperature for amplification is around 60 ℃ within 60 minutes. This LAMP assay was designed to amplify only present in V. dahliae. When this LAMP assay applied to the DNAs for four other soilborne fungi and host plants, no amplification was detected. Therefore, this LAMP assay we developed for V. dahliae is expected to do detection at the early stage of its infection. The fast and reliable detection method will allow us to develop effective management system to monitor and control infection of this pathogen in chrysanthemum plant.

키워드

detection; Loop-mediated isothermal amplification; Verticillium dahliae; wilt disease
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