Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Fusarium oxysporum f. sp niveum in soil
文献类型： 外文期刊
作者： Peng, Jun 1 ; Zhan, Yuanfeng 2 ; Zeng, Fanyun 1 ; Long, Haibo 1 ; Pei, Yuelin 1 ; Guo, Jianrong 1 ;
作者机构： 1.Chinese Acad Trop Agr Sci, Inst Environm & Plant Protect, Key Lab, Minist Agr Integrated Pest Management Trop Crops, Haikou 571101, Hainan, Peoples R China
2.Chinese Acad Trop Agr Sci, Trop Crops Genet Resources Inst, Danzhou, Punjab, Peoples R China
关键词： Fusarium wilt of watermelon;detection of fungi;RealAmp assay;real-time PCR
期刊名称：FEMS MICROBIOLOGY LETTERS （ 影响因子：2.742； 五年影响因子：2.856 ）
ISSN： 0378-1097
年卷期： 2013 年 349 卷 2 期
页码：
收录情况： SCI
摘要： Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (Fon) is one of the major limiting factors for watermelon production worldwide. Rapid and accurate detection of the causal pathogen is the cornerstone of integrated disease management. In this paper, a real-time fluorescence loop-mediated isothermal amplification (RealAmp) assay was developed for the rapid and quantitative detection of Fon in soil. Positive products were amplified only from Fon isolates and not from any other species or formae speciales of F.oxysporum tested, showing a high specificity of the primer sets. The detection limit of the RealAmp assay was 1.2pgL(-1) genomic DNA or 10(3)sporesg(-1) of artificially inoculated soil, whereas real-time PCR could detect as low as 12fgL(-1) or 10(2)sporesg(-1). The RealAmp assay was further applied to detect eight artificially inoculated and 85 field soil samples. No significant differences were found between the results tested by the RealAmp and real-time PCR assays. The RealAmp assay is a simple, rapid and effective technique for the quantitative detection and monitoring of Fon in soil under natural conditions.
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