Development and evaluation of specific PCR and LAMP assays for the rapid detection of Phytophthora melonis
文献类型： 外文期刊
作者： Chen, Qinghe 1 ; Li, Benjin 1 ; Liu, Peiqing 1 ; Lan, Chengzhong 1 ; Zhan, Zhixiong 1 ; Weng, Qiyong 1 ;
作者机构： 1.Fujian Acad Agr Sci, Inst Plant Protect, Fuzhou 350003, Peoples R China
关键词： Loop-mediated isothermal amplification (LAMP);Molecular detection;Phytophthora melonis;Ras-related protein (Ypt1) gene
期刊名称：EUROPEAN JOURNAL OF PLANT PATHOLOGY （ 影响因子：1.907； 五年影响因子：2.022 ）
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收录情况： SCI
摘要： Phytophthora melonis is a widespread and devastating pathogen for the Cucurbitaceae family. Early and accurate detection of P. melonis is essential to control the disease in the field. To establish a simple, visual, and rapid detection system for P. melonis, we developed nested polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) systems based on the Ras-related protein (Ypt1) gene. All 36 isolates of P. melonis, from geographically distinct counties in China, yielded positive detection results on LAMP or nested PCR assays. No cross reaction was observed with other oomycetes or fungal pathogens. A sensitivity assay showed that both methods had a detection limit of 10 fg genomic DNA. We also detected P. melonis in diseased cucumber tissues and soils, and evaluated positive detection rates using LAMP, nested PCR, and conventional isolation methods. The results suggest that the LAMP assay has the greatest potential for active detection of P. melonis in regions that are at risk of contracting the disease, and for use in resource-poor settings.
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