Rapid detection of the phytopathogenic oomycete Phytopythium helicoides with a visualized loop-mediated isothermal amplification assay

文献类型： 外文期刊

第一作者： Ren, Le-Yao

作者： Ren, Le-Yao;Wen, Ke;Jin, Jing-Hao;Chen, Xiao-Ren;Cheng, Bao-Ping;Srivastava, Vaibhav

作者机构：

关键词： Phytopythium helicoides; LAMP; Detection; Leaf blight; Stem rot

期刊名称：BRAZILIAN JOURNAL OF MICROBIOLOGY （ 影响因子：1.9； 五年影响因子：2.4 ）

ISSN： 1517-8382

年卷期： 2025 年 56 卷 1 期

页码：

收录情况： SCI

摘要： The phytopathogenic oomycete Phytopythium helicoides, previously known as Pythium helicoides, has emerged as a new threat to the Shatangju citrus cultivar (Citrus reticulata cv. Shatangju; '(sic)(sic)(sic)' in Chinese) in southern China. To enable rapid diagnosis and control of the leaf blight and stem rot caused by P. helicoides, we developed a visualized loop-mediated isothermal amplification (LAMP) assay targeting the beta-tubulin gene of the pathogen. Using a carefully selected set of primers and optimized reaction conditions at 60 degrees C for 60 min, the LAMP assay can specifically detect the pathogen among various citrus-associated phytopathogens. The assay demonstrated high sensitivity, detecting as little as 30 pg of the pathogen DNA, comparable to the sensitivity of quantitative polymerase chain reaction (qPCR). We validated the practical application of this LAMP assay for diagnosing infections in citrus leaves and strawberry crowns. This newly developed LAMP assay offers a specific, sensitive, and rapid detection tool for P. helicoides, aiding in the control of Shatangju citrus leaf blight and stem rot, as well as other related crop diseases like strawberry crown rot.

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