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

IF 2.1 4区 生物学 Q3 MICROBIOLOGY Brazilian Journal of Microbiology Pub Date : 2024-12-06 DOI:10.1007/s42770-024-01578-6
Le-Yao Ren, Ke Wen, Bao-Ping Cheng, Jing-Hao Jin, Vaibhav Srivastava, Xiao-Ren Chen
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Abstract

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; '' 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 β-tubulin gene of the pathogen. Using a carefully selected set of primers and optimized reaction conditions at 60 °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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用可视化环介导等温扩增法快速检测植物致病卵菌螺旋藻。
植物致病卵菌helicoides (Phytopythium helicoides)已成为沙塘菊柑橘(citrus reticulata cv.)的新威胁。Shatangju;在中国南方。为了快速诊断和控制由螺旋藻引起的叶枯病和茎腐病,我们建立了一种针对病原菌β-微管蛋白基因的可视化环介导等温扩增(LAMP)方法。LAMP实验采用精心挑选的引物和优化的反应条件,在60°C下反应60 min,可以在各种柑橘相关的植物病原体中特异性检测出病原菌。该试验显示出高灵敏度,检测到30pg的病原体DNA,与定量聚合酶链反应(qPCR)的灵敏度相当。我们验证了LAMP检测在柑橘叶和草莓冠感染诊断中的实际应用。新建立的LAMP检测方法为螺旋螺旋体提供了一种特异、灵敏、快速的检测工具,有助于防治沙唐菊柑橘叶枯病和茎腐病,以及草莓冠腐病等其他相关作物病害。
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来源期刊
Brazilian Journal of Microbiology
Brazilian Journal of Microbiology 生物-微生物学
CiteScore
4.10
自引率
4.50%
发文量
216
审稿时长
1.0 months
期刊介绍: The Brazilian Journal of Microbiology is an international peer reviewed journal that covers a wide-range of research on fundamental and applied aspects of microbiology. The journal considers for publication original research articles, short communications, reviews, and letters to the editor, that may be submitted to the following sections: Biotechnology and Industrial Microbiology, Food Microbiology, Bacterial and Fungal Pathogenesis, Clinical Microbiology, Environmental Microbiology, Veterinary Microbiology, Fungal and Bacterial Physiology, Bacterial, Fungal and Virus Molecular Biology, Education in Microbiology. For more details on each section, please check out the instructions for authors. The journal is the official publication of the Brazilian Society of Microbiology and currently publishes 4 issues per year.
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