Establishment of real-time PCR for quantitatively monitoring Pseudoperonospora cubensis in cucumber (Cucumis sativus)
Establishment of real-time PCR for quantitatively monitoring Pseudoperonospora cubensis in cucumber (Cucumis sativus)
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DOI：
10.30848/pjb2021-3(16
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发表时间：
2021-06
影响因子：
1.2
通讯作者：
Dong Liu;Chunqing Pan;Lei Tao;Di Zhang;Chun-lin Wang;Ziwei He;Yanju Zhang
中科院分区：
文献类型：
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作者：
Dong Liu;Chunqing Pan;Lei Tao;Di Zhang;Chun-lin Wang;Ziwei He;Yanju Zhang
由黄瓜霜霉病菌引起的黄瓜霜霉病是我国重要的气传病害，严重威胁着黄瓜生产。早期准确的定量检测是预报和控制的基础。本研究根据GenBank中已发表的古巴青霉rDNA-ITS序列设计了一对特异性引物。结果表明，所设计的引物对黄瓜霜霉病菌的常规PCR检测结果为阳性，对其他病原菌的PCR检测结果为阴性，而对黄瓜霜霉病菌的PCR检测结果为阴性，荧光定量PCR的灵敏度高于常规PCR，且检测结果与病情指数和接种量呈显著正相关。总之，实时荧光PCR检测古巴青霉是快速，高度特异性，灵敏度和重现性。因此，我们成功地建立了一种适用于P. cubensis的SYBR绿色I荧光定量PCR方法。该方法可用于黄瓜霜霉病菌的快速检测，对黄瓜霜霉病的预测预报和防治具有指导意义。
Cucumber downy mildew caused by Pseudoperonospora cubensis is an important airborne disease in China and greatly threatens cucumber production. Early accurate quantitative detection is the foundation of forecasting and control. Here, we designed a pair of specific primers according to the rDNA-ITS sequence of P. cubensis in GenBank. These primers could amplify the DNA of P. cubensis isolates by conventional PCR but not the DNA of other pathogens.The sensitivity of realtime PCR was higher than that of conventional PCR.The real-time PCR detection results of infected cucumber samples were significantly positively correlated with disease indexes and inocula. In conclusion, real-time PCR assays for P. cubensis are fast, highly specific, sensitive, and reproducible. We therefore successfully established a SYBR Green I real-time PCR method for P. cubensis. This method can be used to detect P. cubensis in cucumber and help guide the prediction and control of cucumber downy mildew.