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| Development of SYBR GreenⅠReal-time RT-PCR for the Detection of Cucumber green mottle mosaic virus |
| TIAN Yi-Min1, WU Jian-Xiang2, CHEN Lei3, LUO Jin-Yan3, TENG Kai4, YE Jun1, DU Wei5, YU Cui1,* |
1 Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs District, Shanghai 200135, China;
2 College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China;
3 Shanghai Agricultural Technology Extension Center, Shanghai 201103, China;
4 Division of Animal and Plant Quarantine and Inspection of Shanghai Customs District, Shanghai 200082, China;
5 Agricultural Technology Extension Station of Ningxia, Yinchuan 750001, China |
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Abstract Cucumber green mottle mosaic virus (CGMMV) is an important quarantine plant virus and is also transmitted by seeds which often causes seriously damages to productions of watermelon (Citrullus lanatus), melon (Cucumis melo), cucumber (Cucumis sativus), and other cucurbits. To guarantee the smooth proceeding of seed trade, it is necessary to establish more rapidly and precisely detection methods. This study developed a SYBR GreenⅠreal-time RT-PCR technique for detection of CGMMV in seeds. It is crucial to design and screen a pair of specific primers based on the conserved CGMMV gene sequences. The result showed that the development of SYBR GreenⅠreal time RT-PCR method in this study could detect CGMMV specially, and there was no cross-reactivity with other common cucurbits viruses. Sensitivity assay showed that the sensitivity of this method was 1 000 times that of RT-PCR and 100 times that of TaqMan RT-PCR. And the whole detection process could be completed within 2.5 h, indicating that the SYBR GreenⅠreal-time RT-PCR was a rapid, sensitive and simple detection method. Cucurbitaceae seeds, seedlings, and fruits imported and purchased from domestic market were employed to be detected by SYBR GreenⅠreal-time RT-PCR and TaqMan real-time RT-PCR simultaneously. The result showed that the value of kappa of the two method was above 0.82, indicating that the novel SYBR GreenⅠreal-time RT-PCR method could be effective and reliable. The present study provides new technical means for rapid clearance of seeds.
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Received: 30 September 2021
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Corresponding Authors:
*yucuicn@163.com
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