Use of Reverse Transcription Loop-Mediated Isothermal Amplification for the Detection of Zucchini yellow mosaic virus
| Autor: | T. C. Deng, Yi-Lin Lu, Cheng-Ping Kuan, Hung-Chang Huang, Hsiang‐Hui Chi |
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| Rok vydání: | 2013 |
| Předmět: |
Gel electrophoresis
Zucchini yellow mosaic virus genetic structures Physiology Melon Loop-mediated isothermal amplification food and beverages Plant Science Biology biology.organism_classification Virology Molecular biology eye diseases Staining chemistry.chemical_compound chemistry Genetics SYBR Green I sense organs Agronomy and Crop Science Reverse Transcription Loop-mediated Isothermal Amplification Squash |
| Zdroj: | Journal of Phytopathology. 162:238-244 |
| ISSN: | 0931-1785 |
| Popis: | A Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) assay was employed to develop a simple and efficient system for the detection of Zucchini yellow mosaic virus (ZYMV) in squash and melon plants. The RT-LAMP assay took 30 min under isothermal condition at 64°C by employing a set of four primers targeting ZYMV. The sensitivity of RT-LAMP was 10-fold greater than that of the RT-PCR assay in the detection of ZYMV in infected tissues of squash and melon. No reaction was detected from the tissues of healthy plants by either RT-LAMP or RT-PCR assay. The RT-LAMP product of the tested samples can be visualized by staining directly in the tube with SYBR Green I dye. The sensitivity of SYBR Green I staining method is similar to that analyzed by gel electrophoresis. Field-grown squash and melon plants were tested using RT-PCR and RT-LAMP. Both RT-LAMP and PCR could detect ZYMV in symptomatic or symptomless tissues of infected plants. However, the RT-LAMP assay is superior to RT-PCR because it is rapid, simple, and highly sensitive; therefore, RT-LAMP is a useful and practical method for detection of ZYMV in cucurbits. |
| Databáze: | OpenAIRE |
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