Using homologous cloning method, partial fragments of coat protein (CP) gene of WMV, CMV and ZYMV were cloned from virus-infected melon in Xinjiang. The reaction system of multiplex RT-PCR was optimized based on sin...Using homologous cloning method, partial fragments of coat protein (CP) gene of WMV, CMV and ZYMV were cloned from virus-infected melon in Xinjiang. The reaction system of multiplex RT-PCR was optimized based on singleplex RT-PCR amplification conditions, using single factor analysis. Forth-eight samples were tested separately with multiplex RT-PCR. The results showed that both assays run to consistent results. The optimized multiplex RT-PCR system had certain accuracy and stability, and could be used for quick detection, pathogen identification and positive screening of WMV ( Watermelon mosaic virus), CMV ( Cucumber mosaic virus), and ZYMV (Zucchini yellow mosaic virus). The distribution status and infection form of three kinds of viruses was determined in main melon growing area of Xinjiang, providing theoretical foundation and experimental evidence for virus diseases control, field testing, epidemiological investigation and melon virus-resistant breeding in Xinjiang.展开更多
基金Supported by Research on Cultivating WMV-resistant Hami Melons using RNA Interference Technology(xjnkq-2013033)Research on Cultivating Antiviral Hami Melons using RNA Interference Technology(YB201307)Integration and Application of Systematical Prevention and Control Technology against Main Diseases in Xinjiang Melon(SY12007)
文摘Using homologous cloning method, partial fragments of coat protein (CP) gene of WMV, CMV and ZYMV were cloned from virus-infected melon in Xinjiang. The reaction system of multiplex RT-PCR was optimized based on singleplex RT-PCR amplification conditions, using single factor analysis. Forth-eight samples were tested separately with multiplex RT-PCR. The results showed that both assays run to consistent results. The optimized multiplex RT-PCR system had certain accuracy and stability, and could be used for quick detection, pathogen identification and positive screening of WMV ( Watermelon mosaic virus), CMV ( Cucumber mosaic virus), and ZYMV (Zucchini yellow mosaic virus). The distribution status and infection form of three kinds of viruses was determined in main melon growing area of Xinjiang, providing theoretical foundation and experimental evidence for virus diseases control, field testing, epidemiological investigation and melon virus-resistant breeding in Xinjiang.