文献类型： 外文期刊
作者： Xu, Y. 1 ; Yang, L. 1 ; Zhou, J. 1 ; Yang, Y. 2 ; Lu, M. 1 ; Li, S. 1 ;
作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China
2.Chinese Acad Trop Agr Sci, Environm & Plant Protect Inst, Haikou, Hainan, Peoples R China
关键词： multiplex RT-PCR; peach; peach-associated luteovirus; peach virus D; nectarine stem-pitting-associated virus; peach
期刊名称：LETTERS IN APPLIED MICROBIOLOGY （ 影响因子：2.858； 五年影响因子：2.776 ）
ISSN： 0266-8254
年卷期： 2019 年 69 卷 5 期
页码：
收录情况： SCI
摘要： Peach is a major crop in China, and like any other stone fruit, virus and virus-like diseases can reduce the yield and quality of the fruit. Herein, we developed a multiplex RT-PCR (mRT-PCR) assay for simultaneously detecting three viruses known to infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). Plant nad5 mRNA was used as the internal control. Field samples that were co-infected with PaLV, PeVD and NSPaV were used; we identified three primer pairs to be the most specific for detecting these viruses, followed by determining the ideal concentration of each primer pair and optimizing the annealing temperature for mRT-PCR. We also assessed the detection limit using serial dilutions of RNA and cDNA. The newly developed mRT-PCR assay could simultaneously detect PaLV, PeVD and NSPaV. To validate the reliability of mRT-PCR for virus detection, mRT-PCR was used to detect viruses in the leaves of 21 peach plants collected in Liaoning Province, China. The obtained results revealed the presence of single and co-infections. To conclude, the mRT-PCR assay developed herein is sensitive, reliable and economical, and we believe that it can thus be used for large-scale surveys of PaLV, PeVD and NSPaV. Significance and Impact of the Study In this study, we developed a multiplex reverse transcriptase PCR (mRT-PCR) assay for simultaneously detecting three viruses that infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). This assay is simple, easy to perform, reliable and cost-effective, and can thus be applied for large-scale surveys of PaLV, PeVD and NSPaV.
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