State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming 650201, China.
Modern Education Technology Center, Yunnan Agricultural University, Kunming 650201, China.
J Virol Methods. 2018 Sep;259:122-128. doi: 10.1016/j.jviromet.2018.06.012. Epub 2018 Jun 23.
A generic RT-PCR assay was developed for the universal detection of viruses of the genus Tobamovirus using a novel pair of degenerate primers designed based on conserved regions on replicase genes of 32 tobamoviruses. The assay detected nine tobamoviruses, including six Solanaceae-infecting subgroup tobamoviruses of Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Tomato mottle mosaic virus (ToMMV), Tobacco mottle green mosaic virus (TMGMV), Pepper mild mottle virus (PMMoV), Paprika mild mottle virus (PaMMV), one Orchidaceae-infecting tobamovirus of Odontoglossum ringspot virus (ORSV) and two Cucurbitaceae-infecting subgroup tobamoviruses of Cucumber green mottle mosaic virus (CGMMV) and Zucchini green mottle mosaic virus (ZGMMV), with high amplification efficiency, specificity and sensitivity. The assay was applied to detect tobamoviruses in pepper and tomato fields. Five tobamoviruses, PMMoV, TMV, ToMV, ToMMV and TMGMV, were detected from the pepper fields in single and mixed infections. Single infections of PMMoV, ToMV and ToMMV and mix-infection of ToMV + PMMoV were detected from the tomato fields. Among these viruses, PMMoV was first detected from tomato worldwide, while ToMMV was first detected from tomato plants in China. This generic assay is simple, cost-effective and has great potential to detect more tobamoviruses in the field.
一种通用的 RT-PCR 检测方法被开发出来，用于使用基于 32 种烟草花叶病毒复制酶基因保守区设计的一对新型简并引物，对烟草花叶病毒属病毒进行通用检测。该检测方法检测到 9 种烟草花叶病毒，包括 6 种茄科感染亚组烟草花叶病毒（TMV）、番茄花叶病毒（ToMV）、番茄斑驳花叶病毒（ToMMV）、烟草斑驳绿花叶病毒（TMGMV）、辣椒轻斑驳病毒（PMMoV）、辣椒斑驳病毒（PaMMV），一种兰科感染的 Odontoglossum 环斑病毒（ORSV）和两种葫芦科感染亚组的黄瓜绿斑驳花叶病毒（CGMMV）和南瓜绿斑驳花叶病毒（ZGMMV），具有高效的扩增效率、特异性和灵敏度。该检测方法应用于检测辣椒和番茄田中烟草花叶病毒。从辣椒田中检测到 5 种烟草花叶病毒，PMMoV、TMV、ToMV、ToMMV 和 TMGMV，存在单一和混合感染。从番茄田中检测到 PMMoV、ToMV 和 ToMMV 的单一感染和 ToMV+PMMoV 的混合感染。在这些病毒中，PMMoV 是首次在全球范围内从番茄中检测到的，而 ToMMV 是首次在中国从番茄植株中检测到的。这种通用的检测方法简单、经济高效，具有在田间检测更多烟草花叶病毒的巨大潜力。
