Wu Rui, Wu Gentu, He Muyao, Zhang Haolan, Shen Xinyi, Huang Qian, Li Jiying, Wu Menglin, Ghanem Hussein, Li Mingjun, Qing Ling
Chongqing Key Laboratory of Plant Disease Biology, College of Plant Protection, Southwest University, Chongqing, 400715, China.
National Citrus Engineering Research Center, Southwest University, Chongqing, 400712, China.
New Phytol. 2025 Mar;245(5):2170-2185. doi: 10.1111/nph.20359. Epub 2024 Dec 25.
Virus-derived small interfering RNAs (vsiRNAs) play an important role in viral infection by regulating the expression of host genes. At present, research on the regulation of plant primary metabolic pathways by vsiRNAs is very limited. TvsiRNA24 derived from tobacco curly shoot virus (TbCSV) was amplified by reverse transcription polymerase chain reaction, and its target gene NbTPI (triosephosphate isomerase) was verified using reverse transcription quantitative polymerase chain reaction and GFP fluorescence observation. The effect of the interaction between TvsiRNA24 and NbTPI on TbCSV infection was analyzed by virus mediated, genetic transformation, western blotting, and quantitative detection. The expression of TvsiRNA24 retards the growth of Nicotiana benthamiana and enhances TbCSV accumulation within N. benthamiana. The overexpression of NbTPI attenuates the accumulation of TbCSV, and the silencing of NbTPI leads to the growth retardation of N. benthamiana and intensifies symptoms post-TbCSV infection. Moreover, the expression of some genes related to photosynthesis, primary metabolism and immune response is regulated by NbTPI. Our results unveil the specific role of TvsiRNA24-NbTPI in the pathogenicity of TbCSV, resulting in hindrance to plant growth and facilitation of viral infection. The identification of this regulatory pathway provides valuable insights that can be utilized to devise novel antiviral approaches targeting the reduction of viral pathogenicity.
病毒衍生的小干扰RNA（vsiRNA）通过调节宿主基因的表达在病毒感染中发挥重要作用。目前，关于vsiRNA对植物初级代谢途径调控的研究非常有限。通过逆转录聚合酶链反应扩增了源自烟草曲茎病毒（TbCSV）的TvsiRNA24，并使用逆转录定量聚合酶链反应和绿色荧光蛋白（GFP）荧光观察验证了其靶基因NbTPI（磷酸丙糖异构酶）。通过病毒介导、遗传转化、蛋白质免疫印迹和定量检测分析了TvsiRNA24与NbTPI之间的相互作用对TbCSV感染的影响。TvsiRNA24的表达会延缓本氏烟草的生长，并增强TbCSV在本氏烟草中的积累。NbTPI的过表达会减弱TbCSV的积累，而NbTPI的沉默会导致本氏烟草生长迟缓，并加剧TbCSV感染后的症状。此外，一些与光合作用、初级代谢和免疫反应相关的基因表达受NbTPI调控。我们的结果揭示了TvsiRNA24-NbTPI在TbCSV致病性中的特定作用，导致植物生长受阻并促进病毒感染。对这一调控途径鉴定提供了有价值的见解，可用于设计针对降低病毒致病性的新型抗病毒方法。
