Zhao Ming-Min, An De-Rong, Huang Guang-Hua, He Zu-Hua, Chen Jiang-Ye
College of Plant Protection, North West Science and Technology University of Agriculture and Forestry, Shaanxi 712100, China.
Acta Biochim Biophys Sin (Shanghai). 2005 Apr;37(4):248-53. doi: 10.1111/j.1745-7270.2005.00036.x.
Plant viruses encode suppressors of post-transcriptional gene silencing (PTGS), an adaptive defense response that limits virus replication and its spread in plants. The helper component proteinase (HC-Pro) of the potato virus A (PVA, genus Potyvirus) suppresses PTGS of silenced transgenes. Here, the effect of HC-Pro on siRNA-directed interference in the tobacco mosaic virus (TMV) was examined by using a transient Agrobacterium tumefaciens-based delivery system in intact tissues. It was shown that the interference effect was completely blocked by co-infiltration with HC-Pro plus siRNA constructs in both systemic and hypersensitive hosts. In the system host, all plants agro-infiltrated with HC-Pro plus siRNA constructs displayed the same symptoms as the negative control. Meanwhile, TMV RNA accumulation was found to be abundant in the upper leaves using reverse transcriptase-PCR (RT-PCR) and Northern blot assays. On the contrary, plants agro-infiltrated with the siRNA construct alone were free of symptoms. Therefore, our study suggests that the transient expression of HC-Pro inhibited the siRNA-directed host defenses against TMV infection.
植物病毒编码转录后基因沉默(PTGS)抑制子,PTGS是一种适应性防御反应,可限制病毒在植物中的复制及其传播。马铃薯A病毒(PVA,马铃薯Y病毒属)的辅助成分蛋白酶(HC-Pro)可抑制沉默转基因的PTGS。在此,通过在完整组织中使用基于根癌农杆菌的瞬时递送系统,研究了HC-Pro对烟草花叶病毒(TMV)中siRNA介导的干扰的影响。结果表明,在系统性宿主和过敏反应宿主中,HC-Pro与siRNA构建体共浸润可完全阻断干扰效应。在系统宿主中,所有用HC-Pro加siRNA构建体进行农杆菌浸润的植物都表现出与阴性对照相同的症状。同时,使用逆转录聚合酶链反应(RT-PCR)和Northern印迹分析发现,TMV RNA在上部叶片中大量积累。相反,仅用siRNA构建体进行农杆菌浸润的植物没有症状。因此,我们的研究表明,HC-Pro的瞬时表达抑制了siRNA介导的宿主对TMV感染的防御。