Zheng Luping, Du Zhenguo, Lin Chen, Mao Qianzhuo, Wu Kangcheng, Wu Jianguo, Wei Taiyun, Wu Zujian, Xie Lianhui
Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.
Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Plant Protection Research Institute, GAAS, Guangzhou, 510640, China.
Mol Plant Pathol. 2015 Dec;16(9):921-30. doi: 10.1111/mpp.12220. Epub 2015 Mar 16.
Rice stripe virus (RSV) is the type species of the genus Tenuivirus and represents a major viral pathogen affecting rice production in East Asia. In this study, RSV p2 was fused to yellow fluorescent protein (p2-YFP) and expressed in epidermal cells of Nicotiana benthamiana. p2-YFP fluorescence was found to move to the nucleolus initially, but to leave the nucleolus for the cytoplasm forming numerous distinct bright spots there at later time points. A bimolecular fluorescence complementation (BiFC) assay showed that p2 interacted with fibrillarin and that the interaction occurred in the nucleus. Both the nucleolar localization and cytoplasmic distribution of p2-YFP fluorescence were affected in fibrillarin-silenced N. benthamiana. Fibrillarin depletion abolished the systemic movement of RSV, but not that of Tobacco mosaic virus (TMV) and Potato virus X (PVX). A Tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) method was used to diminish RSV NS2 (encoding p2) or NS3 (encoding p3) during RSV infection. Silencing of NS3 alleviated symptom severity and reduced RSV accumulation, but had no obvious effects on virus movement and the timing of symptom development. However, silencing of NS2 abolished the systemic movement of RSV. The possibility that RSV p2 may recruit or manipulate nucleolar functions to promote virus systemic infection is discussed.
水稻条纹病毒(RSV)是纤细病毒属的代表种,是影响东亚水稻生产的主要病毒病原体。在本研究中,RSV p2与黄色荧光蛋白融合(p2-YFP)并在本氏烟草的表皮细胞中表达。发现p2-YFP荧光最初移动到核仁,但在随后的时间点离开核仁进入细胞质,在那里形成许多明显的亮点。双分子荧光互补(BiFC)分析表明p2与纤维蛋白原相互作用,且这种相互作用发生在细胞核中。在纤维蛋白原沉默的本氏烟草中,p2-YFP荧光的核仁定位和细胞质分布均受到影响。纤维蛋白原缺失消除了RSV的系统移动,但不影响烟草花叶病毒(TMV)和马铃薯X病毒(PVX)的系统移动。在RSV感染期间,使用基于烟草脆裂病毒(TRV)的病毒诱导基因沉默(VIGS)方法来减少RSV NS2(编码p2)或NS3(编码p3)。沉默NS3减轻了症状严重程度并减少了RSV积累,但对病毒移动和症状发展时间没有明显影响。然而,沉默NS2消除了RSV的系统移动。本文讨论了RSV p2可能招募或操纵核仁功能以促进病毒系统感染的可能性。