Jiang Yaoyao, Zheng Weiran, Li Jing, Liu Peng, Zhong Kaili, Jin Peng, Xu Miaoze, Yang Jian, Chen Jianping
College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou, China.
State Key Laboratory for Managing Biotic and Chemical Treats to the Quality and Safety of Agro-Products, Institute of Plant Virology, Ningbo University, Ningbo, China.
Front Plant Sci. 2021 Jan 15;11:603518. doi: 10.3389/fpls.2020.603518. eCollection 2020.
WRKY transcription factors play important roles in plants, including responses to stress; however, our understanding of the function of genes in plant responses to viral infection remains limited. In this study, we investigate the role of in resistance to tomato mosaic virus (ToMV). is significantly downregulated by ToMV infection, and subcellular localization analysis indicates that NbWRKY40 is targeted to the nucleus. In addition, NbWRKY40 activates W-box-dependent transcription in plants and shows transcriptional activation in yeast cells. Overexpressing (OEWRKY40) inhibits ToMV infection, whereas silencing confers susceptibility. The level of salicylic acid (SA) is significantly higher in OEWRKY40 plants compared with that of wild-type plants. In addition, transcript levels of the SA-biosynthesis gene () and SA-signaling genes ( and ) are dramatically higher in OEWRKY40 plants than in the control but lower in -silenced plants than in the control. Furthermore, electrophoretic mobility shift assays show that NbWRKY40 can bind the W-box element of . Callose staining reveals that the plasmodesmata is decreased in OEWRKY40 plants but increased in -silenced plants. Exogenous application of SA also reduces viral accumulation in -silenced plants infected with ToMV. RT-qPCR indicates that NbWRKY40 does not affect the replication of ToMV in protoplasts. Collectively, our findings suggest that NbWRKY40 likely regulates anti-ToMV resistance by regulating the expression of SA, resulting in the deposition of callose at the neck of plasmodesmata, which inhibits viral movement.
WRKY转录因子在植物中发挥着重要作用,包括对胁迫的响应;然而,我们对植物基因在应对病毒感染时功能的了解仍然有限。在本研究中,我们调查了NbWRKY40在番茄对番茄花叶病毒(ToMV)抗性中的作用。ToMV感染显著下调了NbWRKY40的表达,亚细胞定位分析表明NbWRKY40定位于细胞核。此外,NbWRKY40在植物中激活W-box依赖的转录,并在酵母细胞中表现出转录激活。过表达NbWRKY40(OENbWRKY40)抑制ToMV感染,而NbWRKY40沉默则导致感病。与野生型植物相比,OENbWRKY40植物中水杨酸(SA)水平显著更高。此外,SA生物合成基因(ICS1)和SA信号基因(NPR1和PR1)的转录水平在OENbWRKY40植物中比对照显著更高,但在NbWRKY40沉默植物中比对照更低。此外,电泳迁移率变动分析表明NbWRKY40可以结合ICS1的W-box元件。胼胝质染色显示,OENbWRKY40植物中胞间连丝减少,而NbWRKY40沉默植物中胞间连丝增加。外源施用SA也减少了感染ToMV的NbWRKY40沉默植物中的病毒积累。RT-qPCR表明NbWRKY40不影响ToMV在原生质体中的复制。总体而言,我们的研究结果表明,NbWRKY40可能通过调节SA的表达来调控对ToMV的抗性,导致胼胝质在胞间连丝颈部沉积,从而抑制病毒移动。