Li Zhenggang, Du Zhenguo, Tang Yafei, She Xiaoman, Wang Xiaomei, Zhu Yanhua, Yu Lin, Lan Guobing, He Zifu
Plant Protection Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, China.
Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Guangdong Academy of Agricultural Sciences, Guangzhou, China.
Front Microbiol. 2020 May 5;11:851. doi: 10.3389/fmicb.2020.00851. eCollection 2020.
(ToLCGdV) is a begomovirus associated with a Tomato yellow leaf curl disease (TYLCD) epidemic in Guangdong province, China. Being the least conserved protein among geminivirus proteins, the function of C4 during ToLCGdV infection has not been elucidated. In this study, the infectious clones of ToLCGdV and a ToLCGdV mutant (ToLCGdV) with disrupted C4 ORF were constructed. Although ToLCGdV and ToLCGdV could infect and tomato plants, ToLCGdV elicited much milder symptoms compared with ToLCGdV. To further verify the role of C4 in viral pathogenesis, C4 was expressed in from (PVX) vector. The results showed that ToLCGdV C4 enhanced the pathogenicity of PVX and induced more severe developmental abnormalities in plants compared with PVX alone or PVX-mC4. In addition, ToLCGdV C4 suppresses systemic gene silencing in the transgenic line 16c, but not local gene silencing induced by sense GFP in wild-type plants. Moreover, C4 suppresses transcriptional gene silencing (TGS) by reducing the DNA methylation level of 35S promoter in 16c-TGS plants. Furthermore, C4 could also interact with the receptor-like kinase (RLK) BARELY ANY MERISTEM 1 (BAM1), suggesting that C4 may suppress gene silencing by interfering with the function of BAM1 in the cell-to-cell spread of RNAi. All these results suggest that C4 is a pathogenic determinant of ToLCGdV, and C4 may suppress post-transcriptional gene silencing (PTGS) by interacting with BAM1.
番茄黄化曲叶广东病毒(ToLCGdV)是一种与中国广东省番茄黄化曲叶病(TYLCD)流行相关的双生病毒。作为双生病毒蛋白中保守性最低的蛋白,C4在ToLCGdV侵染过程中的功能尚未阐明。在本研究中,构建了ToLCGdV及其C4开放阅读框(ORF)被破坏的ToLCGdV突变体(ToLCGdV)的感染性克隆。尽管ToLCGdV和ToLCGdV能够感染烟草和番茄植株,但与ToLCGdV相比,ToLCGdV引发的症状要轻得多。为了进一步验证C4在病毒致病过程中的作用,通过马铃薯X病毒(PVX)载体在烟草中表达C4。结果表明,与单独的PVX或PVX-mC4相比,ToLCGdV C4增强了PVX的致病性,并在植物中诱导了更严重的发育异常。此外,ToLCGdV C4抑制转基因烟草品系16c中的系统基因沉默,但不抑制野生型烟草植株中由正义绿色荧光蛋白(GFP)诱导的局部基因沉默。此外,C4通过降低16c-TGS烟草植株中35S启动子的DNA甲基化水平来抑制转录基因沉默(TGS)。此外,C4还可以与类受体激酶(RLK)几乎无分生组织1(BAM1)相互作用,这表明C4可能通过干扰BAM1在RNA干扰细胞间传播中的功能来抑制基因沉默。所有这些结果表明,C4是ToLCGdV的致病决定因素,并且C4可能通过与BAM1相互作用来抑制转录后基因沉默(PTGS)。