Abstract
RNA silencing (or RNA interference, RNAi) plays direct roles in plant host defenses against viruses. Viruses encode suppressors of RNAi (VSRs) to counteract host antiviral defenses. The generation of transgenic plants expressing VSRs facilitates the understanding of the mechanisms of VSR-mediated interference with the endogenous silencing pathway. However, studying VSRs independent of other viral components simplifies the complex roles of VSRs during natural viral infection. While suppression of transgene silencing by the VSR 2b protein encoded by cucumber mosaic virus (CMV) requires 2b-small RNA (sRNA) binding activity, suppression of host antiviral defenses requires the binding activity of both sRNAs and AGOs proteins. This study, aimed to understand the functions of 2b in the context of CMV infection; thus, we performed genome-wide analyses of differential DNA methylation regions among wild-type CMV-infected, CMVΔ2b-infected, and 2b-transgenic Arabidopsis plants. These analyses, together with transcriptome sequencing and RT-qPCR analyses, show that while the majority of induced genes in 2b-transgenic plants were involved in extensive metabolic pathways, CMV-infection 2b-dependent induced genes were enriched in plant immunity pathways, including salicylic acid (SA) signaling. Together with infection with CMV mutants that expressed the 2b functional domains of sRNA or AGO binding, our data demonstrate that CMV-accelerated SA signaling depends on 2b-sRNA binding activity which is also responsible for virulence.
Highlights
RNA silencing is an evolutionarily conserved and inducible defense pathway that targets and inactivates invading nucleic acids [1,2,3,4]
To identify the function of 2b protein in inhibiting host DNA methylation during cucumber mosaic virus (CMV) infection, we first performed bisulfite sequencing on CMV- and CMV∆2b-infected plants and identified the DNA methylation regions (DMRs) compared with wild-type Col-0 plants as previously described [31]
The 2b protein encoded by CMV exhibited complex activities, including the suppression of PTGS
Summary
RNA silencing (or RNA interference, RNAi) is an evolutionarily conserved and inducible defense pathway that targets and inactivates invading nucleic acids [1,2,3,4]. RNAi-mediated antiviral plant immunity is triggered by double-stranded RNA (dsRNA) derived from the replication intermediates of RNA viruses, bidirectional transcription of DNA viruses and imperfectly base-paired intermolecular hairpins in viral genomes or transcripts [2,3,4,6,7]. During virus infection, these dsRNAs are processed by the RNase III-type enzyme Dicer-like (DCL). RNA or suppress the transcription of DNA viruses via the RdDM pathway [2,7,8,9,10]. Beyond the limitation of viral RNA accumulation in initially infected cells by processing viral dsRNAs into viRNAs, RNAi-mediated antiviral activity contributes to achieving a robust systemic defense response [2,3]
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