N6-Methyladenosine Modification Regulates Prunus Necrotic Ringspot Virus Infection in <i>Cucumis sativus</i>

N6-methyladenosine (m⁶A) is a common epigenetic modification found in eukaryotic RNA. Recent studies have increasingly highlighted the importance of m⁶A modification in plant defense against viruses. In this investigation, we found that prunus necrotic ringspot virus (PNRSV) infection affected the m⁶A modification process. Plant transcriptomes and epitranscriptomes were sequenced to coanalyze the dynamic changes of m⁶A modifications after PNRSV infection. Further studies revealed that the silencing of evolutionarily conserved C-terminal region (ECTs), encoding m⁶A readers, led to increased PNRSV accumulation, indicating that ECTs confer resistance to PNRSV. Additionally, we demonstrated that <i>UPF3</i> and <i>SMG7</i>, which are involved in non-sense-mediated mRNA decay pathways, as well as phenylalanine ammonia-lyase (PAL), a well-known key enzyme in plant defense and an identified m⁶A-modified gene following PNRSV infection, play crucial roles in regulating PNRSV infection. These findings provide new insights into understanding PNRSV infection and further elucidate the role of m⁶A in modulating viral infection in plants.