Abstract: Plant viruses frequently reprogram conserved growth-defense regulatory hubs to promote infection. Here, we show that the rice grassy stunt virus (RGSV) suppresses salicylic acid (SA)-mediated antiviral immunity by targeting the miR156–SPL–ICS1 module. The viral effector P3 directly binds a conserved 12-bp cis-element in the miR156a promoter, activating its transcription and increasing miR156 accumulation. Increased miR156 represses SPL14 and SPL17 transcripts, while RGSV infection is also associated with a pronounced reduction in SPL14/17 protein abundance. P3 physically associates with SPL14 and SPL17, indicating an additional post-transcriptional layer contributing to SPL attenuation. Genetic analyses demonstrate that SPL14 and SPL17 positively regulate ICS1, a key enzyme in SA biosynthesis, and that loss of SPL14/17 function compromises SA accumulation and antiviral defense. Conversely, overexpression of SPL14 or SPL17 mitigates RGSV symptoms and restricts viral accumulation, whereas exogenous SA restores immunity and partially rescues disease-associated architectural defects. Together, our findings reveal a dual-layer virulence strategy in which RGSV P3 coordinately suppresses the miR156–SPL14/17–ICS1 pathway at transcriptional and post-transcriptional levels, uncovering a central regulatory node that links rice development and antiviral immunity and providing actionable targets for engineering RGSV-resistant rice.

Key words: antiviral immunity, miR156–SPL module, RGSV, rice, salicylic acid