In addition to its functions in antiviral RNA silencing, dsRNA elicits pattern-triggered immunity (PTI), contributing to plant resistance against virus infections. However, the mode-of-action and signalling pathway of dsRNA-induced defence remain poorly characterised. Using multicolor in vivo imaging, analysis of GFP mobility, callose staining, and plasmodesmal marker lines in Arabidopsis thaliana and Nicotiana benthamiana, we show that dsRNA-induced PTI restricts the progression of virus infection by triggering callose deposition at plasmodesmata, thereby limiting the macromolecular transport through these cell-to-cell communication channels. The dsRNA-induced signalling leading to callose deposition at plasmodesmata involves plasma membrane-resident SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE 1, the BOTRYTIS INDUCED KINASE1/AVRPPHB SUSCEPTIBLE1-LIKE KINASE1 kinase module, PLASMODESMATA-LOCATED PROTEINs 1/2/3, as well as CALMODULIN-LIKE 41 and Ca2+ signals. Interestingly, unlike the well-known PTI elicitor flg22, dsRNA does not trigger a detectable reactive oxygen species (ROS) burst. Likely as a counter strategy, viral movement proteins from different viruses suppress the dsRNA-induced host response leading to callose deposition to achieve infection. Plants infected with tobacco mosaic virus show callose levels at plasmodesmata specifically reduced at the infection front where viral movement protein is expressed and known to modify the function of plasmodesmata. Thus, our data support a model in which plant immune signaling constrains virus movement by inducing callose deposition at plasmodesmata and how viruses counteract this layer of immunity. Importantly, the results maay challenge the classical view that viral movement proteins “open” plasmodesmata. Rather, these proteins act as viral effectors that prevent plasmodesmata closure by interfering with a dsRNA-induced immunity response.