We have previously shown the deregulation of various miRNAs post Tomato leaf curl New Delhi virus (ToLCNDV) infection. Host miRNAs can potentially bind to viral DNA and/or virus-encoded mRNAs thus acting as hosts’ antiviral defense. The goal of the present study was to investigate the mechanism of action of ToLCNDV encoded RNA silencing suppressor (RSS) AC2 on the host miRNA machinery. Here, we report the possible role of AC2 in miRNA dysregulation and found that AC2 associates with miR319 and miR172 which are deregulated during ToLCNDV infection. We furthermore confirmed this association was mediated by direct interaction of AC2 with AGO1 using in vitro and ex vivo assays. These results provide mechanistic insight on understanding of AC2-mediated deregulation of host miRNA pathway.