This study investigates the antiviral potential of Ipomoea nil leaf extract in Nicotiana benthamiana plants against the turnip mosaic virus. I. nil, a plant species rich in resin glycosides, which are known for their metabolic, anticancer, and antimicrobial effects. Ipomoea nil is a member of the Convolvulaceae family along with Operculina turpethum, whose extract was demonstrated to exhibit broad-spectrum antiviral properties. Preliminary observations of a restricted Virus-Induced Gene Silencing phenotype to silence the phytoene desaturase gene in Ipomoea nil plants motivated this study of I. nil’s antiviral potential. The primary objectives of this study were to assess the antiviral potential of Ipomoea nil leaf extract by comparing viral titer in extract-treated and non-treated N. benthamiana plants and investigate the impact of the extract on the induction of defense pathways in the N. benthamiana plants. The study also compared the effects of I. nil leaf extract with the N. benthamiana leaf extract to determine the specificity of I. nil leaf extract. Previous results also suggest that the expression of pathogenesis-related protein 1 is increased and light-harvesting chlorophyll a/b complex protein 3 is downregulated as a defense mechanism in N. benthamiana plants. Therefore, we specifically investigated the expression of these two genes in response to I. nil and N. benthamiana leaf extracts. By elucidating the antiviral mechanisms of I. nil, this research contributes to the understanding of plant-virus interactions and the development of novel antiviral strategies.