Coccinia grandisis an interesting model system to understand dioecy in Cucurbitaceae family. Recent transcriptomics and proteomics studies carried out to understand the sex expression inC. grandishave resulted in identification of many candidate sex-biased genes. In absence of an efficient genetic transformation protocol forC. grandis, virus-induced gene silencing (VIGS) would be a powerful tool to enable gene functional analysis. In current study, we explored the apple latent spherical virus (ALSV) for gene knockdown inC. grandis.The viral infection was achieved through mechanical inoculation of ALSV-infectedChenopodium quinoaleaf extract onto the cotyledons ofC. grandis. ALSV-VIGS mediated knockdown ofCgPDSgene was successfully achieved inC. grandisby mechanical inoculation method resulting in characteristic photobleaching. Subsequently, we developed agroinfiltration compatible vectors for direct infection ofC. grandisand shortened the time-frame by skipping viral propagation inC. quinoa. Typical yellow-leaf phenotype was observed inC. grandisplants agroinfiltrated with ALSV-CgSUconstructs, indicating robust silencing ofCgSUgene. In addition, we improved the infection efficiency of ALSV by co-infiltration of P19 viral silencing suppressor. These results suggest that ALSV-VIGS is suitable for characterization of gene function in dioeciousC. grandisand it can help us understand the mechanism of sex expression.