Tomato (Solanum lycopersicum L.) is a globally significant vegetable crop, with India ranking second in production. However, tomato cultivation is prone to several challenges, most notably, the tomato leaf curl virus (ToLCV) disease, causing severe yield losses. Thisstudy was focused on validation of Ty-2gene among 14 tomato genotypes. DNA was extracted and PCR amplification using the TES0344 marker associated with the Ty-2 gene was performed. Gel electrophoresis revealed distinct patterns: genotypes SL159, SL160, SL165, SL167, SL168, and SL169 exhibited a 190 bp band, indicating the presence of the Ty-2 resistance allele. In contrast, genotypes SL113, SL115, SL124, SL142, SL144, SL147, SL149, and SL154 displayed a 205 bp band, signifying susceptible alleles. Using genotypes with Ty-2 genotypes in breeding can boost tomato crop resilience against ToLCV, aiding agricultural sustainability and food security.