During a field survey at the Indian Agricultural Research Institute, New Delhi, the bitter gourd ( L.) cv. Pusa Do Mausami showed prominent symptoms characterized by severe yellow mosaic and leaf curling. Through leaf-dip transmission electron microscopy and PCR assays, the presence of bhendi yellow vein mosaic virus, BYVMV () was confirmed. Sequencing of Pusa Do Mausami (PDM-1, PDM-2) isolates (OR712889, OR712890) revealed 97% similarity to an Indian BYVMV isolate. Comparative analysis revealed a close genetic relationship (92-97%) between the PDM BYVMV isolates and other strains found in vegetable crops from different countries. The Ramachandran plot analysis confirmed that the predicted βC1 region of BYVMV exhibits a structurally reliable conformation, with most residues occupying favoured stereochemical regions. Structural analysis of the βC1 region of the BYVMV revealed moderate model reliability for the sequence OR712889 (GMQE: 0.47, QMEANDisCo Global: 0.46 ± 0.12). Motif analysis of the βC1 region of the BYVMV revealed critical functional domains, including the Geminivirus movement protein domain, that are essential for viral movement and systemic infection. Phylogenetic analysis revealed distinct evolutionary relationships, with isolates (OR712889, OR712890 respectively) clustering with related strains globally, particularly with a bitter gourd yellow mosaic virus, BgYMV () isolate from India, which was supported by a strong 94% bootstrap confidence level. Biological validation revealed successful BYVMV transmission to okra via whiteflies, with 100% infection in 14-day-old seedlings, resembling natural symptoms. The present findings are the first to reveal the association of BYVMV with bitter gourd in India, highlighting its genetic similarities and evolutionary links. These results emphasize the need for effective disease management to increase crop productivity.