<p>Tomato leaf curl New Delhi virus (ToLCNDV) is the most destructive whitefly transmitted “single-stranded DNA begomovirus” causing severe economic loss to cucumber (<i>Cucumis sativus</i> L.) production worldwide. In the present study we have molecularly characterized ToLCNDV infecting cucumber showing symptoms of chlorotic spots, interveinal chlorosis and severe leaf curling. Molecular confirmation of the virus was achieved through sequencing (GenBank accession no. PQ824188). These infected plants were subsequently used as the source of inoculum for artificial screening. Screening of diverse parthenocarpic, gynoecious and monoecious lines of cucumber using whitefly (<i>Bemisia tabaci</i> biotype Asia II) mediated artificial inoculation confirmed the line ‘PCG-7’ was highly resistant against identified ToLCNDV strain. To elucidate the genetic basis of resistance against ToLCNDV, F₁ (including reciprocal cross) and F₂ (segregating) populations derived from a cross between the highly susceptible genotype ‘PBRK-11’ and the highly resistant genotype ‘PCG-7’ were mass inoculated with ToLCNDV using viruliferous whiteflies under artificial inoculation conditions in 2024. Disease scoring of ToLCNDV at different days of post inoculation (dpi) rendered F₁ and its reciprocal cross progenies susceptible, indicating recessive nature of resistance. Chi-square analysis of F₂ population of cross combination “PBRK-11 X PCG-7” showed a segregation ratio of 12(Susceptible): 3(Resistant): 1(Highly Resistant), suggesting digenic recessive inheritance with dominant epistatis for resistance against ToLCNDV in cucumber.</p> Graphical abstract <p></p>