Yellow vein mosaic disease symptoms occur frequently in pumpkin in India. Diseased plants show vein yellowing, which sometimes coalesces to form chlorotic patches. Infected plants are stunted and flowers drop prematurely, greatly reducing yields. Diseased plants are infected by a begomovirus, designated pumpkin yellow vein mosaic virus (PYVMV), which is transmitted readily and in a persistent manner by the whitefly, Bemisia tabaci. Transmission of PYVMV requires minimum acquisition and inoculation access periods of 30 min and 10 min, respectively. The minimum latent period in the insect is 6 h and the virus persists in the vector for at least 8 days. PYVMV has a narrow host range consisting of a small number of cucurbit species and some tobacco cultivars. It was detected serologically in diseased plants and in viruliferous B. tabaci using polyclonal antibodies in a double-antibody sandwich enzyme-linked immunosorbent assay. Reactions with monoclonal antibodies in a triple-antibody sandwich ELISA showed that PYVMV has an epitope profile distinct from those of other begomoviruses from the Indian sub-continent. Polymerase chain reaction amplified fragments from the putative viral coat and movement protein genes. Based on comparative phylogeny of complete coat protein gene sequences, PYVMV was most similar to the bipartite Tomato leaf curl New Delhi virus from India and appears to be a new strain of this virus.