Kenaf (Hibiscus cannabinus L.) is a malvaceous bast fibre crop and the most promising source of "tree-free" newsprint in the world. For the last few years a begomovirus associated leaf curl disease of kenaf, characterized by upward leaf curling and stunted growth, has been observed at different locations in India. In this study biological and molecular analyses of kenaf leaf curl disease-associated begomovirus isolates collected from three different locations were carried out. Whitefly transmission efficiency was different for the three geographical isolates which possessed very narrow host ranges. Except kenaf, the isolates from northern India could not infect other experimental hosts studied. Cloning and sequencing of the complete genomes of these isolates revealed the presence of three different monopartite begomoviruses. In eastern and southern India Mesta yellow vein mosaic virus (MeYVMV) and Tomato leaf curl Joydebpur virus (ToLCJV) were associated with the disease, respectively, while in northern India a new begomovirus species was involved, tentatively named Kenaf leaf curl virus (KLCuV). Nucleic acid spot hybridization tests using non-radiolabeled probes developed against these viruses detected the presence of begomoviruses from symptomatic samples collected from the fields at the different locations.