rDNA internal transcribed spacer (ITS) region from V. mungo var silvestris, Vigna trilobata, Vigna glabrescens and diverse cultivars of Vigna mungo, were amplified and digested with twelve restriction enzymes. There was no size ariation in the ITS region of the diverse cultivars and other Vigna species studied. The approximate length of the amplified product of the entire rDNA ITS region was found to be 650 bp. ITS1 consisted of 250 bp and ITS2 was 300 bp long. Restriction fragment length polymorphisms within species could not be detected in cultivated accessions of V. mungo for all the restriction enzymes tested whereas interspecific variation was found among V. mungo var silvestris, V. trilobataand V. glabrescens.Of the eleven restriction enzymes (EcoRI, HindIII, PstI, SmaI, Sau3AI, TaqI, SacI, MspI, AluI, BamHI and HaeIII) tested, seven endonucleases (Sau3AI, TaqI, SacI, MspI, AluI, BamHI and HaeIII) had restriction site in the ITS region, of which five were in ITS1 and two in ITS2 of cultivated species. Presence of BamHI restriction site which is unique to Vigna was not found to be methylated. The ITS2 of V. glabrescens and V. trilobata had restriction sites for Sau3AI and AluI, which are not found in V. mungo var silvestris and the cultivated varieties studied. MspI enzyme had restriction site specifically present in ITS2 of V. trilobata. No intraspecific variation as observed among widely distributed Indian cultivars of V. mungo and V. mungo var silvestris.