Isozyme variation at six enzyme loci has been studied involving nine members ofDrosophila nasuta subgroup, by employing polyacrylamide gel electrophoretic technique. Alleles at three loci namely Acph, Aph and α-Est are found to be highly polymorphic; whereas at β-Est locus the alleles are less variable while at α-Gpdh and To loci, the alleles are found to be least variable. Null alleles are encountered in low frequencies at α-Est, β-Est and α-Gpdh enzyme loci. The allelic frequencies obtained at the six enzyme loci have been utilised as a yardstick to measure the extent of genetic relationship between the species studied. The genetic identity and genetic distance between these closely-related species have been estimated by adopting the Nei’s formula. These findings have been discussed with reference to earlier cytogenetic and hybridisation studies made on this subgroup.