Isofemale lines derived from Indian natural populations of Drosophila ananassae, D. takahashii and D. nepalensis were analysed through horizontal starch gel electrophoresis as well as post-electrophoretic heat denaturation technique for genetic variation at acid phosphatase locus. Allozymic data revealed nearly uniform patterns in D. takahashii and D. nepalensis populations while latitudinal variation was observed in D. ananassae. The contrasting patterns of allozymic variation in three Drosophila species concur with the niche-width variation hypothesis.