The Asian rice gall midge, Orseolia oryzae (Wood-Mason) (Diptera: Cecidomyiidae), is a major pest of rice [Oryza sativa L. (Poaceae)] in India. Breeding resistant varieties and their cultivation has been the main approach to manage this pest. However, the breakdown of resistance conferred by the major genes, deployed one at a time, through evolution of virulent biotypes has become a major setback to this approach. Development of polymerase chain reaction-based molecular markers for eight of the 10 resistance genes and their possible use in marker-assisted selection has enabled breeders to pyramid resistance genes for achieving durable resistance. However, the choice of resistance genes needs to be made with a better understanding of the virulence composition of the pest populations in the target area and the genetics of plant resistance and insect virulence, as the rice–gall midge interaction is a gene-for-gene one. We adopted a single-female test and coupled it with a modified F2 screen test to note the virulence composition of gall midge populations and estimated the frequency of virulence alleles for adaptation at three pest endemic locations in India, namely, Warangal, Ragolu, and Raipur. Results on biotype composition showed heterogeneous pest populations in all the tests and at all the locations. Tests at Warangal repeated after 8 years showed a rapid increase in frequency of the virulence allele conferring adaptation to the plant resistance gene Gm2 as compared to that of the allele for adaptation to the resistance gene Gm1. This is probably the first direct measurement of a durability parameter of plant genes conferring insect resistance. Results supported earlier observations that sex-linked virulence against Gm2 makes it less durable. The sex ratio did not deviate from the expected 1:1 ratio at Warangal, but at Ragolu females outnumbered males.