Symbiotic bacteria and hormesis in aphids are the driving forces for pesticide resistance. However, the mechanism remains unclear. In this study, the effects of imidacloprid on the population growth parameters and symbiotic bacterial communities of three successive generations of Acyrthosiphon gossypii were investigated. The bioassay results showed that imidacloprid had high toxicity to A. gossypii with an LC50 of 1.46 mg·L−1. The fecundity and longevity of the G0 generation of A. gossypii decreased when exposed to the LC15 of imidacloprid. The net reproductive rate (R0), intrinsic rate of increase (rm), finite rate of increase (λ), and total reproductive rate (GRR) of G1 and G2 offspring were significantly increased, but those of the control and G3 offspring were not. In addition, sequencing data showed that the symbiotic bacteria of A. gossypii mainly belonged to Proteobacteria, with a relative abundance of 98.68%. The dominant genera of the symbiotic bacterial community were Buchnera and Arsenophonus. After treatment with the LC15 of imidacloprid, the diversity and species number of bacterial communities of A. gossypii decreased for G1–G3 and the abundance of Candidatus-Hamiltonella decreased, but Buchnera increased. These results provide insight into the resistance mechanism of insecticides and the stress adaptation between symbiotic bacteria and aphids.