Bacterial endosymbionts play important roles in ecological traits of aphids. In this study, we characterize the bacterial endosymbionts of A. gossypii collected in Karaj, Iran and their role in the performance of the aphid. Our results indicated that beside Buchnera aphidicola, A. gossypii, also harbors both Hamiltonella defensa and Arsenophonus sp. Quantitative PCR (qPCR) results revealed that the populations of the endosymbionts increased throughout nymphal development up to adult emergence; thereafter, populations of Buchnera and Arsenophonus were diminished while the density of H. defensa constantly increased. Buchnera reduction caused prolonged development and no progeny production. Furthermore, secondary symbiont reduction led to reduction of the total life span and intrinsic rate of natural increase as well as appearance of the deformed dead offspring in comparison with the control insects. Reduction of the secondary symbionts did not affect parasitism rate of the aphid by the parasitic wasp Aphidius matricariae. Together these findings showed that H. defensa and Arsenophonus contributed to the fitness of A. gossypii by enhancing its performance, but not through parasitoid resistance.