The insulin and 20‐hydroxyecdysone (20E) pathways coordinately regulate insect vitellogenesis and ovarian development. However, the detailed molecular mechanisms such as the genes mediating the cooperation of the interaction of these 2 pathways in regulating insect reproductive development are not well understood. In the present study, a small GTPase, Rab40C, was identified from the notorious agricultural pest Bactrocera dorsalis. In addition to the well‐known RAB domain, it also has a unique SOCS‐box domain, which is different from other Rab‐GTPases. Moreover, we found that Rab40C was enriched in the ovaries of sexually mature females. RNA interference (RNAi)‐mediated knockdown of BdRab40C resulted in a decrease in vitellogenin synthesis, underdeveloped ovaries, and low fertility. Furthermore, depletion of insulin receptor InR or the heterodimer receptor of 20E (EcR or USP) by RNAi significantly decreased the transcription of BdRab40C and resulted in lower fecundity. Further studies revealed that the transcription of BdRab40C could be upregulated by the injection of insulin or 20E. These results indicate that Rab40C participates in the insulin and 20E pathways to coordinately regulate reproduction in B. dorsalis. Our results not only provide new insights into the insulin‐ and 20E‐stimulated regulatory pathways controlling female reproduction in insects but also contribute to the development of potential eco‐friendly strategies for pest control.

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