AbstractThe host immunosuppression by parasitic wasps is an important component of the host regulation strategy. The venom injected at the oviposition is one of the key‐factors involved in this host alteration and, in some parasitoids, its immunosuppressive role is complemented by wasp's symbionts. Most studies in this research area are related to hosts belonging to Lepidoptera and Diptera, for which a strong immune response is observed, whereas little is known for hemimetabolous host species, characterized by apparently much weaker defense barriers. To fill this research gap, here we focus on the host–parasitoid system Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae) – Aphidius ervi Haliday (Hymenoptera: Braconidae). We functionally characterized a serine protease homolog (AeSPH) protein in vivo, identified in the venom of the aphid endoparasitoid A. ervi, generating AeSPH‐depleted female wasps by RNA interference and evaluating their capacity to successfully parasitize the host. Parasitism success rate was negatively affected by AeSPH knockdown and associated with an increased phenoloxidase (PO) cascade activation in aphids, scored by measuring PO enzymatic activity and the expression of phenoloxidase activating factor 2, a proPO‐activating gene upregulated in response to A. ervi parasitism. Our results indicate that AeSPH contributes to parasitism success by inhibiting the melanization response of the host, which is therefore an important component of the defense barriers involved in the parasitoid egg suppression. The ongoing studies on other virulence factors in A. ervi venom will allow to further characterize the immunosuppression strategy and its possible broader role in the host regulation through its action on aphid symbiont development.