Abstract Phylogenetic reconstruction of Proctotrupinae, the largest subfamily of Proctotrupidae, was conducted for the first time based on total evidence combining molecular and morphological characters. Based on our analysis, we conclude that the genus Nothoserphus, previously belonging to the tribe Cryptoserphini, should be transferred to the tribe Disogmini. New diagnostic characters for each genus were proposed based on the morphological analysis. The results of the analysis showed that the exceptionally hardened ovipositor sheath is a defining character of Proctotrupidae, and it was estimated that its morphological character states were not reflected in the phylogenetic relationship, probably because of its variability related to parasitic strategy and host habitat.