Bumblebees are important pollinators that have evolved between solitary and advanced eusocial insects. Compared with advanced honeybees, workers of social bumblebee species are prone to laying eggs during the competition phase, which leads to the end of the colony. Therefore, worker reproductive behavior has become a popular research topic for exploring various biological phenomena. Here, we demonstrate a novel reproduction-related function of an immune response protein-encoding gene (Immune Responsive Protein 30, IRP30) in Bombus terrestris by employing RNA interference (RNAi) and a transgenic Drosophila melanogaster system. The results show that worker egg-laying was significantly affected by IRP30 expression levels (P < 0.01). Compared with those in the dsGFP-treated groups, the first egg-laying time was delayed by 3.7 d and the egg number was decreased by 41% in the dsIRP30-treated group. In addition, the average size of the largest oocyte and the relative mRNA expression levels of Vg (vitellogenin) were significantly reduced in the dsIRP30-treated group (P < 0.05). Cellular localization by immunofluorescence demonstrated that IRP30 has important functions in the germ cells of workers' ovarioles. Overexpression of IRP30 was confirmed to increase the reproductive capability of the transgenic D. melanogaster. In conclusion, IRP30 regulates worker egg-laying by affecting the expression of Vg, the size of the ovary and the formation of the oocyte. These findings provide essential information for understanding the mechanisms underlying worker reproductive regulation. Copyright © 2021. Published by Elsevier B.V.