Ancestral Wolbachia lineages are likely donors of ribotoxin genes in Aedes aegypti

Ribosome-inactivating proteins (RIPs) are enzymes that irreversibly inhibit protein synthesis by depurinating a specific adenine residue in the ribosomal RNA. Although members of this gene family are widespread in plants and bacteria, their occurrence in metazoans is rare and restricted to a few insect lineages, including Culicinae mosquitoes. Previous studies suggested that these genes were acquired by mosquitoes via horizontal gene transfer (HGT) from bacteria lineage, but the source lineage remained unidentified. Here, we report the discovery of RIP-encoding genes in two Wolbachia strains. Phylogenetic analyses confirmed the monophyletic relationship between Wolbachia and mosquito RIPs, implying Wolbachia as the donor of these genes. These results shed light on the evolutionary dynamics of RIPs and the dual role of Wolbachia as both a functional contributor and genetic donor. By bridging the gap between endosymbiont and host genomes, this work provides new evidence for HGT as a source of adaptive innovation in insects. The implications of these findings for the ongoing debate on HGT in metazoans are also discussed.