AbstractA great deal of behavioral and molecular interactions between male and female insects takes place during insect reproduction. Here, we comprehensively analyze the reproductive system of the olive fruit fly. Specifically, transcriptomic and genomic analysis of the reproductive tissues from virgin and once mated insects were performed. Genes encoding proteins implicated in immune response, mucins, antigen 5 proteins, proteases inhibitors and proteins with putative secretory activity were identified. Comparison of the transcriptomes between virgin and mated insects resulted in the identification of genes that are up- or down-regulated after mating. In testes 106 genes were up-regulated and 344 genes were down-regulated, whereas in male accessory glands with ejaculatory bulb 1,607 genes were up-regulated and 384 genes were down-regulated in mated male insects. Respectively, in mated females 1,705 genes were up-regulated and 120 genes were down-regulated in mated insects. To get a deeper insight, the expression profiles of selected genes throughout sexual maturation for the male tissues and throughout different time points after mating for the female reproductive tissues were determined. Identification of genes that take part in the mating procedure not only gives an insight in the biology of the insects, but it could also help the identification of new target genes in order to disturb the reproductive success of the olive fly and thus develop alternative pest control method.