In many insect species, the ability of males to inhibit their mates from remating is an important component of fitness. This ability is also essential for the effective management of insect pests, including tephritid fruit flies, using the Sterile Insect Technique. Here we apply transcriptomics and proteomics to male reproductive tissues before and after mating to characterize components of semen that might mediate remating inhibition in Queensland fruit fly. We found 144 genes whose transcripts were enriched, or proteins expressed, in reproductive tissue and which also varied in amount after mating. Some were associated with testes, accessory glands and ejaculatory apodeme, but those from the ejaculatory apodeme were over-represented compared to those not enriched in reproductive tissue or mating responsive. These included 13 related genes clustered within one Mb on chromosome 5. Functional annotations implicated a broad range of biochemical processes in the genes/ proteins enriched in reproductive tissue and mating responsive, with cuticle structure most commonly implicated among the subset of these that were apodeme-enriched and a kinase involved in vitellogenesis implicated for one of the 13 clustered genes. We did not find a homolog of the much studied Drosophila melanogaster Sex Peptide but comparative genomics indicated that some of the tissue-enriched, mating responsive genes/proteins were rapidly evolving in tephritids (including in the Queensland fruit fly lineage), suggesting recent adaptation to new functional niches. Our results provide a set of candidate mediators of remating inhibition for further functional testing.

Modern lipidomics has the power and sensitivity to elucidate the role of insects’ lipidomes in their adaptations to the environment at a mechanistic molecular level. However, few lipidomic studies have yet been conducted on insects beyond model species such as Drosophila melanogaster. Here, we present the lipidome of adult males of another higher dipteran frugivore, Bactrocera tryoni. We describe 421 lipids across 15 classes of ester neutral lipids and phospholipids and ether neutral lipids and phospholipids. Most of the lipids are specified in terms of the carbon and double bond contents of each constituent hydrocarbon chain, and more ether lipids are specified to this degree than in any previous insect lipidomic analyses. Class-specific profiles of chain length and (un)saturation are broadly similar to those reported in D. melanogaster, although we found fewer medium-length chains in ether lipids. The high level of chain specification in our dataset also revealed widespread non-random combinations of different chain types in several ester lipid classes, including deficits of combinations involving chains of the same carbon and double bond contents among four phospholipid classes and excesses of combinations of dissimilar chains in several classes. Large differences were also found in the length and double bond profiles of the acyl vs. alkyl or alkenyl chains of the ether lipids. Work on other organisms suggests some of the differences observed will be functionally consequential and mediated, at least in part, by differences in substrate specificity among enzymes in lipid synthesis and remodelling pathways. Interrogation of the B. tryoni genome showed it has comparable levels of diversity overall in these enzymes but with some gene gain/loss differences and considerable sequence divergence from D. melanogaster.

Supplementary Table S3 in our recent publication