Male is the default sex: functional significance of the sex determination cascade in horned dung beetles.

Sex-specific trait expression represents a striking dimension of morphological variation within and across species. The mechanisms instructing sex-specific organ development have been well studied in a small number of insect model systems, suggesting striking conservation in some parts of the somatic sex determination pathway while hinting at possible evolutionary lability in others. However, further resolution of this phenomenon necessitates additional taxon sampling, particularly in groups in which sexual dimorphisms have undergone significant elaboration and diversification. Here, we functionally investigate the somatic sex determination pathway in the gazelle dung beetle , an emerging model system in the study of the development and evolution of sexual dimorphisms. We find that RNA interference (RNAi) targeting caused chromosomal females to develop morphological traits largely indistinguishable from those normally only observed in males, and that is sufficient to induce splicing of the normally male-specific isoform of in chromosomal females, while leaving males unaffected. Further, was found to phenocopy previously described RNAi phenotypes of in female but not male beetles. These findings match predictions derived from models of the sex determination cascade as developed largely through studies in . In contrast, resulted in larval mortality and was not sufficient to affect splicing, whereas RNAi targeting and two putative orthologs of yielded no obvious phenotypic modifications in either males or females, raising the possibility that the function of a subset of sex determination genes may be derived in Diptera and thus non-representative of their roles in other holometabolous orders. Our results help illuminate how the differential evolutionary lability of the somatic sex determination pathway has contributed to the extraordinary morphological diversification of sex-specific trait expression found in nature.