Chemosystematics Using Cuticular Compounds: A Powerful Tool to Separate Species in Mediterranean Dung Beetles (Coleoptera: Geotrupidae).

The use of chemical characters to infer a phylogeny is known to be promising to ascertain phylogenetic relationships in controversial groups. Dung beetle classifications containing the Geotrupidae family, based on morphological characters and genes, are debated with respect to the subfamilies, such as the Bolboceratids. In our study, we used different approaches to generate and compare the Geotrupidae phylogenies based on genetics and chemotaxonomy. Cuticular compounds were analyzed for 12 species of Mediterranean dung beetles to build a chemical phylogeny. In addition, mitochondrial and nuclear marker concatenation have been used to elaborate the molecular phylogeny. Using the cuticular compound continuous data, our results showed that each species was associated with a specific chemical pattern and that all individuals belonging to the same species displayed a similar chemical blend. The most distant species was Bolbelasmus gallicus, with an evident distinction from the other species due to several compounds. The maximum parsimony tree showed that all genera belonging to a Geotrupidae subfamily were grouped in the same clade, with B. gallicus species isolated in another clade, similar to the chemotaxonomy grouping. A strong positive correlation between chemotaxonomy and genetic phylogeny has been demonstrated, underlying a genetic basis for cuticular hydrocarbon variations. Our results are congruent with previous studies using morphological or genetic data. Our results also showed that only 10 compounds can be used to distinguish at least six species of dung beetle and that chemotaxonomy could become a useful and affordable tool to determine phylogenetic relationships in insects.