Phylogenetics and population structure of the steppe species Hycleus polymorphus (Coleoptera: Meloidae: Mylabrini) reveal multiple refugia in Mediterranean mountain ranges

Many continental species distributed in the Eurasian steppe occur as relict populations in the mountains of Western Europe. Their biogeographical responses to Quaternary climate changes have been poorly studied; however, they could have responded as cold-adapted species. We investigated the biogeographic history of a steppe beetle, Hycleus polymorphus, using mitochondrial and nuclear DNA sequences (COI, CAD, ITS2), and species distribution modelling (SDM) under present and past bioclimatic envelopes. We first performed a phylogenetic assessment to define species boundaries within the H. polymorphus species group. Specimens previously treated as Hycleus humerosus on morphological grounds are assigned to H. polymorphus, and those identified as Hycleus zebraeus assigned to Hycleus atratus. ITS2 data analyses revealed a strong phylogeographical structure of H. polymorphus populations, with four haplogroups corresponding to the (i) Italian Alps, (ii) French Alps and Pyrenees, (iii) South Balkan and Pontic mountains, and (iv) North Dinaric Alps. Based on these analyses and the SDM, we propose that during a glacial period, following the spread of steppic habitat, H. polymorphus underwent a range expansion from Asia to South- West Europe. Within the Mediterranean area, during the last interglacial the climatic suitability for the species was limited to mountains that acted as refugia and prompted allopatric divergence into four main lineages.