All species concepts are rooted in reproductive, and ultimately genealogical, relations. Genetic data are thus the most important source of information for species delimitation. Current ease of access to genomic data and recent computational advances are blooming a plethora of coalescent-based species delimitation methods. Despite their utility as objective approaches to identify species boundaries, coalescent-based methods (1) rely on simplified demographic models that may fail to capture some attributes of biological species, (2) do not make explicit use of the geographic information contained in the data, and (3) are often computationally intensive. In this article, we present a case of species delimitation in the Erebia tyndarus species complex, a taxon regarded as a classic example of problematic taxonomic resolution. Our approach to species delimitation used genomic data to test predictions rooted in the biological species concept and in the criterion of coexistence in sympatry. We (1) obtained restriction-site associated DNA (RAD) sequencing data from a carefully designed sample, (2) applied two genotype clustering algorithms to identify genetic clusters, and (3) performed within-clusters and between-clusters analyses of isolation by distance as a test for intrinsic reproductive barriers. Comparison of our results with those from a Bayes factor delimitation coalescent-based analysis, showed that coalescent-based approaches may lead to overconfident splitting of allopatric populations, and indicated that incorrect species delimitation is likely to be inferred when an incomplete geographic sample is analyzed. While we acknowledge the theoretical justification and practical usefulness of coalescent-based species delimitation methods, our results stress that, even in the phylogenomic era, the toolkit for species delimitation should not dismiss more traditional, biologically grounded, approaches coupling genomic data with geographic information.

Abstract. The genus Eupholidoptera includes 46 Mediterranean species distributed from Turkey to Greece, Italy and southern France. In the eastern part of its range, Eupholidoptera has been considered to consist of several distinct species, while in the Balkans and Italian peninsula only E. chabrieri has been recognized. However, the status of some Italian populations, confined to particular geographic areas, remains uncertain. To investigate the delimitation of the Italian taxa of Eupholidoptera, we performed both morphological and molecular analyses. Morphological analysis was carried out by considering diagnostic characters usually used to distinguish different taxa, such as the shape of titillators in males and the subgenital plate in females. Molecular analysis was performed by sequencing three mitochondrial genes: 12S rRNA, 16S rRNA, partially sequenced and the entire gene of cox1 . Molecular markers were used to infer phylogenetic relationships among the Italian Eupholidoptera species and to reconstruct the historical processes that shaped their current geographic distribution. Results from both morphological and molecular analyses were used to revise the taxonomic arrangement of species. On the whole we were able to distinguish nine lineages of Italian Eupholidoptera, of which E. tyrrhenica sp.n. from Corsica is described as a new species.This published work has been registered in ZooBank, http://zoobank.org/urn:lsid: zoobank.org:pub:EBD181A0-5263-4880-AC80-66F624506E3A.