The effects of Pleistocene glaciations on the phylogeography of Melitaea cinxia (Lepidoptera: Nymphalidae)

Partial (600 bp) sequences of mitochondrial cytochrome oxidase I (COI) gene were used to infer the phylogeography of Melitaea cinxia (Lepidoptera: Nymphalidae) across the entire distributional range of the species, encompassing north Africa and Eurasia. Cladistic analysis of 49 distinct haplotypes (haplotype and nucleotide diversity were 0.95 and 0.027, respectively) revealed strong phylogeographic structure in M cinxia, characterised by four major clades: Morocco; Western (Iberia, France, Italy); Central (central and northern Western Europe, Balkans, Greece, Anatolia, Levant); and Eastern (eastern Baltic, Urals, Iran, Siberia, China); separated by average pairwise distances of beween 2 and 6 percent. This pattern is consistent with the location of southern glacial refugia in the Iberian, Italian and Balkan peninsulas, as well as multiple eastern refugia. The Western clade is further structured into south-central Iberian, northern Iberian (and French) and southern Italian sub-clades; and the Eastern clade into Near Eastern and Far Eastern sub-clades; with weaker phylogeographical concordance within the Central clade, except for a large area in central and northern Western Europe which is monomorphic for COI haplotype. The Baltic and eastern Europe have been primarily colonized by the Far Eastern sub-clade, rather than the Central (Balkan) clade, highlighting the importance of including Near and Far Eastern populations in phylogeographic studies of Palearctic species. Maps showing them extent of clades and sub-clades suggest several regions of secondary contact and possible hybridization. Interspecific comparison of representative M cinxia haplotypes supports a monophyletic origin of all M cinxia.