Abstract

Pleistocene glaciations had significant effects on the distribution and evolution of species inhabiting the Holarctic region. Phylogeographic studies concerning the entire region are still rare. Here, we compared global phylogeographic patterns of one boreo-montane and one boreo-temperate butterflies with largely overlapping distribution ranges across the Northern Hemisphere, but with different levels of range fragmentation and food specialization. We reconstructed the global phylogeographic history of the boreo-montane specialist Boloria eunomia (n = 223) and of the boreo-temperate generalist Boloria selene (n = 106) based on mitochondrial and nuclear DNA markers, and with species distribution modelling (SDM). According to the genetic structures obtained, both species show a Siberian origin and considerable split among populations from Nearctic and Palaearctic regions. According to SDMs and molecular data, both butterflies could inhabit vast areas during the moderate glacials. In the case of B. selene, high haplotype diversity and low geographic structure suggest long-lasting interconnected gene flow among populations. A stronger geographic structuring between populations was identified in the specialist B. eunomia, presumably due to the less widespread, heterogeneously distributed food resources, associated with cooler and more humid climatic conditions. Populations of both species show opposite patterns across major parts of North America and in the case of B. eunomia also across Asia. Our data underline the relevance to cover entire distribution ranges to reconstruct the correct phylogeographic history of species.

Highlights

  • The Quaternary period is characterized by severe fluctuations of long glacial and shorter interglacial periods

  • We evaluated as the best K = 6 (FCT = 0.82) because higher levels of K become less biologically informative

  • B. eunomia is characterised by a high genetic structuring across Europe, but panmixia in Asia and a lack of genetic differentiation across the Nearctic region

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Summary

Methods

We sampled 124 individuals of Boloria eunomia and 60 individuals of B. selene, covering the whole Holarctic region. A further 99 sequences of B. eunomia and 46 of B. selene were added to the dataset from GenBank (http://www.ncbi.nlm.nih.gov/) (S1 Table, including specimen ID numbers, GenBank accession numbers, information about specimen repository). One fast mitochondrial (cytochrome c oxidase subunit I, COI) and two more conservative nuclear (arginine kinase, ArgKin; wingless, WG) genes were amplified by Polymerase Chain Reactions (PCR) in 20 μl volume (10 μl PPP Mastermix Top-Bio, 6.9 μl PCR H2O, 0.8 + 0.8 μl primers, 1.5 μl DNA). Two forward-reverse primer pairs were used for COI: LCO/HCO and Ron/HCO; for ArgKin, one pair: ArginineF/ ArginineR; and for wingless, one pair: LepWG1/LepWG2 [37,38]. Sequencing was provided by Macrogen Inc. on ABI3730XL DNA analysers. Sequences were checked manually and aligned in Geneious v. 8.0.5 [39]

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