The root of understanding speciation lies in determining the forces which drive it. In many closely-related species, including Sphyrapicus varius, S. nuchalis, and S. ruber, it is assumed that speciation occurred due to isolation in multiple Pleistocene refugia. We used genetic data from 457 samples at the control region (CR), cytochrome oxidase I (COI), and chromo-helicase DNA binding protein (CHD1Z) to examine range-wide population genetic structure and differentiation amongst these 3 species across each species' breeding range. In addition, we modeled these species' ecological niches for the Holocene (~6000 years ago), Last Glacial Maximum (~22000 years ago), and Last Interglacial (~120000-140000 years ago) to determine if Pleistocene glaciations could have contributed to allopatric distributions, therefore allowing these groups to differentiate. Population genetic data show a potential Pleistocene refugium in Haida Gwaii, an east-west split among S. varius, and low genetic differentiation within each species. Our CR data show some polyphyly, while COI and CHD1Z data show differentiation among species using composite genotypes. Ecological Niche Modeling shows a large amount of niche overlap at each time period suggesting that S. varius, S. nuchalis, and S. ruber may not have been completely allopatric, and these species likely had repeated intermittent contact. Our data support the growing body of research that suggests differentiation despite gene flow.

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