Hybrid zones have yielded considerable insight into many evolutionary processes, including speciation and the maintenance of species boundaries. Presented here are analyses from a hybrid zone that occurs among three salamanders -Plethodon jordani, Plethodon metcalfi and Plethodon teyahalee -from the southern Appalachian Mountains. Using a novel statistical approach for analysis of non-clinal, multispecies hybrid zones, we examined spatial patterns of variation at four markers: single-nucleotide polymorphisms (SNPs) located in the mtDNA ND2 gene and the nuclear DNA ILF3 gene, and the morphological markers of red cheek pigmentation and white flecks. Concordance of the ILF3 marker and both morphological markers across four transects is observed. In three of the four transects, however, the pattern of mtDNA is discordant from all other markers, with a higher representation of P. metcalfi mtDNA in the northern and lower elevation localities than is expected given the ILF3 marker and morphology. To explore whether climate plays a role in the position of the hybrid zone, we created ecological niche models for P. jordani and P. metcalfi. Modelling results suggest that hybrid zone position is not determined by steep gradients in climatic suitability for either species. Instead, the hybrid zone lies in a climatically is currently studying interactions between the amphibian chytrid fungus and frogs of the eastern United States. K.H.K. is interested in the evolutionary processes of southern Appalachian salamanders, especially as revealed by ecological niche modelling and physiological limits. B.M.F. studies conservation, population genetics, and patterns of hybridization across a variety of salamander taxa. P.K.T.'s research focuses on speciation genetics, as informed by an extensive house mouse hybrid zone in Eastern Europe.Supporting information Additional supporting information may be found in the online version of this article. Table S1 Transects, sites codes, elevation, and latitude and longitude for animals captured in the hybrid zone Table S2 Sites codes, elevation, and latitude and longitude for parental taxa Table S3 Pure parental individuals (Plethodon jordani, P. metcalfi, and P. teyahalee) and marker scores used in panel for marker development Table S4 Samples (arranged by transect) and marker scores for all samples used in analyses Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. Author Manuscript homogenous region that is broadly suitable for both P. jordani and P. metcalfi. We discuss various selective (natural selection associated with climate) and behavioural processes (sex-biased dispersal, asymmetric reproductive isolation) that might explain the discordance in the extent to which mtDNA and nuclear DNA and colour-pattern traits have moved across this hybrid zone.
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