Conservation and genetics of the frosted flatwoods salamander ( Ambystoma cingulatum ) on the Atlantic coastal plain

The federally threatened frosted flatwoods salamander, Ambystoma cingulatum, occurs in isolated populations on the coastal plain of northern Florida, Georgia and South Carolina. An earlier phylogeographic study based primarily on mitochondrial DNA (mtDNA) and morphometrics demonstrated that the previously recognized species A. cingulatum contained two cryptic species, and that two distinct mtDNA clades were contained within the newly restricted A. cingulatum. However, salamanders from the northern extent of the species range in South Carolina were not previously available for analysis. Here, we used individuals from a newly discovered South Carolina breeding site to determine whether A. cingulatum from South Carolina are genetically distinct from their more southerly conspecifics. These analyses included the mitochondrial gene studied previously as well as broad geographic sampling of three rapidly evolving nuclear genes that allowed us to further investigate lineage diversification of flatwoods salamanders. The mitochondrial and nuclear results are largely congruent, yielding strong support for two distinct species of flatwoods salamanders and also two lineages within the eastern species. Further, the South Carolina specimens are closely related to other haplotypes found in eastern Georgia and Florida . Our summary of field surveys over the past 20 years indicates that this South Carolina population may be one of only three remaining in the entire Atlantic coastal plain distribution of this rare and declining amphibian species.