Pseudemys alabamensis is one of the most endangered freshwater turtle species in the United States due to its restricted geographic distribution in coastal Alabama and Mississippi. Populations of P. alabamensis are geographically isolated from one another by land and salt water, which could act as barriers to gene flow. It is currently unknown how differentiated these isolated populations are from one another and whether they have experienced reductions in population size. Previous work found morphological differences between Alabama and Mississippi populations, suggesting that they may be evolutionarily distinct. Other Pseudemys turtles such as P. concinna and P. floridana occur naturally within the same geographic area as P. alabamensis and are known to hybridize with each other. These more abundant species could threaten the unique genetic identity of P. alabamensis through introgression. In order to evaluate the endangered status of P. alabamensis and the level of hybridization with other species, we used mitochondrial and nuclear microsatellite markers to assess genetic variation within and among populations of P. alabamensis throughout its range and estimate admixture with co-occurring Pseudemys species. In P. alabamensis, we found no variation in mitochondrial DNA and observed an excess of homozygosity in the microsatellite data. Our results indicate evidence of genetic differentiation between Alabama and Mississippi populations of P. alabamensis, and low estimated breeding sizes and inbreeding for two populations (Fowl River, Alabama and Biloxi, Mississippi). Our results also support admixture of P. alabamensis with P. concinna/P. floridana. Based on our results, P. alabamensis is highly endangered throughout its range and threatened by both low population sizes and hybridization. In order to improve the species’ chances of survival, focus should be placed on habitat preservation, maintenance of genetic diversity within both Mississippi and Alabama populations, and regular population monitoring activities such as nest surveillance and estimates of recruitment. 

We collected mitochondrial DNA (d-loop) and microsatellite data to obtain estimates of genetic diversity within and among species and inference of hybridization among species.

Supplementary Data 1: Microsatellite genotyping for each individual and each locus. “-9” refers to missing data.

Supplementary Data 2: Q assignment values for each individual with respect to each different species and P. alabamensis clusters (Alabama vs. Mississippi). Each tab indicates the Q values for specific comparisons (e.g., P. alabamensis vs. P. concinna)