Guiding Species Recovery through Assessment of Spatial and Temporal Population Genetic Structure of Two Critically Endangered Freshwater Mussel Species (Bivalvia: Unionidae)

Abstract The Cumberlandian Combshell (Epioblasma brevidens) and Oyster Mussel (E. capsaeformis) are critically endangered freshwater mussel species native to the Tennessee and Cumberland River drainages, major tributaries of the Ohio River in the eastern United States. The Clinch River in northeastern Tennessee (TN) and southwestern Virginia (VA) harbors the only remaining stronghold population for either species, containing tens of thousands of individuals per species; however, a few smaller populations are still extant in other rivers. We collected and analyzed genetic data to assist with population restoration and recovery planning for both species. We used an 888 base-pair sequence of the mitochondrial NADH dehydrogenase 1 (ND1) gene and ten nuclear DNA microsatellite loci to assess patterns of genetic differentiation and diversity in populations at small and large spatial scales, and at a 9-year (2004 to 2013) temporal scale, which showed how quickly these populations can diverge from each other in a short time period. Intraspecific mitochondrial DNA (mtDNA) and microsatellite DNA variation was higher in E. capsaeformis than in E. brevidens. These two species have maintained quite different levels of genetic diversity within their Clinch River stronghold and in their smaller peripheral populations in the Big South Fork Cumberland and Nolichucky rivers, TN. For instance, with only three mtDNA haplotypes detected overall across populations, E. brevidens’ capacity for maintaining genetic diversity appears to be less than that of E. capsaeformis, which had 18 haplotypes. At the relatively small spatial scales (15-30 kilometers) investigated in the Clinch River, demes of both species exhibited minimal genetic differentiation in either the 2004 or 2013 sampling periods, typically <0.02 based on FST and <0.1 based on Jost’s D. Our genetic data suggest that mussels at the numerous shoals in a 32-kilometer section of the Clinch River comprise a single, large population of each respective species with very high gene-flow among individual demes. However, we also observed a high level of genetic differentiation among demes at the 9-year temporal scale, with differentiation metrics for E. brevidens (D = 0.47 and FST = 0.12) and E. capsaeformis (D = 0.31 and FST = 0.05) proving higher than the within-year values. This result strongly suggests that genetic drift is playing an important role in allele frequency change over time in these populations. At the spatial and temporal scales investigated in this study, various demographic, life history, and environmental factors are influencing maintenance of genetic variation and need to be considered during conservation planning for each species.