Phylogenetic and taxonomic assessment of the endangered Cumberland bean, Villosa trabalis and purple bean, Villosa perpurpurea (Bivalvia: Unionidae)

Inadequate understanding of the phylogeography, taxonomy, and historical distribution of two critically imperiled freshwater mussels, Cumberland bean, Villosa trabalis , and purple bean, Villosa perpurpurea , has hindered management and recovery actions related to population restoration within their extant ranges. For more than 100 years, the purple-to-pink nacre of V. perpurpurea and white nacre of V. trabalis have been the only defining phenotypic characteristics used to distinguish each species. Genetic samples were analyzed from 140 individuals collected from 10 streams located in Virginia, Tennessee, and Kentucky, representing all known extant populations of each species. A 784-bp section of the mitochondrial DNA ND1 region was sequenced to assess the phylogeography and taxonomic validity of these taxa. Results of our phylogenetic analyses showed 100 % Bayesian posterior support for two distinct clades, one occurring in the Cumberland River basin and the other in the Tennessee River basin, separated by a mean genetic distance of 4 %. Mean genetic distances between haplotypes within each clade was <1 %. Among individuals from the Cumberland River basin, the nacre of shells was white to bluish-white, but in the Tennessee River basin, nacre graded from white to pink to dark purple; thus, nacre color is a variable and inconsistent character in nominal V. trabalis and V. perpurpurea occurring in the Tennessee River basin. Our data suggest that these morphologically similar species do not co-occur, as was previously believed. Instead, we conclude that the two species most likely share a common ancestor, but became isolated within each basin and experienced allopatric speciation. Updates to nomenclature, taxonomic placement, and recovery plans for the investigated species are needed.