One of the largest impediments to the conservation of freshwater mussels is the absence of host-fish data. Suitable hosts must be present in sufficient numbers and occur at the appropriate time for successful mussel recruitment. Habitat degradation and fragmentation caused by dams and other anthropogenic alterations may reduce host availability. Host data are lacking for the federally threatened Purple Bankclimber mussel (Elliptoideus sloatianus), which is endemic to the Apalachicola–Chattahoochee–Flint basin (ACF) in Alabama, Florida, and Georgia, and the Ochlockonee basin in Florida and Georgia. We tested 29 fish species in 7 families as potential hosts for Purple Bankclimbers and observed high metamorphosis success (79–89%) with 4 species of sturgeon: Gulf (Acipenser oxyrinchus desotoi), Atlantic (Acipenser oxyrinchus oxyrinchus), Lake (Acipenser fulvescens), and Shortnose (Acipenser brevirostrum). Metamorphosis was less successful with Blackbanded Darters (Percina nigrofasciata) and Halloween Darters (Percina crypta) as hosts (34–36% metamorphosis), and the remainder of the fishes we tested were not suitable hosts. The federally threatened Gulf Sturgeon is the only sturgeon species present in the ACF, but access of this migratory fish to most of the basin is blocked by Jim Woodruff Dam on the Apalachicola River. In the absence of sturgeon upstream of Jim Woodruff Dam, darters appear to have facilitated persistence of this mussel species, but at abundances far lower than historical conditions. This relationship between the Purple Bankclimber and Gulf Sturgeon is the first description of a federally protected fish serving as a host for a federally protected mussel and is an archetypal example of the role of habitat fragmentation in the ecology of listed species. Recovery of the Purple Bankclimber and other mussel species probably will require restoration of habitat connectivity for fish passage.