Abstract The broadstripe shiner, Pteronotropis euryzonus is a freshwater minnow endemic to a small area that includes the tributaries of the middle and lower Chattahoochee River in Alabama and Georgia. Populations of P. euryzonus appear fragmented because individuals have not been detected in the main channel. This suggests limited dispersal potential and low gene flow between populations, and previous studies have suggested multiple forms of P. euryzonus in the Chattahoochee. In total, 125 samples of P. euryzonus were collected for genetic analysis from 23 sites in 11 tributaries of the Chattahoochee River, and museum specimens were used for morphometric analyses. The mitochondrial genes cytochrome c oxidase I (COI) and cytochrome b (cytb) were used to assess the genetic structure of P. euryzonus throughout its range. Geometric morphometrics were used to quantify shape variation among populations. Results suggest the presence of three genetically distinct populations: Northern, Pataula, and Southern, which also exhibit distinct morphologies. These populations support previous hypotheses of multiple forms of P. euryzonus in the Chattahoochee River. The presence of three genetically and morphologically distinct populations has significant conservation implications for P. euryzonus, which is currently listed as imperilled in both Alabama and Georgia. Three distinct populations, a restricted range, and recent disturbance to gene flow from local infrastructure may necessitate further protections to prevent extirpation. Copyright © 2015 John Wiley & Sons, Ltd.
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