Strategic conservation planning for broad, multi-species landscapes benefits from a data-driven approach that emphasizes persistence of all priority species’ populations and utilized landscapes, while simultaneously accounting for human uses. This study presents such an assessment for priority fishes of the Great Plains of the United States. Species distribution models for 28 priority fish species were created and incorporated into a prioritization framework using the open source software Zonation, accounting for species-specific connectivity needs and current fish habitat condition. Multiple additional assessments were then produced that i.) identify distinct species management units based on distance and compositional similarity of stream segments containing priority species, ii.) compare results of ranking species’ conservation values at the local (state) and global scale, and iii.) provide ‘bang-for-buck’ perspectives, emphasizing richness of priority species, at state and major basin scales. Together, these analyses are intended to aid managers in effective allocation of conservation action with regards to imperiled fishes of the Great Plains. Implementation of a broad-scale multi-species approach such as this complements traditional reactive management and restoration by encouraging cooperation and coordination among stakeholders and partners, increasing efficiency of future monitoring and management efforts.