To determine the genetic origin of individual sturgeon that are morphologically intermediate to pallid (Scaphirhynchus albus) and shovelnose (Scaphirhynchus platorhynchus) sturgeon, we combined previously published mitochondrial DNA (mtDNA) and microsatellite data with additional microsatellite data. Two sympatric populations of pallid and shovelnose sturgeon from the upper Missouri River and a sympatric population containing pallid, shovelnose, and putative pallid-shovelnose hybrids from the Atchafalaya River were analyzed using an index of hybridization and a principle components analysis of individual relatedness scores. The addition of new microsatellite data improved our ability to genetically differentiate individual pallid and shovelnose sturgeon collected in both areas. Our methods distinguished morphologically intermediate Atchafalaya River sturgeon, which appear to be genetically intermediate between pallid and shovelnose sturgeon. The results support a hybrid origin for morphologically intermediate individuals, although it is unclear whether they are all first-generation hybrids or if some are the result of subsequent backcrossing with the more common shovelnose sturgeon.