Stocking black basses (Micropterus spp.) is a common practice used to increase angling opportunities in impoundments; however, when non-native black basses are introduced they often invade riverine habitats where they threaten the persistence of other fishes, including native black basses. Neosho Smallmouth Bass (M. dolomieu velox) is endemic to portions of the Ozark Highlands and Boston Mountains ecoregions and is threatened by introductions of non-native Smallmouth Bass (“SMB”) forms. Because of recent interest in stocking SMB into Grand Lake o’ the Cherokees, we assessed the suitability of local Neosho SMB populations as potential broodstock sources by assessing introgression with non-native SMB forms, as well as characterizing population structure and genetic diversity. The majority of Neosho SMB populations contained low, but non-negligible, genomic proportions of two genetically distinct non-native SMB forms. Introgression was highest in the Illinois River upstream of Lake Tenkiller, where Tennessee ‘lake strain’ SMB were stocked in the early 1990’s. We recovered three genetically distinct clusters of Neosho SMB at the uppermost hierarchical level of population structure: a distinct Illinois River cluster and two Grand River clusters that appear to naturally mix at some sites. Genetic diversity measures generally increased with stream size, and smaller populations with low diversity measures may benefit from immigration of novel genetic material. Overall, introgression with non-native SMB forms appears to pose a prominent threat to Neosho SMB; however, relatively intact populations of Neosho SMB exist in some Grand Lake o’ the Cherokees tributaries. Results could be used in developing a stocking program that promotes and sustains existing genetic diversity within and among Neosho SMB populations.