The genus Castanea in North America contains multiple tree and shrub taxa of 10 conservation concern. The two species within the group, American chestnut (Castanea dentata) 11 and chinquapin (C. pumila sensu lato), display remarkable morphological diversity across their 12 distributions in the eastern United States and southern Ontario. Previous investigators have 13 hypothesized that hybridization between C. dentata and C. pumila has played an important role 14 in generating morphological variation in wild populations. A putative hybrid taxon, Castanea 15 alabamensis, was identified in northern Alabama in the early 20th century; however, the 16 question of its hybridity has been unresolved. We tested the hypothesized hybrid origin of C. 17 alabamensis using genome-wide sequence-based genotyping of C. alabamensis, all currently 18 recognized North American Castanea taxa, and two Asian Castanea species at >100,000 single-19 nucleotide polymorphism (SNP) loci. With these data, we generated a high-resolution 20 phylogeny, tested for admixture among taxa, and analyzed population genetic structure of the 21 study taxa. Bayesian clustering and principal components analysis provided no evidence of 22 admixture between C. dentata and C. pumila in C. alabamensis genomes. Phylogenetic analysis 23 of genome-wide SNP data indicated that C. alabamensis forms a distinct group within C. pumila 24 sensu lato. Our results are consistent with the model of a nonhybrid origin for C. alabamensis. 25 Our finding of C. alabamensis as a genetically and morphologically distinct group within the 26 North American chinquapin complex provides further impetus for the study and conservation of 27 the North American Castanea species. 28 29