Bermudagrass (Cynodonspp.) breeding and cultivar development is hampered by limited information regarding its genetic and phenotypic diversity. To explore diversity in bermudagrass, a total of 206Cynodonaccessions consisting of 193 common bermudagrass (C. dactylonvar.dactylon) and 13 African bermudagrass (C. transvaalensis) accessions of worldwide origin were assembled for genetic characterization. Genotyping-by-sequencing (GBS) was employed for genetic marker development. With a minor allele frequency of 0.05 and a minimum call rate of 0.5, a total of 37,496 raw single nucleotide polymorphisms (SNPs) were calledde novoand were used in the genetic diversity characterization. Population structure analysis using ADMIXTURE revealed four subpopulations in this germplasm panel, which was consistent with principal component analysis (PCA) and phylogenetic analysis results. The first three principal components explained 15.6%, 10.1%, and 3.8% of the variance in the germplasm panel, respectively. The first subpopulation consisted ofC.dactylonaccessions from various continents; the second subpopulation was comprised mainly ofC. transvaalensisaccessions; the third subpopulation containedC. dactylonaccessions primarily of African origin; and the fourth subpopulation representedC. dactylonaccessions obtained from the Oklahoma State University bermudagrass breeding program. Genetic diversity parameters including Nei’s genetic distance, inbreeding coefficient, and Fst statistic revealed substantial genetic variation in theCynodonaccessions, demonstrating the potential of this germplasm panel for further genetic studies and cultivar development in breeding programs.