The Stippled Darter, Etheostoma punctulatum, has long been recognized as a single species of the subgenus Ozarka, endemic to the clear, high-gradient creeks and rivers of the Arkansas, White, and Missouri river drainages in the Ozark Plateau of the Interior Highlands of North America. Etheostoma punctulatum is redescribed and two new species are described based on morphological and allozyme characters, and phylogenetic analyses. The Stippled Darter, E. punctulatum, is restricted to tributaries of the Missouri River. The Autumn Darter, E. autumnale, new species, is endemic to the White River system, Missouri and Arkansas. The Sunburst Darter, E. mihileze, new species, is known only from the Arkansas River system, Missouri, Arkansas, Oklahoma, and Kansas. The three species are found to be divergent and diagnosable from one another and represent valid species under a variety of species concepts. Phylogenetic analyses indicate that E. punctulatum is sister to E. mihileze, new species, plus E. autumnale, new species.