Abstract
Hybridization is common among minnows and shiners in the family Leuciscidae, and mitonuclear discordance can reveal evidence of historical hybridization and introgression events that have shaped extant species and populations. We have identified a notable case of serial mitogenome replacement in populations of two shiner species, Luxilus zonatus and L. chrysocephalus, which are syntopic in drainages throughout the northern and eastern Ozark Interior Highlands of North America. These mitogenome replacement events involved L. zonatus acquiring the mitogenome of L. chrysocephalus, and populations of L. chrysocephalus acquiring the mitogenome of an allopatric congeneric species, L. cornutus. The latter species has a more northern distribution that was likely shifted southward by glacial advances during the Pleistocene. The geographic extent of mitogenome replacements in both species spans multiple separate drainages encompassing most of the major river systems that comprise the northern and eastern Ozark Highlands. We attribute these patterns to shifting species distributions, which were impacted by multiple glacial advances and coincident geomorphological changes to Ozark Highland drainages throughout the Pleistocene. The serial nature of mitogenome replacements in L. zonatus and L. chrysocephalus may exclude a role for natural selection in these introgression events, but the dynamic shifts in species distributions and gene flow connections throughout the Pleistocene may have favored an invasion-with-hybridization model that predicts massive asymmetric introgression between invading and resident species. These results have applied significance for eDNA metabarcoding methods of biodiversity assessment in Ozark streams, as they are dependent on mitogenome detections.