Limited hybridization and introgression despite stocking among endemic Interior Highlands black basses (Centrarchidae: Micropterus)

<p><strong>Aim</strong></p> <p>Smallmouth Bass (<em>Micropterus</em> <em>dolomieu</em>; SMB) are globally popular among anglers and have been widely introduced (i.e., stocked) for population management and sportfishing. Importantly, stocking was prevalent before cryptic diversity within the SMB complex was known, which now includes three newly elevated species: Neosho Bass (<em>M</em>. <em>velox</em>; NB), Little River Bass (<em>M</em>. sp. cf. <em>dolomieu</em> Little River; LRB), and Ouachita Bass (<em>M</em>. sp. cf. <em>dolomieu</em> Ouachita River; OB). We sought to quantify population structure and hybridization and introgression in these three recently described species.</p> <p><strong>Location</strong></p> <p>Species-level diversity, particularly in the basin-restricted LRB and OB in the Ouachita Mountains within the Central Interior Highlands (CIH), North America, suggests the presence of distinct genetic variation that could be eroded by introgression.  </p> <p><strong>Methods</strong></p> <p>We estimated interspecific introgression and intraspecific population differentiation in the Smallmouth Bass species complex (SMB-C) using 472 specimens comprising SMB, NB, LRB, and OB, including the naturally sympatric Spotted Bass (<em>M</em>. <em>punctulatus</em>; SPB). Genomic samples were genotyped on a SNP panel of 192 loci designed to detect allele-sharing on multiple hierarchical levels. </p> <p><strong>Results</strong></p> <p>We found low range-wide hybridization between species in the SMB-C and SPB (mostly SMB-C backcrosses), and interspecific heterozygosity varied, indicating differential introgression. Range-wide hybridization between species in the CIH and SMB was similar overall (but mostly F₂ and CIH backcrosses) and was observed in streams with known SMB stocking in connected reservoirs. Interspecific heterozygosity in SMB hybrids was also generally lower, indicating later-generation backcrosses. We found strong population structure in the Ouachita Mountains (LRB and OB). </p> <p><strong>Main Conclusions</strong></p> <p>Despite isolated incidences of natural (SPB) and human-mediated (SMB) introgression, genomic identity appears intact in endemic LRB and OB, suggesting potential ecological or behavioral isolating mechanisms preventing cross-species reproduction. Our findings reveal that genetic variation remains in cryptic, basin-restricted species in the Ouachita Mountains ecoregion that may be managed for long-term conservation.</p>