Geographic isolation and habitat specialization has aided in the evolution and maintenance of genetic integrity of the lepomid sunfishes (Lepomis: Centrarchidae) of North America. Our goal was to measure genetic distances between four of the eleven extant sunfish species by using mitochondrial DNA analysis. Mitochondrial DNA restriction fragment length polymorphisms (RFLPs) were examined in bluegill (L. macrochirus), redear sunfish (L. microlophus), longear sunfish (L. megalotis), and green sunfish (L. cyanellus) using 15 restriction endonucleases. The largemouth bass (Micropterus salmoides) was used as an outgroup. The phylogeny inferred from Dollo parsimony cladistic analysis largely concurred with published results from allozyme analyses and the fossil record, yet was inconsistent with published anatomical analyses. Genetic distances between species ranged from 0.1627 to 0.3328. The green sunfish was the basal member of the genus, whereas the bluegill was the most diverged from the largemouth bass. These four species diverged over a broad time frame, with estimated times of speciation occurring during Miocene (8.14 - 16.64 mya).

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