We used 10 microsatellite loci to examine the genetic population structure of cutthroat trout Oncorhynchus clarki within the Lahontan Basin complex. Genetic diversity was analyzed for trout from Nevada, California, and Utah representing three putative subspecies: Lahontan O. c. henshawi, Paiute O. c. seleniris, and Humboldt (an unnamed subspecies) cutthroat trout. We found significant differences in microsatellite diversity among the three putative subspecies found in this area. Analysis of molecular variance partitioned microsatellite variation as 9.8% among subspecies, 27.7% among populations, and 62.5% within populations of Lahontan Basin cutthroat trout. Genetic distance analyses (Cavalli-Sforza-Edwards and F _st) supported unique population structure in cutthroat trout from the Humboldt and Pilot Peak drainages. Pairwise F _st values for Lahontan cutthroat trout were not significantly correlated with geographic distance between population pairs (r ² = 0.008; P < 0.0001), suggesting that they are extremely isolated populations with small effective sizes that are vulnerable to extinction. Two extant hatchery strains of Lahontan cutthroat trout showed genetic associations with different geographic source populations. The Pyramid Lake hatchery strain was most closely associated genetically with fish from Summit Lake. The Pilot Peak hatchery strain was associated genetically with Pilot Peak wild trout (Utah) and Macklin Creek trout (California). The phylogeographic diversity depicted in this study supports unique population structure and suggests important evolutionary relationships needed to evaluate transplanted populations and hatchery supplementation within the basin.