The California Floristic Province (CFP) is considered a global biodiversity hotspot because of its confluence of high species diversity across a wide range of threatened habitats. To understand how biodiversity hotspots such as the CFP maintain and generate diversity, we conducted a phylogeographic analysis of the flightless darkling beetle, Nyctoporis carinata, using multiple genetic markers. Analyses of both nuclear and mitochondrial loci revealed an east–west genetic break through the Transverse Ranges and high genetic diversity and isolation of the southern Sierra Nevada Mountains. Overall, the results obtained suggest that this species has a deep evolutionary history whose current distribution resulted from migration out of a glacial refugium in the southern Sierra Nevada via the Transverse Ranges. This finding is discussed in light of similar genetic patterns found in other taxa to develop a foundation for understanding the biodiversity patterns of this dynamic area. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99, 424–444.