Newly discovered populations of Rhyacotritonidae were investigated for taxonomic identity, hybridization, and sympatry. Species in the genus Rhyacotriton have been historically difficult to identify using morphological characters. Mitochondrial (mtDNA) 16S ribosomal RNA sequences (491 bp) and allozymes (6 loci) were used to identify the distribution of populations occurring intermediate between the previously described ranges of R. variegatus and R. cascadae in the central Cascade Mountain region of Oregon. Allozyme and mitochondrial sequence data both indicated the presence of two distinct evolutionary lineages, with each lineage corresponding to the allopatric distribution of R. cascadae and R. variegatus. Results suggest the Willamette River acts as a phylogeographic barrier limiting the distribution of both species, although we cannot exclude the possibility that reproductive isolation also exists that reinforces species' distributions. This study extends the previously described geographical ranges of both R. cascadae and R. variegatus and defines an eastern range limit for R. variegatus conservation efforts.