Mitochondrial and nuclear DNA variability was examined to assess population genetic structure and phylogeographic relationships in rainbow trout. Single-strand conformation polymorphisms and restriction site differences within 1055 bp of the mitochondrial D-loop region and 1566 bp of nuclear DNA in six single-copy nuclear DNA regions identified 31 mitochondrial genotypes and 50 nuclear alleles. Gene trees were constructed by sequencing each variant allele or mitochondrial genotype identified. Examination of 30 populations in 10 native rainbow trout groups using an analysis of molecular variance (AMOVA) indicated that 65% of mitochondrial variability and 35% of nuclear variability was explained by differences among the 10 groups. Phylogenetic patterns evident in mitochondrial and nuclear DNA were not always concordant. Differences in the evolutionary patterns detected by mitochondrial and nuclear DNA may reflect the differential impact of past introgression events on variability in the two genomes.