Genetic variation at the major histocompatibility (MHC) class I loci was examined in captive and wild samples of three subspecies of tiger (Bengal, Panthera tigris tigris; Siberian, P. t. altaica; Sumatran, P. t. sumatrae) by Southern blot analysis using the domestic cat probe FLA 24. All captive populations showed at least moderate variability compared to other felids. Captive tigers were, on average, significantly more variable than wild tigers, but this was not consistent among subspecies. Significant geographic genetic population structure was found at the MHC class I loci among both captive and wild tigers. Phylogenetic analysis of both captive and wild tigers placed Sumatran tigers basally. In a combined phylogenetic analysis, captive and wild tigers of the same subspecies occurred near one another, indicating that the structure of the captive populations reflects that of the wild populations.