Allozymic variation at 22 gene loci in populations of Peromyscus truei, P. difficilis, P. melanotis, and P. maniculatus is used to examine patterns of geographic differentiation. Samples of P. maniculatus collected throughout most of its immense range, from southern Mexico to northern Canada and from the Pacific to Atlantic coasts, are very similar in genic composition. Evidence from morphology, chromosomal configurations, ecology, and patterns of reproductive isolation argue that contemporary gene flow among P. maniculatus populations is not sufficient to account for the similarity in their allelic configurations. The macrogeographic conservatism in level of genic divergence conceivably results from relatively recent separation of populations, coupled with a genetic inertia resulting from a selected cohesion of the genome. Nonetheless, significant intersample heterogeneity of allele frequencies at polymorphic loci may certainly result from stochastic effects. The relative geographic uniformity in P. maniculatus contrasts somewhat with the geographic differences observed in P. truei and in P. difficilis. Members of the truei and maniculatus species groups are added to a biochemical dendrogram, which now includes 20 named species of Peromyscus. The large and successful rodent genus Peromyscus includes about 60 named species, variously assigned to five to seven subgenera (Osgood, 1909; Hall and Kelson, 1959; Hooper and Musser, 1964; Hooper, 1968). The diversity in size of geographic range and in pattern of recognized geographic variation in morphology among nominal Peromyscus species is striking. Roughly one-half of the species are monotypic, exhibiting no definable geographic variation in morphology (Baker, 1968). Most monotypic species, such as P. stephani and P. dickeyi, are insular endemics, but a few others such as P. melanotis are continental and occupy larger ranges. Similarly, polytypic species consisting of more than one named subspecies may have narrow (P. sitkensis) or wide (P. maniculatus) geographic ranges. In this study, we continue our biochemical survey of the genus Peromyscus by examining geographic variation in electromorphic content of species belonging to the truei and maniculatus species groups of the subgenus Peromyscus. We are particularly interested in the systematic and evolutionary ramifications of contrasting patterns of geographic genetic differentiation. Peromyscus maniculatus is probably the most widespread and abundant small mammal on the North American continent. Populations referable to some 65 named subspecies are distributed from southern Mexico to the Northwest Territories of Canada, and from the Pacific to North Atlantic seaboards. The diversity of occupied habitats ranges from arid grasslands and brushy slopes to humid coniferous and deciduous forests. We have examined allozymic variation in proteins encoded by 21 genetic loci in a total of 716 P. maniculatus collected throughout most of its range. We have also examined several populations of a more narrowly distributed sibling...