G- and C-banded karyotypes from 9 previously unstudied species of Peromyscus are described and compared to the 9 species described in the literature. Additional new data are presented for P. eremicus. A hypothetical evolutionary tree for the 18 species was constructed based on the assumption that, where possible, shared chromosomal rearrangements became established in a common ancestor. A minimum of 60 chromosomal rearrangements (34 heterochromatic additions and 26 pericentric inversions) were needed to construct the most parsimonious tree. Seven of the 18 species have heterochromatin in noncentromeric areas. Three times as many inversions have been identified in the 11 largest pairs as have been observed in the 12 smallest pairs. Peromyscine rodents are characterized by a pattern of chromosomal variation quite distinct from that described for other rodent genera, such as Sigmodon and Mus. Karyotypic orthoselection in rodents appears to be relatively common in closely related species.