This report summarizes a decade of morphological and molecular research on the phylogenetic relationships of didelphid marsupials (opossums), a substantially intact radiation of New World metatherian mammals. We review the comparative morphology of Recent opossums, emphasizing those anatomical systems from which taxonomically useful information is available for the majority of living genera and species, namely the integument, cranium, and dentition. Morphological similarities and differences among didelphids and other plesiomorphic marsupials (caenolestids, microbiotheriids, dasyurids, and peramelids) are also described. These observations, representing evolved differences in diverse functional-morphological systems, together with karyotypic information gleaned from the literature, provide the basis for coding 129 phylogenetic characters that we scored for 44 ingroup and seven outgroup taxa.

Published information about the size, internal organization, chromosomal location, and physiological properties of five nuclear genes (BRCA1, DMP1, IRBP, RAG1, vWF) sequenced for this study suggest that these loci are unlinked, exist as single copies, are active in different tissues, and encode protein products with widely divergent functions. All of the sequenced fragments are long (>900 bp), free of ingroup alignment ambiguities, and translate to open reading frame. Nucleotide data from a total of 7320 aligned sites were obtained from 43 ingroup and seven outgroup taxa.

Separate parsimony, likelihood, and Bayesian analyses of these six data partitions (morphology + karyotypes, five genes) resulted in highly congruent estimates of didelphid phylogeny with few examples of conflict among strongly supported nodes. Analyses of concatenated sequences and combined (nonmolecular + sequence) datasets effectively summarize all of the common signal recovered from separate analyses: a completely resolved ingroup phylogeny with high support statistics at most nodes. Remaining problems (not conclusively resolved in this study) include the position of the ingroup root and the relationships of three genera (Chacodelphys, Cryptonanus, Tlacuatzin) within their respective suprageneric clades.

The history of didelphid classification is reviewed, and all previous systems are found to contain nonmonophyletic groups. A revised phylogenetic classification consistent with our analytic results includes the following higher taxa: Glironiinae (for Glironia), Caluromyinae (Caluromys and Caluromysiops), Hyladelphinae (Hyladelphys), Didelphinae (Marmosini, Metachirini, Didelphini, and Thylamyini), Marmosini (Marmosa, Monodelphis, and Tlacuatzin), Metachirini (Metachirus), Didelphini (Chironectes, Didelphis, Lutreolina, and Philander), and Thylamyini (Chacodelphys, Cryptonanus, Gracilinanus, Lestodelphys, Marmosops, and Thylamys). The probable relationships of several Neogene fossil genera are also discussed. To facilitate identifications, all Recent genera are redescribed, representative crania are illustrated, and a key is provided.