Phylogenies of Trifolium species were constructed using nucleotide sequence data of the internal transcribed spacers (ITS) of nuclear ribosomal DNA and chloroplast DNA restriction site data from PCR-amplified genes and genic regions (rbcL, trnK, and rpoC1-C2). Biogeography, morphological evolution, and the existing classification for Trifolium were examined. The genus Trifolium is strongly supported as monophyletic, however, only one small section (Chronosemium) is monophyletic, although the data are in conflict regarding its placement. The two largest sections of the genus, Sections Lotoidea and Trifolium, are not supported as monophyletic, as currently circumscribed. Section Chronosemium is embedded within Section Trifolium and many members of Section Lotoidea form a basal grade within the genus. This basal position of Section Lotoidea supports previously-proposed hypotheses concerning plesiomorphic morphological characters and a Mediterranean-Mideast biogeographic origin of the genus.

Key words: Fabaceae, phylogeny, Trifolium