Systematic and biogeographical relationships within the Hawaiian clade of the pantropical understory shrub genus Psychotria (Rubiaceae) were investigated using phylogenetic analysis of 18S-26S ribosomal DNA internal (ITS) and external (ETS) transcribed spacers. Phylogenetic analyses strongly suggest that the Hawaiian Psychotria are monophyletic and the result of a single introduction to the Hawaiian Islands. The results of phylogenetic analyses of ITS and ETS partitions alone give slightly different topologies among basal lineages of the Hawaiian clade; however, such differences are not well supported. Relationships in the section Straussia clade in particular are not well resolved because of few nucleotide changes on internal branches, suggesting extremely rapid radiation in the lineage. Parsimony and likelihood reconstructions of ancestral geographical distributions using the topologies inferred from both parsimony and likelihood analysis of combined data and using different combinations of models and branch lengths gave highly congruent results. However, for one internal node (corresponding to the majority of the "greenwelliae" clade), parsimony reconstructions were unable to distinguish between three possible island states, whereas likelihood reconstructions resulted in clear ordering of possible states, with the island of Oàhu slightly more probable than other islands under all but one model and branch length combination considered (the Jukes-Cantor-like model with branch lengths inferred under parsimony, under which conditions Maui Nui is more probable). A pattern of colonization from oldest to youngest islands was inferred from the phylogeny, using maximum parsimony and maximum likelihood. Additionally, a much higher incidence of intraisland versus interisland speciation was inferred.