Restriction fragment-length polymorphisms (RFLPs) of chloroplast DNA (cpDNA) were used to study phylogenetic relationships among twenty-six Fragaria taxa and two closely related species, Potentilla fruticosa L. and Duchesnea indica (Andrews) Focke. Sixteen restriction enzymes and probes of the entire Nicotiana tabacum L. chloroplast genome revealed a very low level of variation among the Fragaria taxa, limiting phylogenetic resolution. However, Fragaria appears to be more closely related to Potentilla than Duchesnea. The diploid taxa, F. iinumae Makino, F. nilgerrensis Schlect. and F. vesca L. were the most divergent Fragaria taxa and F. iinumae appears to be the most ancestral taxon. Little variation was revealed within the economically important octoploid group of taxa, which gave rise to the cultivated strawberry, and no progenitor taxa to the octoploid group could be identified. The lack of variation in the chloroplast genome suggests that these Fragaria species may be of relatively recent evolutionary origin.