A study conducted with the aim of developing a protocol for callus induction and plantlet regeneration in vitro from leaf segments of three accessions of African rice (O. glaberrimaSteud.) indigenous to Ghana. Leaf segments of the accessions namely, Guame, N/4 and SARI 1 were assessed for callus induction and plantlet regeneration ability on different concentrations of plant growth regulators, incorporated into Murashige and Skoog, (1962) (MS) basal medium. Frequency of callus induction which was achieved on MS medium supplemented with (0-10) mg/l 2,4-D differed significantly (p≤0.05) among the accessions, as well as among the levels of 2,4-dichlorophenoxyacetic acid (2,4-D) tested. Highest callus induction frequency was exhibited at a concentration of 6 mg/l 2,4-D for all three accessions.Sub-culturing of callus on regeneration medium, which consisted of MS supplemented with (1:0-5) mg/l NAA:BAP resulted in no plantlet regeneration in all tested accessions. Instead, prolific root formation was observed.