An efficient system for plant regeneration of Senna occidentalis from hypocotyl-derived callus was developed. Callus was induced from leaf and hypocotyl explants on MS medium amended with 9.04 μM 2,4-D + 2.22 μM BAP and 10.74 μM NAA + 2.22 μM BAP. Medium browning due to leaching of compounds from callus was encountered and ameliorated through incorporation of 2.84 μM ascorbic acid. Leaf-derived callus showed no shoot induction ability, while hypocotyl-derived callus produced shoots in all cytokinin-amended treatments and also in combination with 2.68 μM NAA. For shoot formation, BAP-augmented treatments were better than medium with Kin added. Rhizogenesis was better on 1/2 MS basal medium with IBA than in the NAA and IAA treatments. Regenerated plants were acclimatized with 94% survival and showed similar morphology to field-grown plants.