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Ceropegia bulbosa Roxb. (family - Asclepiadaceae), a medicinally important plant, is classified as a ’threatened plant’. An efficient protocol of shoot organogenesis and plant regeneration from seedling derived callus has been developed. Out of different explants i.e. epicotyl, hypocotyl, cotyledonary node and root, epicotyl was found to be the most suitable for establishment of cultures and further shoot regeneration. Callus was induced from epicotyl explants on MS medium containing 1.0 mg l(-1) of 2,4-D. Proliferation of callus was observed on MS + 1.0 mg l(-1) of 2,4-D and 0.5 mg l(-1) of BAP. The shoot regeneration was achieved when proliferated callus was transferred to MS medium supplemented with BAP either alone or in combination with IAA or NAA. Maximum number (25.5) of shoots were regenerated on MS medium containing 1.0 mg l(-1) BAP and 0.1 mg l(-1) NAA. The shoots (100%) were rooted ex vitro when treated with IBA (100 mg l(-1)) for 3 min. Rooted plantlets were hardened successfully and transferred to the field. The described method can be successfully used for the large-scale multiplication and conservation of germplasm of this important and threatened plant. (C) 2012 Elsevier B.V. All rights reserved.