ORIGINAL PAPER In vitro propagation and biosynthesis of steroidal sapogenins from various morphogenetic stages of Moringa oleifera Lam., and their antioxidant potential Manas Mathur • Sunita Yadav • Pawan K. Katariya • Raka Kamal Received: 13 May 2013 / Revised: 16 April 2014 / Accepted: 23 April 2014 Ó Franciszek Go ´rski Institute of Plant Physiology, Polish Academy of Sciences, Krako ´w 2014 Abstract Moringa oleifera is a highly valued medicinal plant. The present research reports callus cultures of M. oleifera Lam., established from seeds and nodal segments on Murashige and Skoog’s (MS) medium using different concentrations and combinations of auxins and cytokinins. Best induction of callus was observed at BAP:IBA (3 mg l -1 each). Shooting and rooting from callus in terms of morphogenesis were observed in MS media supple- mented with BAP:KN (2:0.2 mg l -1 ) and IBA:NAA (3:0.5 mg l -1 ), respectively. Multiple shooting was observed at treatment dose of BAP:NAA:IAA (1:1:0.2 mg l -1 ). Regenerated shoots were rooted and mature plants were established, acclimatized, and thrived in greenhouse conditions. Over 95 % of plantlets survived after transplanting plantlets into trays with a mixture of sand and perlite (2:1) for 20 days. The regeneration pro- tocol developed in this study provides a basis for germ- plasm conservation and for further investigation of bioactive constituents of this medicinal plant. Further qualitative and quantitative production of steroidal sapo- genins (diosgenin and tigogenin) from various morphoge- netic stages was studied using TLC, PTLC, IR spectra, HPLC and GC–MS analysis. Steroidal sapogenins were maximum in the callus associated with rooting. Various stages were further analyzed for their antioxidant potential. Keywords Moringa oleifera  Nodal segment  Morphogenesis  Steroidal sapogenins  PTLC  HPLC  GC–MS  IR spectra  Antioxidant Abbreviations 2, 4 D 2, 4-Dichlorophenoxyacetic acid MS Murashige and Skoog NAA a-Naphthalene acetic acid IBA Indole butyric acid IAA Indole acetic acid Kn Kinetin BAP Benzyl amino purine TLC Thin layer chromatography PTLC Preparative thin layer chromatography IR-Infra Red spectroscopy HPLC High performance thin layer chromatography GC–MS Gas chromatography and mass spectroscopy DPPH 2,2-Diphenyl-1-picrylhydrazyl Introduction Moringa oleifera Lam., commonly called as ‘Sahanjana’ belongs to family Moringaceae, which bears 14 species among which M. oleifera is most commonly found. The plant is native to northern India. The pods and leaves are reported to contain 2.5 and 6.7 g protein/100 g, respec- tively (Verma et al. 1976). The plant parts are used in folk remedies for tumors, abdominal discomfort, boils, cold, conjunctivitis, high blood pressure, hysteria, relapsing fever, skin diseases, etc. (Hartwell 1967–1971). The plant also bears some bioactivities, viz., anti-inflammatory (Su- laiman et al. 2008), anti-asthmatic (Agrawal and Mehta Communicated by K.-Y. Paek. M. Mathur  S. Yadav  P. K. Katariya  R. Kamal (&) Laboratory of Medicinal Plant Biotechnology, Department of Botany, University of Rajasthan, Jaipur 302004, India e-mail: rakakamal@hotmail.com 123 Acta Physiol Plant DOI 10.1007/s11738-014-1549-1 Author's sample copy