Journal of Crop Science and Biotechnology

, Volume 15, Issue 4, pp 297–303 | Cite as

Actions for ex situ conservation of Gloriosa superba L. - an endangered ornamental cum medicinal plant

  • Kuldeep Yadav
  • Ashok Aggarwal
  • Narender Singh
Research Article


Factors affecting in vitro propagation and microtuberization were evaluated for Gloriosa superba L., an endangered ornamental cum medicinal plant having limited reproductive capacity. Surface sterilization of tuber explants with 0.1% mercuric chloride (HgCl2) for 5 min eliminated the contamination effectively with highest survival rate. Among the various combinations used, Murashige and Skoog (MS) medium with 2.0 mg L−1 6-benzylaminopurine (BAP) + 0.5 mg L−1 α-naphthalene acetic acid (NAA) containing 3% sucrose with 16-h photoperiod exhibited the greatest in vitro tuberization (3.2) with the highest shoot regeneration frequency (90%). The longest tuber regeneration occurred on MS media containing 4% sucrose. Transfer of in vitro-regenerated shoots to half-strength MS medium with 1.0 mg L−1 indole-3-butyric acid (IBA) + 0.5 mg L−1 NAA showed maximum root induction (66.6%). The in vitro-grown plantlets were successfully acclimatized and transplanted to sterilized soil and sand mixture (3:1) in the glasshouse with 70% survival. The colchicine content was determined in the tubers of ex vitro plants by HPLC using the same retention time (1.5 min) as that of the standard colchicine. This revealed that the micropropagation protocol developed by us for rapid mass production could be used as raw material for colchicine extraction and provides a basis for germplasm conservation and genetic improvement of G. superba.

Key words

conservation endangered Gloriosa superba HPLC microtuber induction 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ade R, Rai MK. 2009. Review: Current advances in Gloriosa superba L. Biodiversitas 10: 210–214Google Scholar
  2. Aksenova NP, Konstamtinova TN, Golyanovskaya SS, Kossmann J, Willmitzer L, Romanov. 2000. In vitro microtuberization in Potato. Russ. J. Plant Physiol. 133: 23–27Google Scholar
  3. Alali F, Tawaha K, Qasaymch RM. 2004. Determination of colchicine in Colchicum steveni and C. hierosolymitanum (Colchicaceae): Comparison between two analytical methods. Phytochem. Anal. 15: 27–29PubMedCrossRefGoogle Scholar
  4. Amoroso EC. 1935. Colchicine and tumour growth. Nature 135: 266–267CrossRefGoogle Scholar
  5. Ang BH, Chan LK. 2003. Micropropagation of Spilanthes acmella L., a bio-insecticide plant, through proliferation of multiple shoots. J. Appl. Hortic. 5: 65–68Google Scholar
  6. Anonymous. 1956. The Wealth of India: a dictionary of Indian raw material and industrial products. CSIR, Delhi. 4: 139–140Google Scholar
  7. Behera KK, Sahoo S, Prusti A. 2009. Regeneration of plantlet of water yam (Dioscorea oppositifoliaa L.) through in vitro culture from nodal segments. Not. Bot. Hort. Agrobot. Cluj. 37: 94–102Google Scholar
  8. Bernier G, Pe’rilleux C. 2005. A physiological overview of the genetics of flowering time control. Plant Biotechnol. J. 3: 3–16PubMedCrossRefGoogle Scholar
  9. Bhojwani SS, Razdan MK. 2005. Production of secondary metabolites. In Plant Tissue Culture: Theory and Practice, Rev. eds, Amsterdam, Elsevier, pp 537–562Google Scholar
  10. Charles G, Rossignol L, Rossignol M. 1992. Environmental effects on potato plants in vitro. J. Plant Physiol. 139: 708–713CrossRefGoogle Scholar
  11. Chawla HS. 2003. Plant Biotechnology: A Practical Approach. Science Publishers Inc., Enfield (NH), USA, ISBN:157808296X, pp 51–55Google Scholar
  12. Chen FQ, Fu Y, Wang DL, Gao X, Wang L. 2007. The effect of plant growth regulators and sucrose on the micropropagation and microtuberization of Dioscorea nipponica Makino. J. Plant Growth Regul. 26: 38–45CrossRefGoogle Scholar
  13. Fernie AR, Willmitzer L. 2001. Molecular and biochemical triggers of potato tuber development. Plant Physiol. 127: 1459–1465PubMedCrossRefGoogle Scholar
  14. Forsyth C, Van Staden J. 1984. Tuberization of Dioscorea bulbifera stems nodes in culture. J. Plant Physiol. 115: 79–83PubMedCrossRefGoogle Scholar
  15. Ghosh B, Mukherjee S, Jha TB, Jha S. 2002. Enhanced colchicine production in root cultures of Gloriosa superba by direct and indirect precursors of the biosynthetic path ways. Biotechnol. Lett. 24: 231–234CrossRefGoogle Scholar
  16. Ghosh S, Ghosh B, Jha S. 2006. Aluminium chloride enhances colchicine production in root cultures of Gloriosa superba. Biotechnol. Lett. 28: 497–503PubMedCrossRefGoogle Scholar
  17. Goyal D, Bhadauria S. 2006. In vitro propagation of Ceropegia bulbosa using nodal segments. Ind. J. Biotech. 5: 565–568Google Scholar
  18. Grison C. 1991. Influence des facteurs d’environnement sur le cycle ve’ge’tatif de la pomme de terre. La pomme de terre française 462: 7–15Google Scholar
  19. Hara M, Tanaka S, Tabata M. 1994. Induction of a specific methyltransferase activity regulating berberine biosynthesis by cytokinin in Thalictrum minus cell cultures. Phytochemistry 36: 327–332CrossRefGoogle Scholar
  20. Hassan AKMS, Roy SK. 2005. Micropropagation of Gloriosa superba L. through high frequency shoot proliferation. Plant Tiss. Cult. 15: 67–74Google Scholar
  21. Jana S, Shekhawat GS. 2011a. Critical review on medicinally potent plant species: Gloriosa superba. Fitoterapia 82: 293–301PubMedCrossRefGoogle Scholar
  22. Jana S, Shekhawat GS. 2011b. Plant growth regulators, adenine sulfate and carbohydrates regulate organogenesis and in vitro flowering of Anethum graveolens. Acta Physiol. Plant. 33: 305–311CrossRefGoogle Scholar
  23. Jo EA, Tewari RK, Hahn EJ, Paek KY. 2009. In vitro sucrose concentration affects growth and acclimatization of Alocasia amazonica plantlets. Plant Cell Tiss. Organ Cult. 96: 307–315CrossRefGoogle Scholar
  24. Kala C, Farooquee N, Dhar U. 2004. Prioritization of medicinal plants on the basis of available knowledge, existing practices and use value status in Uttaranchal, India. Biodivers. Conserv. 13: 453–469CrossRefGoogle Scholar
  25. Komalavalli N, Rao MV. 2000. In vitro micropropagation of Gymnema sylvestre — A multipurpose medicinal plant. Plant Cell Tiss. Org. Cult. 61: 97–105CrossRefGoogle Scholar
  26. Krause J. 1986. Production of Gloriosa tubers from seeds. Acta Hortic. 177: 353–360Google Scholar
  27. Kumar S, Mangal M, Dhawan AK, Singh N. 2011. Assessment of genetic fidelity of micropropagated plants of Simmondsia chinensis (Link) Schneider using RAPD and ISSR markers. Acta Physiol. Plant. 33: 2541–2545CrossRefGoogle Scholar
  28. Lawrence CH, Barker WG. 1963. A study of tuberization in the potato, Solanum tuberosum. Amer. Potato J. 40: 349–356CrossRefGoogle Scholar
  29. Maiti CK, Sen S, Paul AK, Acharya K. 2007. First report of leaf blight disease of Gloriosa superba L. caused by Alternaria alternata (Fr.) Keissler. Ind. J. Plant Pathol. 73: 377–378Google Scholar
  30. Mantell SH, Hugo SA. 1989. Effect of photoperiod, mineral medium strength, ionic ammonium and cytokinin on root, shoot and microtuber development in shoot cultures of D. alata and D. bulbifera yam. Plant Cell Tiss. Organ Cult. 16: 23–27CrossRefGoogle Scholar
  31. Medina RD, Flachsland EA, Gonzalez AM, Terada G, Faloci MM, Mroginski LA. 2009. In vitro tuberization and plant regeneration from multimodal segment culture of Habenaria bractescens Lindl., an Argentinean wetland orchid. Plant Cell Tiss. Organ Cult. 97: 91–101CrossRefGoogle Scholar
  32. Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497CrossRefGoogle Scholar
  33. Ohyama K. 1970. Tissue culture in mulberry tree. Jpn. Agric. Res. Quart. 5: 30–34Google Scholar
  34. Omokolo ND, Boudjeko T, Tsafack TJJ. 2003. In vitro tuberization of Xanthosoma sagittifolium (L.) Schott: Effects of phytohormones, sucrose, nitrogen and photoperiod. Sci. Hortic. 98: 337–345CrossRefGoogle Scholar
  35. Omokolo ND, Tsala NG, Kanmegne G, Balange AP. 1995. Production of multiple shoots, callus, plant regeneration and tuberization in Xanthosoma sagittifolium cultured in vitro. C R Acad. Sci. 318: 773–778Google Scholar
  36. Ovono PO, Kevers C, Dommes J. 2007. Axillary proliferation and tuberization of Dioscorea cayenensis-D. rotundata complex. Plant Cell Tiss. Org. Cult. 91: 107–114CrossRefGoogle Scholar
  37. Ovono PO, Kevers C, Dommes J. 2009. Effects of reducing sugar concentration on in vitro tuber formation and sprouting in yam (Dioscorea cayenensis-D. rotundata complex). Plant Cell Tiss. Organ Cult. 99: 55–59CrossRefGoogle Scholar
  38. Pan RC. 2001. Phytophysiology 4th edn. Higher Education Press, Beijing, China, pp 176–186Google Scholar
  39. Pelacho AM, Mingo-Castel AM. 1991. Effects of photoperiod on kinetin-induced tuberization of isolated potato stolons cultured in vitro. Am. Potato J. 68: 533–541CrossRefGoogle Scholar
  40. Perl A, Aviv D, Willmitzer L, Galun E. 1991. In vitro tuberization in transgenic potatoes harboring-glucuronidase linked to a patatin promoter: effects of sucrose levels and photoperiods. Plant Sci. 73: 87–95CrossRefGoogle Scholar
  41. Pierik RLM. 1967. Factors affecting the induction and initiation of flower buds in vitro in plant species with a cold requirement for flowering. In G Bernier, eds, Cellular and Molecular Aspects of Floral Induction, Longman Grp Ltd., London, pp 408–4115Google Scholar
  42. Poornima GN, Rai VR. 2007. In vitro propagation of wild yams, Dioscorea oppositifolia (Linn) and Dioscorea pentaphylla (Linn). Afr. J. Biotechnol. 6: 2348–2352Google Scholar
  43. Rai VR. 2002. Rapid clonal propagation of Nothapodytes foetida (Weight) Sleumer — a threatened medicinal tree. In Vitro Cell Dev. Biol. Plant 38: 347–351Google Scholar
  44. Robins RJ, Parr AJ, Payne J, Walton NJ, Rhodes MJC. 1990. Factors regulating tropane-alkaloid production in a trans formed root culture of a Datura candida X D. aurea hybrid. Planta 181: 414–422CrossRefGoogle Scholar
  45. Sahoo Y, Chand PK. 1998. Micropropagation of Vitex negundo L., a woody aromatic medicinal shrub, through high frequency axillary shoot proliferation. Plant Cell Rep. 18: 301–307CrossRefGoogle Scholar
  46. Sarkar D. 2008. The signal transduction pathways controlling in planta tuberization in potato: An emerging synthesis. Plant Cell Rep. 27: 1–8PubMedCrossRefGoogle Scholar
  47. Satyavati GV, Raina MK, Sharma M. 1976. Medicinal Plants of India. ICMR, Delhi, Vol.1, pp 433–434Google Scholar
  48. Seabrook JEA, Coleman S, Levy D. 1993. Effect of photoperiod on in vitro tuberization of potato (Solanum tuberosum L.). Plant Cell Tiss. Org. Cult. 34: 43–51CrossRefGoogle Scholar
  49. Sivakumar G, Krishnamurthy KV. 2000. Micropropagation of Gloriosa superba L-an endangered species of Asia and Africa. Curr. Sci. 78: 31–32Google Scholar
  50. Sivakumar G, Krishnamurthy KV. 2002. Gloriosa superba L. — a very useful medicinal plant. In VK Singh, JN Govil, S Hashmi, G Singh, eds, Recent Progress in Medicinal Plants, Vol. 7, Ethnomedicine and Pharmacognosy, Part II. Series Sci. Tech. Pub., Texas, USA, pp 465–82Google Scholar
  51. Sivakumar G, Krishnamurthy KV. 2004. In vitro organogenetic responses of Gloriosa superba. Russ. J. Plant Physiol. 51: 790–798CrossRefGoogle Scholar
  52. Tiwari KN, Sharma NC, Tiwari V, Singh BD. 2000. Micropropagation of Centella asiatica (L.), a valuable medicinal herb. Plant Cell Tiss. Organ Cult. 63: 179–185CrossRefGoogle Scholar
  53. Vaillant V, Bade P, Constant C. 2005. Photoperiod affects the growth and development of yam plantlets obtained by in vitro propagation. Biol. Plant. 49: 355–359CrossRefGoogle Scholar
  54. Yadav K, Singh N. 2011. Effect of seed harvesting season and sterilization treatments on germination and In vitro propagation of Albizia lebbeck (L.) Benth. Analele din Oradea - Fascicula Biologie 18: 151–156Google Scholar
  55. Yadav K, Singh N. 2012. Factors influencing in vitro plant regeneration of Liquorice (Glycyrrhiza glabra L.). Iranian J. Biotechnol. 10: 161–167Google Scholar

Copyright information

© Korean Society of Crop Science and Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  1. 1.Department of BotanyKurukshetra UniversityHaryanaIndia

Personalised recommendations