Physiology and Molecular Biology of Plants

, Volume 16, Issue 3, pp 249–257 | Cite as

In vitro propagation of an endangered medicinal herb Chlorophytum borivilianum Sant. et Fernand. through somatic embryogenesis

  • Mohd Zahid RizviEmail author
  • Arun Kumar Kukreja
  • Narendra Singh Bisht
Research Article


Tuberous roots of Chlorophytum borivilianum Sant. et Fernand. which are a source of steroidal saponins, possess immunomodulatory, adaptogenic, aphrodisiac, antipyretic, diuretic, hemostatic and anti-tumour properties. Poor seed setting and germination and slow growth in conventional vegetative propagation are major constraints in the large-scale cultivation of this commercially important medicinal plant. In the present study, a procedure for in vitro propagation of this endangered herb through somatic embryogenesis has been established. Seeds of Chlorophytum borivilianum were germinated on MS medium supplemented with 57.74 μM gibberellic acid and hypocotyl portion from germinated seedling was used as explant for callus induction. Moderate to good callus induction was observed on MS medium containing 1.16 μM kinetin and 1.13–2.26 μM 2,4-dichlorophenoxyacetic acid. Regular subculturing of callus on kinetin (1.16 μM) and 2,4-dichlorophenoxyacetic acid (1.13 μM) supplemented medium induced somatic embryogenesis. In modified MS medium, 1.79 mM NH4NO3 and 10.72 mM KNO3 was optimal for somatic embryogenesis. 7.38 μM 2-isopentenyladenine supplemented to modified MS medium, showed best response for somatic embryogenesis while proline (0.76 mM) as an amino acid supplement gave better response than glutamine. 30% germination of mature somatic embryos was achieved on MS medium supplemented with 15.54 μM 6-benzylaminopurine. Multiplication of C. borivilianum through somatic embryogenesis may offer a better approach compared to organogenesis for developing scale-up technology employing bioreactors for its mass propagation.


Liliaceae Safed musli Saponins Somatic embryogenesis Proline 



We are thankful to Director, Central Institute of Medicinal and Aromatic Plants for providing necessary facilities during the work period. Grant of Senior Research Fellowship to M. Z. Rizvi by Council of Scientific and Industrial Research, New Delhi is duly acknowledged.


  1. Armstrong CL, Green CE (1985) Establishment and maintenance of friable embryogenic maize callus and the involvement of L-proline. Planta 164:207–214CrossRefGoogle Scholar
  2. Arora DK (1999) Micropropagation and selection of variants in certain medicinal plants. PhDThesis, Mohan Lal Sukhadia University, Udaipur, India, pp 1–96Google Scholar
  3. Bordia PC, Joshi A, Simlot MM (1995) Safed musli. In: Chadha KL, Gupta R (eds) Advances in horticulture: medicinal and aromatic plants, vol II. Malhotra Pub House, New Delhi, pp 429–451Google Scholar
  4. Choi YE, Jeong JH, Shin CK (2003) Hormone independent embryogenic callus production from ginseng cotyledons using high concentrations of NH4NO3 and progress towards bioreactor production. Plant Cell Tissue Organ Cult 72:229–235CrossRefGoogle Scholar
  5. Debergh PC, Maene LJ (1981) A scheme for commercial propagation of ornamental plants by tissue culture. Sci Hortic 14:335–345CrossRefGoogle Scholar
  6. Duque A, Pires AS, Santos DMD, Fevereiro P (2006) Efficient somatic embryogenesis and plant regeneration from long-term cell suspension cultures of Medicago truncatula cv. Jemalong. In Vitro Cell Dev Biol Plant 42:270–273CrossRefGoogle Scholar
  7. Feher A, Pasternak TP, Dudits D (2003) Transition of somatic plant cells to an embryogenic state. Plant Cell Tissue Organ Cult 74:201–228CrossRefGoogle Scholar
  8. Feng GH, Ouyang J (1988) The effects of KNO3 concentration in callus induction medium for wheat anther culture. Plant Cell Tissue Organ Cult 12:3–12CrossRefGoogle Scholar
  9. Finer JJ, McMullen D (1991) Transformation of soybean via particle bombardment of embryogenic suspension culture tissue. In Vitro Cell Dev Biol Plant 27:175–182CrossRefGoogle Scholar
  10. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158CrossRefPubMedGoogle Scholar
  11. Ho CW, Jian WT, Lai HC (2006) Plant regeneration via somatic embryogenesis from suspension cell cultures of Lilium x formolongi Hort. using a bioreactor system. In Vitro Cell Dev Biol Plant 42:240–246Google Scholar
  12. Jat RD, Bordia PC (1990) Propagation studies in safed musli (Chlorophytum species). In: Chaudhary BL, Aery NC, Katewa SS (eds) Proceedings of national symposium on advances in plant sciences: current status and emerging challenges. Department of Botany, Mohan Lal Sukhadia University, Udaipur, p 46Google Scholar
  13. Jha S, Mitra GC, Sen S (1984) In vitro regeneration from bulb explants of Indian squill. Plant Cell Tissue Organ Cult 3:91–100CrossRefGoogle Scholar
  14. Kirtikar KR, Basu BD (1975) Liliaceae: Chlorophytum. In: Kirtikar KR, Basu BD (eds) Indian medicinal plants. Bishan Singh Mahendra Pal Singh, Dehradun, pp 2508–2509Google Scholar
  15. Krikorian AD, Kann RP (1986) Regeneration in Liliaceae, Iridaceae and Amaryllidaceae. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants. Academic, New York, pp 187–205Google Scholar
  16. Kukda G, Purohit SD, Dave A (1994) Somatic embryogenesis and plantlet regeneration in safed musli. Indian J Plant Genet Resour 7(1):65–71Google Scholar
  17. Kumar JV, Ranjitha Kumari BD, Castano E (2008) Cyclic somatic embryogenesis and efficient plant regeneration from callus of safflower. Biol Plant 52(3):429–436CrossRefGoogle Scholar
  18. Li XC, Wang DZ, Yang CR (1990) Steroidal saponins from Chlorophytum malayense. Phytochem 29:3893–3898CrossRefGoogle Scholar
  19. Menke-Milczarek I, Zimny J (2001) NH4+ and NO3 requirement for wheat somatic embryogenesis. Acta Physiol Plant 23(1):37–42CrossRefGoogle Scholar
  20. Merkle SA, Parrott WA, Flinn BS (1995) Morhogenetic aspects of somatic embryognesis. In: Thorpe TA (ed) In vitro embryogenesis in plants. Kluwer Acad, Dordrecht, pp 155–203Google Scholar
  21. Mimaki Y, Kanmoto T, Sashida Y, Nishino A, Satomi Y, Nishino H (1996) Steroidal saponins from the underground parts of Chlorophytum comosum and their inhibitory activity on tumour promoter-induced phospholipids metabolism of He La cells. Phytochem 41:1405–1410CrossRefGoogle Scholar
  22. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  23. Nayar MP, Sastry ARK (1988) Chlorophytum borivilianum. In: Nayar MP, Sastry ARK (eds) Red Data Book of Indian Plants, Vol 2. Botanical Survey of India, p 412Google Scholar
  24. Nuti RV, Caligo MA, Nozzolini M, Luccarine G (1984) Stimulation of carrot somatic embryogenesis by proline and serine. Plant Cell Rep 3:210–213CrossRefGoogle Scholar
  25. Ojha A (1987) Botanical study of safed musli (aphrodisiac effects). MD Thesis, MMR Ayurvedic College, Udaipur, IndiaGoogle Scholar
  26. Purohit SD, Dave A, Kukda G (1994) Soamtic embryogenesis and plantlet regeneration in ‘safed musli’ (Chlorophytum borivilianum). Indian J Plant Genet Resour 7(1):65–71Google Scholar
  27. Ramawat KG, Jain S, Suri SS, Arora DK (1988) Aphrodisiac plants of Aravalli Hills with special reference to safed musli. In: Khan I, Khanum A (eds) Role of biotechnology in medicinal and aromatic plants. Ukaz, Hyderabad, pp 210–223Google Scholar
  28. Rangasamy NS (1986) Somatic embryogenesis in angiosperm cell, tissue and organ cultures. Proc Indian Acad Sci 95:247–271CrossRefGoogle Scholar
  29. Sagare AP, Lee YL, Lin TC, Chen CC, Tsay SS (2000) Cytokinin induced somatic embryogenesis and plant regeneration in Corydalis yanhusuo-a medicinal plant. Plant Sci 160(1):139–147CrossRefPubMedGoogle Scholar
  30. Shibli RA, Ajlouni MM (2000) Somatic embryogenesis in the endemic black iris. Plant Cell Tissue Organ Cult 61(1):15–21CrossRefGoogle Scholar
  31. Shibli RA, Shatnawi M, Abu-Ein, Al-Juboory KH (2001) Somatic embryogenesis and plant recovery from callus of ‘Nabali’ Olive (Olea europea L.). Sci Hortic 88(3):243–256CrossRefGoogle Scholar
  32. Siriwardana S, Nabros MW (1983) Tryptophan enhancement of somatic embryogenesis in rice. Plant Physiol 73:142–146CrossRefPubMedGoogle Scholar
  33. Sozinov A, Lukyanyuk S, Ignatova S (1981) Anther cultivation and induction of haploid plant in triticale. Z. Pflanzenzuchtg 86:272–285Google Scholar
  34. Tandon M, Shukla YN (1992) Sapogenins from Asparagus adscendens and Chlorophytum arundinaceum. J Indian Chem Soc 69:893Google Scholar
  35. Tandon M, Shukla YN (1993) A bibenzyl xyloside from Chlorophytum arundinaceum. Phytochem 32:1624–1625CrossRefGoogle Scholar
  36. Tandon M, Shukla YN (1995) Phytoconstituents of Asparagus adscendens, Chlorophytum arundinaceum and Curculigo orchioides: a review. Curr Res Med Aromat Plants 17:42–50Google Scholar
  37. Tandon M, Shukla YN, Thakur RS (1992) 4-hydroxyl-8, 11-oxidohemicosanol and other constituents from Chlorophytum arundinaceum roots. Phytochem 31:2525–2526CrossRefGoogle Scholar
  38. Varisai MS, Wang CS, Thiruvengadam M, Jayabalan N (2004) In vitro plant regerneration via somatic embryogenesis through cell suspension cultures of horsegram [Macrotyloma uniflorum (Lam.) Verdc.]. In Vitro Cell Dev Biol Plant 40:284–289CrossRefGoogle Scholar
  39. Wang W, Cui SX, Zhang CL (2001) Plant regeneration from embryogenic suspension cultures of dune reed. Plant Cell Tissue Organ Cult 67:11–17CrossRefGoogle Scholar
  40. Xu ZH, Sunderland N (1981) Glutamine, inositol and conditioning factor in the production of barley pollen callus in vitro. Plant Sci Lett 23:161–168CrossRefGoogle Scholar
  41. Yeung EC (1995) Structural and developmental patterns in somatic embryogenesis. In: Thorpe TA (ed) In vitro embryogenesis in plants. Kluwer Acad, Dordrecht, pp 205–247Google Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2010

Authors and Affiliations

  • Mohd Zahid Rizvi
    • 1
    Email author
  • Arun Kumar Kukreja
    • 1
  • Narendra Singh Bisht
    • 2
  1. 1.Plant Tissue Culture Division, Central Institute of Medicinal and Aromatic Plants (CIMAP)Council of Scientific and Industrial ResearchLucknow P.O. CIMAPIndia
  2. 2.Department of BotanyHemwati Nandan Bahuguna Garhwal UniversityUttrakhandIndia

Personalised recommendations