Comparison of secondary plant metabolite production in cell suspension, callus culture and temporary immersion system

  • Dirk Wilken
  • Elio Jiménez González
  • Annette Hohe
  • Miguel Jordan
  • Rafael Gomez Kosky
  • Guillermo Schmeda Hirschmann
  • André Gerth


Cell and organ cultures of Lavandula officinalis, Hypericum perforatum, Cymbopogon citratus and Fabiana imbricata were established for the production of secondary metabolites in vitro. Shoot multiplication was performed by conventional micropropagation on agar-solidified medium as well as in temporary immersion systems (TIS), the latter resulted in higher multiplication rates compared to the culture in microcontainers for all plant species tested. The concentration of bioactive compounds was determined in different in vitro cell and organ cultures and was compared to field grown plants. For Lavandula the highest content of rosmarinic acid was found in cell cultures, for the other three species in field grown plants. Concentrations of bioactive compounds were always higher in plant material grown in TIS compared to cell suspension and callus cultures.

Key words

bioactive compounds bioreactor culture Cymbopogon Fabiana Hypericum in vitro culture Lavandula pharmaceuticals suspension culture 


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  1. Akula A, Becker D & Bateson M (2000) High-yielding somatic embryogenesis and plant recovery in a selected tea clone, ‘TRI-2025’, by temporary immersion. Plant Cell Rep. 19: 1140–1145Google Scholar
  2. Berlin J, Sieg S, Strack D, Bokern M & Harms H (1986) Production of betalains by suspension cultures of Chenopodium rubrum. Plant Cell, Tiss. Org. Cult. 5: 163–174Google Scholar
  3. Bhojwani SS & Razdan MK (1996) Plant Tissue Culture: Theory and Practice, a revised edition. Elsevier, Amsterdam, Lausanne, New York, Oxford, Shannon, TokyoGoogle Scholar
  4. Charlwood BV & Rhodes MJC (1990) Secondary Products from Plant Tissue Culture. Clarendon Press, OxfordGoogle Scholar
  5. De-Eknamkul W & Ellis B (1984) Rosmarinic acid production and growth characterization of Anchusa officinalis cell suspension cultures. Planta Med. 50: 346–350PubMedGoogle Scholar
  6. Drapeau D, Blanch HW & Wilke CR (1987) Ajmalicine, serpentine, and catharanthine accumulation in Catharanthus roseus bioreactor cultures. Planta Med. 53: 373–376Google Scholar
  7. European Pharmacopeia (2002) 4th Edition, Council of Europe, 67075 Strasbourg Cedex, France 2001, ISBN: 92-871-4587-3: 183–184Google Scholar
  8. Fowler MW (1992) Plant cell culture, process systems and product synthesis. In: Fowler MW, Warren GS & Moo-Young M (eds) Plant Biotechnology: Comprehensive Biotechnology, Second Supplement (pp. 79–88)Google Scholar
  9. Fujita Y (1988) Shikonin: production by plant (Lithospermum erythrorizon) cell cultures. In: Bajaj YPS (ed) Biotechnology in Agriculture and Forestry Vol. 4. Medicinal and Aromatic Plants 1 (pp. 225–236). Springer, BerlinGoogle Scholar
  10. Fujita Y (1990) The production of industrial compounds. In: Bhojwani SS (ed) Plant Tissue Culture: Applications and Limitations (pp. 259–275). Elsevier, AmsterdamGoogle Scholar
  11. Fujita Y, Tabata M, Nishi A & Yamada Y (1982) New medium and production of secondary compounds with the two-staged culture method. In: Fujiwara A (ed) Plant Tissue Culture (pp. 399–400) Jpn. Assoc. Plant Tissue Cult. TokyoGoogle Scholar
  12. Gracza L & Ruff P (1984) Rosmarinsäure in Arzneibuchdrogen und ihre HPLC-Bestimmung. Arch. Pharm. 317: 339–345Google Scholar
  13. Halkes SBA (1998) Filipendula ulmaria: a study on the immunomodulatory activity of extracts and constituents. PhD Thesis, Chapter 5 (pp. 91–92). University Utrecht, The NetherlandsGoogle Scholar
  14. Jardin B, Tom R, Chavarie C, Rho D & Archambault J (1991) Stimulated indole alkaloid release from Catharanthus roseus immobilized cultures. Initial studies. J. Biotechnol. 21: 43–62Google Scholar
  15. Jiménez E, Pérez N, de Feria M, Barbón R, Capote A, Chávez M, Quiala E & Pérez JC (1999) Improved production of potato microtubers using a temporary immersion system. Plant Cell, Tiss. Org. Cult. 59: 19–23Google Scholar
  16. Kartnig T, Gröbel I & Heydel B (1996) Production of hypericin, pseudohypericin and flavanoids in cell cultures of various Hypericum species and their chemotypes. Planta Med. 62: 51–53PubMedGoogle Scholar
  17. Lambie AJ (1990) Commercial aspects of the production of secondary compounds by immobilized plant cells. In: Charlwood BV & Rhodes MJC (eds) Secondary Products from Plant Tissue Culture (pp. 265–278). Clarendon Press, OxfordGoogle Scholar
  18. Lopez-Arnaldos T, Lopez-Serrano M, Ros Barcelo A, Calderon AA & Zapata JM (1994) Tentative evidence of a rosmarinic acid peroxidase in cell cultures from lavandin (Lavandula intermediata) flowers. Biochem. Molecular Biol. Internat. 34: 809–816Google Scholar
  19. Lorenzo JC, González BL, Escalona M, Teisson C, Espinosa P & Borroto C (1998) Sugarcane shoot formation in an improved temporary immersion system. Plant Cell, Tiss. Org. Cult. 54: 197–200Google Scholar
  20. Moreno PRH, van der Heijden R & Verpoorte R (1995) Cell and tissue culture of Catharanthus roseus: a literature survey. II. Updating from 1988 to 1993. Plant Cell, Tiss. Org. Cult. 42: 1–25Google Scholar
  21. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15: 473–497Google Scholar
  22. Payne GF, Payne NN, Shuler ML & Asada M (1988) In situ adsorption for enhanced alkaloid production by Catharanthus roseus. Biotechnol. Lett. 10: 187–192Google Scholar
  23. Schiel O & Berlin J (1987) Large scale fermentation and alkaloid production of cell suspension cultures of Catharanthus roseus. Plant Cell, Tiss. Org. Cult. 8: 153–161Google Scholar
  24. Schlatmann JE, Moreno PRH, Ten Hoopen HJG, Verpoorte R & Heijnen JJ (1994) Effect of oxygen and nutrient limitation on ajmalicine production and related enzyme activities in high density cultures of Catharanthus roseus. Biotechnol. Bioeng. 44: 461–468Google Scholar
  25. Schlatmann JE, Ten Hoopen HJG & Heijnen JJ (1992) Optimization of the medium composition for alkaloid production of Catharanthus roseus using statistical experimental designs. Med. Fac. Landbouw., Univ. Gent 57: 1567–1569Google Scholar
  26. Scragg AH (1986) The economics of mass cell culture. In: Morris P (ed) Secondary Metabolism in Plant Cell Cultures (pp. 202–207). Cambridge University Press, CambridgeGoogle Scholar
  27. Tisserat B & Vaughn SF (2001) Essential oils enhanced by ultra-high carbon dioxide levels from Lamiaceae species grown in vitro and in vivo. Plant Cell Rep. 20: 361–368Google Scholar
  28. Ulbrich B, Weisner W & Arens H (1985) Large scale production of rosmarinic acid from plant cell culture of Coleus blumei. In: Neumann KH (ed) Primary and Secondary Metabolism of Plant Cell Cultures (pp. 293–303). Springer, BerlinGoogle Scholar
  29. Zenk MH, El-Shagi H, Arens H, Stockigt J, Weiler EW & Deus D (1977) Formation of the indole alkaloids serpentine and ajmalicine in cell suspension cultures of Catharanthus roseus. In: Barz WRE Reinhard E & Zenk MH (ed) Plant Tissue Culture and its Biotechnological Application (pp. 27–44). Springer, BerlinGoogle Scholar
  30. Zobayed SMA, Murch SJ, Rupasinghe HPV & Saxena PK (2003) Elevated carbon supply altered hypericin and hyperforin contents of St. John’s wort (Hypericum perforatum) grown in bioreactors. Plant Cell, Tiss. Org. Cult. 75: 143–149Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Dirk Wilken
    • 1
  • Elio Jiménez González
  • Annette Hohe
    • 1
  • Miguel Jordan
    • 3
  • Rafael Gomez Kosky
    • 2
  • Guillermo Schmeda Hirschmann
    • 4
  • André Gerth
    • 1
  1. 1.BioPlanta GmbHLeipzigGermany
  2. 2.Instituto de BiotecnologÍa de las PlantasSanta ClaraCuba
  3. 3.Departamento de EcologÍaPontificia Universidad Católica de ChileSantiagoChile
  4. 4.Instituto de Quimica de Recursos NaturalesUniversidad de TalcaTalcaChile

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