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Production of asiaticoside and madecassoside in Centella asiatica in vitro and in vivo

  • Original Papers
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Biologia Plantarum

Abstract

The localization was determined of the triterpenoids, asiaticoside and madecassoside, in different organs of glasshouse-grown plants and cultured material, including transformed roots, of two phenotypes of Centella asiatica (L.) Urban of Malaysian origin. Methanolic extracts of asiaticoside and madecassoside were prepared for gradient HPLC analysis. The two phenotypes of C. asiatica exhibited differences in terpenoid content that were tissue specific and varied between glasshouse-grown plants and tissue culture-derived material. Terpenoid content was highest in leaves, with asiaticoside (0.79 ± 0.03 and 1.15 ± 0.10 % of dry mass) and madecassoside [0.97 ± 0.06 and 1.65 ± 0.01 %(d.m.)] in the fringed (F) and smooth leaf (S) phenotypes, respectively. Roots of the F-phenotype contained the lowest content of asiaticoside [0.12 ± 0.01 %(d.m.)], whereas petioles of S-phenotype plants contained the lowest content of asiaticoside [0.16 ± 0.01 %(d.m.)] and madecassoside [0.18 ± 0.14 %(d.m.)]. Transformed roots were induced using Agrobacterium rhizogens and their growth was maximal on Murashige and Skoog basal medium supplemented with 60 g dm−3 sucrose. However, asiaticoside and madecassoside were undetectable in transformed roots and undifferentiated callus.

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Abbreviations

ANOVA:

analysis of variance

BAP:

6-benzylaminopurine

d.m.:

dry mass

2,4-D:

2,4-dichlorophenoxyacetic acid

ELISA:

enzyme-linked immunosorbent assay

f.m.:

fresh mass

IBA:

indolebutyric acid

MS:

Murashige and Skoog (1962)

LB:

Luria broth

NAA:

α-naphthaleneacetic acid

nptII:

neomycin phosphotransferase gene

NPTII:

neomycin phosphotransferase protein

SEM:

standard error of the mean

References

  • Al-Forkan, M., Power, J.B., Anthony, P., Lowe, K.C., Davey, M.R.: Agrobacterium-mediated transformation of Bangladeshi Indica rices.-Cell. mol. Biol. Lett. 9: 287–300, 2004.

    PubMed  CAS  Google Scholar 

  • Baek, Y.W.: Micropropagation of Centella asiatica (L.) Urban by In Vitro Cultures and Production of Triterpene Glycosides.-Ph.D. Thesis, Chonnan University, Gwangju 1997.

  • Balz, J.P., Courtois, D., Drieu, J., Drieu, K., Reynoird, J.P., Sohier, C., Teng, B.P., Touche, A., Petiard, V.: Production of ginkgolides and bilobalide by Ginkgo biloba plants and tissue cultures.-Planta med. 65: 620–626, 1999.

    Article  PubMed  CAS  Google Scholar 

  • Banerjee, S., Zehra, M., Kumar, S.: In vitro multiplication of Centella asiatica, a medicinal herb from leaf explants.-Curr. Sci. 7: 147–148, 1999.

    Google Scholar 

  • Booncong, P., Eshbaugh, W.H., Ogzewalla, C.D.: Centella asiatica (L.) (Apiaceae): Morphological and cytological characterization.-In: Nahrstedt, A. (ed.): Abstracts of Lectures and Poster Presentations in the 43rd Annual Congress on Medicinal Plant Research. Pp. 92–93. Halle 1995.

  • Cartayrade, A., Neau, E. Sohier, C., Balz, J.P., Carde, J.P., Walter, J.: Ginkgolide and bilobalide in Ginkgo biloba: 1. Sites of synthesis, translocation and accumulation of ginkgolides and bilobalide.-Plant Physiol. Biochem. 35: 859–868, 1997.

    CAS  Google Scholar 

  • Collin, H.A.: Secondary product formation in plant tissue cultures.-Plant Growth Regul. 34: 119–134, 2001.

    Article  CAS  Google Scholar 

  • Curtis, I.S., Power, J.P., Blackhall, N.W., De Laat, A.M.M., Davey, M.R.: Genotype-independent transformation of lettuce using Agrobacterium tumefaciens.-J. exp. Bot. 45: 1441–1449, 1994.

    Article  CAS  Google Scholar 

  • Das, A., Mallick, R.: Correlation between genomic diversity and asiaticoside content in Centella asiatica (L.) Urban.-Bot. Bull. Acad. sin. 32: 1–8, 1991.

    CAS  Google Scholar 

  • Dutta, T., Basu, U.P.: Isothankunic acid — a new triterpene acid from Centella asiatica (URB).-Bull. nat. Inst. Sci. India 37: 178–184, 1968.

    Google Scholar 

  • Flück, H., Jaspersen-Schib, R.: Medicinal Plants and Their Uses.-Foulsham, New York 1976.

    Google Scholar 

  • Giri, A., Narasu, M.L.: Transgenic hairy roots: recent trends and applications.-Biotechnol. Adv. 18: 1–22, 2000.

    Article  PubMed  CAS  Google Scholar 

  • Granicher, F., Christen, P. Kamalaprija, P., Burger, U.: An iridoid diester from Valeriana officinalis var. sambucifolia hairy roots.-Phytochemistry 38: 103–105, 1995b.

    Article  Google Scholar 

  • Granicher, F., Christen, P., Kapetanidis, I.: Production of valepotriates by hairy root cultures of Centranthus ruber DC.-Plant Cell Rep. 14: 294–298, 1995a.

    Article  Google Scholar 

  • Gupta, A.P., Gupta, M.M., Kumar, S.: High performance thin layer chromatography of asiaticoside in Centella asiatica.-J. indian chem. Soc. 76: 321–322, 1999.

    CAS  Google Scholar 

  • Inamdar, P.K., Yoele, R.D., Ghogare, A.B., De Souza, N.J.: Determination of biologically active constituents in Centella asiatica.-J. Chromatogr. A 742: 127–130, 1996.

    Article  CAS  Google Scholar 

  • Josekutty, P.C.: Callus culture and micropropagation of Hydrocotyle asiatica [Centella asiatica (L.) Urban], a medicinal plant.-Int. J. exp. Bot. 63: 275–278, 1998.

    Google Scholar 

  • Kim, O.T., Kim, M.Y., Hong, M.H., Ahn, J.C., Hwang, B.: Stimulation of asiaticoside accumulation in the whole plant cultures of Centella asiatica (L.) Urban by elicitors.-Plant Cell Rep. 23: 339–344, 2004.

    Article  PubMed  CAS  Google Scholar 

  • Kim, Y., Wyslouzil, B.E., Weathers, P.J.: Secondary metabolism of hairy root cultures in bioreactors.-In Vitro cell. dev. Biol. Plant 38: 1–10, 2002.

    CAS  Google Scholar 

  • Mahagamasekera, M.G.P., Doran, P.M.: Intergeneric co-culture of genetically transformed organs for the production of scopolamine.-Phytochemistry 47: 17–25, 1998.

    Article  CAS  Google Scholar 

  • Massot, B., Milesi, S., Gontier, E., Bourgaud, F., Guckert, A.: Optimised culture conditions for the production of furanocoumarins by micropropagated shoots of Ruta graveolens.-Plant Cell Tissue Organ Cult. 62: 11–19, 2000.

    Article  CAS  Google Scholar 

  • Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures.-Physiol. Plant. 15: 473–497, 1962.

    Article  CAS  Google Scholar 

  • Nath, S., Buragohain A.K.: Establishment of callus and cell suspension cultures of Centella asiatica.-Biol. Plant. 49: 411–414, 2005.

    Article  Google Scholar 

  • Opitz, S., Schnitzler, J.P., Hause, B., Schneider, B.: Histochemical analysis of phenylphenalenone-related compounds in Xiphidium caeruleum (Haemodoraceae).-Planta 216: 881–889, 2003.

    PubMed  CAS  Google Scholar 

  • Pythoud, F., Sinkar, V.P., Nester, E.W., Gordon, M.P.: Increased virulence of Agrobacterium rhizogenes conferred by the vir region of pTiBo542: Application to genetic engineering of poplar.-Bio/Technology 5: 1323–1327, 1987.

    Article  Google Scholar 

  • Rouillard-Guellec, F., Robin, J.R., Ratsimamanga, A.R., Ratsimamanga, S., Rasaoanaivo, P.: Comparative study of Centella asiatica of Madagascar origin and Indian origin.-Acta bot. gallica 144: 489–493, 1997.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F., Maniatis, T.: Molecular Cloning: A Laboratory Manual. 2nd Edition. Vol. 3.-Cold Spring Harbor Laboratory Press, New York 1989.

    Google Scholar 

  • Samuelsson, G.: Drugs of Natural Origin, a Textbook of Pharmacognosy. Pp. 17–26. Swedish Pharmaceutical Press, Stockholm 1992.

    Google Scholar 

  • Singh, B., Rastogi, R.P.: A reinvestigation of the triterpenes of Centella asiatica III.-Phytochemistry 8: 917–921, 1969.

    Article  CAS  Google Scholar 

  • Soldati, F.: Panax ginseng: standardization and biological activity.-In: Cutler, S.J., Cutler, H.G. (ed.): Biologically Active Natural Products: Pharmaceuticals. Pp. 209–232. CRC Press, Boca Raton 2000.

    Google Scholar 

  • Solet, J.M., Bistermiel, F., Galons, H., Spagnoli, R., Guignard, J.L., Cosson, L.: Glucocosylation of thiocolchicine by a cell-suspension culture of Centella asiatica.-Phytochemistry 33: 817–820, 1993.

    Article  CAS  Google Scholar 

  • Snedecor, G.W., Cochran, W.G.: Statistical Methods. 8th Edition.-Iowa State Univ. Press, Ames 1989.

    Google Scholar 

  • Subroto, A.M., Kwok, K.H., Hamill, J.D., Doran, P.M.: Co-culture of genetically transformed roots and shoots for synthesis, translocation and biotransformation of secondary metabolites.-Biotechnol. Bioeng. 49: 481–494, 1996.

    Article  CAS  PubMed  Google Scholar 

  • Xu, T.F., Zhang, L., Sun, X.F., Zhang, H.M., Tang, K.X.: Production and analysis of organic acids in hairy-root cultures of Isatis indigotica Fort. (Indigo Woad).-Biotechnol. Appl. Biochem. 39: 123–128, 2004.

    Article  PubMed  Google Scholar 

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Correspondence to M. R. Davey.

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Aziz, Z.A., Davey, M.R., Power, J.B. et al. Production of asiaticoside and madecassoside in Centella asiatica in vitro and in vivo . Biol Plant 51, 34–42 (2007). https://doi.org/10.1007/s10535-007-0008-x

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  • DOI: https://doi.org/10.1007/s10535-007-0008-x

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