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Callus production, suspension culture and in vitro alkaloid yields of Ephedra

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Abstract

The effects of a range of plant growth regulators on callus production in various Ephedra species were examined. Species examined were E. andina, E. distachya, E. equisitina, E. fragilis var, camplyopoda, E. gerardiana, E. intermedia, E. major ssp procera, E. minima and E. saxatilis. All species produced callus on modified MS medium supplemented with 0.25 μM kinetin and 5.0 μM 2,4-dichlorophenoxyacetic acid or 1-naphthaleneacetic acid. Neither indole-3-acetic acid nor 3-indolebutyric acid induced significant callus formation but the latter maintained growth of established callus cultures in several species. Suspension cultures of several species were established in MS medium supplemented with 0.25 μM kinetin and 5.0 μM 2,4-dichlorophenoxyacetic acid or 1-naphthaleneacetic acid. Sustained fresh weight doubling times of 70±7h were recorded for cell suspension cultures of E. andina grown in a semi-continous air-lift bubble bioreactor and a minimum doubling time of 56 h was recorded for E. andina in batch culture. It also proved possible to immobilise E. andina batch cultures in sodium alginate beads.

Neither parent plants or in vitro cultures of E. distachya, E. fragilis or E. saxatilis produced alkaloids. Trace quantities of 1-ephedrine and trace-0.14% dwt d-pseudoephedrine were produced by in vitro cultures of other species. The ability to produce alkaloid diminished to zero with successive subcultures.

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Abbreviations

Eph:

1-ephedrine

Peph:

d-pseudoephedrine

RGR:

relative growth rate

KIN:

kinetin

2,4-d :

2,4-dichlorophenoxyacetic acid

NAA:

1-naphthaleneacetic acid

IBA:

3-in-dolebutyric acid

IAA:

indole-3-acetic acid

References

  • Barkan S, Weber JD & Smith E (1981) Determination of cross-contamination of the diastereomers ephedrine and pseudoephedrine by high performance liquid chromatography, thin layer chromatography and carbon-13 nuclear magnetic resonance spectroscopy. J. Chromatogr. 219: 81–88

    Google Scholar 

  • Bhatnagar SP & Singh MN (1984) Organogenesis in the cultured female gametophyte of Ephedra foliata. J. Exp. Bot. 35: 268–278

    Google Scholar 

  • Darlington CD & Wylie AP (1955) Chromosome Atlas of Flowering Plants (p. 7) (2nd edition). George Allen and Unwin, London

    Google Scholar 

  • Dixon RA (1985) Plant Cell Culture-a practical approach (p.18) IRL Press, Oxford

    Google Scholar 

  • Evans WC (1989) Alkaloids. In: Trease and Evan's Pharmacognosy (13th edition, p. 99). Balliere Tindall, London

    Google Scholar 

  • Feldman DS, Gagnon J, Hofmann R & Simpson J (1987) Statview II. Abacus Concepts Inc., Berkeley, California

    Google Scholar 

  • Fidgeon C & Wilson G (1987) Growth regulation of Galium mollugo L. cell suspensions by α-naphthaleneacetic acid. J. Exp. Bot. 38: 1491–1500

    Google Scholar 

  • Khanna P & Uddin A (1976) Production of ephedrine from in vitro tissue cultures of Ephedra foliata Boiss. Proceedings of the Indian Science Congress 62: 93

    Google Scholar 

  • Konar RN & Singh MN (1979) Production of plantlets from female gametophytes of Ephedra foliata Boiss. Z. Pflanzenphysiol. 95: 87–90

    Google Scholar 

  • Lee TT (1971) Cytokinin controlled indoleacetic acid oxidase isoenzymes in tobacco callus cultures. Plant Physiol. (Bethesda) 47: 181–185

    Google Scholar 

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissues cultures. Physiol. Plant. 15: 473–497

    Google Scholar 

  • Nissen SJ & Sutter EG (1990) Stability of IAA and IBA in nutrient medium to several tissue culture procedures. HortScience. 25: 800–802

    Google Scholar 

  • O'Dowd N (1991) In vitro culture, micropropagation and secondary metabolite production of Ephedra. PhD thesis. Trinity College, University of Dublin

  • Owens JN & Hardev V (1990) Sex expression in gymnosperms. Crit. Rev. Plant Sci. 9: 281–294

    Google Scholar 

  • Ramawat KG & Arya HC (1976) Growth and morphogenesis in callus cultures of Ephedra gerardiana, Phytomorphology 26: 395–403

    Google Scholar 

  • Ramawat KG & Arya HC (1977) Carbohydrate nutrition of Ephedra tissues grown in culture. Indian J. Exp. Bot. 15: 524–527

    Google Scholar 

  • Ramawat KG & Arya HC (1979a) Alkaloid content of Ephedra in vivo and in vitro. Indian J. Exp. Bot. 17: 106–107

    Google Scholar 

  • Ramawat KG & Arya HC (1979b) Effect of some growth regulators on ephedrine production in Ephedra gerardiana callus cultures. Indian J. Exp. Bot. 17: 227–228

    Google Scholar 

  • Ramawat KG & Arya HC (1980) Nitrogen nutrition of Ephedra tissues grown in culture. Phytomorphology 29: 15–26

    Google Scholar 

  • Sankhla N, Sankhla D & Chatterji UN (1967a) In vitro induction of proliferation in female gametophytic tissue of E. foliata Boiss. Naturwissenschaften 54: 203

    Google Scholar 

  • Shukla RM (1980) Effect of light quality on ephedrine production in Ephedra foliata callus culture. Indian Drugs 17: 392–393

    Google Scholar 

  • Singer SR (1986) Analysing growth in cell cultures I: calculating growth rates. Can. J. Bot. 64: 233–237

    Google Scholar 

  • Straus J & Gerding RK (1963) Auxin oxidase and growth control in tissue cultures of Ephedra. Plant Physiol. (Bethesda) 38: 621–627

    Google Scholar 

  • Syono K & Furuya T (1972) Effects of cytokinins on the auxin requirement and auxin content of tobacco calluses. Plant Cell Physiol. 13: 843–856

    Google Scholar 

  • Teasdale RD, Dawson PA & Woolhouse HW (1986) Mineral nutrient requirements of a loblolly pine (Pinus taeda) cell suspension culture. Plant Physiol. (Bethesda) 82: 942–945

    Google Scholar 

  • Uddin A (1977) Production of amino acids in Ephedra foliata suspension cultures. Curr. Sci. (Bangalore). 46: 825–826

    Google Scholar 

  • Wilson G (1990) Screeing and selection of cultured plant cells for increased yields of secondary metabolites. In: Dix P (Ed) Plant Cell Line Selection. V.C.H. Weinheim

    Google Scholar 

  • Yeomann MM (1987) Bypassing the plant. Ann. Bot. (London) 4: 157–174

    Google Scholar 

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Author notes

  1. Niamh A. O'Dowd

    Present address: Department of Biology, Carleton University, 1125 Colonel By Drive, K1S 5B6, Ottawa, Canada

Authors and Affiliations

  1. School of Botany, Trinity College, University of Dublin, 2, Dublin, Ireland

    Niamh A. O'Dowd, Patrick G. McCauley & David H. S. Richardson

  2. Department of Botany, University College Dublin, Belfield, 4, Dublin, Ireland

    Graham Wilson

Authors
  1. Niamh A. O'Dowd
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  2. Patrick G. McCauley
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  3. David H. S. Richardson
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  4. Graham Wilson
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O'Dowd, N.A., McCauley, P.G., Richardson, D.H.S. et al. Callus production, suspension culture and in vitro alkaloid yields of Ephedra . Plant Cell Tiss Organ Cult 34, 149–155 (1993). https://doi.org/10.1007/BF00036095

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  • DOI: https://doi.org/10.1007/BF00036095

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