, Volume 86, Issue 4, pp 405–410 | Cite as

Isolation, culture, and induction of embryogenesis in protoplasts from cell-suspensions ofAtropa belladonna

  • G. Gosch
  • Y. P. S. Bajaj
  • J. Reinert
Brief Report


Protoplasts isolated from actively growing cell-suspensions ofAtropa belladonna have been induced to divide repeatedly, and to undergo embryogenesis. An optimal protoplast yield of up to 80% was obtained in 4–5 hours by treating cell-suspensions with an enzyme mixture of cellulase R 10 (1%) and macerozyme R 10 (0.5%) in 0.6 M sorbitol at 30 °C. The protoplasts cultured at a density of 6 · 104/ml in a modifiedMurashige andSkoog's (1962) liquid medium supplemented with NAA (2 mg/l), kinetin (0.1 mg/l) and 0.5 M sorbitol, and incubated in the dark at 28 °C regenerated cell walls within 48 hours. They underwent first division in 3–4 days and formed cell clumps and colonies in 10 days, which when plated on an agar-solidified medium developed into masses of calli. After transfer to an auxin-free liquid medium these calli underwent embryogenesis within the next two weeks and eventually developed into plantlets.


Cell Wall Cellulase Liquid Medium Sorbitol Kinetin 
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  1. Bajaj, Y. P. S., 1976: Protoplast isolation, culture and somatic hybridization. In: Applied and fundamental aspects of plant cell, tissue and organ culture (J. Reinert, andY. P. S. Bajaj, eds.). Berlin-Heidelberg-New York: Springer-Verlag.Google Scholar
  2. Cocking, E. C., and J. F.Peberdy (eds.), 1974: The use of protoplasts from fungi and higher plants as genetic systems.—A practical handbook. Nottingham Univ.Google Scholar
  3. Gosch, G., Y. P. S. Bajaj, andJ. Reinert, 1975: Isolation, culture, and fusion studies on protoplasts from different species. Protoplasma85, 327–336.Google Scholar
  4. Grambow, H. J., K. N. Kao, R. A. Miller, andO. L. Gamborg, 1972: Cell division and plant development from protoplasts of carrot cell suspension cultures. Planta103, 348–355.Google Scholar
  5. Murashige, T., andF. Skoog, 1962: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plantarum15, 473–497.Google Scholar
  6. Reinert, J., G.Gosch, and Y. P. S.Bajaj, 1976: Induction of interspecific and intergeneric fusion of protoplasts from higher plants. (Under preparation.)Google Scholar
  7. Thomas, E., andH. E. Street, 1970: Organogenesis in cell suspension cultures ofAtropa belladonna L. andAtropa belladonna cultivar lutea Döll. Ann. Bot.34, 657–669.Google Scholar
  8. Vardi, A., P. Spiegel-Roy, andE. Galun, 1975: Citrus cell culture: Isolation of protoplasts, plating densities, effect of mutagens, and regeneration of embryos. Plant Sci. Letters4, 231–236.Google Scholar
  9. Wood, H. N., andA. C. Braun, 1961: Studies on the regulation of certain essential biosynthetic systems in normal and crown-gall tumor. Proc. nat. Acad. Sci. (U.S.A.)47, 1907–1913.Google Scholar

Copyright information

© Springer-Verlag 1975

Authors and Affiliations

  • G. Gosch
    • 1
  • Y. P. S. Bajaj
    • 1
  • J. Reinert
    • 1
  1. 1.Institut für Pflanzenphysiologie und ZellbiologieFreie Universität BerlinGermany

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