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Protoplasma

, 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

Summary

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.

Keywords

Cell Wall Cellulase Liquid Medium Sorbitol Kinetin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

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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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