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The isolation and culture of protoplasts from an embryogenic cell suspension culture of Picea glauca (moench) voss

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Abstract

Conditions were standardized for the isolation and culture of protoplasts from an embryogenic cell suspension culture of Picea glauca. A combination of 0.5% Cellulase R-10, 0.25% Macerozyme, 0.25% Driselase, 0.25% Rhozyme HP-150 with 0.5M mannitol and 5 mM CaCl2.2H2O produced an average of 4.5 × 106 protoplasts per gram fresh weight of cells. Of the several protoplast culture media tested, von Arnold and Eriksson and Kao and Michayluk (KM8P) media best supported mitotic divisions of protoplasts. A density of 105 protoplasts per ml and the addition of 5 mM glutamine to the culture medium was necessary to induce sustained divisions and microcallus formation. Microcalli grew into subculturable callus using a nurse culture technique.

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Abbreviations

BAP:

benzylaminopurine

2,4-D:

2,4-dichlorophenoxy-acetic acid

FDA:

fluorescein diacetate

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Authors and Affiliations

  1. Plant Biotechnology Institute, National Research Council, 110 Gymnasium Road, S7N 0W9, Saskatoon, SK, Canada

    Faouzi Bekkaoui & David I. Dunstan

  2. Department of Biology, University of Saskatchewan, S7N 0W0, Saskatoon, SK, Canada

    Praveen K. Saxena, Stephen M. Attree & Larry C. Fowke

Authors
  1. Faouzi Bekkaoui
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  2. Praveen K. Saxena
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  3. Stephen M. Attree
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  4. Larry C. Fowke
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  5. David I. Dunstan
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Additional information

Communicated by F. Constabel

NRCC No. 27937

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Bekkaoui, F., Saxena, P.K., Attree, S.M. et al. The isolation and culture of protoplasts from an embryogenic cell suspension culture of Picea glauca (moench) voss. Plant Cell Reports 6, 476–479 (1987). https://doi.org/10.1007/BF00272787

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  • DOI: https://doi.org/10.1007/BF00272787

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