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Induction of efficient cell division in alfalfa protoplasts

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Abstract

Alfalfa (Medicago sativa L.) protoplasts derived from cell suspension cultures divided inefficiently in liquid culture. The onset of cell division activity occurred synchronously among the protoplasts; however, many were blocked at cytokinesis and therefore did not complete first division. Very few of the cells that began to divide continued to do so. Immobilization of protoplasts in agarose after 1 to 4 days in liquid culture overcame this inhibition of division. Continuous growth in agarose was restricted and therefore microcolonies were transferred to agar medium to complete callus development. Plating efficiencies of 2–10% were achieved within 30 days of protoplast isolation. The agarose treatment was responsible for a 5- to 30-fold improvement in plating efficiency.

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Authors and Affiliations

  1. Ottawa Research Station, Agriculture Canada, K1A OC6, Ottawa, Canada

    Larry A. Holbrook, Margaret Haffner & Brian L. Miki

  2. Department of Biology, Carleton University, K1S 5B6, Ottawa, Canada

    Terrence J. Reich & V. N. Iyer

Authors
  1. Larry A. Holbrook
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  2. Terrence J. Reich
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  3. V. N. Iyer
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  4. Margaret Haffner
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  5. Brian L. Miki
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Additional information

Communicated by F. Constabel

Contribution No. 774 Ottawa Research Station

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Holbrook, L.A., Reich, T.J., Iyer, V.N. et al. Induction of efficient cell division in alfalfa protoplasts. Plant Cell Reports 4, 229–232 (1985). https://doi.org/10.1007/BF00269364

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  • DOI: https://doi.org/10.1007/BF00269364

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