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High plating efficiency and plant regeneration frequency in low density protoplast cultures derived from an embryogenic Brassica napus cell suspension

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Abstract

Protoplasts were isolated from an embryogenic cell suspension culture derived from microspores of Brassica napus cv. Jet Neuf. Protoplast yield varied with the cell suspension growth medium. Optimization of protoplast plating density, manipulation of culture medium, carbon source and medium matrix, and inclusion of Ficoll resulted in protoplast plating efficiencies close to 30%. Placement of the protoplasts close to the gas interface contributed greatly to the elevated plating efficiency. Low density cultures could be induced to regenerate calli at optimum plating efficiencies if grown in the presence of nurse culture. This is of great advantage for manipulation of individual protoplasts or for microinjection. Plants were regenerated directly from the cell suspension or from the protoplast cultures.

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Abbreviations

BA:

N6-benzyladenine

2,4-D:

2,4-dichlorophenoxyacetic acid

IAA:

indole-3-acetic acid

NAA:

naphthaleneacetic acid

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Author notes

  1. Wilfred A. Keller

    Present address: Plant Biotechnology Institute, National Research Council of Canada, S7N 0W9, Saskatoon, Sask

Authors and Affiliations

  1. Plant Biotechnology Program, Plant Research Centre, Agriculture Canada, K1A OC6, Ottawa, Ontario, (Canada)

    Daina H. Simmonds, Nancy E. Long & Wilfred A. Keller

Authors
  1. Daina H. Simmonds
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  2. Nancy E. Long
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  3. Wilfred A. Keller
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Simmonds, D.H., Long, N.E. & Keller, W.A. High plating efficiency and plant regeneration frequency in low density protoplast cultures derived from an embryogenic Brassica napus cell suspension. Plant Cell Tiss Organ Cult 27, 231–241 (1991). https://doi.org/10.1007/BF00157586

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  • DOI: https://doi.org/10.1007/BF00157586

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