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Stable Plastid Transformation in PEG-treated Protoplasts of Nicotiana tabacum

Bio/Technology volume 11, pages 95–97 (1993)Cite this article

Abstract

Plastid engineering currently relies on DNA delivery by the biolistic process. We report here stable plastid transformation in tobacco by an alternate direct transformation protocol that is based on polyethylene glycol (PEG) treatment of leaf protoplasts in the presence of the transforming DNA. Clones with transformed plastid genomes were selected by spectinomycin resistance encoded by a mutant 16S ribosomal RNA gene. Incorporation of the transforming DNA into the plastid genome was confirmed by two unselected markers, streptomycin resistance and a novel PstI site that flank the spectinomycin resistance mutation in plasmid pZS148. Our simple and inexpensive protocol eliminates the dependence on the particle gun for chloroplast transformation and should facilitate applications of plastome engineering in crops.

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Author notes

  1. Hans-Ulrich Koop: Corresponding author.

Authors and Affiliations

  1. Laboratory of Plant Cell Biology and Cell Culture, Ludwig-Maximilians-University of Munich, W-8000, Munich, F.R.G.

    Timothy Golds & Hans-Ulrich Koop

  2. Waksman Institute, Rutgers, The State University of New Jersey, Piscataway, NJ, 08855–0759

    Pal Maliga

Authors
  1. Timothy Golds
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  2. Pal Maliga
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  3. Hans-Ulrich Koop
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Golds, T., Maliga, P. & Koop, HU. Stable Plastid Transformation in PEG-treated Protoplasts of Nicotiana tabacum. Nat Biotechnol 11, 95–97 (1993). https://doi.org/10.1038/nbt0193-95

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