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Transformation of the Plastid Genome in Tobacco: The Model System for Chloroplast Genome Engineering

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Chloroplast Biotechnology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2317))

Abstract

The protocol we report here is based on biolistic delivery of transforming DNA to tobacco leaves, selection of transplastomic clones by spectinomycin or kanamycin resistance and regeneration of plants with uniformly transformed plastid genomes. Because the plastid genome of Nicotiana tabacum derives from Nicotiana sylvestris, and the two genomes are highly conserved, vectors developed for N. tabacum can be used in N. sylvestris. The tissue culture responses of N. tabacum cv. Petit Havana and N. sylvestris accession TW137 are similar. Plastid transformation in a subset of N. tabacum cultivars and in Nicotiana benthamiana requires adjustment of the tissue culture protocol. We describe updated vectors targeting insertions in the unique and repeated regions of the plastid genome, vectors suitable for regulated gene expression by the engineered PPR10 RNA binding protein as well as systems for marker gene excision.

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Acknowledgments

This work was supported by grants from the USDA Biotechnology Risk Assessment Research Grant Program Award No. 2005-33120-16524, 2008-03012, and 2010-2716, The USDA NIFA Foundational Program Award No. 2014-67013-21600 and NSF MCB Grant 1716102.

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Authors and Affiliations

  1. Waksman Institute of Microbiology, Rutgers University, Piscataway, NJ, USA

    Pal Maliga & Tarinee Tungsuchat-Huang

  2. Biology Department, Farmingdale State College, Farmingdale, NY, USA

    Kerry Ann Lutz

Authors
  1. Pal Maliga
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  2. Tarinee Tungsuchat-Huang
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  3. Kerry Ann Lutz
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Correspondence to Pal Maliga .

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Editors and Affiliations

  1. Waksman Institute of Microbiology, Rutgers University, Piscataway, NJ, USA

    Pal Maliga

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Maliga, P., Tungsuchat-Huang, T., Lutz, K.A. (2021). Transformation of the Plastid Genome in Tobacco: The Model System for Chloroplast Genome Engineering. In: Maliga, P. (eds) Chloroplast Biotechnology. Methods in Molecular Biology, vol 2317. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1472-3_6

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  • DOI: https://doi.org/10.1007/978-1-0716-1472-3_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1471-6

  • Online ISBN: 978-1-0716-1472-3

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