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Transplastomics in Arabidopsis: Progress Toward Developing an Efficient Method

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Chloroplast Research in Arabidopsis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 774))

Abstract

Protocols developed for plastome engineering in Nicotiana tabacum rely on biolistic delivery of the transforming DNA to chloroplasts in intact leaf tissue; integration of the foreign DNA into the plastid genome by homologous recombination via flanking plastid DNA (ptDNA) targeting regions; and gradual dilution of non-transformed ptDNA during cultivation in vitro. Plastid transformation in Arabidopsis was obtained by combining the tobacco leaf transformation protocol with Arabidopsis-specific tissue culture and plant regeneration protocols. Because the leaf cells in Arabidopsis are polyploid, this protocol yielded sterile plants. Meristematic cells in a shoot apex or cells of a developing embryo are diploid. Therefore, we developed a regulated embryogenic root culture system that will generate diploid tissue for plastid transformation. This embryogenic culture system is created by steroid-inducible expression of the BABY BOOM transcription factor. Plastid transformation in Arabidopsis will enable the probing of plastid gene function, and the characterization of posttranscriptional mechanisms of gene regulation and the regulatory interactions of plastid and nuclear genes.

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Acknowledgments

This research was supported by Grant MCB-039958 from the NSF Eukaryotic Genetics Program. K.A.L. was the recipient of a Charles and Johanna Busch Predoctoral Fellowship.

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Authors and Affiliations

  1. Waksman Institute of Microbiology Rutgers, The State University of New Jersey, Piscataway, NJ, USA

    Kerry Ann Lutz, Arun Azhagiri & Pal Maliga

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  1. Kerry Ann Lutz
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  2. Arun Azhagiri
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  3. Pal Maliga
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Correspondence to Pal Maliga .

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Editors and Affiliations

  1. , Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, United Kingdom

    R. Paul Jarvis

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Lutz, K.A., Azhagiri, A., Maliga, P. (2011). Transplastomics in Arabidopsis: Progress Toward Developing an Efficient Method. In: Jarvis, R. (eds) Chloroplast Research in Arabidopsis. Methods in Molecular Biology, vol 774. Humana Press. https://doi.org/10.1007/978-1-61779-234-2_9

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  • DOI: https://doi.org/10.1007/978-1-61779-234-2_9

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-233-5

  • Online ISBN: 978-1-61779-234-2

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