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Plant Chromosome Engineering pp 37–50Cite as

Engineering the Plastid Genome of Nicotiana sylvestris, a Diploid Model Species for Plastid Genetics

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 701))

Abstract

The plastids of higher plants have their own ∼120–160-kb genome that is present in 1,000–10,000 copies per cell. Engineering of the plastid genome (ptDNA) is based on homologous recombination between the plastid genome and cloned ptDNA sequences in the vector. A uniform population of engineered ptDNA is obtained by selection for marker genes encoded in the vectors. Manipulations of ptDNA include (1) insertion of transgenes in intergenic regions; (2) posttransformation excision of marker genes to obtain marker-free plants; (3) gene knockouts and gene knockdowns, and (4) cotransformation with multiple plasmids to introduce nonselected genes without physical linkage to marker genes. Most experiments on plastome engineering have been carried out in the allotetraploid Nicotiana tabacum. We report here for the first time plastid transformation in Nicotiana sylvestris, a diploid ornamental species. We demonstrate that the protocols and vectors developed for plastid transformation in N. tabacum are directly applicable to N. sylvestris with the advantage that the N. sylvestris transplastomic lines are suitable for mutant screens.

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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 and 2008-03012.

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Authors and Affiliations

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

    Pal Maliga & Zora Svab

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  1. Pal Maliga
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  2. Zora Svab
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Correspondence to Pal Maliga .

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Editors and Affiliations

  1. , Division of Biological Sciences, University of Missouri, Tucker Hall 117, Columbia, 65211, Missouri, USA

    James A. Birchler

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Maliga, P., Svab, Z. (2011). Engineering the Plastid Genome of Nicotiana sylvestris, a Diploid Model Species for Plastid Genetics. In: Birchler, J. (eds) Plant Chromosome Engineering. Methods in Molecular Biology, vol 701. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61737-957-4_2

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  • DOI: https://doi.org/10.1007/978-1-61737-957-4_2

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  • Publisher Name: Humana Press, Totowa, NJ

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