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Chloroplast Genetic Engineering Via Organogenesis or Somatic Embryogenesis

  • Protocol
Arabidopsis Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 323))

Abstract

Chloroplast genetic engineering offers a number of unique advantages, including high-level transgene expression, multigene engineering in a single transformation event, transgene containment via maternal inheritance, lack of gene silencing, position and pleiotropic effects and undesirable foreign DNA. More than 40 transgenes have been stably integrated and expressed via the tobacco chloroplast genome to confer desired agronomic traits or express high levels of vaccine antigens and biopharmaceuticals. Despite such significant progress, this technology has not been extended to other important plant species. For example, Arabidopsis may be an ideal model system for chloroplast functional genomics. The employment of chloroplast transformation technology in Arabidopsis has been hampered by the lack of an efficient and reproducible protocol that provides fertile chloroplast transgenic plants. Transformation of the Arabidopsis chloroplast genome was achieved via organogenesis but the efficiency was at least a 100-fold lower than in tobacco and had the drawback of polyploidy in the leaf tissue that resulted in sterile transgenic plants. This problem can be overcome by adapting procedures that are now available to regenerate plants from both diploid and tetraploid explants via callus. In addition, it is feasible to regenerate Arabidopsis via somatic embryogenesis. Recent breakthroughs in highly efficient plastid transformation of recalcitrant crops such as cotton and soybean have opened the possibility of engineering Arabidopsis plastid genome via somatic embryogenesis. Therefore, protocols of recent improvements in tissue culture, DNA delivery, and the novel vector designs are provided here in order to achieve highly efficient plastid transformation in Arabidopsis.

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Authors and Affiliations

  1. Department of Molecular Biology and Microbioloty, University of Central Florida, Orlando, FL

    Amit Dhingra & Henry Daniell

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  1. Amit Dhingra
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  2. Henry Daniell
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Editors and Affiliations

  1. Dept. of Biotechnology, Instituto Nacional de Investigacion y TecnologĂ­a Agraria y Alimentaria (INIA), Madrid, Spain

    Julio Salinas PhD

  2. Dept. de Genetica Molecular de Plantas, Centro Nacional de BiotecnologĂ­a-CSIC, Campus de Cantoblanco, Madrid, Spain

    Jose J. Sanchez-Serrano PhD

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Dhingra, A., Daniell, H. (2006). Chloroplast Genetic Engineering Via Organogenesis or Somatic Embryogenesis. In: Salinas, J., Sanchez-Serrano, J.J. (eds) Arabidopsis Protocols. Methods in Molecular Biology™, vol 323. Humana Press. https://doi.org/10.1385/1-59745-003-0:245

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  • DOI: https://doi.org/10.1385/1-59745-003-0:245

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-395-4

  • Online ISBN: 978-1-59745-003-4

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