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Agrobacterium Persistence in Plant Tissues After Transformation

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Book cover Transgenic Plants: Methods and Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 286))

Summary

Agrobacterium spp. are routinely used in plant transformation to introduce genes of interest in valuable economic species. However, several agrobacteria species are also plant pathogens with ability to survive in different environments including the inner part of the plants. To avoid the release of genetic modified bacteria a successful plant transformation protocol must include the total elimination of agrobacteria by the use of antibiotics. Because sometimes these antibiotics failed in removing the bacteria entirely, confirmation of agrobacteria absence after plant transformation and regeneration is required. Different methodologies can be used for this purpose: isolation techniques followed by identification are used if detection of viable and culturable bacteria is necessary and techniques based on the polymerase chain reaction can be used to detect agrobacteria independently of their physiological state. Here we present several protocols to detect Agrobacterium in tissues of transformed plants as well as methods to identify the strains isolated. These identification methods can help to elucidate if they are the engineered bacteria used in the transformation process or just part of the natural endophytic microbiota.

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Author information

Authors and Affiliations

  1. Department of Plant Protection, INIA, Madrid, Spain

    Jaime Cubero

  2. Department of Plant Protection and Biotechnology, Instituto Valenciano de Investigaciones Agrarias, Valencia, Spain

    María M. López

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  1. Jaime Cubero
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  2. María M. López
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Editor information

Editors and Affiliations

  1. Instituto Valenciano de Investigaciones Agrarias, Valencia, Spain

    Leandro Peña

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© 2005 Humana Press Inc.

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Cubero, J., López, M.M. (2005). Agrobacterium Persistence in Plant Tissues After Transformation. In: Peña, L. (eds) Transgenic Plants: Methods and Protocols. Methods in Molecular Biology™, vol 286. Humana Press. https://doi.org/10.1385/1-59259-827-7:351

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  • DOI: https://doi.org/10.1385/1-59259-827-7:351

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-263-6

  • Online ISBN: 978-1-59259-827-4

  • eBook Packages: Springer Protocols

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