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AGROBEST: A Highly Efficient Agrobacterium-Mediated Transient Expression System in Arabidopsis Seedlings

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Plant Synthetic Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2379))

Abstract

Agrobacterium-mediated transient transformation for gene expression is a simple and fast method to analyze transgene functions in plants. Agroinfiltration in leaves of Nicotiana benthamiana is a common method for transient expression. However, agroinfiltration in leaves of Arabidopsis thaliana is challenging due to the low and variable efficiency. Here, we describe procedures of a highly efficient and robust Agrobacterium-mediated transient expression system, named AGROBEST (Agrobacterium-mediated enhanced seedling transformation) for gene expression in A. thaliana seedlings. High efficiency of AGROBEST has been achieved by virulence (vir) gene pre-induction of a specific disarmed Agrobacterium tumefaciens strain C58C1(pTiB6S3ΔT)H followed by co-cultivation with Arabidopsis seedlings in an optimized medium with AB salts and buffered acidic plant culture medium. The stable acidic medium largely increases Agrobacterium-mediated transient expression levels and reduces plant defense responses, suggesting that AGROBEST enables high transient expression efficiency by compromising plant immunity. In summary, AGROBEST is a simple, fast, reliable, and robust transient expression system offering a quick and convenient method to observe protein localization, protein–protein interactions, promoter activities, and gene functional studies in Arabidopsis seedlings.

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Authors and Affiliations

  1. Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan

    Hung-Yi Wu & Erh-Min Lai

  2. Department of Plant Pathology and Microbiology, National Taiwan University, Taipei, Taiwan

    Hung-Yi Wu

Authors
  1. Hung-Yi Wu
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  2. Erh-Min Lai
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Correspondence to Erh-Min Lai .

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Editors and Affiliations

  1. Institute of Synthetic Biology and CEPLAS, University of Düsseldorf, Düsseldorf, Germany

    Matias D. Zurbriggen

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Wu, HY., Lai, EM. (2022). AGROBEST: A Highly Efficient Agrobacterium-Mediated Transient Expression System in Arabidopsis Seedlings. In: Zurbriggen, M.D. (eds) Plant Synthetic Biology. Methods in Molecular Biology, vol 2379. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1791-5_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1791-5_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1790-8

  • Online ISBN: 978-1-0716-1791-5

  • eBook Packages: Springer Protocols

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