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Characterization of Phytoplasmal Effector Protein Interaction with Proteinaceous Plant Host Targets Using Bimolecular Fluorescence Complementation (BiFC)

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Phytoplasmas

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1875))

Abstract

Elucidating the molecular mechanisms underlying plant disease development has become an important aspect of phytoplasma research in the last years. Especially unraveling the function of phytoplasma effector proteins has gained interesting insights into phytoplasma-host interaction at the molecular level. Here, we describe how to analyze and visualize the interaction of a phytoplasma effector with its proteinaceous host partner using bimolecular fluorescence complementation (BiFC) in Nicotiana benthamiana mesophyll protoplasts. The protocol comprises a description of how to isolate protoplasts from leaves and how to transform these protoplasts with BiFC expression vectors containing the phytoplasma effector and the host interaction partner, respectively. If an interaction occurs, a fluorescent YFP-complex is reconstituted in the protoplast, which can be visualized using fluorescence microscopy.

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Author notes

  1. Katrin Janik and Hagen Stellmach contributed equally to this work.

Authors and Affiliations

  1. Functional Genomics, Laimburg Research Centre, Auer/Ora (BZ), Italy

    Katrin Janik & Cecilia Mittelberger

  2. Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Halle, Germany

    Hagen Stellmach & Bettina Hause

Authors
  1. Katrin Janik
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  2. Hagen Stellmach
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  3. Cecilia Mittelberger
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  4. Bettina Hause
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Corresponding author

Correspondence to Katrin Janik .

Editor information

Editors and Affiliations

  1. Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy

    Rita Musetti

  2. Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy

    Laura Pagliari

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Janik, K., Stellmach, H., Mittelberger, C., Hause, B. (2019). Characterization of Phytoplasmal Effector Protein Interaction with Proteinaceous Plant Host Targets Using Bimolecular Fluorescence Complementation (BiFC). In: Musetti, R., Pagliari, L. (eds) Phytoplasmas. Methods in Molecular Biology, vol 1875. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8837-2_24

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  • DOI: https://doi.org/10.1007/978-1-4939-8837-2_24

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8836-5

  • Online ISBN: 978-1-4939-8837-2

  • eBook Packages: Springer Protocols

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