Phytoplasmas pp 321-331 | Cite as

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

  • Katrin Janik
  • Hagen Stellmach
  • Cecilia Mittelberger
  • Bettina Hause
Part of the Methods in Molecular Biology book series (MIMB, volume 1875)


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.

Key words

Bimolecular fluorescence complementation (BiFC) Effector protein Fluorescence microscopy Phytoplasma Protoplast isolation Protoplast transfection Protoplast transformation 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Katrin Janik
    • 1
  • Hagen Stellmach
    • 2
  • Cecilia Mittelberger
    • 1
  • Bettina Hause
    • 2
  1. 1.Functional GenomicsLaimburg Research CentreAuer/Ora (BZ)Italy
  2. 2.Department of Cell and Metabolic BiologyLeibniz Institute of Plant BiochemistryHalleGermany

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