Abstract
Agroinfiltration is a versatile, rapid and simple technique that is widely used for transient gene expression in plants. In this chapter we focus on its use in molecular plant pathology, and especially for the expression of plant resistance (R) and fungal avirulence (Avr) (effector) genes in leaves of Nicotiana benthamiana. Co-expression of an R gene with the corresponding Avr gene triggers host-defence responses that often culminate in a hypersensitive response (HR). This HR is visible as a necrotic sector in the infiltrated leaf area. Staining of the infiltrated leaves with trypan blue allows visual scoring of the HR. Furthermore, fusion of a fluorescent tag to the recombinant protein facilitates determination of its sub-cellular localization by confocal microscopy. The matching gene pair I-2 and Avr2, respectively from tomato and the fungal root-pathogen Fusarium oxysporum f. sp. lycopersici, is presented as a typical example.
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Acknowledgements
We thank Anna Pietraszewska (CAM, University of Amsterdam) for the assistance using the confocal microscope. We are grateful to Ludek Tikovsky, Harold Lemereis and Thijs Hendrix for plant care.
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Ma, L., Lukasik, E., Gawehns, F., Takken, F.L.W. (2012). The Use of Agroinfiltration for Transient Expression of Plant Resistance and Fungal Effector Proteins in Nicotiana benthamiana Leaves. In: Bolton, M., Thomma, B. (eds) Plant Fungal Pathogens. Methods in Molecular Biology, vol 835. Humana Press. https://doi.org/10.1007/978-1-61779-501-5_4
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DOI: https://doi.org/10.1007/978-1-61779-501-5_4
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