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Measurement of Cyclic GMP During Plant Hypersensitive Disease Resistance Response

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Plant Programmed Cell Death

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1743))

Abstract

Cyclic guanosine-3′,5′-monophosphate (cGMP) is recognized as an important second messenger in plants, mediating intracellular signal in important physiological processes, including the hypersensitive disease resistance response induced by avirulent pathogens. In this context, the analysis of cGMP levels in infected plants requires an accurate and specific detection method allowing its quantification. Here, we describe an assay based on the Alphascreen technology, developed for animal cells and further adapted and optimized for the detection of cGMP in plants. The method is applied for the measurement of cGMP in Arabidopsis thaliana plants challenged with an avirulent strain of Pseudomonas syringae pv. tomato. This protocol includes the extraction of cGMP, the assay procedure and the calculation of cGMP concentration.

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Acknowledgments

E.V. was supported by the Future in Research Program (FIRB 2010—RBFR10S1LJ_004) funded by the Italian Ministry of Education, University and Research.

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Authors and Affiliations

  1. Department of Biological Sciences, University of South Carolina, Columbia, SC, USA

    Jian Chen

  2. Department of Biotechnology, University of Verona, Verona, Italy

    Jian Chen, Diana Bellin & Elodie Vandelle

Authors
  1. Jian Chen
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  2. Diana Bellin
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  3. Elodie Vandelle
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Corresponding author

Correspondence to Elodie Vandelle .

Editor information

Editors and Affiliations

  1. Food Sciences and Human Nutrition Unit, Università Campus Bio-Medico di Roma, Roma, Italy

    Laura De Gara

  2. Food Sciences and Human Nutrition Unit, Università Campus Bio-Medico di Roma, Roma, Italy

    Vittoria Locato

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Chen, J., Bellin, D., Vandelle, E. (2018). Measurement of Cyclic GMP During Plant Hypersensitive Disease Resistance Response. In: De Gara, L., Locato, V. (eds) Plant Programmed Cell Death. Methods in Molecular Biology, vol 1743. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7668-3_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7668-3_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7667-6

  • Online ISBN: 978-1-4939-7668-3

  • eBook Packages: Springer Protocols

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