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Quantitative Analysis of Microbe-Associated Molecular Pattern (MAMP)-Induced Ca2+ Transients in Plants

  • Fabian Trempel
  • Stefanie Ranf
  • Dierk Scheel
  • Justin LeeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1398)

Abstract

Ca2+ is a secondary messenger involved in early signaling events triggered in response to a plethora of biotic and abiotic stimuli. In plants, environmental cues that induce cytosolic Ca2+ elevation include touch, reactive oxygen species, cold shock, and salt or osmotic stress. Furthermore, Ca2+ signaling has been implicated in early stages of plant–microbe interactions of both symbiotic and antagonistic nature. A long-standing hypothesis is that there is information encoded in the Ca2+ signals (so-called Ca2+ signatures) to enable plants to differentiate between these stimuli and to trigger the appropriate cellular response. Qualitative and quantitative measurements of Ca2+ signals are therefore needed to dissect the responses of plants to their environment. Luminescence produced by the Ca2+ probe aequorin upon Ca2+ binding is a widely used method for the detection of Ca2+ transients and other changes in Ca2+ concentrations in cells or organelles of plant cells. In this chapter, using microbe-associated molecular patterns (MAMPs), such as the bacterial-derived flg22 or elf18 peptides as stimuli, a protocol for the quantitative measurements of Ca2+ fluxes in apoaequorin-expressing seedlings of Arabidopsis thaliana in 96-well format is described.

Key words

Ca2+ measurements Aequorin Microbe-associated molecular pattern (MAMP) 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Fabian Trempel
    • 1
  • Stefanie Ranf
    • 1
    • 2
  • Dierk Scheel
    • 1
  • Justin Lee
    • 1
    Email author
  1. 1.Department of Stress & Developmental BiologyLeibniz Institute of Plant BiochemistryHalle/SaaleGermany
  2. 2.Phytopathology, Center of Life and Food Sciences WeihenstephanTechnische Universität MünchenFreising-WeihenstephanGermany

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