title: Quantitative Analysis of Microbe-Associated Molecular Pattern (MAMP)-Induced Ca(2+) Transients in Plants.
abstract: Ca(2+) 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 Ca(2+) elevation include touch, reactive oxygen species, cold shock, and salt or osmotic stress. Furthermore, Ca(2+) 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 Ca(2+) signals (so-called Ca(2+) signatures) to enable plants to differentiate between these stimuli and to trigger the appropriate cellular response. Qualitative and quantitative measurements of Ca(2+) signals are therefore needed to dissect the responses of plants to their environment. Luminescence produced by the Ca(2+) probe aequorin upon Ca(2+) binding is a widely used method for the detection of Ca(2+) transients and other changes in Ca(2+) 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 Ca(2+) fluxes in apoaequorin-expressing seedlings of Arabidopsis thaliana in 96-well format is described.
authors: Department of Stress & Developmental Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle/Saale, Germany.
authors: Fabian
authors: F
authors: Trempel
authors: Department of Stress & Developmental Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle/Saale, Germany.|Phytopathology, Center of Life and Food Sciences Weihenstephan, Technische Universität München, Emil-Ramann-Str. 2, Freising-Weihenstephan, Germany.
authors: Stefanie
authors: S
authors: Ranf
authors: Department of Stress & Developmental Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle/Saale, Germany.
authors: Dierk
authors: D
authors: Scheel
authors: Department of Stress & Developmental Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle/Saale, Germany. jlee@ipb-halle.de.
authors: Justin
authors: J
authors: Lee
mesh_terms: D000331:Aequorin / Q000235:genetics / Q000378:metabolism*; D001059:Apoproteins / Q000235:genetics / Q000378:metabolism*; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism / Q000382:microbiology; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D036226:Seedlings / Q000235:genetics / Q000378:metabolism / Q000382:microbiology
chemical_list: D001059:Apoproteins; D029681:Arabidopsis Proteins; D011994:Recombinant Proteins; C060014:apoaequorin; D000331:Aequorin; D002118:Calcium
keywords: Aequorin; Ca2+ measurements; Microbe-associated molecular pattern (MAMP)