Journal
MOLECULAR PLANT
Volume 8, Issue 8, Pages 1188-1200Publisher
CELL PRESS
DOI: 10.1016/j.molp.2015.05.006
Keywords
calcium imaging; R-GECO1; flg22; chitin; sensor; Arabidopsis
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Funding
- Deutsche Forschungsgemeinschaft [KE 1719/2-1, FOR964]
- National Institute of Health [GM060396-ES010337]
- National Science Foundation [MCB1414339]
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1414339] Funding Source: National Science Foundation
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Intracellular Ca2+ transients are an integral part of the signaling cascade during pathogen-associated molecular pattern (PAMP)-triggered immunity in plants. Yet, our knowledge about the spatial distribution of PAMP-induced Ca2+ signals is limited. Investigation of cell-and tissue-specific properties of Ca2+ dependent signaling processes requires versatile Ca2+ reporters that are able to extract spatial information from cellular and subcellular structures, as well as from whole tissues over time periods from seconds to hours. Fluorescence-based reporters cover both a broad spatial and temporal range, which makes them ideally suited to study Ca2+ signaling in living cells. In this study, we compared two fluorescence-based Ca2+ sensors: the Forster resonance energy transfer (FRET)-based reporter yellow cameleon NES-YC3.6 and the intensity-based sensor R-GECO1. We demonstrate that R-GECO1 exhibits a significantly increased signal change compared with ratiometric NES-YC3.6 in response to several stimuli. Due to its superior sensitivity, R-GECO1 is able to report flg22- and chitin-induced Ca2+ signals on a cellular scale, which allowed identification of defined [Ca2+](cyt) oscillations in epidermal and guard cells in response to the fungal elicitor chitin. Moreover, we discovered that flg22- and chitin-induced Ca2+ signals in the root initiate from the elongation zone.
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