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Mol Plant. 2015 Aug;8(8):1188-200. doi: 10.1016/j.molp.2015.05.006. Epub 2015 May 19.

Live Cell Imaging with R-GECO1 Sheds Light on flg22- and Chitin-Induced Transient [Ca(2+)]cyt Patterns in Arabidopsis.

Author information

1
Centre for Organismal Studies, Plant Developmental Biology, University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany.
2
Centre for Organismal Studies, Plant Developmental Biology, University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany; Division of Biological Sciences, Cell and Developmental Biology Section, University of California San Diego, 92093 La Jolla, USA.
3
Centre for Organismal Studies, University of Heidelberg, 69120 Heidelberg, Germany.
4
Division of Biological Sciences, Cell and Developmental Biology Section, University of California San Diego, 92093 La Jolla, USA.
5
Centre for Organismal Studies, Plant Developmental Biology, University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany. Electronic address: melanie.krebs@cos.uni-heidelberg.de.

Abstract

Intracellular Ca(2+) 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 Ca(2+) signals is limited. Investigation of cell- and tissue-specific properties of Ca(2+)-dependent signaling processes requires versatile Ca(2+) 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 Ca(2+) signaling in living cells. In this study, we compared two fluorescence-based Ca(2+) sensors: the Förster 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 Ca(2+) signals on a cellular scale, which allowed identification of defined [Ca(2+)]cyt oscillations in epidermal and guard cells in response to the fungal elicitor chitin. Moreover, we discovered that flg22- and chitin-induced Ca(2+) signals in the root initiate from the elongation zone.

KEYWORDS:

Arabidopsis; R-GECO1; calcium imaging; chitin; flg22; sensor

PMID:
26002145
PMCID:
PMC5134422
DOI:
10.1016/j.molp.2015.05.006
[Indexed for MEDLINE]
Free PMC Article

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