Live Cell Imaging with R-GEC01 Sheds Light on fig22- and Chitin-Induced Transient [Ca2＋]cyt Patterns in Arabidopsis  

作　　者：Nana F. Keinath Rainer Waadt Rik Brugman Julian I. Schroeder Guido Grossmann Karin Schumacher Melanie Krebs 

机构地区：[1]Centre for Organismal Studies, Plant Developmental Biology, University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany [2]Centre for Organismal Studies, University of Heidelberg, 69120 Heidelberg, Germany [3]Division of Biological Sciences, Cell and Developmental Biology Section, University of California San Diego, 92093 La Jolla, USA [4]These authors contributed equally to this article.

出　　处：《Molecular Plant》2015年第8期1188-1200,共13页分子植物（英文版）

摘　　要：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 F6rster 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 fig22- 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 fig22- and chitin-induced Ca2＋ signals in the root initiate from the elongation zone.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 F6rster 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 fig22- 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 fig22- and chitin-induced Ca2＋ signals in the root initiate from the elongation zone.

关 键 词：calcium imaging R-GECO1 fig22 CHITIN sensor ARABIDOPSIS 

分 类 号：Q943.1[生物学—植物学] TB873[一般工业技术—摄影技术]