title: Live Cell Imaging with R-GECO1 Sheds Light on flg22- and Chitin-Induced Transient [Ca(2+)]cyt Patterns in Arabidopsis.
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.
authors.affiliation: Centre for Organismal Studies, Plant Developmental Biology, University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany.
authors.forename: Nana F
authors.initials: NF
authors.lastname: Keinath
authors.affiliation: 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.
authors.forename: Rainer
authors.initials: R
authors.lastname: Waadt
authors.affiliation: Centre for Organismal Studies, University of Heidelberg, 69120 Heidelberg, Germany.
authors.forename: Rik
authors.initials: R
authors.lastname: Brugman
authors.affiliation: Division of Biological Sciences, Cell and Developmental Biology Section, University of California San Diego, 92093 La Jolla, USA.
authors.forename: Julian I
authors.initials: JI
authors.lastname: Schroeder
authors.affiliation: Centre for Organismal Studies, University of Heidelberg, 69120 Heidelberg, Germany.
authors.forename: Guido
authors.initials: G
authors.lastname: Grossmann
authors.affiliation: Centre for Organismal Studies, Plant Developmental Biology, University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Schumacher
authors.affiliation: 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.
authors.forename: Melanie
authors.initials: M
authors.lastname: Krebs
mesh_terms: D017360:Arabidopsis / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium; D020013:Calcium Signaling / Q000187:drug effects*; D002686:Chitin / Q000494:pharmacology*; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism*; D005408:Flagellin / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D008164:Luminescent Proteins / Q000378:metabolism*; D057054:Molecular Imaging / Q000379:methods*; D000069452:Pathogen-Associated Molecular Pattern Molecules / Q000378:metabolism; D019441:Plant Epidermis / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018515:Plant Leaves / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018517:Plant Roots / Q000187:drug effects / Q000378:metabolism; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D000069452:Pathogen-Associated Molecular Pattern Molecules; D011993:Recombinant Fusion Proteins; D005408:Flagellin; D002686:Chitin; D002118:Calcium
keywords: Arabidopsis; R-GECO1; calcium imaging; chitin; flg22; sensor