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Multiparameter imaging of calcium and abscisic acid and high‐resolution quantitative calcium measurements using R‐GECO1‐mTurquoise in Arabidopsis
Author(s) -
Waadt Rainer,
Krebs Melanie,
Kudla Jörg,
Schumacher Karin
Publication year - 2017
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/nph.14706
Subject(s) - calcium , abscisic acid , calcium imaging , calcium signaling , arabidopsis , arabidopsis thaliana , auxin , chemistry , biophysics , biochemistry , microbiology and biotechnology , biology , mutant , organic chemistry , gene
Summary Calcium signals occur in specific spatio‐temporal patterns in response to various stimuli and are coordinated with, for example, hormonal signals, for physiological and developmental adaptations. Quantification of calcium together with other signalling molecules is required for correlative analyses and to decipher downstream calcium‐decoding mechanisms. Simultaneous in vivo imaging of calcium and abscisic acid has been performed here to investigate the interdependence of the respective signalling processes in Arabidopsis thaliana roots. Advanced ratiometric genetically encoded calcium indicators have been generated and in vivo calcium calibration protocols were established to determine absolute calcium concentration changes in response to auxin and ATP . In roots, abscisic acid induced long‐term basal calcium concentration increases, while auxin triggered rapid signals in the elongation zone. The advanced ratiometric calcium indicator R‐ GECO 1‐ mT urquoise exhibited an increased calcium signal resolution compared to commonly used Förster resonance energy transfer‐based indicators. Quantitative calcium measurements in Arabidopsis root tips using R‐ GECO 1‐ mT urquoise revealed detailed maps of absolute calcium concentration changes in response to auxin and ATP. Calcium calibration protocols using R‐ GECO 1‐ mT urquoise enabled high‐resolution quantitative imaging of resting cytosolic calcium concentrations and their dynamic changes that revealed distinct hormonal and ATP responses in roots.