Analysis of Plant Mitochondrial Function Using Fluorescent Protein Sensors
Mitochondrial physiology sets the basis for function of the organelle and vice versa. While a limited range of in vivo parameters, such as oxygen consumption, has been classically accessible for measurement, a growing collection of fluorescent protein sensors can now give insights into the physiology of plant mitochondria. Nevertheless, the meaningful application of these sensors in mitochondria is technically challenging and requires rigorous experimental standards. Here we exemplify the application of three genetically encoded sensors to monitor glutathione redox potential, pH, and calcium in the matrix of mitochondria in intact plants. We describe current methods for quantitative imaging and analysis in living root tips by confocal microscopy and discuss methodological limitations.
Key wordsPlant mitochondria Fluorescent protein sensors In vivo imaging Confocal microscopy Respiratory physiology roGFP Cameleon cpYFP
M.S. was supported by the Deutsche Forschungsgemeinschaft through the Emmy Noether Programme (SCHW1719/1-1).
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