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Analysis of Plant Mitochondrial Function Using Fluorescent Protein Sensors

  • Stephan Wagner
  • Thomas Nietzel
  • Isabel Aller
  • Alex Costa
  • Mark D. Fricker
  • Andreas J. Meyer
  • Markus Schwarzländer
Part of the Methods in Molecular Biology book series (MIMB, volume 1305)

Abstract

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 words

Plant mitochondria Fluorescent protein sensors In vivo imaging Confocal microscopy Respiratory physiology roGFP Cameleon cpYFP 

Notes

Acknowledgement

M.S. was supported by the Deutsche Forschungsgemeinschaft through the Emmy Noether Programme (SCHW1719/1-1).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Stephan Wagner
    • 1
  • Thomas Nietzel
    • 1
  • Isabel Aller
    • 2
  • Alex Costa
    • 3
  • Mark D. Fricker
    • 4
  • Andreas J. Meyer
    • 2
  • Markus Schwarzländer
    • 5
  1. 1.Plant Energy Biology Lab, INRES – Chemical SignallingUniversity of BonnBonnGermany
  2. 2.INRES – Chemical SignallingUniversity of BonnBonnGermany
  3. 3.Department of BiosciencesUniversity of MilanMilanItaly
  4. 4.Department of Plant SciencesUniversity of OxfordOxfordUK
  5. 5.Plant Energy Biology Lab, INRES – Chemical SignallingUniversity of BonnBonnGermany

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