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Measurements of Electron Partitioning Between Cytochrome and Alternative Oxidase Pathways in Plant Tissues

  • Nestor Fernandez Del-Saz
  • Miquel Ribas-Carbo
  • Gabriel Martorell
  • Alisdair R. Fernie
  • Igor Florez-SarasaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1670)

Abstract

Plant respiration is characterized by the existence of the alternative oxidase pathway (AOP) that competes with cytochrome oxidase pathway (COP) for the electrons of the ubiquinone pool of the mitochondrial electron transport chain, thus reducing ATP synthesis. The oxygen (O2) isotope fractionation technique is the only available to determine the electron partitioning between the two pathways and their in vivo activities in plant tissues. In this chapter, the basis of the O2 isotope fractionation technique and its derived calculations are carefully explained together with a detailed description of the dual-inlet isotope ratio mass spectrometry (DI-IRMS) system and the protocol developed at the University of Balearic Islands. The key advantages of the DI-IRMS over other systems are highlighted as well as the potential problems of this technique. Among these problems, those associated with leakage, diffusion, and inhibitor treatments are noted and solutions to prevent, detect, and repair these problems are detailed.

Key words

Oxygen isotope fractionation Respiration In vivo mitochondrial electron partitioning Cytochrome oxidase pathway Alternative oxidase pathway 

Notes

Acknowledgments

We would like to thank all the staff at the Serveis Cientifico-Tecnics of the Universitat de les Illes Balears for their help while running the experiments on the IRMS.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Nestor Fernandez Del-Saz
    • 1
  • Miquel Ribas-Carbo
    • 1
  • Gabriel Martorell
    • 2
  • Alisdair R. Fernie
    • 4
  • Igor Florez-Sarasa
    • 3
    Email author
  1. 1.Grup de Recerca en Biologia de les Plantes en Condicions MediterraniesUniversitat de les Illes BalearsPalma de MallorcaSpain
  2. 2.Serveis Científico-TècnicsUniversitat de les Illes BalearsPalma de MallorcaSpain
  3. 3.Max-Planck-Institut für Molekulare PflanzenphysiologiePotsdam-GolmGermany
  4. 4.Max Planck Institute of Molecular Plant PhysiologyPotsdam-GolmGermany

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