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

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Plant Respiration and Internal Oxygen

Part of the book series: Methods in Molecular Biology ((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.

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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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Authors and Affiliations

  1. Grup de Recerca en Biologia de les Plantes en Condicions Mediterranies, Universitat de les Illes Balears, Carretera de Valldemossa Km 7.5, 07122, Palma de Mallorca, Spain

    Nestor Fernandez Del-Saz & Miquel Ribas-Carbo

  2. Serveis Científico-Tècnics, Universitat de les Illes Balears, Carretera de Valldemossa Km 7.5, 07122, Palma de Mallorca, Spain

    Gabriel Martorell

  3. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476, Potsdam-Golm, Germany

    Igor Florez-Sarasa

  4. Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476, Potsdam-Golm, Germany

    Alisdair R. Fernie

Authors
  1. Nestor Fernandez Del-Saz
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  2. Miquel Ribas-Carbo
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  3. Gabriel Martorell
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  4. Alisdair R. Fernie
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  5. Igor Florez-Sarasa
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Corresponding author

Correspondence to Igor Florez-Sarasa .

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Editors and Affiliations

  1. National Institute of Plant Genome Research, New Delhi, Delhi, India

    Kapuganti Jagadis Gupta

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Del-Saz, N.F., Ribas-Carbo, M., Martorell, G., Fernie, A.R., Florez-Sarasa, I. (2017). Measurements of Electron Partitioning Between Cytochrome and Alternative Oxidase Pathways in Plant Tissues. In: Jagadis Gupta, K. (eds) Plant Respiration and Internal Oxygen. Methods in Molecular Biology, vol 1670. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7292-0_17

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  • DOI: https://doi.org/10.1007/978-1-4939-7292-0_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7291-3

  • Online ISBN: 978-1-4939-7292-0

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