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Photosynthesis pp 141-154 | Cite as

Measurement of O2 Uptake and Evolution in Leaves In Vivo Using Stable Isotopes and Membrane Inlet Mass Spectrometry

  • Steven M. Driever
  • Neil R. Baker
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1770)

Abstract

Oxygen is both product and substrate of photosynthesis and metabolism in plants, by oxygen evolution through water splitting and uptake by photorespiration and respiration. It is important to investigate these processes simultaneously in leaves, especially in response to environmental variables, such as light and temperature. To distinguish between processes that evolve or take up O2 in leaves in the light, in vivo gas exchange of stable isotopes of oxygen and membrane inlet mass spectrometry is used. A closed-cuvette system for gas exchange of leaf disks is described, using the stable isotopes 16O2 and 18O2, with a semipermeable membrane gas inlet and isotope mass separation and detection by mass spectrometry. Measurement of evolution and uptake, as well as CO2 uptake, at a range of light levels allows composition of a light–response curve, here described for French bean and maize leaf disks.

Key words

Oxygen evolution Oxygen uptake Stable isotopes Membrane inlet mass spectrometry MIMS 

Notes

Acknowledgments

This work was initiated and done under the supervision of Prof. Neil R. Baker (University of Essex, UK) and supported by a research studentship from the Department of Biological Sciences at the University of Essex to Dr. Steven M. Driever. We thank Prof. Suzanne von Caemmerer (Australian National University) for providing details on the leaf cuvette as used by Maxwell et al. [15] and Ruuska et al. [12].

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Steven M. Driever
    • 1
  • Neil R. Baker
    • 2
  1. 1.Centre for Crop System AnalysisWageningen University and ResearchWageningenThe Netherlands
  2. 2.School of Biological SciencesUniversity of EssexColchesterUK

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