Skip to main content
SpringerLink
Log in
Book cover

Photosynthesis pp 141–154Cite as

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

  • Protocol
  • First Online:

  • 2435 Accesses

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

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Hunt S (2003) Measurements of photosynthesis and respiration in plants. Physiol Plantarum 117(3):314–325. https://doi.org/10.1034/j.1399-3054.2003.00055.x

    Article  CAS  Google Scholar 

  2. Delieu T, Walker DA (1981) Polarographic measurement of photosynthetic oxygen evolution by leaf-disks. New Phytol 89(2):165–178. https://doi.org/10.1111/j.1469-8137.1981.tb07480.x

    Article  CAS  Google Scholar 

  3. Delieu TJ, Walker DA (1983) Simultaneous measurement of oxygen evolution and chlorophyll fluorescence from leaf pieces. Plant Physiol 73(3):534–541. https://doi.org/10.1104/Pp.73.3.534

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. van Gorkom HJ, Gast P (1996) Measurement of photosynthetic oxygen evolution. In: Biophysical techniques in photosynthesis. Springer, Dordrecht, pp 391–405

    Chapter  Google Scholar 

  5. Davey PA, Hunt S, Hymus GJ, DeLucia EH, Drake BG, Karnosky DF, Long SP (2004) Respiratory oxygen uptake is not decreased by an instantaneous elevation of [CO2], but is increased with long-term growth in the field at elevated [CO2]. Plant Physiol 134(1):520–527. https://doi.org/10.1104/pp.103.030569

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Willms JR, Dowling AN, Dong ZM, Hunt S, Shelp BJ, Layzell DB (1997) The simultaneous measurement of low rates of CO2 and O2 exchange in biological systems. Anal Biochem 254(2):272–282. https://doi.org/10.1006/abio.1997.2416

    Article  PubMed  CAS  Google Scholar 

  7. Cousins AB, Pracharoenwattana I, Zhou WX, Smith SM, Badger MR (2008) Peroxisomal malate dehydrogenase is not essential for photorespiration in arabidopsis but its absence causes an increase in the stoichiometry of photorespiratory CO2 release. Plant Physiol 148(2):786–795. https://doi.org/10.1104/pp.108.122622

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  8. Cousins AB, Walker BJ, Pracharoenwattana I, Smith SM, Badger MR (2011) Peroxisomal hydroxypyruvate reductase is not essential for photorespiration in arabidopsis but its absence causes an increase in the stoichiometry of photorespiratory CO2 release. Photosynth Res 108(2–3):91–100. https://doi.org/10.1007/s11120-011-9651-3

    Article  PubMed  CAS  Google Scholar 

  9. Walker BJ, Cousins AB (2013) Influence of temperature on measurements of the CO2 compensation point: differences between the Laisk and O2-exchange methods. J Exp Bot 64(7):1893–1905. https://doi.org/10.1093/jxb/ert058

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Biehler K, Haupt S, Beckmann J, Fock H, Becker TW (1997) Simultaneous CO2- and 16O2/18O2-gas exchange and fluorescence measurements indicate differences in light energy dissipation between the wild type and the phytochrome-deficient aurea mutant of tomato during water stress. J Exp Bot 48(312):1439–1449. https://doi.org/10.1093/jxb/48.7.1439

    Article  CAS  Google Scholar 

  11. Driever SM, Baker NR (2011) The water-water cycle in leaves is not a major alternative electron sink for dissipation of excess excitation energy when CO2 assimilation is restricted. Plant Cell Environ 34(5):837–846. https://doi.org/10.1111/j.1365-3040.2011.02288.x

    Article  PubMed  CAS  Google Scholar 

  12. Ruuska SA, Badger MR, Andrews TJ, von Caemmerer S (2000) Photosynthetic electron sinks in transgenic tobacco with reduced amounts of Rubisco: little evidence for significant Mehler reaction. J Exp Bot 51:357–368. https://doi.org/10.1093/jexbot/51.suppl_1.357

    Article  PubMed  CAS  Google Scholar 

  13. Haupt-Herting S, Fock HP (2000) Exchange of oxygen and its role in energy dissipation during drought stress in tomato plants. Physiol Plant 110(4):489–495. https://doi.org/10.1111/j.1399-3054.2000.1100410.x

    Article  CAS  Google Scholar 

  14. Haupt-Herting S, Fock HP (2002) Oxygen exchange in relation to carbon assimilation in water-stressed leaves during photosynthesis. Ann Bot 89:851–859. https://doi.org/10.1093/aob/mcf023

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Maxwell K, Badger MR, Osmond CB (1998) A comparison of CO2 and O2 exchange patterns and the relationship with chlorophyll fluorescence during photosynthesis in C3 and CAM plants. Aust J Plant Physiol 25(1):45–52

    Article  Google Scholar 

  16. Shirao M, Kuroki S, Kaneko K, Kinjo Y, Tsuyama M, Forster B, Takahashi S, Badger MR (2013) Gymnosperms have increased capacity for electron leakage to oxygen (Mehler and PTOX reactions) in photosynthesis compared with angiosperms. Plant Cell Physiol 54(7):1152–1163. https://doi.org/10.1093/pcp/pct066

    Article  PubMed  CAS  Google Scholar 

  17. Siebke K, Ghannoum O, Conroy JP, Badger MR, von Caemmerer S (2003) Photosynthetic oxygen exchange in C4 grasses: The role of oxygen as electron acceptor. Plant Cell Environ 26(12):1963–1972. https://doi.org/10.1046/j.1365-3040.2003.01112.x

    Article  CAS  Google Scholar 

  18. Arnon D, Hoagland D (1940) Crop production in artificial culture solutions and in soils with special reference to factors influencing yields and absorption of inorganic nutrients. Soil Sci 50:463–485

    CAS  Google Scholar 

  19. Coe RA, Lin H (2018) Light-response curves in land plants. In: Covshoff S (ed) Photosynthesis: methods and protocols, Methods in molecular biology, vol 1770. Springer, New York

    Chapter  Google Scholar 

  20. Radmer RJ, Kok B (1976) Photoreduction of O2 primes and replaces CO2 assimilation. Plant Physiol 58(3):336–340. https://doi.org/10.1104/Pp.58.3.336

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

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].

Author information

Authors and Affiliations

  1. Centre for Crop System Analysis, Wageningen University and Research, Wageningen, The Netherlands

    Steven M. Driever

  2. School of Biological Sciences, University of Essex, Colchester, UK

    Neil R. Baker

Authors
  1. Steven M. Driever
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Neil R. Baker
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Niceville, Florida, USA

    Sarah Covshoff

Rights and permissions

Reprints and permissions

Copyright information

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

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Driever, S.M., Baker, N.R. (2018). Measurement of O2 Uptake and Evolution in Leaves In Vivo Using Stable Isotopes and Membrane Inlet Mass Spectrometry. In: Covshoff, S. (eds) Photosynthesis. Methods in Molecular Biology, vol 1770. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7786-4_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7786-4_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7785-7

  • Online ISBN: 978-1-4939-7786-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation