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.
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References
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
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
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
van Gorkom HJ, Gast P (1996) Measurement of photosynthetic oxygen evolution. In: Biophysical techniques in photosynthesis. Springer, Dordrecht, pp 391–405
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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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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
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DOI: https://doi.org/10.1007/978-1-4939-7786-4_9
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