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Quantitative Local Photosynthetic Flux Measurements at Isolated Chloroplasts and Thylakoid Membranes Using Scanning Electrochemical Microscopy (SECM)

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Department of Chemistry, Molecular Assembly and Organisation in Cells (MOAC) Doctoral Training Centre, and §School of Life Sciences, University of Warwick, Coventry, U.K. CV4 7AL
*Address: Department of Chemistry, University of Warwick, Coventry, U.K. CV4 7AL. Phone: +44 (0)2476 523264. E-mail: [email protected]
Cite this: J. Phys. Chem. B 2013, 117, 26, 7878–7888
Publication Date (Web):June 10, 2013
https://doi.org/10.1021/jp403048f
Copyright © 2013 American Chemical Society

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    Abstract

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    Scanning electrochemical microscopy (SECM) offers a fast and quantitative method to measure local fluxes within photosynthesis. In particular, we have measured the flux of oxygen and ferrocyanide (Fe(CN)64–), from the artificial electron acceptor ferricyanide (Fe(CN)63–), using a stationary ultramicroelectrode at chloroplasts and thylakoid membranes (sourced from chloroplasts). Oxygen generation at films of chloroplasts and thylakoid membranes was detected directly during photosynthesis, but in the case of thylakoid membranes, this switched to sustained oxygen consumption at longer illumination times. An initial oxygen concentration spike was detected over both chloroplast and thylakoid membrane films, and the kinetics of the oxygen generation were extracted by fitting the experimental data to a finite element method (FEM) simulation. In contrast to previous work, the oxygen generation spike was attributed to the limited size of the plastoquinone pool, a key component in the linear electron transport pathway and a contributing factor in photoinhibition. Finally, the mobile nature of the SECM probe, and its high spatial resolution, also allowed us to detect ferrocyanide produced from a single thylakoid membrane. These results further demonstrate the power of SECM for localized flux measurements in biological processes, in this case photosynthesis, and that the high time resolution, combined with FEM simulations, allows the elucidation of quantitative kinetic information.

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    This article is cited by 11 publications.

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