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bioRxiv · 10.1101/2025.08.05.668659

Assessing the redox state of the plastoquinone pool in algae and cyanobacteria via OJIP fluorescence: perspectives and limitations

Zavrel, T.·Pohland, A. C.·Pfennig, T.·Matuszynska, A. B.·Toth, S. Z.·Bernat, G.·Cerveny, J.
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Abstract

The redox state of the plastoquinone pool (PQ-redox) acts as a central element in a variety of intracellular signal pathways. Several methods for determining PQ-redox have been established. Although some of these methods may be quantitative, such as those based on liquid chromatography, they are typically sensitive to sample preparation. Here, we critically evaluate the use of fast chlorophyll a fluorescence induction kinetics (the so-called OJIP transient) for semi-quantitative PQ-redox estimation in green algae (Chlorella vulgaris) and cyanobacteria (Synechocystis sp. PCC 6803). The method, based on the evaluation of relative fluorescence yield at the J-step of the OJIP transient (VJ, VJ), has already been reported; however, thus far, it has been used mostly for studying dark-acclimated leaves, which limits its range of application. Here, we show that the OJIP transient can be used for semi-quantitative estimation of PQ-redox in algal and cyanobacterial cell cultures, in addition to plants. We further show that it can reflect PQ-redox in both dark-acclimated and light-acclimated samples. Our systematic comparison of Multi-Color PAM, AquaPen, and FL 6000 fluorometers demonstrates that accurate measurement of VJ and VJ parameters in suspension cultures requires low culture density and a high-intensity saturation pulse. We further show that with increasing light intensity to which the cells are exposed, the state of photosystem II (PSII) changes due to light-induced reduction of quinone A (QA-) and conformational changes, which in turn influence both the sensitivity and dynamic range of the VJ parameter towards PQ-redox estimation. A comparison of fluorescence transients in Chlorella and Synechocystis revealed high homeostatic control over PQ-redox in Synechocystis, maintained by terminal oxidases present at the thylakoid membrane. While we discuss certain limitations, our systematic assessment suggests that the OJIP method has great potential to become a routine tool for semi-quantitative PQ-redox estimation under a wide range of experimental conditions in green algae and cyanobacteria.

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Zavrel, T., Pohland, A. C., Pfennig, T., Matuszynska, A. B., Toth, S. Z., Bernat, G., Cerveny, J.. 2025-08-05. Assessing the redox state of the plastoquinone pool in algae and cyanobacteria via OJIP fluorescence: perspectives and limitations. https://doi.org/10.1101/2025.08.05.668659

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