A four state parametric model for the kinetics of the non-photochemical quenching in Photosystem II

Joris J. Snellenburg, Matthew P. Johnson, Alexander V. Ruban, Rienk van Grondelle, Ivo H. M. van Stokkum

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Abstract

The phenomenon of non-photochemical quenching (NPQ) was studied in spinach chloroplasts using pulse amplitude modulated (PAM) fluorometry. We present a new analysis method which describes the observed fluorescence quantum yield as the sum of the product of four different states of PSII and their corresponding quantum yields. These four distinct states are PSII in the quenched or unquenched state, and with its reaction center either open or closed depending upon the reduction of the QA site. With this method we can describe the dynamics of the NPQ induction and recovery as well as quantify the percentage of photoinactivated RC throughout the measurement. We show that after one cycle of quenching followed by a period of recovery, approximately 8-9% of the RC are photoinactivated, after two cycles of illumination this number becomes 1-5-17%. The recovery from the quenching appeared with rates of (50 s)(-1) and (1 hr)(-1). The new analysis method presented here is flexible, allowing it to be applied to any type of PAM fluorometry protocol. The method allows to quantitatively compare qualitatively different PAM curves on the basis of statistically relevant fitting parameters and to quantify quenching dynamics and photoinactivation. Moreover, the results presented here demonstrate that the analysis of a single PAM fluorometry quenching experiment can already provide information on the relative quantum yield of the four different states of PSII for the intact chloroplasts- something no other form of spectroscopy could provide in a single measurement.
Original languageEnglish
Pages (from-to)854-864
Number of pages11
JournalBiochimica et Biophysica Acta (BBA) - Bioenergetics
Volume1858
Issue number10
Early online date8 Aug 2017
DOIs
Publication statusPublished - Oct 2017

Funding

This research is performed as part of the BioSolar Cells research programme, sponsored by the Dutch Ministry of Economic Affairs. IHMvS and RvG acknowledge financial support of the European Research Council (Advanced Grant Proposal 267333 (PHOTPROT) to RvG). RvG gratefully acknowledges his Academy Professor grant from the Netherlands Royal Academy of Sciences (KNAW). AVR acknowledges the Leverhulme Trust grant RPG-2012-478 and UK Biotechnology and Biological Sciences Research Council grant BB/L019027/1. AVR would like to acknowledge The Royal Society for the Wolfson Research Merit Award and the NWO visiting scholarship grant 040.11.433. MPJ would like to acknowledge the Leverhulme Trust grant ECF-2012-398\2. JM Gruber and LW Bielczynski are thanked for helpful discussions regarding the interpretation of the modeling.

FundersFunder number
Dutch Ministry of Economic Affairs
Koninklijke Nederlandse Akademie van Wetenschappen
Biotechnology and Biological Sciences Research CouncilBB/L019027/1
Leverhulme TrustRPG-2012-478
Royal Society
European Research Council267333
Koninklijke Nederlandse Akademie van Wetenschappen
Nederlandse Organisatie voor Wetenschappelijk Onderzoek040.11.433, ECF-2012-398\2

    Keywords

    • Fluorescence quenching analysis
    • Non-photochemical quenching
    • Parameter estimation
    • Time-resolved fluorescence spectroscopy

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