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On the polyphasic quenching kinetics of chlorophyll a fluorescence in algae after light pulses of variable length

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Abstract

This study reports on kinetics of the fluorescence decay in a suspension of the alga Scenedesmus quadricauda after actinic illumination. These are monitored as the variable fluorescence signal in the dark following light pulses of variable intensity and duration. The decay reflects the restoration of chlorophyll fluorescence quenching of the photosystem II (PSII) antennas and shows a polyphasic pattern which suggests the involvement of different processes. The overall quenching curve after a fluorescence-saturating pulse (SP) of 250-ms duration, commonly used in pulse amplitude modulation applications as the tool for estimating the maximal fluorescence (F m), has been termed P–O, in which P and O have the same meaning as used in the OJIP induction curve in the light. Deconvolution of this signal shows at least three distinguishable exponential phases with reciprocal rate constants of the order of 10, 102, and 103 ms. The size of the long (>103 ms) and moderate (~102 ms) lasting components relative to the complete quenching signal after an SP increases with the duration of the actinic pulse concomitantly with an increase in the reciprocal rate constants of the fast (~10 ms) and moderate quenching phases. Fluorescence responses upon single turnover flashes of 30-μs duration (STFs) given at discrete times during the P–O quenching were used as tools for identifying the quencher involved in the P–O quenching phase preceding the STF excitation. Results are difficult to interpret in terms of a single-hit two-state trapping mechanism with distinguishable quenching properties of open and closed reaction centers only. They give support for an earlier hypothesis on a double-hit three-state trapping mechanism in which the so-called semi-closed reaction centers of PSII are considered. In these trapping-competent centers the single reduced acceptor pair [PheQ A]1−, depending on the size of photoelectrochemically induced pH effects on the Q B-binding site, functions as an efficient fluorescence quencher.

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Abbreviations

β :

Variable fraction of Q B-nonreducing RCs

DCMU:

3(3,4-Dichlorophenyl)-1,1-dimethylurea

F 0 :

Fluorescence level of dark-adapted system with 100 % open RCs

F m :

Fluorescence level of dark-adapted system with 100 % closed RCs after fluorescence-saturating pulse excitation

F PP(E)m :

Fluorescence level with 100 % semi-closed RCs after release of photo-(electro)chemical quenching (q PP(E) = 0, see for definition below)

FIA:

Fluorescence induction algorithm

k AB :

Rate constant of Q −A oxidation

k β :

Rate constant of oxidation of [PheQ A]2− in reduced Q B-nonreducing RCs

k L :

Excitation rate of photosystem in light pulse

rF v :

Relative variable fluorescence (F − F 0)/(F m − F 0)

OEC:

Oxygen evolving complex

Ph(e):

Pheophytin, primary electron electron acceptor of PSII

PSII:

Photosystem II

Q A :

Primary quinone electron acceptor of PSII

Q B :

Secondary quinone electron acceptor of PSII

q PP :

Degree of photochemical quenching with 0 ≤ q PP ≤ 1

q PE :

Degree of photoelectrochemical quenching with 0 ≤ q PE ≤ 1

RC:

Reaction center of photosystem

SP:

Fluorescence-saturating pulse

STF:

Single turnover flash (excitation)

TSTM:

Three-state trapping model

YZ :

Secondary electron donor of PSII

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Acknowledgments

The authors thank Dr. Ondrej Komarek for performing the variable fluorescence measurements and Mrs. Jana Hofhanzlova for growth of Scenedesmus cultures. The research of O.P. has been supported by the project Algatech (CZ.1.05/2.1.00/03.0110).

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Authors and Affiliations

  1. Department of Plant Physiology, Wageningen University and Research, Wageningen, The Netherlands

    Wim Vredenberg

  2. Laboratory of Photosynthesis-Algatech, Institute of Microbiology, Academy of Sciences Czech Republic, Trebon, Czech Republic

    Ondrej Prasil

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  1. Wim Vredenberg
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  2. Ondrej Prasil
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Correspondence to Wim Vredenberg.

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Vredenberg, W., Prasil, O. On the polyphasic quenching kinetics of chlorophyll a fluorescence in algae after light pulses of variable length. Photosynth Res 117, 321–337 (2013). https://doi.org/10.1007/s11120-013-9917-z

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