Esa Tyystjärvi

Photoinhibition of Photosystem II.

Photoinhibition of Photosystem II (PSII) is the light-induced loss of PSII electron-transfer activity. Although photoinhibition has been studied for a long time, there is no consensus about its mechanism. On one hand, production of singlet oxygen ((1)O(2)) by PSII has promoted models in which this reactive oxygen species (ROS) is considered to act as the agent of photoinhibitory damage. These chemistry-based models have often not taken into account the photophysical features of photoinhibition-like light response and action spectrum. On the other hand, models that reproduce these basic photophysical features of the reaction have not considered the importance of data about ROS. In this chapter, it is shown that the evidence behind the chemistry-based models and the photophysically oriented models can be brought together to build a mechanism that confirms with all types of experimental data. A working hypothesis is proposed, starting with inhibition of the manganese complex by light. Inability of the manganese complex to reduce the primary donor promotes recombination between the oxidized primary donor and Q(A), the first stable quinone acceptor of PSII. (1)O(2) production due to this recombination may inhibit protein synthesis or spread the photoinhibitory damage to another PSII center. The production of (1)O(2) is transient because loss of activity of the oxygen-evolving complex induces an increase in the redox potential of Q(A), which lowers (1)O(2) production.

Excess Copper Predisposes Photosystem II to Photoinhibition in Vivo by Outcompeting Iron and Causing Decrease in Leaf Chlorophyll

Photoinhibition of photosystem II was studied in vivo with bean (Phaseolus vulgaris) plants grown in the presence of 0.3 (control), 4, or 15 μm Cu2+. Although photoinhibition, measured in the presence of lincomycin to block concurrent recovery, is faster in leaves of Cu2+-treated plants than in control leaves, thylakoids isolated from Cu-treated plants did not show high sensitivity to photoinhibition. Direct effects of excess Cu2+ on chloroplast metabolism are actually unlikely, because the Cu concentration of chloroplasts of Cu-treated plants was lower than that of their leaves. Excess Cu in the growth medium did not cause severe oxidative stress, collapse of antioxidative defenses, or loss of photoprotection. Thus, these hypothetical effects can be eliminated as causes for Cu-enhanced photoinhibition in intact leaves. However, Cu treatment lowered the leaf chlorophyll (Chl) concentration and reduced the thylakoid membrane network. The loss of Chl and sensitivity to photoinhibition could be overcome by adding excess Fe together with excess Cu to the growth medium. The addition of Fe lowered the Cu2+ concentration of the leaves, suggesting that Cu outcompetes Fe in Fe uptake. We suggest that the reduction of leaf Chl concentration, caused by the Cu-induced iron deficiency, causes the high photosensitivity of photosystem II in Cu2+-treated plants. A causal relationship between the susceptibility to photoinhibition and the leaf optical density was established in several plant species. Plant species adapted to high-light habitats apparently benefit from thick leaves because the rate of photoinhibition is directly proportional to light intensity, but photosynthesis becomes saturated by moderate light.

Linking chlorophyll a fluorescence to photosynthesis for remote sensing applications: mechanisms and challenges

Chlorophyll a fluorescence (ChlF) has been used for decades to study the organization, functioning, and physiology of photosynthesis at the leaf and subcellular levels. ChlF is now measurable from remote sensing platforms. This provides a new optical means to track photosynthesis and gross primary productivity of terrestrial ecosystems. Importantly, the spatiotemporal and methodological context of the new applications is dramatically different compared with most of the available ChlF literature, which raises a number of important considerations. Although we have a good mechanistic understanding of the processes that control the ChlF signal over the short term, the seasonal link between ChlF and photosynthesis remains obscure. Additionally, while the current understanding of in vivo ChlF is based on pulse amplitude-modulated (PAM) measurements, remote sensing applications are based on the measurement of the passive solar-induced chlorophyll fluorescence (SIF), which entails important differences and new challenges that remain to be solved. In this review we introduce and revisit the physical, physiological, and methodological factors that control the leaf-level ChlF signal in the context of the new remote sensing applications. Specifically, we present the basis of photosynthetic acclimation and its optical signals, we introduce the physical and physiological basis of ChlF from the molecular to the leaf level and beyond, and we introduce and compare PAM and SIF methodology. Finally, we evaluate and identify the challenges that still remain to be answered in order to consolidate our mechanistic understanding of the remotely sensed SIF signal.

Parameterization of photosystem II photoinactivation and repair.

The photoinactivation (also termed photoinhibition or photodamage) of Photosystem II (PSII) and the counteracting repair reactions are fundamental elements of the metabolism and ecophysiology of oxygenic photoautotrophs. Differences in the quantification, parameterization and terminology of Photosystem II photoinactivation and repair can erect barriers to understanding, and particular parameterizations are sometimes incorrectly associated with particular mechanistic models. These issues lead to problems for ecophysiologists seeking robust methods to include photoinhibition in ecological models. We present a comparative analysis of terms and parameterizations applied to photoinactivation and repair of Photosystem II. In particular, we show that the target size and quantum yield approaches are interconvertible generalizations of the rate constant of photoinactivation across a range of incident light levels. Our particular emphasis is on phytoplankton, although we draw upon the literature from vascular plants. This article is part of a Special Issue entitled: Photosystem II.

ATP and light regulate D1 protein modification and degradation Role of D1* in photoinhibition

We have recently shown that during in vivo photoinhibition the D1 protein is degraded via a modified form, designated D1*. Depending on light conditions, the amount of D1* varies in leaves between 0 and 50% of total D1 content. By isolating thylakoids from leaves acclimated to different light levels, and performing photoinhibition experiments on these thylakoids, the following results on D1 protein degradation were obtained: (1) the protease involved in D1 degradation requires activation by light; (ii) neither acceptor nor donor side photoinhibition of PSII induces formation of D1* in vitro; (iii) in isolated thylakoids, the transformation of D1 to D1* can be induced in low light in the presence of ATP, which suggests that D1* is a phosphorylated form of the D1 protein; (iv) D1*, induced either in vivo or in vitro, is much less susceptible to degradation during illumination of isolated thylakoids than the original D1 protein. We suggest that the modification to D1* is a means to prevent disassembly of photodamaged photosystem II complex in appressed membranes.

Molecular mechanism of high-temperature-induced inhibition of acceptor side of Photosystem II

High-temperature-induced inhibition of the acceptor side of Photosystem II (PS II) was studied in tobacco thylakoids using oxygen evolution, chlorophyll a (Chl a) fluorescence and redox potential measurements. When thylakoids were heated at 2 °C/min from 25 to 50 °C, the oxygen evolving complex became inhibited between 32 and 45 °C, whereas the acceptor side of PS II tolerated higher temperatures. Variable Chl a fluorescence decreased more slowly than oxygen evolution, suggesting that transitions between some S-states occurred even after heat-induced inhibition of the oxygen evolving activity. 77 K emission spectroscopy reveals that heating does not cause detachment of the light-harvesting complex II from PS II, and thus the heat-induced increase in the initial F0 fluorescence is due to loss of exciton trapping in the heated PS II centers. Redox titrations showed a heat-induced increase in the midpoint potential of the QA/QA-) couple from the control value of –80 mV to +40 mV at 50 °C, indicating a loss of the reducing power of QA-). When its driving force thus decreased, electron transfer from QA-) to QB in the PS II centers that still could reduce QA became gradually inhibited, as shown by measurements of the decay of Chl a fluorescence yield after a single turnover flash. Interestingly, the heat-induced loss of variable fluorescence and inhibition of electron transfer from QA-) to QB could be partially prevented by the presence of 5 mM bicarbonate during heating, suggesting that high temperatures cause release of the bicarbonate bound to PS II. We speculate that both the upshift in the redox potential of the QA/QA-) couple and the release of bicarbonate may be caused by a heat-induced structural change in the transmembrane D1 or D2 proteins. This structural change may, in turn, be caused by the inhibition of the oxygen evolving complex during heating.

D1 protein degradation during photoinhibition of intact leaves a modification of the D1 protein precedes degradation

Illumination of intact pumpkin leaves with high light led to severe photoinhibition of photosystem II with no net degradation of the D1 protein. Instead, however, a modified form of D1 protein with slightly slower electrophoretic mobility was induced with corresponding loss in the original form of the D1 protein. When the leaves were illuminated in the presence of chloramphenicol the modified form was degraded, which led to a decrease in the total amount of the D1 protein. Subfractionation of the thylakoid membranes further supported the conclusion that the novel form of the D1 protein was not a precursor but a high‐light modified form that was subsequently degraded.

A microcomputer program and fast analog to digital converter card for the analysis of fluorescence induction transients.

A microcomputer program and an analog to digital conversion card were developed for the analysis of fluorescence induction curves. The program and the analog to digital conversion card are compatible with all commercially available fluorometers. Most of the current analysis methods for fluorescence induction curves are implemented in the program, including analysis of OIDPSMT-kinetics, dissection of fluorescence quenching into two components, measurement of the slope of the fluorescence curve, complementary area analysis and analysis of energy ‘spillover’ from PS II to PS I. The program can also do basic statistical calculations from the measured parameters. The architecture of the program is open, allowing the user to add new methods to the main body of the program. Split time-scale is used in data capture and analysis. A new procedure facilitates accurate determination of F0.

Protection by α-tocopherol of the repair of photosystem II during photoinhibition in Synechocystis sp. PCC 6803

α-Tocopherol is a lipophilic antioxidant that is an efficient scavenger of singlet oxygen. We investigated the role of α-tocopherol in the protection of photosystem II (PSII) from photoinhibition using a mutant of the cyanobacterium Synechocystis sp. PCC 6803 that is deficient in the biosynthesis of α-tocopherol. The activity of PSII in mutant cells was more sensitive to inactivation by strong light than that in wild-type cells, indicating that lack of α-tocopherol enhances the extent of photoinhibition. However, the rate of photodamage to PSII, as measured in the presence of chloramphenicol, which blocks the repair of PSII, did not differ between the two lines of cells. By contrast, the repair of PSII from photodamage was suppressed in mutant cells. Addition of α-tocopherol to cultures of mutant cells returned the extent of photoinhibition to that in wild-type cells, without any effect on photodamage. The synthesis de novo of various proteins, including the D1 protein that plays a central role in the repair of PSII, was suppressed in mutant cells under strong light. These observations suggest that α-tocopherol promotes the repair of photodamaged PSII by protecting the synthesis de novo of the proteins that are required for recovery from inhibition by singlet oxygen.

Action spectrum of psbA gene transcription is similar to that of photoinhibition in Synechocystis sp. PCC 6803.

The photosystem II (PSII) reaction center protein D1 undergoes rapid light‐dependent turnover, which is caused by photoinhibition. To identify the photoreceptor(s) involved in the light‐dependent expression of the psbA gene encoding the D1 protein, we determined the action spectra of psbA transcription, PSII activity, photosynthesis and photoinhibition in Synechocystis sp. PCC 6803. In accordance with its phycobilisome antenna, PSII showed the highest activity in the spectral region from yellow to red and only low activity in the ultraviolet‐A (UV‐A) to green region. Photoinhibition, in turn, was fastest in UV‐A to violet light and a minor peak was found in the orange region. The action spectrum of psbA transcription resembled closely that of photoinhibition, suggesting that photoinhibition creates a signal for up‐regulation of the psbA gene.