Intervening dark periods negatively affect the photosynthetic performance of Chlamydomonas reinhardtii during growth under fluctuating high light.

The acclimation of the green algae Chlamydomoas reinhardtii to high light (HL) has been studied predominantly under continuous illumination of the cells. Here, we investigated the impact of fluctuating HL in alternation with either low light (LL) or darkness on photosynthetic performance and on photoprotective responses. Compared to intervening LL phases, dark phases led to (1) more pronounced reduction of the photosystem II quantum efficiency, (2) reduced degradation of the PsbS protein, (3) lower energy dissipation capacity and (4) an increased pool size of the xanthophyll cycle pigments. These characteristics indicate increased photo-oxidative stress when HL periods are interrupted by dark phases instead of LL phases. This overall trend was similar when comparing long (8 h) and short (30 min) HL phases being interrupted by long (16 h) and short (60 min) phases of dark or low light, respectively. Only the degradation of PsbS was clearly more efficient during long (16 h) LL phases when compared to short (60 min) LL phases.

Less water in agriculture? Potential and challenges in optimizing water use efficiency.

This article comments on: Turc B, Sahay S, Haupt J, de Oliveira Santos T, Bai G, Glowacka K. 2024. Up-regulation of non-photochemical quenching improves water use efficiency and reduces whole-plant water consumption under drought in Nicotianatabacum. Journal of Experimental Botany 75, 3959-3972.

Specific thylakoid protein phosphorylations are prerequisites for overwintering of Norway spruce (Picea abies) photosynthesis.

Coping of evergreen conifers in boreal forests with freezing temperatures on bright winter days puts the photosynthetic machinery in great risk of oxidative damage. To survive harsh winter conditions, conifers have evolved a unique but poorly characterized photoprotection mechanism, a sustained form of nonphotochemical quenching (sustained NPQ). Here we focused on functional properties and underlying molecular mechanisms related to the development of sustained NPQ in Norway spruce (Picea abies). Data were collected during 4 consecutive years (2016 to 2019) from trees growing in sun and shade habitats. When day temperatures dropped below -4 °C, the specific N-terminally triply phosphorylated LHCB1 isoform (3p-LHCII) and phosphorylated PSBS (p-PSBS) could be detected in the thylakoid membrane. Development of sustained NPQ coincided with the highest level of 3p-LHCII and p-PSBS, occurring after prolonged coincidence of bright winter days and temperatures close to -10 °C. Artificial induction of both the sustained NPQ and recovery from naturally induced sustained NPQ provided information on differential dynamics and light-dependence of 3p-LHCII and p-PSBS accumulation as prerequisites for sustained NPQ. Data obtained collectively suggest three components related to sustained NPQ in spruce: 1) Freezing temperatures induce 3p-LHCII accumulation independently of light, which is suggested to initiate destacking of appressed thylakoid membranes due to increased electrostatic repulsion of adjacent membranes; 2) p-PSBS accumulation is both light- and temperature-dependent and closely linked to the initiation of sustained NPQ, which 3) in concert with PSII photoinhibition, is suggested to trigger sustained NPQ in spruce.

Mechanisms shaping the synergism of zeaxanthin and PsbS in photoprotective energy dissipation in the photosynthetic apparatus of plants.

Safe operation of photosynthesis is vital to plants and is ensured by the activity of processes protecting chloroplasts against photo-damage. The harmless dissipation of excess excitation energy is considered to be the primary photoprotective mechanism and is most effective in the combined presence of PsbS protein and zeaxanthin, a xanthophyll accumulated in strong light as a result of the xanthophyll cycle. Here we address the problem of specific molecular mechanisms underlying the synergistic effect of zeaxanthin and PsbS. The experiments were conducted with Arabidopsis thaliana, using wild-type plants, mutants lacking PsbS (npq4), and mutants affected in the xanthophyll cycle (npq1), with the application of molecular spectroscopy and imaging techniques. The results lead to the conclusion that PsbS interferes with the formation of densely packed aggregates of thylakoid membrane proteins, thus allowing easy exchange and incorporation of xanthophyll cycle pigments into such structures. It was found that xanthophylls trapped within supramolecular structures, most likely in the interfacial protein region, determine their photophysical properties. The structures formed in the presence of violaxanthin are characterized by minimized dissipation of excitation energy. In contrast, the structures formed in the presence of zeaxanthin show enhanced excitation quenching, thus protecting the system against photo-damage.

Light-harvesting complex stress-related proteins play crucial roles in the acclimation of Physcomitrella patens under fluctuating light conditions.

Photosynthetic organisms have evolved photoprotective mechanisms to acclimate to light intensity fluctuations in their natural growth environments. Photosystem (PS) II subunit S (PsbS) and light-harvesting complex (LHC) stress-related proteins (LhcSR) are essential for triggering photoprotection in vascular plants and green algae, respectively. The activity of both proteins is strongly enhanced in the moss Physcomitrella patens under high-light conditions. However, their role in regulating photosynthesis acclimation in P. patens under fluctuating light (FL) conditions is still unknown. Here, we compare the responses of wild-type (WT) P. patens and mutants lacking PsbS (psbs KO) or LhcSR1 and 2 (lhcsr KO) to FL conditions in which the low-light phases were periodically interrupted with high-light pulses. lhcsr KO mutant showed a strong reduction in growth with respect to WT and psbs KO under FL conditions. The lack of LhcSR not only decreased the level of non-photochemical quenching, resulting in an over-reduced plastoquinone pool, but also significantly increased the PSI acceptor limitation values with respect to WT and psbs KO under FL conditions. Moreover, in lhcsr KO mutant, the abundance of PSI core and PSI-LHCI complex decreased greatly under FL conditions compared with the WT and psbs KO. We proposed that LhcSR in P. patens play a crucial role in moss acclimation to dynamic light changes.

Chilling Upregulates Expression of the PsbS and LhcSR Genes in the Chloroplasts of the Green Microalga Lobosphaera incisa IPPAS C-2047.

Non-photochemical quenching (NPQ) of excited chlorophyll states is essential for protecting the photosynthetic apparatus (PSA) from the excessive light-induced damage in all groups of oxygenic photosynthetic organisms. The key component of the NPQ mechanism in green algae and some other groups of algae and mosses is the LhcSR protein of the light harvesting complex (LHC) protein superfamily. In vascular plants, LhcSR is replaced by PsbS, another member of the LHC superfamily and a subunit of photosystem II (PSII). PsbS also performs the photoprotective function in mosses. For a long time, PsbS had been believed to be nonfunctional in green algae, although the corresponding gene was discovered in the genome of these organisms. The first evidence of the PsbS accumulation in the model green alga Chlamydomonas reinhardtii in response to the increase in irradiance was obtained only six years ago. However, the observed increase in the PsbS content was short-termed (on an hour-timescale). Here, we report a significant (more than three orders of magnitude) and prolonged (four days) upregulation of PsbS expression in response to the chilling-induced high-light stress followed by a less significant (~ tenfold) increase in the PsbS expression for nine days. This is the first evidence for the long-term upregulation of the PsbS expression in green alga (Chlorophyta) in response to stress. Our data indicate that the role of PsbS in the PSA of Chlorophyta is not limited to the first-line defense against stress, as it was previously assumed, but includes full-scale participation in the photoprotection of PSA from the environmental stress factors.

High Light Acclimation Mechanisms Deficient in a PsbS-Knockout Arabidopsis Mutant.

The photosystem II PsbS protein of thylakoid membranes is responsible for regulating the energy-dependent, non-photochemical quenching of excess chlorophyll excited states as a short-term mechanism for protection against high light (HL) stress. However, the role of PsbS protein in long-term HL acclimation processes remains poorly understood. Here we investigate the role of PsbS protein during long-term HL acclimation processes in wild-type (WT) and npq4-1 mutants of Arabidopsis which lack the PsbS protein. During long-term HL illumination, photosystem II photochemical efficiency initially dropped, followed by a recovery of electron transport and photochemical quenching (qL) in WT, but not in npq4-1 mutants. In addition, we observed a reduction in light-harvesting antenna size during HL treatment that ceased after HL treatment in WT, but not in npq4-1 mutants. When plants were adapted to HL, more reactive oxygen species (ROS) were accumulated in npq4-1 mutants compared to WT. Gene expression studies indicated that npq4-1 mutants failed to express genes involved in plastoquinone biosynthesis. These results suggest that the PsbS protein regulates recovery processes such as electron transport and qL during long-term HL acclimation by maintaining plastoquinone biosynthetic gene expression and enhancing ROS homeostasis.

Non-Photochemical Quenching under Drought and Fluctuating Light.

Plants grow in a variable environment in regard to soil water and light driving photochemical reactions. Light energy exceeding plant capability to use it for photochemical reactions must be dissipated by processes of non-photochemical quenching (NPQ). The aim of the study was to evaluate the impact of various components of NPQ on the response of Arabidopsis thaliana to fluctuating light and water availability. A laboratory experiment with Arabidopsis thaliana wild type (WT) and mutants npq1 and npq4 grown under optimum or reduced water availability was conducted. Dark-adapted plants were illuminated with fluctuating light (FL) of two intensities (55 and 530 µmol m-2 s-1) with each of the phases lasting for 20 s. The impact of water availability on the role of zeaxanthin and PsbS protein in NPQ induced at FL was analysed. The water deficit affected the dynamics of NPQ induced by FL. The lack of zeaxanthin or PsbS reduced plant capability to cope with FL. The synergy of both of these components was enhanced in regard to the amplitude of NPQ in the drought conditions. PsbS was shown as a component of primary importance in suiting plant response to FL under optimum and reduced water availability.

The Mechanism of Non-Photochemical Quenching in Plants: Localization and Driving Forces.

Non-photochemical chlorophyll fluorescence quenching (NPQ) remains one of the most studied topics of the 21st century in photosynthesis research. Over the past 30 years, profound knowledge has been obtained on the molecular mechanism of NPQ in higher plants. First, the largely overlooked significance of NPQ in protecting the reaction center of photosystem II (RCII) against damage, and the ways to assess its effectiveness are highlighted. Then, the key in vivo signals that can monitor the life of the major NPQ component, qE, are presented. Finally, recent knowledge on the site of qE and the possible molecular events that transmit ΔpH into the conformational change in the major LHCII [the major trimeric light harvesting complex of photosystem II (PSII)] antenna complex are discussed. Recently, number of reports on Arabidopsis mutants lacking various antenna components of PSII confirmed that the in vivo site of qE rests within the major trimeric LHCII complex. Experiments on biochemistry, spectroscopy, microscopy and molecular modeling suggest an interplay between thylakoid membrane geometry and the dynamics of LHCII, the PsbS (PSII subunit S) protein and thylakoid lipids. The molecular basis for the qE-related conformational change in the thylakoid membrane, including the possible onset of a hydrophobic mismatch between LHCII and lipids, potentiated by PsbS protein, begins to unfold.

An underlying mechanism of qE deficiency in marine angiosperm Zostera marina.

Non-photochemical quenching (NPQ) of photosystem II (PSII) fluorescence is one of the most important protective mechanisms enabling the survival of phototropic organisms under high-light conditions. A low-efficiency NPQ, characterized by weak NPQ induction capacity and a low level of protective NPQ, was observed in the marine angiosperm Zostera marina, which inhabits the shallow water regions. Furthermore, chlorophyll fluorescence and Western blot analysis verified that the fast-inducted component of NPQ, i.e., the energy-dependent quenching (qE), was not present in this species. In contrast with the lack of PSII antenna quenching sites for qE induction in brown algae and the lack of functional XC in Ulvophyceae belonging to green algae, all the antenna proteins and the functional XC are present in Z. marina. A novel underlying mechanism was observed that the limited construction of the trans-thylakoid proton gradient (ΔpH) caused by photoinactivation of the oxygen evolving complex (OEC) did not induce protonation of PsbS, thus explaining the inability to form quenching sites for qE induction. Although the ΔpH established under light exposure activated violaxanthin (V) de-epoxidase enzyme to catalyze conversion of V via antheraxanthin (A) and then to zeaxanthin (Z), the quenching capacity of de-epoxidized pigment was weak in Z. marina. We suggest that the low-efficiency NPQ was conducive to efficiently utilize the limited electrons to perform photosynthesis, resisting the adverse effect of OEC photoinactivation on the photosynthetic rate.

Role of Thylakoid Protein Phosphorylation in Energy-Dependent Quenching of Chlorophyll Fluorescence in Rice Plants.

Under natural environments, light quality and quantity are extremely varied. To respond and acclimate to such changes, plants have developed a multiplicity of molecular regulatory mechanisms. Non-photochemical quenching of chlorophyll fluorescence (NPQ) and thylakoid protein phosphorylation are two mechanisms that protect vascular plants. To clarify the role of thylakoid protein phosphorylation in energy-dependent quenching of chlorophyll fluorescence (qE) in rice plants, we used a direct Western blot assay after BN-PAGE to detect all phosphoproteins by P-Thr antibody as well as by P-Lhcb1 and P-Lhcb2 antibodies. Isolated thylakoids in either the dark- or the light-adapted state from wild type (WT) and PsbS-KO rice plants were used for this approach to detect light-dependent interactions between PsbS, PSII, and LHCII proteins. We observed that the bands corresponding to the phosphorylated Lhcb1 and Lhcb2 as well as the other phosphorylated proteins were enhanced in the PsbS-KO mutant after illumination. The qE relaxation became slower in WT plants after 10 min HL treatment, which correlated with Lhcb1 and Lhcb2 protein phosphorylation in the LHCII trimers under the same experimental conditions. Thus, we concluded that light-induced phosphorylation of PSII core and Lhcb1/Lhcb2 proteins is enhanced in rice PsbS-KO plants which might be due to more reactive-oxygen-species production in this mutant.

On the PsbS-induced quenching in the plant major light-harvesting complex LHCII studied in proteoliposomes.

Non-photochemical quenching (NPQ) in photosynthetic organisms provides the necessary photoprotection that allows them to cope with largely and quickly varying light intensities. It involves deactivation of excited states mainly at the level of the antenna complexes of photosystem II using still largely unknown molecular mechanisms. In higher plants the main contribution to NPQ is the so-called qE-quenching, which can be switched on and off in a few seconds. This quenching mechanism is affected by the low pH-induced activation of the small membrane protein PsbS which interacts with the major light-harvesting complex of photosystem II (LHCII). We are reporting here on a mechanistic study of the PsbS-induced LHCII quenching using ultrafast time-resolved chlorophyll (Chl) fluorescence. It is shown that the PsbS/LHCII interaction in reconstituted proteoliposomes induces highly effective and specific quenching of the LHCII excitation by a factor ≥ 20 via Chl-Chl charge-transfer (CT) state intermediates which are weakly fluorescent. Their characteristics are very broad fluorescence bands pronouncedly red-shifted from the typical unquenched LHCII fluorescence maximum. The observation of PsbS-induced Chl-Chl CT-state emission from LHCII in the reconstituted proteoliposomes is highly reminiscent of the in vivo quenching situation and also of LHCII quenching in vitro in aggregated LHCII, indicating a similar quenching mechanism in all those situations. The PsbS mutant lacking the two proton sensing Glu residues induced significant, but much smaller, quenching than wild type. Added zeaxanthin had only minor effects on the yield of quenching in the proteoliposomes. Overall our study shows that PsbS co-reconstituted with LHCII in liposomes represents an excellent in vitro model system with characteristics that are reflecting closely the in vivo qE-quenching situation.

Photosystem II 22kDa protein level - a prerequisite for excess light-inducible memory, cross-tolerance to UV-C and regulation of electrical signalling.

It is well known that PsbS is a key protein for the proper management of excessive energy in plants. Plants without PsbS cannot trigger non-photochemical quenching, which is crucial for optimal photosynthesis under variable conditions. Our studies showed wild-type plants had enhanced tolerance to UV-C-induced cell death (CD) upon induction of light memory by a blue or red light. However, npq4-1 plants, which lack PsbS, as well as plants overexpressing this protein (oePsbS), responded differently. Untreated oePsbS appeared more tolerant to UV-C exposure, whereas npq4-1 was unable to adequately induce cross-tolerance to UV-C. Similarly, light memory induced by episodic blue or red light was differently deregulated in npq-4 and oePsbS, as indicated by transcriptomic analyses, measurements of the trans-thylakoid pH gradient, chlorophyll a fluorescence parameters, and measurements of foliar surface electrical potential. The mechanism of the foliar CD development seemed to be unaffected in the analysed plants and is associated with chloroplast breakdown. Our results suggest a novel, substantial role for PsbS as a regulator of chloroplast retrograde signalling for light memory, light acclimation, CD, and cross-tolerance to UV radiation.

Photoprotective energy dissipation is greater in the lower, not the upper, regions of a rice canopy: a 3D analysis.

High light intensities raise photosynthetic and plant growth rates but can cause damage to the photosynthetic machinery. The likelihood and severity of deleterious effects are minimised by a set of photoprotective mechanisms, one key process being the controlled dissipation of energy from chlorophyll within PSII known as non-photochemical quenching (NPQ). Although ubiquitous, the role of NPQ in plant productivity is important because it momentarily reduces the quantum efficiency of photosynthesis. Rice plants overexpressing and deficient in the gene encoding a central regulator of NPQ, the protein PsbS, were used to assess the effect of protective effectiveness of NPQ (pNPQ) at the canopy scale. Using a combination of three-dimensional reconstruction, modelling, chlorophyll fluorescence, and gas exchange, the influence of altered NPQ capacity on the distribution of pNPQ was explored. A higher phototolerance in the lower layers of a canopy was found, regardless of genotype, suggesting a mechanism for increased protection for leaves that experience relatively low light intensities interspersed with brief periods of high light. Relative to wild-type plants, psbS overexpressors have a reduced risk of photoinactivation and early growth advantage, demonstrating that manipulating photoprotective mechanisms can impact both subcellular mechanisms and whole-canopy function.

Dynamics of regulated YNPQ and non-regulated YNO energy dissipation in sunflower leaves exposed to sinusoidal lights.

Better understanding of photosynthetic efficiency under fluctuating light requires a specific approach to characterize the dynamics of energy dissipation in photosystem II. In this study, we characterized the interaction between the regulated YNPQ and non-regulated YNO energy dissipation in outdoor- and indoor-grown sunflower leaves exposed to repetitive cycles of sinusoidal lights of five amplitudes (200, 400, 600, 800, 1000 µmol m-2 s-1) and periods (20, 40, 60, 90, 120 s). The different light cycles induced various patterns of ChlF emission, from which were calculated the complementary quantum yields of photochemical energy conversion YII, light-regulated YNPQ, and non-regulated YNO non-photochemical energy dissipation. During the light cycles, YNO varied in complex but small patterns relative to those of YNPQ, whose variations were mostly mirrored by changes in YII. The YNO patterns could be decomposed by fast Fourier transform into a main (MH) and several upper harmonics (UH). Concerning YNPQ dynamics, they were described by sinusoidal regressions with two components, one constant during the light cycles but increasing with the average light intensity (YNPQc), and one variable (YNPQv). Formation and relaxation of YNPQv followed the intensity of the sinusoidal lights, with lags ranging from 5 to 13 s. These lags decreased with the amplitude of the incident light, and were shorter by 37% in outdoor than indoor leaves. YNPQv and UHs responses to the growth conditions, amplitudes, and the periods of the sinusoidal light were closely correlated (r = 0.939), whereas MH and YNPQc varied similarly (r = 0.803). The analysis of ChlF induced by sinusoidal lights may be a useful tool to better understand the dynamics of energy dissipation in PSII under fluctuating lights.

The rise and fall of Light-Harvesting Complex Stress-Related proteins as photoprotection agents during evolution.

Photosynthesis depends on light. However, excess light can be harmful for the photosynthetic apparatus because it produces reactive oxygen species (ROS) that cause photoinhibition. Oxygenic organisms evolved photoprotection mechanisms to counteract light-dependent ROS production, including preventive dissipation of excited states of chlorophyll (1Chl*) into heat in the process termed non-photochemical quenching (NPQ). This consists in the activation of 1Chl* quenching reactions when the thylakoid luminal pH drops below 5.2. In turn, acidification occurs when the rate of the CO2 reducing cycle is saturated and cannot regenerate ADP+Pi, thus inhibiting ATPase activity and the return of protons (H+) to the stromal compartment. The major and fastest component of NPQ is energy quenching, qE, which in algae depends on the Light-Harvesting Complex Stress-Related (LHCSR) proteins. In mosses, LHCSR proteins have remained the major catalysts of energy dissipation, but a minor contribution also occurs via a homologous protein, Photosystem II Subunit S (PSBS). In vascular plants, however, LHCSR has disappeared and PSBS is the only pH-sensitive trigger of qE. Why did PSBS replace LHCSR in the later stages of land colonization? Both PSBS and LHCSR belong to the Light Harvesting Complex superfamily (LHC) and share properties such as harboring protonatable residues that are exposed to the chloroplast lumen, which is essential for pH sensing. However, there are also conspicuous differences: LHCSR binds chlorophylls and xanthophylls while PSBS does not, implying that the former may well catalyse quenching reactions while the latter needs pigment-binding partners for its quenching function. Here, the evolution of quenching mechanisms for excess light is reviewed with a focus on the role of LHCSR versus PSBS, and the reasons for the redundancy of LHCSR in vascular plants as PSBS became established.

Photoprotection and growth under different lights of Arabidopsis single and double mutants for energy dissipation (npq4) and state transitions (pph1).

KEY MESSAGE: Arabidopsis single and double mutants for energy dissipation (npq4) and state transitions (pph1, blocked in State II) show enhanced growth and flowers + siliques production under controlled low-light conditions. Non-photochemical quenching (NPQ) is a short-term regulation important to maintain efficient photosynthesis and to avoid photooxidative damages by dissipation of excess energy. Full activation of NPQ in plants requires the protonation of the PsbS protein, which is the sensor of the low lumenal pH triggering the thermal dissipation. State transitions are a second important photosynthetic regulation to respond to changes in light quality and unbalanced excitation of photosystems. State transitions allow energy redistribution between PSI and PSII through the reversible exchange of LHCII antenna complexes between photosystems thanks to the opposite action of the STN7 kinase and PPH1 phosphatase: phosphorylation of LHCII promotes its mobilization from PSII to PSI, while dephosphorylation has the opposite effect. In this work, we produced the pph1/npq4 double mutant and characterized some photosynthetic, growth and reproduction properties in comparison with wild-type and single-mutant plants in high- and low-light conditions. Results indicate that in high light, the pph1 mutant maintains good photoprotection ability, while npq4 plants show more susceptibility to photodamages. The pph1/npq4 double mutant showed a resistance to high-light stress similar to that of the single npq4 mutant. In low-light condition, the single mutants showed a significant increase of growth and flowering compared to wild-type plants and this effect was further enhanced in the pph1/npq4 double mutant. Results suggest that photosynthetic optimisation to improve crop growth and productivity might be possible, at least under controlled low-light conditions, by modifying NPQ and regulation of state transitions.

UV-B photoreceptor-mediated protection of the photosynthetic machinery in Chlamydomonas reinhardtii.

Life on earth is dependent on the photosynthetic conversion of light energy into chemical energy. However, absorption of excess sunlight can damage the photosynthetic machinery and limit photosynthetic activity, thereby affecting growth and productivity. Photosynthetic light harvesting can be down-regulated by nonphotochemical quenching (NPQ). A major component of NPQ is qE (energy-dependent nonphotochemical quenching), which allows dissipation of light energy as heat. Photodamage peaks in the UV-B part of the spectrum, but whether and how UV-B induces qE are unknown. Plants are responsive to UV-B via the UVR8 photoreceptor. Here, we report in the green alga Chlamydomonas reinhardtii that UVR8 induces accumulation of specific members of the light-harvesting complex (LHC) superfamily that contribute to qE, in particular LHC Stress-Related 1 (LHCSR1) and Photosystem II Subunit S (PSBS). The capacity for qE is strongly induced by UV-B, although the patterns of qE-related proteins accumulating in response to UV-B or to high light are clearly different. The competence for qE induced by acclimation to UV-B markedly contributes to photoprotection upon subsequent exposure to high light. Our study reveals an anterograde link between photoreceptor-mediated signaling in the nucleocytosolic compartment and the photoprotective regulation of photosynthetic activity in the chloroplast.

A siphonous morphology affects light-harvesting modulation in the intertidal green macroalga Bryopsis corticulans (Ulvophyceae).

MAIN CONCLUSION: The macroalga Bryopsis corticulans relies on a sustained protective NPQ and a peculiar body architecture to efficiently adapt to the extreme light changes of intertidal shores. During low tides, intertidal algae experience prolonged high light stress. Efficient dissipation of excess light energy, measured as non-photochemical quenching (NPQ) of chlorophyll fluorescence, is therefore required to avoid photodamage. Light-harvesting regulation was studied in the intertidal macroalga Bryopsis corticulans, during high light and air exposure. Photosynthetic capacity and NPQ kinetics were assessed in different filament layers of the algal tufts and in intact chloroplasts to unravel the nature of NPQ in this siphonous green alga. We found that the morphology and pigment composition of the B. corticulans body provides functional segregation between surface sunlit filaments (protective state) and those that are underneath and undergo severe light attenuation (light-harvesting state). In the surface filaments, very high and sustained NPQ gradually formed. NPQ induction was triggered by the formation of transthylakoid proton gradient and independent of the xanthophyll cycle. PsbS and LHCSR proteins seem not to be active in the NPQ mechanism activated by this alga. Our results show that B. corticulans endures excess light energy pressure through a sustained protective NPQ, not related to photodamage, as revealed by the unusually quick restoration of photosystem II (PSII) function in the dark. This might suggest either the occurrence of transient PSII photoinactivation or a fast rate of PSII repair cycle.

The evolution of the photoprotective antenna proteins in oxygenic photosynthetic eukaryotes.

Photosynthetic organisms require rapid and reversible down-regulation of light harvesting to avoid photodamage. Response to unpredictable light fluctuations is achieved by inducing energy-dependent quenching, qE, which is the major component of the process known as non-photochemical quenching (NPQ) of chlorophyll fluorescence. qE is controlled by the operation of the xanthophyll cycle and accumulation of specific types of proteins, upon thylakoid lumen acidification. The protein cofactors so far identified to modulate qE in photosynthetic eukaryotes are the photosystem II subunit S (PsbS) and light-harvesting complex stress-related (LHCSR/LHCX) proteins. A transition from LHCSR- to PsbS-dependent qE took place during the evolution of the Viridiplantae (also known as 'green lineage' organisms), such as green algae, mosses and vascular plants. Multiple studies showed that LHCSR and PsbS proteins have distinct functions in the mechanism of qE. LHCX(-like) proteins are closely related to LHCSR proteins and found in 'red lineage' organisms that contain secondary red plastids, such as diatoms. Although LHCX proteins appear to control qE in diatoms, their role in the mechanism remains poorly understood. Here, we present the current knowledge on the functions and evolution of these crucial proteins, which evolved in photosynthetic eukaryotes to optimise light harvesting.