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Highlights • Plants utilize super/megacomplexes to survive severe light conditions on land. • Several LHCII trimers can bind to the PSI–PSII megacomplex via the LHCI belt. • Excitation energy is diverted automatically to PSI in the megacomplex. • Megacomplexes help regulate the excitation energy in photosystems. Traditionally, two types of photosystem reaction centers (PSI and PSII) are thought to be spatially dispersed in the plant thylakoid membrane. In this model, PSI and PSII independently accept excitation energy from their own peripheral light-harvesting complexes, LHCI and LHCII, respectively, and form supercomplexes (PSI–LHCI and PSII–LHCII). However, recent studies using a combination of mild detergent treatment and spectroscopic analysis have revealed the existence of various megacomplexes such as a PSI–PSII megacomplex and a PSII megacomplex. Flexibility in the formation of supercomplexes and megacomplexes is important for land plants to regulate excitation energy to survive under strong and fluctuating sunlight on land. [ABSTRACT FROM AUTHOR]

Recently, we proposed a simple conceptual fluctuating antenna model (FAM), describing excitation diffusion and trapping in a continuous medium, where variations of the excitation transfer pathways are taken into account by the introduced fractional space dimension. Since then, this model has been successfully applied to simulate multi-exponential excitation quenching kinetics in a series of plant photosynthetic systems, purified from the thylakoid membranes, without invoking a radical pair state in the reaction centre. Here, we overview this model and its parameters obtained for various systems, and extend the area of its applications to several pigment-protein supercomplexes containing the photosystem I (PSI). We show that while the diffusion in the PSI core is virtually three-dimensional, the PSI core aggregates interconnected with other light-harvesting complexes (LHCI and/or LHCII) are characterized by a substantially reduced dimension, which indicates a smaller number of energy transfer links from LHCI to the PSI core. We also suggest that in vivo both PSI and PSII antennae are substantially larger than those observed in the isolated systems: PSII antenna contains in total about 6 LHCII trimers while PSI is aggregated with at least one LHCII trimer. The obtained results show that FAM can be a very useful tool to follow photosynthetic apparatus transformations during short- and long-term adaptation to varying light, monitored by kinetic fluorescence spectroscopy. [ABSTRACT FROM AUTHOR]

Light-harvesting-like (LIL) proteins are a group of proteins that share a consensus amino acid sequence with light-harvesting Chl-binding (LHC) proteins. We hypothesized that they might be involved in photosynthesis-related processes. In order to gain a better understanding of a potential role in photosynthesis-related processes, we examined the most recently identified LIL protein, LIL8/PSB33. Recently, it was suggested that this protein is an auxiliary PSII core protein which is involved in organization of the PSII supercomplex. However, we found that the majority of LIL8/PSB33 was localized in stroma lamellae, where PSI is predominant. Moreover, the PSI antenna sizes measured under visible light were slightly increased in the lil8 mutants which lack LIL8/PSB33 protein. Analysis of fluorescence decay kinetics and fluorescence decay-associated spectra indicated that energy transfer to quenching sites within PSI was partially hampered in these mutants. On the other hand, analysis of the steady-state fluorescence spectra in these mutants indicates that a population of LHCII is energetically disconnected from PSII. Taken together, we suggest that LIL8/PSB33 is involved in the fine-tuning of light harvesting and/or energy transfer around both photosystems. [ABSTRACT FROM AUTHOR]

Comparative proteome analysis of subcellular compartments like thylakoid membranes and their associated supercomplexes can deliver important in-vivo information on the molecular basis of physiological functions which go far beyond to that what can be learnt from transcriptional-based gene expression studies. For instance, the finding that light intensity influences mainly the relative stoichiometry of subunits could be obtained only by high resolution proteome analysis. The high sensitivity of LC-ESI–MS/MS based proteome analysis allows the determination of proteins in very small subfractions along with their non-labeled semi quantitative analysis. This provides insights in the protein-protein interactions of supercomplexes that are the operative units in intact cells. Here, we have focused on functional proteome approaches for the identification of microalgal light-harvesting complex proteins in chloroplasts and the eyespot in general and in detail for those of diatoms that are exposed to varying light conditions [ABSTRACT FROM AUTHOR]

The study investigated the effect of the thylakoid membrane lipids monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), sulphoquinovosyldiacylglycerol (SQDG) and phosphatidylglycerol (PG) on the structure of two algal light-harvesting complexes (LHCs). In contrast to higher plants whose thylakoid membranes are characterized by an enrichment of the neutral galactolipids MGDG and DGDG, both the green alga Mantoniella squamata and the centric diatom Thalassiosira pseudonana contain membranes with a high content of the negatively charged lipids SQDG and PG. The algal thylakoids do not show the typical grana-stroma differentiation of higher plants but a regular arrangement. To analyze the effect of the membrane lipids, the fucoxanthin chlorophyll protein (FCP) complex of T. pseudonana and the LHC of M. squamata (MLHC) were prepared by successive cation precipitation using Triton X-100 as detergent. With this method, it is possible to isolate LHCs with a reduced amount of associated lipids in an aggregated state. The results from 77 K fluorescence and photon correlation spectroscopy show that neither the neutral galactolipids nor the negatively charged lipids are able to significantly alter the aggregation state of the FCP or the MLHC. This is in contrast to higher plants where SQDG and PG lead to a strong disaggregation of the LHCII whereas MGDG and DGDG induce the formation of large macroaggregates. The results indicate that LHCs which are integrated into thylakoid membranes with a high amount of negatively charged lipids and a regular arrangement are less sensitive to lipid-induced structural alterations than their counterparts in membranes enriched in neutral lipids with a grana-stroma differentiation. [ABSTRACT FROM AUTHOR]

The acclimation to high light, elevated temperature, and combination of both factors was evaluated in tomato ( Solanum lycopersicum cv. M82) by determination of photochemical activities of PSI and PSII and by analyzing 77 K fluorescence of isolated thylakoid membranes. Developed plants were exposed for six days to different combinations of temperature and light intensity followed by five days of a recovery period. Photochemical activities of both photosystems showed different sensitivity towards the heat treatment in dependence on light intensity. Elevated temperature exhibited more negative impact on PSII activity, while PSI was slightly stimulated. Analysis of 77 K fluorescence emission and excitation spectra showed alterations in the energy distribution between both photosystems indicating alterations in light-harvesting complexes. Light intensity affected the antenna complexes of both photosystems stronger than temperature. Our results demonstrated that simultaneous action of high-light intensity and high temperature promoted the acclimation of tomato plants regarding the activity of both photosystems in thylakoid membranes. [ABSTRACT FROM AUTHOR]

Plants are sessile organisms and need to acclimate to ever-changing light conditions in order to survive. These changes trigger a dynamic reorganization of the membrane protein complexes in the thylakoid membranes. Photosystem II (PSII) and its light harvesting system (LHCII) are the major target of this acclimation response, and accumulating evidences indicate that the amount and composition of PSII–LHCII supercomplexes in thylakoids are dynamically adjusted in response to changes in light intensity and quality. In this study, we characterized the PSII–LHCII supercomplexes in thylakoid membranes of pea plants in response to long-term acclimation to different light intensities. We provide evidence of a reorganization of the PSII–LHCII supercomplexes showing distinct changes in their antenna moiety. Mass spectrometry analysis revealed a specific reduction of Lhcb3, Lhcb6 and M -LHCII trimers bound to the PSII cores, while the Lhcb4.3 isoform increased in response to high light intensities. The modulation of Lhcb protein content correlates with the reduction of the functional PSII antenna size. These results suggest that the Lhcb3, Lhcb4.3 and Lhcb6 antenna subunits are major players in modulation of the PSII antenna size upon long-term acclimation to increased light levels. PsbS was not detected in the isolated PSII–LHCII supercomplexes at any light condition, despite an increased accumulation in thylakoids of high light acclimated plants, suggesting that PsbS is not a constitutive component of PSII–LHCII supercomplexes. [ABSTRACT FROM AUTHOR]

The group of homoiochlorophyllous resurrection plants evolved the unique capability to survive severe drought stress without dismantling the photosynthetic machinery. This implies that they developed efficient strategies to protect the leaves from reactive oxygen species ( ROS) generated by photosynthetic side reactions. These strategies, however, are poorly understood. Here, we performed a detailed study of the photosynthetic machinery in the homoiochlorophyllous resurrection plant Craterostigma pumilum during dehydration and upon recovery from desiccation. During dehydration and rehydration, C. pumilum deactivates and activates partial components of the photosynthetic machinery in a specific order, allowing for coordinated shutdown and subsequent reinstatement of photosynthesis. Early responses to dehydration are the closure of stomata and activation of electron transfer to oxygen accompanied by inactivation of the cytochrome b 6 f complex leading to attenuation of the photosynthetic linear electron flux ( LEF). The decline in LEF is paralleled by a gradual increase in cyclic electron transport to maintain ATP production. At low water contents, inactivation and supramolecular reorganization of photosystem II becomes apparent, accompanied by functional detachment of light-harvesting complexes and interrupted access to plastoquinone. This well-ordered sequence of alterations in the photosynthetic thylakoid membranes helps prepare the plant for the desiccated state and minimize ROS production. [ABSTRACT FROM AUTHOR]

Non-photochemical quenching ( NPQ) is a photoprotective mechanism in light-harvesting antennae. NPQ is triggered by chloroplast thylakoid lumen acidification and is accompanied by violaxanthin de-epoxidation to zeaxanthin, which further stimulates NPQ. In the present study, we show that violaxanthin can act in the opposite direction to zeaxanthin because an increase in the concentration of violaxanthin reduced NPQ in the light-harvesting antennae of Chromera velia. The correlation overlapped with a similar relationship between violaxanthin and NPQ as observed in isolated higher plant light-harvesting complex II. The data suggest that violaxanthin in C. velia can act as an inhibitor of NPQ, indicating that violaxanthin has to be removed from the vicinity of the protein to reach maximal NPQ. [ABSTRACT FROM AUTHOR]

Chloroplasts utilize photons from solar radiation to synthesize energy-rich molecules of ATPs and NADPHs, which are further used in active cellular processes. Multiprotein complexes (MPCs), including photosystems (PSII and PSI), and the cellular architecture responsible for generation of the proton motive force and the subsequent photophosphorylation, mediate the task of ATP and NADPH synthesis. Both photosystems and other multiprotein assemblies are embedded in thylakoid membranes. Advances in techniques used to study structural biology, biophysics, and comparative genomics and proteomics have enabled us to gain insights of structure, function, and localization of each individual component of the photosynthetic apparatus. An efficient coordination among MPCs is essential for normal functioning of photosynthesis, but there are various stressors that might directly or indirectly interact with photosynthetic components and processes. Cadmium is one of the toxic heavy metals that interact with photosynthetic components and damage photosystems and other MPCs in thylakoids. In plants, iron deficiency shows similar symptoms as those caused by Cd. Our article provides a general overview of chloroplast structure and a critical account of Cd-induced changes in photosystems and other MPCs in thylakoids, and suggests the possible mechanisms involved in mediating these changes. The connection between Cd-induced Fe deficiency and the elevated Cd toxicity under the Fe-deficient condition was also discussed. [ABSTRACT FROM AUTHOR]

Styrene-maleic acid copolymer was used to effect a non-detergent partial solubilization of thylakoids from spinach. A high density membrane fraction, which was not solubilized by the copolymer, was isolated and was highly enriched in the Photosystem (PS) I-light-harvesting chlorophyll (LHC) II supercomplex and depleted of PS II, the cytochrome b6/f complex, and ATP synthase. The LHC II associated with the supercomplex appeared to be energetically coupled to PS I based on 77 K fluorescence, P700 photooxidation, and PS I electron transport light saturation experiments. The chlorophyll (Chl) a/b ratio of the PS I-LHC II membranes was 3.2 ± 0.9, indicating that on average, three LHC II trimers may associate with each PS I. The implication of these findings within the context of higher plant PS I antenna organization is discussed. [ABSTRACT FROM AUTHOR]

Light-induced infrared difference spectroscopy (IR-DS) has been used, especially in the last decade, to investigate early photophysics, energy transfer and photoprotection mechanisms in isolated and membrane-bound light harvesting complexes (LHCs). The technique has the definite advantage to give information on how the pigments and the other constituents of the biological system (proteins, membranes, etc.) evolve during a given photoreaction. Different static and time-resolved approaches have been used. Compared to the application of IR-DS to photosynthetic Reaction Centers (RCs), however, IR-DS applied to LHCs is still in an almost pioneering age: very often sophisticated techniques (step-scan FTIR, ultrafast IR) or data analysis strategies (global analysis, target analysis, multivariate curve resolution) are needed. In addition, band assignment is usually more complicated than in RCs. The results obtained on the studied systems (chromatophores and RC-LHC supercomplexes from purple bacteria; Peridinin-Chlorophyll-a-Proteins from dinoflagellates; isolated LHCII from plants; thylakoids; Orange Carotenoid Protein from cyanobacteria) are summarized. A description of the different IR-DS techniques used is also provided, and the most stimulating perspectives are also described. Especially if used synergically with other biophysical techniques, light-induced IR-DS represents an important tool in the investigation of photophysical/photochemical reactions in LHCs and LHC-containing systems. [ABSTRACT FROM AUTHOR]

Leaves from three different Arabidopsis lines with different expression levels of PsbS protein showed different levels of non-photochemical quenching. The PsbS deficient plant npq4 showed remarkable reduction of electron transport rate, while the other two lines with a moderate amount (wild type) or an overexpression of PsbS ( L17 ) presented unchanged electron transport rates under the same range of high light intensities. Biochemical investigation revealed that the plant with the highest PsbS content ( L17 ) sustained the highest level of stable PSII–LHCII supercomplex structure, and displayed the smallest fluorescence quenching in the thylakoid membranes, the most efficient linear electron transport and the smallest cyclic electron transport. Based on these observations, it is proposed that the remodeling of PSII–LHCII supercomplexes affected by PsbS plays important roles in regulating the energy balance in thylakoid membrane and in ensuring the sophisticated coordination between energy excitation and dissipation. [ABSTRACT FROM AUTHOR]