Trimeric organization of photosystem I is required to maintain the balanced photosynthetic electron flow in cyanobacterium Synechocystis sp. PCC 6803.

In Synechocystis sp. PCC 6803 and some other cyanobacteria photosystem I reaction centres exist predominantly as trimers, with minor contribution of monomeric form, when cultivated at standard optimized conditions. In contrast, in plant chloroplasts photosystem I complex is exclusively monomeric. The functional significance of trimeric organization of cyanobacterial photosystem I remains not fully understood. In this study, we compared the photosynthetic characteristics of PSI in wild type and psaL knockout mutant. The results show that relative to photosystem I trimer in wild-type cells, photosystem I monomer in psaL- mutant has a smaller P700+ pool size under low and moderate light, slower P700 oxidation upon dark-to-light transition, and slower P700+ reduction upon light-to-dark transition. The mutant also shows strongly diminished photosystem I donor side limitations [quantum yield Y(ND)] at low, moderate and high light, but enhanced photosystem I acceptor side limitations [quantum yield Y(NA)], especially at low light (22 µmol photons m-2 s-1). In line with these functional characteristics are the determined differences in the relative expression genes encoding of selected electron transporters. The psaL- mutant showed significant (ca fivefold) upregulation of the photosystem I donor cytochrome c6, and downregulation of photosystem I acceptors (ferredoxin, flavodoxin) and proteins of alternative electron flows originating in photosystem I acceptor side. Taken together, our results suggest that photosystem I trimerization in wild-type Synechocystis cells plays a role in the protection of photosystem I from photoinhibition via maintaining enhanced donor side electron transport limitations and minimal acceptor side electron transport limitations at various light intensities.

Opposite domination of cyclic and pseudocyclic electron flows in short-illuminated dark-adapted leaves of angiosperms and gymnosperms.

The present work was aimed to explain the recently reported higher O2-dependent electron flow capacity in gymnosperms than in angiosperms and to search for other differences in the electron transport processes by simultaneous characterization of the relative capacities of pseudocyclic (direct or Flavodiiron proteins (Flv)-mediated O2-reduction, Mehler(-like) reactions) and cyclic electron flows around photosystem I (CEF-PSI). To this end, a comparative multicomponent analysis was performed on the fluorescence decay curves of dark-adapted leaves after illumination with a 1-s saturating light pulse. In both gymnosperms and angiosperms, two or three exponential decay components were resolved: fast (t 1/21 ~ 170-260 ms), middle (~1.0-2.3 s), and slow (>4.2 s). The sensitivity of the decay parameters (amplitudes A1-3, halftimes t 1/2 1-3) to the alternative electron flows was assessed using Arabidopsis pgr5 and ndhM mutants, defective in CEF-PSI, Synechocystis sp. PCC 6803 Δflv1 mutant, defective in Flv-mediated O2-photoreduction, different O2 concentrations, and methyl viologen treatment. A1 reflected the part of electrons involved in linear and O2-photoreduction pathways after PSI. The middle component appeared in pgr5 (but not in ndhM), in gymnosperms under low O2, and in Δflv1, and reflected limitations at the PSI acceptor side. The slow component was sensitive to CEF-PSI. The comparison of decay parameters provided evidence that Flv mediate O2-photoreduction in gymnosperms, which explains their higher O2-dependent electron flow capacity. The concomitant quantification of relative electrons branching in O2-photoreduction and CEF-PSI pathways under the applied non-steady-state photosynthetic conditions reveals that CEF-PSI capacity significantly exceeds that of O2-photoreduction in angiosperms while the opposite occurs in gymnosperms.

Action and target sites of nitric oxide in chloroplasts.

Nitric oxide (NO) is an important signalling molecule in plants under physiological and stress conditions. Here we review the influence of NO on chloroplasts which can be directly induced by interaction with the photosynthetic apparatus by influencing photophosphorylation, electron transport activity and oxido-reduction state of the Mn clusters of the oxygen-evolving complex or by changes in gene expression. The influence of NO-induced changes in the photosynthetic apparatus on its functions and sensitivity to stress factors are discussed.

Bilayer structural destabilization by low amounts of chlorophyll a.

The present study shows that small admixtures of one chlorophyll a (Chla) molecule per several hundred lipid molecules have strong destabilizing effect on lipid bilayers. This effect is clearly displayed in the properties of the L(alpha)-H(II) transformations and results from a Chla preference for the H(II) relative to the L(alpha) phase. Chla disfavors the lamellar liquid crystalline phase L(alpha) and induces its replacement with inverted hexagonal phase H(II), as is consistently demonstrated by DSC and X-ray diffraction measurements on phosphatidylethanolamine (PE) dispersions. Chla lowers the L(alpha)-H(II) transition temperature (42 degrees C) of the fully hydrated dipalmitoleoyl PE (DPoPE) by approximately 8 degrees C and approximately 17 degrees C at Chla/DPoPE molar ratios of 1:500 and 1:100, respectively. Similar Chla effect was recorded also for dielaidoyl PE dispersions. The lowering of the transition temperature and the accompanying significant loss of transition cooperativity reflect the Chla repartitioning and preference for the H(II) phase. The reduction of the H(II) phase lattice constant in the presence of Chla is an indication that Chla favors H(II) phase formation by decreasing the radius of spontaneous monolayer curvature, and not by filling up the interstitial spaces between the H(II) phase cylinders. The observed Chla preference for H(II) phase and the substantial bilayer destabilization in the vicinity of a bilayer-to-nonbilayer phase transformation caused by low Chla concentrations can be of interest as a potential regulatory or membrane-damaging factor.

Photoelectron transport ability of chloroplast thylakoid membranes treated with NO donor SNP: changes in flash oxygen evolution and chlorophyll fluorescence.

The nitric oxide (NO) donor sodium nitroprusside (SNP) is frequently used in plant science in vivo. The present in vitro study reveals its effects on the photosynthetic oxygen evolution and the chlorophyll fluorescence directly on isolated pea thylakoid membranes. It was found that even at very low amounts of SNP (chlorophyll/SNP molar ratio∼67:1), the SNP-donated NO stimulates with more than 50% the overall photosystem II electron transport rate and diminishes the evolution of molecular oxygen. It was also found that the target site for SNP-donated NO is the donor side of photosystem II. Compared with other NO-donors used in plant science, SNP seems to be the only one exhibiting stimulation of electron transport through photosystem II.

Photosystem I oligomerization affects lipid composition in Synechocystis sp. PCC 6803.

In cyanobacteria, increasing growth temperature decreases lipid unsaturation and the ratio of monomer/trimer photosystem I (PSI) complexes. In the present study we applied Fourier-transform infrared (FTIR) spectroscopy and lipidomic analysis to study the effects of PSI monomer/oligomer ratio on the physical properties and lipid composition of thylakoids. To enhance the presence of monomeric PSI, a Synechocystis sp. PCC6803/ΔpsaL mutant strain (PsaL) was used which, unlike both trimeric and monomeric PSI-containing wild type (WT) cells, contain only the monomeric form. The protein-to-lipid ratio remained unchanged in the mutant but, due to an increase in the lipid disorder in its thylakoids, the gel to liquid-crystalline phase transition temperature (Tm) is lower than in the WT. In thylakoid membranes of the mutant, digalactosyldiacylglycerol (DGDG), the most abundant bilayer-forming lipid is accumulated, whereas those in the WT contain more monogalactosyldiacylglycerol (MGDG), the only non-bilayer-forming lipid in cyanobacteria. In PsaL cells, the unsaturation level of sulphoquinovosyldiacylglycerol (SQDG), a regulatory anionic lipid, has increased. It seems that merely a change in the oligomerization level of a membrane protein complex (PSI), and thus the altered protein-lipid interface, can affect the lipid composition and, in addition, the whole dynamics of the membrane. Singular value decomposition (SVD) analysis has shown that in PsaL thylakoidal protein-lipid interactions are less stable than in the WT, and proteins start losing their native secondary structure at much milder lipid packing perturbations. Conclusions drawn from this system should be generally applicable for protein-lipid interactions in biological membranes.

Chlorophyll a is the crucial redox sensor and transmembrane signal transmitter in the cytochrome b6f complex. Components and mechanisms of state transitions from the hydrophobic mismatch viewpoint.

The cytochrome (cyt) b6f complex is involved in the transmembrane redox signaling that triggers state transitions in cyanobacteria and chloroplasts. However, the components and molecular mechanisms are still unclear. In an attempt to solve this long-standing problem, we first focused on the unknown role of a single chlorophyll a (Chla) in cyt b6f with a new approach based on Chla structural properties. Various b6f X-ray crystal structures were analyzed to identify their differences, which correlate with differences in Chla molecular volume. We found that the distance of the Rieske [2Fe-2S] cluster to Chla correlates with the distance between a pair of residues at the Qo-site and the distance between a pair of residues at the opposite membrane side. These correlations were accompanied by the rotation of a key peripheral residue and by changes in the hydrophobic thickness of cyt b6f. Parallel analysis of cyt bc1 crystal structures allowed us to conclude that Chla acts as the crucial redox sensor and transmembrane signal transmitter in b6f for changes in the plastoquinone pool redox state. The hydrophobic mismatch induced by the changed hydrophobic thickness of cyt b6f is the driving force for the structural reorganizations of the photosynthetic apparatus during induction and the progression of state transitions in cyanobacteria and chloroplasts. A mechanism for LHCII kinase activation in chloroplasts is also proposed. Our understanding of the dynamic structural changes in bc-complexes during turnover at the Qo-site and state transitions is augmented by the time-sequence ordering of 56 bc crystal structures.

Effects of exogenous 24-epibrassinolide on the photosynthetic membranes under non-stress conditions.

In the present work the effects of exogenous 24-epibrassinolide (EBR) on functional and structural characteristics of the thylakoid membranes under non-stress conditions were evaluated 48 h after spraying of pea plants with different concentrations of EBR (0.01, 0.1 and 1.0 mg.L(-1)). The results show that the application of 0.1 mg.L(-1) EBR has the most pronounced effect on the studied characteristics of the photosynthetic membranes. The observed changes in 540 nm light scattering and in the calorimetric transitions suggest alterations in the structural organization of the thylakoid membranes after EBR treatment, which in turn influence the kinetics of oxygen evolution, accelerate the electron transport rate, increase the effective quantum yield of photosystem II and the photochemical quenching. The EBR-induced changes in the photosynthetic membranes are most probably involved in the stress tolerance of plants.