The role of nitric oxide (NO) in photosynthesis is poorly understood as indicated by a number of studies in this field with often conflicting results. As various NO donors may be the primary source of discrepancies, the aim of this study was to apply a set of NO donors and its scavengers, and examine the effect of exogenous NO on photosynthetic electron transport in vivo as determined by chlorophyll fluorescence of pea (Pisum sativum) leaves. Sodium nitroprusside-induced changes were shown to be mediated partly by cyanide, and S-nitroso-N-acetylpenicillinamine provided low yields of NO. However, the effects of S-nitrosoglutathione are inferred exclusively by NO, which made it an ideal choice for this study. Q A 2 reoxidation kinetics show that NO slows down electron transfer between Q A and Q B , and inhibits charge recombination reactions of Q A 2 with the S 2 state of the water-oxidizing complex in photosystem II. Consistent with these results, chlorophyll fluorescence induction suggests that NO also inhibits steady-state photochemical and nonphotochemical quenching processes. NO also appears to modulate reaction-center-associated nonphotochemical quenching.Plants, as well as animals, respond to ambient levels of nitric oxide (NO), and also generate NO themselves via various enzymatic and nonenzymatic pathways (Yamasaki, 2000;Neill et al., 2003;Río et al., 2004). Indeed, in the past years, a growing amount of research has provided evidence for the multiple physiological roles of this gaseous free radical in plants (for review, see Wendehenne et al., 2004;Delledonne, 2005). The turnover of NO depends on its concentration, the ambient redox status, and the concentration of target molecules. In biological systems, NO is capable of targeting thiol-and metal-containing proteins (Lamattina et al., 2003). Photosynthetic and mitochondrial electron transport chains are abundant in transition metalcontaining complexes, and NO and its derivative peroxynitrite are known to inhibit the mitochondrial electron transport chain (Millar and Day, 1996;Yamasaki et al., 2001). Yet, the effect of exogenous NO on photosynthetic activity in intact leaves has so far been poorly addressed, with often conflicting results.Previous research suggests that NO gas decreases net photosynthesis rates in oat (Avena sativa) and alfalfa (Medicago sativa) leaves (Hill and Bennett, 1970). Lum et al. (2005) have identified a number of intracellular targets of NO signalization including mitochondria, peroxisomes, and chloroplasts. They found that the NO donor sodium nitroprusside (SNP) decreases the amount of Rubisco activase and the b-subunit of the Rubisco subunit-binding protein in mung bean (Phaseolus aureus).NO is also able to influence the photosynthetic electron transport chain directly. An important action site of NO is PSII. Electron paramagnetic resonance and chlorophyll fluorescence measurements using NO gas treatment of isolated thylakoid membrane complexes have clearly demonstrated that NO can reversibly bind to several sites in PSII and...

We have studied the pH effect on the S(0) and S(2) multiline electron paramagnetic resonance (EPR) signals from the water-oxidizing complex of photosystem II. Around pH 6, the maximum signal intensities were detected. On both the acidic and alkaline sides of pH 6, the intensities of the EPR signals decreased. Two pKs were determined for the S(0) multiline signal; pK(1) = 4.2 +/- 0.2 and pK(2) = 8.0 +/- 0.1, and for the S(2) multiline signal the pKs were pK(1) = 4.5 +/- 0.1 and pK(2) = 7.6 +/- 0.1. The intensity of the S(0)-state EPR signal was partly restored when the pH was changed from acidic or alkaline pH back to pH approximately 6. In the S(2) state we observed partial recovery of the multiline signal when going from alkaline pH back to pH approximately 6, whereas no significant recovery of the S(2) multiline signal was observed when the pH was changed from acidic pH back to pH approximately 6. Several possible explanations for the intensity changes as a function of pH are discussed. Some are ruled out, such as disintegration of the Mn cluster or decay of the S states and formal Cl(-) and Ca(2+) depletion. The altered EPR signal intensities probably reflect the protonation/deprotonation of ligands to the Mn cluster or the oxo bridges between the Mn ions. Also, the possibility of decreased multiline signal intensities at alkaline pH as an effect of changed redox potential of Y(Z) is put forward.

Fluorescence yield relaxation following a light pulse was studied in various cyanobacteria under aerobic and microaerobic conditions. In Synechocystis PCC 6803 fluorescence yield decays in a monotonous fashion under aerobic conditions. However, under microaerobic conditions the decay exhibits a wave feature showing a dip at 30-50 ms after the flash followed by a transient rise, reaching maximum at ~1s, before decaying back to the initial level. The wave phenomenon can also be observed under aerobic conditions in cells preilluminated with continuous light. Illumination preconditions cells for the wave phenomenon transiently: for few seconds in Synechocystis PCC 6803, but up to one hour in Thermosynechocystis elongatus BP-1. The wave is eliminated by inhibition of plastoquinone binding either to the QB site of Photosystem-II or the Qo site of cytochrome b6f complex by 3-(3',4'-dichlorophenyl)-1,1-dimethylurea or 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, respectively. The wave is also absent in mutants, which lack either Photosystem-I or the NAD(P)H-quinone oxidoreductase (NDH-1) complex. Monitoring the redox state of the plastoquinone pool revealed that the dip of the fluorescence wave corresponds to transient oxidation, whereas the following rise to re-reduction of the plastoquinone pool. It is concluded that the unusual wave feature of fluorescence yield relaxation reflects transient oxidation of highly reduced plastoquinone pool by Photosystem-I followed by its re-reduction from stromal components via the NDH-1 complex, which is transmitted back to the fluorescence yield modulator primary quinone electron acceptor via charge equilibria. Potential applications of the wave phenomenon in studying photosynthetic and respiratory electron transport are discussed. This article is part of a special issue entitled: photosynthesis research for sustainability: keys to produce clean energy.

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SummaryCoral bleaching is an important environmental phenomenon, whose mechanism has not yet been clarified. The involvement of reactive oxygen species (ROS) has been implicated, but direct evidence of what species are involved, their location and their mechanisms of production remains unknown.Histidine-mediated chemical trapping and singlet oxygen sensor green (SOSG) were used to detect intra-and extracellular singlet oxygen (

The rapid turn-over of the D1 polypeptide of the photosystem two complex has been suggested to be due to the presence of a "PEST"-like sequence located between putative transmembrane helices IV and V of D1 (Greenberg, B. M., Gaba, V., Mattoo, A. K. and Edelman, M. (1987) EMBO J. 6, 2865-2869). We have tested this hypothesis by constructing a deletion mutant (delta 226-233) of the cyanobacterium Synechocystis sp. PCC 6803 in which residues 226-233 of the D1 polypeptide, containing the PEST-like sequence, have been removed. The resulting mutant, delta PEST, is able to grow photoautotrophically and give light-saturated rates of oxygen at wild type levels. However electron transfer on the acceptor side of the complex is perturbed. Analysis of cells by thermoluminescence and by monitoring the decay in quantum yield of variable fluorescence following saturating flash excitation indicates that Q-B, but not Q-A, is destabilized in this mutant. Electron transfer on the donor side of photosystem two remains largely unchanged in the mutant. Turnover of the D1 polypeptide as examined by pulse-chase experiments using [35S]methionine was enhanced in the delta PEST mutant compared to strain TC31 which is the wild type control. We conclude that the PEST sequence is not absolutely required for turnover of the D1 polypeptide in vivo although deletion of residues 226-233 does have an effect on the redox equilibrium between QA and QB.

The green algae Chlamydomonas sp. MACC-549 and Chlamydomonas reinhardtii cc124 were investigated for their hydrogen-evolution capability in mixed algal-bacterial cultures. Stable bacterial contaminations were identified during the cultivation of Chlamydomonas sp. 549. The bacterial symbionts belonged to various genera, mostly Brevundimonas, Rhodococcus, and Leifsonia, each of which enhanced the algal hydrogen production. This phenomenon was not limited to natural associations. Increased algal hydrogen evolution was achieved by simple artificial algal-bacterial communities as well.Algal-bacterial cocultures were designed and tested in hydrogen evolution experiments. The highest hydrogen yields were obtained when hydrogenase-deficient Escherichia coli was used as a symbiotic bacterium (Chlamydomonas sp. 549 generated 1196.06 ± 4.42 μL H 2 L −1 , while C. reinhardtii cc124 produced 5800.54 ± 65.73 μL H 2 L −1 ). The results showed that oxygen elimination is the most crucial factor for algal hydrogen production and that efficient bacterial respiration is essential for the activation of algal Fe-hydrogenase. The algae-based hydrogen evolution method described represents a novel combination of fermentative and photolytic hydrogen generation processes. Active photosynthesis was maintained during the entire hydrogen evolution process, which contributes to the sustainability of hydrogen production.

In Thermosynechococcus elongatus BP-1, which is the preferred organism in recent structural studies of PSII, three psbA and two psbD genes code for three D1 and one D2 protein isoforms, respectively. The regulation and function of these genes and protein products is largely unknown. Therefore, we used quantitative RT-PCR to follow changes in the mRNA level of the respective genes, in combination with biophysical measurements to detect changes in the electron transport activity of Photosystem II under exposure to different visible and UV light, and temperature conditions. In cells which are acclimated to 40 micromol m(-2)s(-1) growth light conditions at 40 degrees C the main populations of the psbA and psbD transcripts arise from the psbA1 and psbD1 genes, respectively. When the temperature is raised to 60 degrees C psbA1 becomes the single dominating psbA mRNA species. Upon exposure of the cells to 500 micromol m(-2)s(-1) intensity visible light psbA3 replaces psbA1 as the dominating psbA mRNA species, and psbD2 increases at the expense of psbD1. UV-B radiation also increases the abundance of psbA3, and psbD2 at the expense of psbA1 and psbD1, respectively. From the different extent of total D1 protein loss in the absence and presence of lincomycin it was estimated that the PsbA3 protein isoform replaces PsbA1 in about 65% of PSII centers after 2 h of high light acclimation. Under the conditions of different psbA transcript distributions chlorophyll fluorescence and thermoluminescence measurements were applied to monitor charge recombination characteristics of the S2Q(A)(-) and S2Q(B)(-) states. We obtained faster decay of flash-induced chlorophyll fluorescence in the presence of DCMU, as well as lower peak temperature of the Q and B thermoluminescence bands when PsbA3 replaced PsbA1 as the main D1 protein isoform. The relevance of dynamic changes in the abundance of psbA and psbD transcript levels, as well as D1 protein isoforms in the acclimation of T. elongatus to changing environmental conditions is discussed.