Photosynthetic Apparatus Of Cyanobacteria Research Articles

Light wavelength as a contributory factor of environmental fitness in the cyanobacterial circadian clock.

The circadian clock is an essential system that drives the 24-hour expression rhythms for adaptation to day-night cycles. The molecular mechanism of the circadian clock has been extensively studied in cyanobacteria harboring the KaiC-based timing system. Nevertheless, our understanding of the physiological significance of the cyanobacterial circadian clock is still limited. In this study, we cultured wild-type Synechococcus elongatus PCC 7942 and circadian clock mutants in day-night cycles at different light qualities and found that the growth of the circadian clock mutants was specifically impaired during 12-hour blue light/12-hour dark cycles (BD) for the first time. The arrhythmic mutant kaiCAA was further analyzed by photosynthetic measurements. Compared with the wild type, the mutant exhibited decreases in the chlorophyll content, the ratio of photosystem I to II, net O2 evolution rate, efficiency of photosystem II photochemistry during BD cycles. These results indicate that the circadian clock is necessary for the growth and the maintenance of the optimum function of the photosynthetic apparatus in cyanobacteria under blue photoperiodic conditions.

Excited-State Dynamics of All the Mono-cis and the Major Di-cis Isomers of β-Apo-8'-Carotenal as Revealed by Femtosecond Time-Resolved Transient Absorption Spectroscopy.

Cis isomers of carotenoids play important roles in light harvesting and photoprotection in photosynthetic bacteria, such as the reaction center in purple bacteria and the photosynthetic apparatus in cyanobacteria. Carotenoids containing carbonyl groups are involved in efficient energy transfer to chlorophyll in light-harvesting complexes, and their intramolecular charge-transfer (ICT) excited states are known to be important for this process. Previous studies, using ultrafast laser spectroscopy, have focused on the central-cis isomer of carbonyl-containing carotenoids, revealing that the ICT excited state is stabilized in polar environments. However, the relationship between the cis isomer structure and the ICT excited state has remained unresolved. In this study, we performed steady-state absorption and femtosecond time-resolved absorption spectroscopy on nine geometric isomers (7-cis, 9-cis, 13-cis, 15-cis, 13'-cis, 9,13'-cis, 9,13-cis, 13,13'-cis, and all-trans) of β-apo-8'-carotenal, whose structures are well-defined, and discovered correlations between the decay rate constant of the S1 excited state and the S0-S1 energy gap, as well as between the position of the cis-bend and the degree of stabilization of the ICT excited state. Our results demonstrate that the ICT excited state is stabilized in polar environments in cis isomers of carbonyl-containing carotenoids and suggest that the position of the cis-bend plays an important role in the stabilization of the excited state.

Specific Features of Technogenic Pollutants Impact on Photosynthetic Activity of Unicellular Cyanobacteria

The effect of heavy-metal ions (Cd2+, Cr6+, and Cu2+) on cyanobacteria sp. Microcystis and sp. Synechocystis has been studied. Since heavy metals impact causes blocking of light-energy transmission channels and arrest of electron transport in photosynthetic apparatus of cyanobacteria, their influence can be detected within a few hours by fluorescence techniques. In this investigation three standard fluorescence techniques are used: fluorescence spectroscopy, pulsed amplitude modulation fluorimetry, and confocal microscopic spectroscopy. All three used methods showed a decrease in photosynthetic activity for cyanobacterial cells exposed to heavy metals. However, the mechanisms of action for different heavy metals are slightly different. In this paper, we analyze variations in spectroscopic and kinetic properties of emmitted fluorescence for cyanobacterial cultures and single living cells in order to identify a specific mechanism of heavy metals influence.

Boronic Acids of Pharmaceutical Importance Affect the Growth and Photosynthetic Apparatus of Cyanobacteria in a Dose-Dependent Manner.

The dynamic increase in the commercial application of antimicrobial derivatives of boronic acids, and potential impact of their presence in aquatic systems, supports the necessity to study the toxicity of these substances towards microorganisms of crucial meaning in the environment. One example of the mentioned derivatives is tavaborole (5-fluoro-substituted benzoxaborole), a pharmaceutical agent with antifungal activity. Cyanobacteria were used as model organisms, which are photoautotrophic prokaryotes, as representative aquatic bacteria and photoautotrophs associated with the plant kingdom. To the best of our knowledge, we investigated this issue for the first time. In order to recognize the under-stress response of those microorganisms, the concentration of photopigments—a key factor in the activity of photosynthetic apparatus—was measured spectrophotometrically. We found that the 3-piperazine bis(benzoxaborole) significantly suppressed the growth of halophilic and freshwater cyanobacteria, at a concentration 3.0 mM and 0.3 mM, respectively. Our results also showed that the tested substances at micromolar concentrations stimulated the growth of cyanobacteria, particularly in the freshwater strain Chroococcidiopsis thermalis. The tested substances acted with various strengths, depending on their structure and concentration; nevertheless, they had a greater influence on the synthesis of phycobiliproteins (e.g., lowered their concentration) than on the formation of chlorophyll and carotenoids.

Roles of CpcF and CpcG1 in Peroxiredoxin-Mediated Oxidative Stress Responses and Cellular Fitness in the Cyanobacterium Synechocystis sp. PCC 6803.

As a component of the photosynthetic apparatus in cyanobacteria, the phycobilisome (PBS) plays an important role in harvesting and transferring light energy to the core photosynthetic reaction centers. The size, composition (phycobiliprotein and chromophore), and assembly of PBSs can be dynamic to cope with tuning photosynthesis and associated cellular fitness in variable light environments. Here, we explore the role of PBS-related stress responses by analyzing deletion mutants of cpcF or cpcG1 genes in Synechocystis sp. PCC 6803. The cpcF gene encodes a lyase that links the phycocyanobilin (PCB) chromophore to the alpha subunit of phycocyanin (PC), a central phycobiliprotein (PBP) in PBSs. Deletion of cpcF (i.e., ΔcpcF strain) resulted in slow growth, reduced greening, elevated reactive oxygen species (ROS) levels, together with an elevated accumulation of a stress-related Peroxiredoxin protein (Sll1621). Additionally, ΔcpcF exhibited reduced sensitivity to a photosynthesis-related stress inducer, methyl viologen (MV), which disrupts electron transfer. The cpcG1 gene encodes a linker protein that serves to connect PC to the core PBP allophycocyanin. A deletion mutant of cpcG1 (i.e.,ΔcpcG1) exhibited delayed growth, a defect in pigmentation, reduced accumulation of ROS, and insensitivity to MV treatment. By comparison, ΔcpcF and ΔcpcG1 exhibited similarity in growth, pigmentation, and stress responses; yet, these strains showed distinct phenotypes for ROS accumulation, sensitivity to MV and Sll1621 accumulation. Our data emphasize an importance of the regulation of PBS structure in ROS-mediated stress responses that impact successful growth and development in cyanobacteria.

Direct single-molecule measurements of phycocyanobilin photophysics in monomeric C-phycocyanin

Phycobilisomes are highly organized pigment-protein antenna complexes found in the photosynthetic apparatus of cyanobacteria and rhodophyta that harvest solar energy and transport it to the reaction center. A detailed bottom-up model of pigment organization and energy transfer in phycobilisomes is essential to understanding photosynthesis in these organisms and informing rational design of artificial light-harvesting systems. In particular, heterogeneous photophysical behaviors of these proteins, which cannot be predicted de novo, may play an essential role in rapid light adaptation and photoprotection. Furthermore, the delicate architecture of these pigment-protein scaffolds sensitizes them to external perturbations, for example, surface attachment, which can be avoided by study in free solution or in vivo. Here, we present single-molecule characterization of C-phycocyanin (C-PC), a three-pigment biliprotein that self-assembles to form the midantenna rods of cyanobacterial phycobilisomes. Using the Anti-Brownian Electrokinetic (ABEL) trap to counteract Brownian motion of single particles in real time, we directly monitor the changing photophysical states of individual C-PC monomers from Spirulina platensis in free solution by simultaneous readout of their brightness, fluorescence anisotropy, fluorescence lifetime, and emission spectra. These include single-chromophore emission states for each of the three covalently bound phycocyanobilins, providing direct measurements of the spectra and photophysics of these chemically identical molecules in their native protein environment. We further show that a simple Förster resonant energy transfer (FRET) network model accurately predicts the observed photophysical states of C-PC and suggests highly variable quenching behavior of one of the chromophores, which should inform future studies of higher-order complexes.

Association of High Light-Inducible HliA/HliB Stress Proteins with Photosystem 1 Trimers and Monomers of the Cyanobacterium Synechocystis PCC 6803.

Hlip (high light-inducible proteins) are important for protection of the photosynthetic apparatus of cyanobacteria from light stress. However, the interaction of these proteins with chlorophyll-protein complexes of thylakoids remains unclear. The association of HliA/HliB stress proteins with photosystem 1 (PS1) complexes of the cyanobacterium Synechocystis PCC 6803 was studied to understand their function. Western blotting demonstrated that stress-induced HliA/HliB proteins are associated with PS1 trimers in wild-type cells grown under moderate light condition (40 µmol photons/m(2) per sec). The content of these proteins increased 1.7-fold after light stress (150 µmol photons/m(2) per sec) for 1 h. In the absence of PS1 trimers (ΔpsaL mutant), the HliA/HliB proteins are associated with PS1 monomers and the PS2 complex. HliA/HliB proteins are associated with PS1 monomers but not with PS1 trimers in Synechocystis PS2-deficient mutant grown at 5 µmol photons/m(2) per sec; the content of Hli proteins associated with PS1 monomers increased 1.2-fold after light stress. The HliA/HliB proteins were not detected in wild-type cells of cyanobacteria grown in glucose-supplemented medium at 5 µmol photons/m(2) per sec, but light stress induces the synthesis of stress proteins associated with PS1 trimers. Thus, for the first time, the association of HliA/HliB proteins not only with PS1 trimers, but also with PS1 monomers is shown, which suggests a universal role of these proteins in the protection of the photosynthetic apparatus from excess light.

Evolutionary origins, molecular cloning and expression of carotenoid hydroxylases in eukaryotic photosynthetic algae

BackgroundXanthophylls, oxygenated derivatives of carotenes, play critical roles in photosynthetic apparatus of cyanobacteria, algae, and higher plants. Although the xanthophylls biosynthetic pathway of algae is largely unknown, it is of particular interest because they have a very complicated evolutionary history. Carotenoid hydroxylase (CHY) is an important protein that plays essential roles in xanthophylls biosynthesis. With the availability of 18 sequenced algal genomes, we performed a comprehensive comparative analysis of chy genes and explored their distribution, structure, evolution, origins, and expression.ResultsOverall 60 putative chy genes were identified and classified into two major subfamilies (bch and cyp97) according to their domain structures. Genes in the bch subfamily were found in 10 green algae and 1 red alga, but absent in other algae. In the phylogenetic tree, bch genes of green algae and higher plants share a common ancestor and are of non-cyanobacterial origin, whereas that of red algae is of cyanobacteria. The homologs of cyp97a/c genes were widespread only in green algae, while cyp97b paralogs were seen in most of algae. Phylogenetic analysis on cyp97 genes supported the hypothesis that cyp97b is an ancient gene originated before the formation of extant algal groups. The cyp97a gene is more closely related to cyp97c in evolution than to cyp97b. The two cyp97 genes were isolated from the green alga Haematococcus pluvialis, and transcriptional expression profiles of chy genes were observed under high light stress of different wavelength.ConclusionsGreen algae received a β-xanthophylls biosynthetic pathway from host organisms. Although red algae inherited the pathway from cyanobacteria during primary endosymbiosis, it remains unclear in Chromalveolates. The α-xanthophylls biosynthetic pathway is a common feature in green algae and higher plants. The origination of cyp97a/c is most likely due to gene duplication before divergence of green algae and higher plants. Protein domain structures and expression analyses in green alga H. pluvialis indicate that various chy genes are in different manners response to light. The knowledge of evolution of chy genes in photosynthetic eukaryotes provided information of gene cloning and functional investigation of chy genes in algae in the future.

Protective dissipation of excess absorbed energy by photosynthetic apparatus of cyanobacteria: role of antenna terminal emitters

Two mechanisms of photoprotective dissipation of the excessively absorbed energy by photosynthetic apparatus of cyanobacteria are described that divert energy from reaction centers. Energy dissipation, monitored as nonphotochemical fluorescence quenching, occurs at different steps of energy transfer within the phycobilisomes or core antenna of photosystem I. Although these mechanisms differ significantly, in both cases, energy dissipates mainly from terminal emitters: allophycocyanin B or core membrane linker protein (L(CM)) in phycobilisomes, or the longest-wavelength chlorophylls in photosystem I antenna. It is supposed that carotenoid-induced energy dissipation in phycobilisomes is triggered by light-induced transformation of the nonquenched state of antenna into quenched state due to conformation changes caused by orange carotinoid-binding protein (OCP)-phycobilisome interaction. Fluorescence of the longest-wavelength chlorophylls of photosystem I antenna is strongly quenched by P700 cation radical or by P700 triplet state, dependent on redox state of the acceptor side cofactors of photosystem I.

Phycobiliprotein fluorescence of Nostoc punctiforme changes during the life cycle and chromatic adaptation: characterization by spectral confocal laser scanning microscopy and spectral unmixing

ABSTRACTMany cyanobacteria are highly adaptable to light quality, and many species undergo a complex life cycle. In this study we show that adaptive changes in the photosynthetic apparatus of cyanobacteria are not only caused by environmental, but also by developmental factors. Spectral confocal laser scanning microscopy (CLSM) was used to analyse in vivo the fluorescence spectra of the photosynthetic pigments chlorophyll a (Chl a), allophycocyanin (APC), phycocyanin (PC) and phycoerythrin (PE) of two Nostoc punctiforme strains. Changes in pigment fluorescence emission occurred in different developmental stages. Strain 1:1‐26 showed an emission maximum at 674 nm in motile hormogonia stages, whereas vegetative stages showed maxima at 658 and 575 nm. These changes were not caused by chromatic adaptation. In contrast, the second strain (1:1‐26lg) showed distinct fluorescence spectra, pigment localization and clear chromatic adaptation in red light. When these properties are known, both strains can be easily distinguished by the spectral CLSM method, which also allows the localization of the pigments within single cells. To calculate the contribution of individual phycobiliproteins to the observed changes, fluorescence spectra were analysed by spectral unmixing. This allowed the mathematical estimation of fluorescence shares for the individual phycobiliproteins in different developmental stages and both before and after chromatic adaptation. It is concluded that care should be taken when characterizing cyanobacteria by differences in pigment fluorescence, because these differences are influenced not only by chromatic adaptation, but also developmental stages. Spectral CLSM offers a powerful method to study the phycobiliprotein composition in vivo.

Biosynthesis of chromophores for phytochrome and related photoreceptors

A broad group of open tetrapyrroles derived from heme are collectively called bilins. Phycocyanobilin and phycoerythrobilin are utilized as accessory chromophore for light harvesting by phycobiliproteins that comprise the photosynthetic apparatus in cyanobacteria and algae. Phytochromes, known as a major photoreceptor in plants, contain a phytochromobilin chromophore as an essential prosthetic group for photo-sensing. The possible ancestral proteins of phytochromes, as found in cyanobacteria and eubacteria, possess phycocyanobilin and/or biliverdin as a prosthetic group. In this mini-review, the diverse functions of bilin chromophores and their biosynthesis in photosynthetic organisms are summarized and the potential applications to plant biotechnology will be discussed.

Spectroscopic investigation on the energy transfer process in photosynthetic apparatus of cyanobacteria

Spectroscopic investigation on the energy transfer process in photosynthetic apparatus of cyanobacteria

Spectroscopic investigation on the energy transfer process in photosynthetic apparatus of cyanobacteria

In this work, we employ cyanobacteria, Spirulina platensis, and separate their photosynthetic apparatus, phycobilisome (PBS), thylakoid membrane and phycobilisome–thylakoid membrane complex. The steady state absorption spectra, fluorescence spectra and corresponding deconvoluted spectra and picosecond time-resolved spectra are used to investigate the energy transfer process in phycobilisome–thylakoid membrane complex. The results on steady state spectra show chlorophylls of the photosystem II are able to transfer excitation energy to phycobilisome with Chl a molecules selectively excited. The decomposition of the steady state spectra further suggest the uphill energy transfer originate from chlorophylls of photosystem II to cores of phycobilisome, while rods and cores of phycobilisome cannot receive energy from the chlorophylls of photosystem I. The time constant for the back energy transfer process is 18 ps.

Separation and quantitation of phycobiliproteins using phytic acid in capillary electrophoresis with laser-induced fluorescence detection

The similar electrophoretic mobilities and sizes of several of the phycobiliproteins, which are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae, render their separation and quantitation a challenging problem. However, we have developed a suitable capillary electrophoresis (CE) method that employs a phytic acid–boric acid buffer and laser-induced fluorescence (LIF) detection with a single 594 nm He–Ne laser. This method takes advantage of the remarkably high quantum yields of these naturally fluorescent proteins, which can be attributed to their linear tetrapyrrole chromophores covalently bound to cysteinyl residues. As such, limits of detection of 1.18·10 −14, 5.26·10 −15, and 2.38·10 −15 mol/l were obtained for R-phycoerythrin, C-phycocyanin, and allophycocyanin proteins, respectively, with a linear dynamic range of eight orders of magnitude in each case. Unlike previously published CE–LIF methods, this work describes the separation of all three major classes of phycobiliproteins in under 5 min. Very good recoveries, ranging from 93.2 to 105.5%, were obtained for a standard mixture of the phycobiliproteins, based on seven-point calibration curves for both peak height and peak area. It is believed that this development will prove useful for the determination of phycobiliprotein content in naturally occurring cyanobacteria populations, thus providing a useful tool for understanding biological and chemical oceanographic processes.

Determination of phycobiliproteins by capillary electrophoresis with laser-induced fluorescence detection.

Phycobiliproteins are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae. They are composed of a protein backbone to which linear tetrapyrrole chromophores are covalently bound. Furthermore, they are water-soluble highly fluorescent, and relatively stable at room temperature and neutral pH. For this reason, capillary electrophoresis-laser induced fluorescence (CE-LIF) seems the idea method for determination of these important proteins. The effects of buffer additives such as sodium dodecyl sulfate (SDS)and putrescine on the separation of the three major phycobiliprotein types, namely allophycocyanin, phycocyanin, and phycoerythrin, with excitation and emission maxima at 652/660, 615/647, and 565(494)/575 nm, respectively, are considered. Detection limits for these proteins by CE-LIF are some 60-500 times better than by absorbance detection. The development of a fast and sensitive CE-LIF assay such as this is of potential significance to our understand ing of chemical and biological oceanographic processes.

Constitutive expression of a small heat-shock protein confers cellular thermotolerance and thermal protection to the photosynthetic apparatus in cyanobacteria

The role of a small heat-shock protein (Hsp) in the acquisition of thermotolerance in cyanobacteria was investigated. Synechococcus sp. PCC 7942 was transformed with an expression vector carrying the coding sequence of the hspA gene encoding a small heat-shock protein from Synechococcus vulcanus under the control of the tac promoter. The transformant which was shown to constitutively express HspA displayed improved viability compared with the reference strain upon transfer from 30 to 50°C in the light. When the heat shock was given in darkness, the survival rate in the reference strain increased greatly, approaching a level similar to that for the HspA expressing strain after heat shock in the light. Expression of HspA increased thermal resistance of photosystem II (PS II) and protected phycocyanin from heat-induced photobleaching. Our results are indicative of a central role for HspA in amelioration of the harmful effect of light during heat stress and identified the possible sites of action of the small Hsp in vivo to be the PS II complex and the light-harvesting phycobilisomes.

Ultraviolet-B Effects onSpirulina platensisCells: Modification of Chromophore–Protein Interaction and Energy Transfer Characteristics of Phycobilisomes

Exposure of ultraviolet-B (280–320 nm, 1.9 mW m−2s−1) radiation of intactSpirulina platensisfor 9 h caused specific loss of the 85.5 KDa anchor protein of phycobilisomes, the major light-harvesting antenna complex of photosystem II. Associated with the loss of 85.5 KDa protein, the UV-B irradiation also caused photobleaching of phycobilins and alteration in the chromophore protein interactions, as evidenced from the visible circular dichroic measurements, and it also affected the energy transfer process within the phycobilisomes, as inferred from the low-temperature, 77 K, fluorescence spectral analysis. Our results, thus, clearly demonstrate for the first time that the phycobilisomes effectively act as targets for UV-B induced damage of photosynthetic apparatus in cyanobacteria.

Electrophoretic applications of phycobiliproteins.

Phycobiliproteins are homologous chromoproteins which constitute the phycobilisomes, the light harvesting complexes of the photosynthetic apparatus in cyanobacteria, rhodophyta and cryptophyta. In the present work, phycocyanin (PC) and phycoerythrin (PE) from a Nostoc species are proposed as protein markers for electrophoretic techniques. Phycocyanin is a blue-colored phycobiliprotein; it carries phycocyanobilin as chromophoric group and is composed of two subunits, alpha and beta, with Mr of 14000 and 17000, respectively. In contrast, the PE subunits, having a similar Mr of 21000, are deep rose chromoproteins and carry phycoerythrobilin residues. Both low molecular weight phycobiliproteins are also suitable for monitoring protein blotting and the focusing time of protein samples during isoelectric focusing as internal markers. The PE subunits which form a single broad band after sodium dodecyl sulfate-polyacrylamide gel electrophoresis have different isoelectric points, and they form two visible bands when they reach their isoelectric point. The phycobilisomes constitute up to 50% of the total protein in cyanobacteria and their content in PC or PE can be up- or down-regulated by using different light conditions (chromatic adaptation).

Effect of light intensity on the ultrastructure of the filamentous cyanobacterium, Anabaena variabilis

Cyanobacteria are Gram negative prokaryotes that are capable of oxygenic photosynthesis. Although there are many similarities between eukaryotes and cyanobacteria in electron transfer and phosphorylation during photosynthesis, there are two features of the photosynthetic apparatus in cyanobacteria which distinguishes them from plants. Cyanobacteria contain phycobiliproteins organized in phycobilisomes on the surface of photosynthetic membrane. Another difference is in the organization of the photosynthetic membranes. Instead of stacked thylakolds within a chloroplast envelope membrane, as seen In eukaryotes, IntracytopIasmlc membranes generally are arranged in three to six concentric layers. Environmental factors such as temperature, nutrition and light fluency can significantly affect the physiology and morphology of cells. The effect of light Intensity shifts on the ultrastructure of Internal membrane in Anabaena variabilis grown under controlled environmental conditions was examined. Since a major constituent of cyanobacterial thylakolds are lipids, the fatty acid content also was measured and correlated with uItrastructural changes. The regulation of fatty acid synthesis in cyanobacteria ultimately can be studied if the fatty acid content can be manipulated.

Adaptation to salt of the photosynthetic apparatus in cyanobacteria

Cyanobacteria grow under a wide range of environmental conditions. Their distribution in environments of highly diversified osmotica is well documented [ 11. Hence, the photosynthetic apparatus must adapt to the different degrees of salinity in the habitat. This is achieved by means of either osmoregulation or on the level of the organism [2] or modification of the enzyme structure [3]. Here we correlated the salt tolerance of the whole organism with the salt tolerance of activities of the photosynthetic apparatus and NADPH-cytochrome c reductase in 4 cyanobacteria characterized by different degrees of salt tolerance: marine Spirulina tolerates up to 3-times sea water [4] ; Gzlothrix scopulorum, sea water [S]; Spirulina platensis, brackish water [6]; Nostoc muscorum 7119, fresh water. We show that the soluble components of the final steps of photosynthetic electron transport constitute a salt-sensitive site and that salt tolerance may be acquired during growth in saline water.