Makio Yokono

Identification of the special pair of photosystem II in a chlorophyll d-dominated cyanobacterium

The composition of photosystem II (PSII) in the chlorophyll (Chl) d-dominated cyanobacterium Acaryochloris marina MBIC 11017 was investigated to enhance the general understanding of the energetics of the PSII reaction center. We first purified photochemically active complexes consisting of a 47-kDa Chl protein (CP47), CP43′ (PcbC), D1, D2, cytochrome b559, PsbI, and a small polypeptide. The pigment composition per two pheophytin (Phe) a molecules was 55 ± 7 Chl d, 3.0 ± 0.4 Chl a, 17 ± 3 α-carotene, and 1.4 ± 0.2 plastoquinone-9. The special pair was detected by a reversible absorption change at 713 nm (P713) together with a cation radical band at 842 nm. FTIR difference spectra of the specific bands of a 3-formyl group allowed assignment of the special pair. The combined results indicate that the special pair comprises a Chl d homodimer. The primary electron acceptor was shown by photoaccumulation to be Phe a, and its potential was shifted to a higher value than that in the Chl a/Phe a system. The overall energetics of PSII in the Chl d system are adjusted to changes in the redox potentials, with P713 as the special pair using a lower light energy at 713 nm. Taking into account the reported downward shift in the potential of the special pair of photosystem I (P740) in A. marina, our findings lend support to the idea that changes in photosynthetic pigments combine with a modification of the redox potentials of electron transfer components to give rise to an energetic adjustment of the total reaction system.

Identification of a Novel Vinyl Reductase Gene Essential for the Biosynthesis of Monovinyl Chlorophyll in Synechocystis sp. PCC6803

The vast majority of oxygenic photosynthetic organisms use monovinyl chlorophyll for their photosynthetic reactions. For the biosynthesis of this type of chlorophyll, the reduction of the 8-vinyl group that is located on the B-ring of the macrocycle is essential. Previously, we identified the gene encoding 8-vinyl reductase responsible for this reaction in higher plants and termed it DVR. Among the sequenced genomes of cyanobacteria, only several Synechococcus species contain DVR homologues. Therefore, it has been hypothesized that many other cyanobacteria producing monovinyl chlorophyll should contain a vinyl reductase that is unrelated to the higher plant DVR. To identify the cyanobacterial gene that is responsible for monovinyl chlorophyll synthesis, we developed a bioinformatics tool, correlation coefficient calculation tool, which calculates the correlation coefficient between the distributions of a certain phenotype and genes among a group of organisms. The program indicated that the distribution of a gene encoding a putative dehydrogenase protein is best correlated with the distribution of the DVR-less cyanobacteria. We subsequently knocked out the corresponding gene (Slr1923) in Synechocystis sp. PCC6803 and characterized the mutant. The knock-out mutant lost its ability to synthesize monovinyl chlorophyll and accumulated 3,8-divinyl chlorophyll instead. We concluded that Slr1923 encodes the vinyl reductase or a subunit essential for monovinyl chlorophyll synthesis. The function and evolution of 8-vinyl reductase genes are discussed.

Identification of the primary electron donor in PS II of the Chl d-dominated cyanobacterium Acaryochloris marina

The primary electron donor of photosystem (PS) II in the chlorophyll (Chl) d‐dominated cyanobacterium Acaryochloris marina was confirmed by delayed fluorescence (DF) and further proved by pigment contents of cells grown under several light intensities. The DF was found only in the Chl a region, identical to Synechocystis sp. PCC 6803, and disappeared following heat treatment. Pigment analyses indicated that at least two Chl a molecules were present per each two pheophytin a molecules, and these Chl a molecules are assigned to PD1 and PD2. These findings clearly indicate that Chl a is required for water oxidation in PS II.

A megacomplex composed of both photosystem reaction centres in higher plants

Throughout the history of oxygen evolution, two types of photosystem reaction centres (PSI and PSII) have worked in a coordinated manner. The oxygen evolving centre is an integral part of PSII, and extracts an electron from water. PSI accepts the electron, and accumulates reducing power. Traditionally, PSI and PSII are thought to be spatially dispersed. Here, we show that about half of PSIIs are physically connected to PSIs in Arabidopsis thaliana. In the PSI-PSII complex, excitation energy is transferred efficiently between the two closely interacting reaction centres. PSII diverts excitation energy to PSI when PSII becomes closed-state in the PSI-PSII complex. The formation of PSI-PSII complexes is regulated by light conditions. Quenching of excess energy by PSI might be one of the physiological functions of PSI-PSII complexes.

Excitation energy transfer between photosystem II and photosystem I in red algae: Larger amounts of phycobilisome enhance spillover

We examined energy transfer dynamics from the photosystem II reaction center (PSII-RC) in intact red algae cells of Porphyridium cruentum, Bangia fuscopurpurea, Porphyra yezoensis, Chondrus giganteus, and Prionitis crispata. Time resolved fluorescence measurements were conducted in the range of 0-80ns at -196°C. The delayed fluorescence spectra were then determined, where the delayed fluorescence was derived from the charge recombination between P680(+) and pheophytin a in PSII-RC. Therefore, the delayed fluorescence spectrum reflected the energy migration processes including PSII-RC. All samples examined showed prominent distribution of delayed fluorescence in PSII and PSI, which suggests that a certain amount of PSII attaches to PSI to share excitation energy in red algae. The energy transfer from PSII to PSI was found to be dominant when the amount of phycoerythrobilin was increased.

Deregulated Chlorophyll b Synthesis Reduces the Energy Transfer Rate Between Photosynthetic Pigments and Induces Photodamage in Arabidopsis thaliana

Chl b is one of the major light-harvesting pigments in land plants. The synthesis of Chl b is strictly regulated in response to light conditions in order to control the antenna size of photosystems. Regulation of Chl b also affects its distribution as it occurs preferentially in the peripheral antenna complexes. However, it has not been experimentally shown how plants respond to environmental conditions when they accumulate excess Chl b. Previously, we produced an Arabidopsis transgenic plant (referred to as the BC plant) in which Chl b biosynthesis was enhanced. In this study, we analyzed the photosynthetic properties and genome-wide gene expression in this plant under high light conditions in order to understand the effects of deregulated Chl b biosynthesis. The energy transfer rates between Chl a molecules in PSII decreased and H(2)O(2) accumulated extensively in the BC plant. Microarray analysis revealed that a group of genes involved in anthocyanin biosynthesis was down-regulated and that another group of genes, reported to be sensitive to H(2)O(2), was up-regulated in the BC plant. We also found that anthocyanin levels were low, which was consistent with the results of the microarray analysis. These results indicate that deregulation of Chl b caused severe photodamage and altered gene expression profiles under strong illumination. The importance of the regulation of Chl b synthesis is discussed in relation to the correct localization of Chl b and gene expression.

Adaptation of light-harvesting systems of Arthrospira platensis to light conditions, probed by time-resolved fluorescence spectroscopy.

Cyanobacteria change the quantity and/or quality of their pigment-protein complexes in response to light conditions. In the present study, we analyzed excitation relaxation dynamics in the cyanobacterium, Arthrospira (Spirulina) platensis, grown under lights exhibiting different spectral profiles, by means of steady-state absorption and picosecond time-resolved fluorescence spectroscopies. It was found that F760, which is the PSI red-chlorophyll characteristic of A. platensis, contributes to slower energy-transfer phase in the PSI of A. platensis. Excitation energy transfers in phycobilisome and those from PSII to PSI were modified depending on the light quality. Existence of quencher was suggested in PSI of the blue-light grown cells. Phycobilisomes in the green-light grown cells and the far-red-light grown cells transferred excitation energy from phycobilisome to chlorophyll without loss of energy. In these cells, excitation energy was shared between two photosystems. Fast energy transfer was established in phycobilisome under the yellow-light condition where only the phycobilisome can absorb the cultivation light. Differences in light-harvesting and energy-transfer processes under different cultivation-light conditions are discussed. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.

Specific Gene bciD for C7-Methyl Oxidation in Bacteriochlorophyll e Biosynthesis of Brown-Colored Green Sulfur Bacteria

The gene named bciD, which encodes the enzyme involved in C7-formylation in bacteriochlorophyll e biosynthesis, was found and investigated by insertional inactivation in the brown-colored green sulfur bacterium Chlorobaculum limnaeum (previously called Chlorobium phaeobacteroides). The bciD mutant cells were green in color, and accumulated bacteriochlorophyll c homologs bearing the 7-methyl group, compared to C7-formylated BChl e homologs in the wild type. BChl-c homolog compositions in the mutant were further different from those in Chlorobaculum tepidum which originally produced BChl c: (31 S)-8-isobutyl-12-ethyl-BChl c was unusually predominant.

Variations in photosystem I properties in the primordial cyanobacterium Gloeobacter violaceus PCC 7421.

We compared the optical properties of the trimeric photosystem (PS) I complexes of the primordial cyanobacterium Gloeobacter violaceus PCC 7421 with those of Synechocystis sp. PCC 6803. Gloeobacter violaceus PS I showed (1) a shorter difference maximum of P700 by approximately 2 nm, (2) a smaller antenna size by approximately 10 chlorophyll (Chl) a molecules and (3) an absence of Red Chls. The energy transfer kinetics in the antennae at physiological temperatures were very similar between the two species due to the thermal equilibrium within the antenna; however, they differed at 77 K where energy transfer to Red Chls was clearly observed in Synechocystis sp. PCC 6803. Taken together with the lower P700 redox potential in G. violaceus by approximately 60 mV, we discuss differences in the optical properties of the PS I complexes with respect to the amino acid sequences of core proteins and further to evolution of cyanobacteria.

Visualizing structural dynamics of thylakoid membranes

To optimize photosynthesis, light-harvesting antenna proteins regulate light energy dissipation and redistribution in chloroplast thylakoid membranes, which involve dynamic protein reorganization of photosystems I and II. However, direct evidence for such protein reorganization has not been visualized in live cells. Here we demonstrate structural dynamics of thylakoid membranes by live cell imaging in combination with deconvolution. We observed chlorophyll fluorescence in the antibiotics-induced macrochloroplast in the moss Physcomitrella patens. The three-dimensional reconstruction uncovered the fine thylakoid membrane structure in live cells. The time-lapse imaging shows that the entire thylakoid membrane network is structurally stable, but the individual thylakoid membrane structure is flexible in vivo. Our observation indicates that grana serve as a framework to maintain structural integrity of the entire thylakoid membrane network. Both the structural stability and flexibility of thylakoid membranes would be essential for dynamic protein reorganization under fluctuating light environments.