Igor N. Stadnichuk

Carotenoid-induced quenching of the phycobilisome fluorescence in photosystem II-deficient mutant of Synechocystis sp.

Brief – 10‐second long – irradiation of a photosystem II‐deficient mutant of cyanobacterium Synechocystis sp. PCC 6803 with intense blue or UV‐B light causes an about 40% decrease of phycobilisome (PBS) fluorescence, slowly reversible in the dark. The registered action spectrum of PBS fluorescence quenching only shows bands at 500, 470 and 430 nm, typical of carotenoids, and an additional UV‐B band; no peaks in the region of chlorophyll or PBS absorption have been found. We propose that quenching induced by carotenoids, possibly protein‐bound or glycoside, reveals a new regulatory mechanism protecting photosynthetic apparatus of cyanobacteria against photodamage.

Rhodobaca barguzinensis sp. nov., a new alkaliphilic purple nonsulfur bacterium isolated from a soda lake of the Barguzin Valley (Buryat Republic, Eastern Siberia)

A novel strain, alga-05, of alkaliphilic purple nonsulfur bacteria was isolated from sediments of a small saline (60 g/l) soda lake near Lake Algin (Barguzin Valley, Buryat Republic, Russia). These bacteria contain bacteriochlorophyll a and carotenoids of the alternative spirilloxanthin group with predominating demethylspheroidenone. They are facultative anaerobes; their photosynthetic structures are of the vesicular type and arranged along the cell periphery. Growth of this strain is possible in a salinity range of 5–80 g/l NaCl, with an optimum at 20 g/l NaCl. Best growth occurred at 20–35°C. Analysis of the 16S rRNA gene sequences demonstrated that the studied isolate is closely related to the alkaliphilic purple nonsulfur bacterium Rhodobaca bogoriensis (99% similarity) isolated from soda lakes of the African Rift Zone. According to the results of DNA-DNA hybridization, strain alga-05 has a 52% similarity with the type species of the genus Rhodobaca. On the basis of the obtained genotypic data and some phenotypic properties (dwelling in a hypersaline soda lake of Siberia, moderate halophily, ability to grow at relatively low temperatures, etc.), the isolated strain of purple bacteria was described as a new species of the genus Rhodobaca, Rca. barguzinensis sp. nov.

Fluorescence quenching of the phycobilisome terminal emitter LCM from the cyanobacterium Synechocystis sp. PCC 6803 detected in vivo and in vitro

The fluorescence emission of the phycobilisome (PBS) core-membrane linker protein (L(CM)) can be directly quenched by photoactivated orange carotenoid protein (OCP) at room temperature both in vitro and in vivo, which suggests the crucial role of the OCP-L(CM) interaction in non-photochemical quenching (NPQ) of cyanobacteria. This implication was further supported (i) by low-temperature (77K) fluorescence emission and excitation measurements which showed a specific quenching of the corresponding long-wavelength fluorescence bands which belong to the PBS terminal emitters in the presence of photoactivated OCP, (ii) by systematic investigation of the fluorescence quenching and recovery in wild type and L(CM)-less cells of the model cyanobacterium Synechocystis sp. PCC 6803, and (iii) by the impact of dephosphorylation of isolated PBS on the quenching. The OCP binding site within the PBS and the most probable geometrical arrangement of the OCP-allophycocyanin (APC) complex was determined in silico using the crystal structures of OCP and APC. Geometrically modeled attachment of OCP to the PBS core is not at variance with the OCP-L(CM) interaction. It was concluded that besides being a very central element in the PBS to reaction center excitation energy transfer and PBS assembly, L(CM) also has an essential role in the photoprotective light adaptation processes of cyanobacteria.

Unique Properties vs. Common Themes: The Atypical Cyanobacterium Gloeobacter violaceus PCC 7421 is Capable of State Transitions and Blue-Light-Induced Fluorescence Quenching

The atypical unicellular cyanobacterium Gloeobacter violaceus PCC 7421, which diverged very early during the evolution of cyanobacteria, can be regarded as a key organism for understanding many structural, functional, regulatory and evolutionary aspects of oxygenic photosynthesis. In the present work, the performance of two basic photosynthetic adaptation/protection mechanisms, common to all other oxygenic photoautrophs, had been challenged in this ancient cyanobacterium which lacks thylakoid membranes: state transitions and non-photochemical fluorescence quenching. Both low temperature fluorescence spectra and room temperature fluorescence transients show that G. violaceus is capable of performing state transitions similar to evolutionarily more recent cyanobacteria, being in state 2 in darkness and in state 1 upon illumination by weak blue or far-red light. Compared with state 2, variable fluorescence yield in state 1 is strongly enhanced (almost 80%), while the functional absorption cross-section of PSII is only increased by 8%. In contrast to weak blue light, which enhances fluorescence yield via state 1 formation, strong blue light reversibly quenches Chl fluorescence in G. violaceus. This strongly suggests regulated heat dissipation which is triggered by the orange carotenoid protein whose presence was directly proven by immunoblotting and mass spectrometry in this primordial cyanobacterium. The results are discussed in the framework of cyanobacterial evolution.

The new alkaliphilic bacteriochlorophyll a-containing bacterium Roseinatronobacter monicus sp. nov. from the hypersaline Soda Mono Lake (California, United States)

Two strains of pink-colored aerobic bacteriochlorophyll a-containing bacteria were isolated from aerobic (strain ROS 10) and anaerobic (strain ROS 35) zones of the water column of Mono Lake (California, United States). Cells of the bacteria were nonmotile oval gram-negative rods multiplying by binary fission by means of a constriction. No intracellular membranes were detected. Polyphosphates and poly-β-hydroxybutyric acid were the storage compounds. Pigments were represented by bacteriochlorophyll a and carotenoids of the spheroidene series. The strains were obligately aerobic, mesophilic (temperature optimum of 25–30°C), alkaliphilic (pH optimum of 8.5–9.5), and moderately halophilic (optimal NaCl concentration of 40 g/l). They were obligately heterotrophic and grew aerobically in the dark and in the light. Respiration was inhibited by light at wavelengths corresponding to the absorption of the cellular pigments. The substrate utilization spectra were strain-specific. In the course of organotrophic growth, the bacteria could oxidize thiosulfate to sulfate; sulfide and polysulfide could also be oxidized. The DNA G+C content was 59.4 mol % in strain ROS 10 and 59 mol % in strain ROS 35. In their phenotypic properties, the new strains were close but not identical to the alkaliphilic bacterium Roseinatronobacter thiooxidans. The distinctions in the nucleotide sequences of the 16S rRNA genes (2%) and low DNA-DNA hybridization level with Rna. thiooxidans (22–25%) allow the new strains to be assigned to a new species of the genus Roseinatronobacter, Roseinatronobacter monicus sp. nov. with the type strain ROS 35T (=UNIQEM U-251T = VKM B-2404T).

Chromosome numbers and nuclear DNA contents in the red microalgae Cyanidium caldarium and three Galdieria species

An image analysis system was used to count and measure acetoorcein-stained mitotic chromosomes of the acidothermophilic unicellular red algae Galdieria maxima, G. partita, G. sulphuraria and Cyanidium caldarium (Cyanidiophyceae). Chromosome numbers fell into two groups: n = 2 for all three species of Galdieria and n = 5(7) for C. caldarium. It seems that a separation of C. caldarium from Galdieria is karyologically justified. A chromosome number of 2 appears to be indicative of the genus Galdieria and could be used as a marker to distinguish this taxon from Cyanidium. The two smaller chromosomes in the karyotype of C. caldarium were about 0.4 µm long whereas the other three were 0.5–0.7 µm long. In karyotypes of Galdieria species, the two chromosomes differed in length, the smaller chromosome ranging from 0.8 to 1.8 µm and the larger one from 1.2 to 2.3 µm. The visualization of these extremely small chromosomes was possible due to pretreatment of the cells with the DNA intercalator 9-aminoacridine. The mean absolute length of each chromosome of the three members of Galdieria had statistically significant interspecies differences. Nuclear 1C DNA contents were estimated in the algal cells by the Feulgen technique. All species investigated had genome sizes of 1.50–2.25 × 10−2 pg. Thus it seems that the members of Cyanidiophyceae have the smallest known genomes of all photosynthetic eukaryotes.

Hybrid systems of quantum dots mixed with the photosensitive protein phycoerythrin

It is shown that semiconductor nanocrystals (or quantum dots) can be used to increase the absorbability of a pigment protein. In the mixture of phycoerythrin with quantum dots, the fluorescence of the quantum dots is suppressed several times due to the transfer of absorbed energy to phycoerythrin. The Forster resonance energy transfer is discussed as a possible mechanism of energy transfer in quantum dot-phycoerythrin donor-acceptor pairs. Calculations based on experimental data show that the efficiency of energy migration from quantum dots to phycoerythrin is 88% and the corresponding rate constant is 1.17 × 109 s−1.

Inhibition by glucose of chlorophyll a and phycocyanobilin biosynthesis in the unicellular red alga Galdieria partita at the stage of coproporphyrinogen III formation

Abstract The repression effect of d -glucose on the pigment apparatus of photosynthesis in the unicellular acidophilic red alga Galdieria partita capable of heterotrophic growth was studied. The rate of cell growth under heterotrophic conditions was higher by more than one order of magnitude than that in the autotrophic ones, and the size of those cells was larger. The addition of 1% d -glucose to the growth medium caused an increase in the number of mitochondrion profiles in the cell and a decline of thylakoid number in the chloroplast of G. partita . The loss of thylakoid membranes is accompanied by a reduction of the contents of chlorophyll a . C-phycocyanin and allophycocyanin in the heterotrophic cells. Simultaneously, switching G. partita from autotrophic growth to heterotrophic nutrition led to the excretion of coproporphyrin(ogen) III into the growth medium. Thus, the presence of d -glucose caused a reduction of pigment contents in the cell, due to the inhibition of chlorophyll a and phycocyanobilin biosynthesis at the stage of the transformation of their universal precursor, coproporphyrinogen III, to protoporphyrinogen IX. In the mixotrophic culture of G. partita light induces both the biosynthesis of photosynthetic pigments and the excretion of coproporphyrin(ogen) III. It is supposed that light affects an earlier stage in the biosynthetic pathway of chlorophyll a and phycocyanobilin than does d -glucose.

Cyanobacterial phycobilisomes and phycobiliproteins

In cyanobacteria, phycobilisomes (PBS) act as antenna of the photosynthetic pigment apparatus. They contain brightly colored phycobiliproteins (PBP) and form giant supramolecular complexes (up to 3000–7000 kDa) containing 200 to 500 phycobilin chromophores covalently bound to the proteins. There are over ten various PBP known, which falls into one of three groups: phycoerythrins, phycocyanins, and allophycocyanins. Hollow disks of PBP trimers and hexamers are arranged into cylinders by colorless linker proteins; the cylinders are then assembled into PBS. Typical semidiscoidal PBS consists of a central core formed by three allophycocyanin cylinders and of six lateral cylinders consisting of other PBP and attached as a fan to the nucleus. The PBS number, size, and pigment composition in cyanobacteria depend on light conditions and other ambient factors. While PBSs have certain advantages compared to other antennae, these pigment-protein complexes require more energy for their biosynthesis than the chlorophyll a/b and chlorophyll a/c proteins of oxygenic photosynthetic organisms.

Far-red light-regulated efficient energy transfer from phycobilisomes to photosystem I in the red microalga Galdieria sulphuraria and photosystems-related heterogeneity of phycobilisome population

Phycobilisomes (PBS) are the major photosynthetic antenna complexes in cyanobacteria and red algae. In the red microalga Galdieria sulphuraria, action spectra measured separately for photosynthetic activities of photosystem I (PSI) and photosystem II (PSII) demonstrate that PBS fraction attributed to PSI is more sensitive to stress conditions and upon nitrogen starvation disappears from the cell earlier than the fraction of PBS coupled to PSII. Preillumination of the cells by actinic far-red light primarily absorbed by PSI caused an increase in the amplitude of the PBS low-temperature fluorescence emission that was accompanied by the decrease in PBS region of the PSI 77 K fluorescence excitation spectrum. Under the same conditions, fluorescence excitation spectrum of PSII remained unchanged. The amplitude of P700 photooxidation in PBS-absorbed light at physiological temperature was found to match the fluorescence changes observed at 77 K. The far-red light adaptations were reversible within 2-5min. It is suggested that the short-term fluorescence alterations observed in far-red light are triggered by the redox state of P700 and correspond to the temporal detachment of the PBS antenna from the core complexes of PSI. Furthermore, the absence of any change in the 77 K fluorescence excitation cross-section of PSII suggests that light energy transfer from PBS to PSI in G. sulphuraria is direct and does not occur through PSII. Finally, a novel photoprotective role of PBS in red algae is discussed.