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DPA membranes from Allochromatium vinosum cells, in which carotenoid biosynthesis was inhibited using diphenylamine (DPA) were obtained, into which rhodopin was incorporated. The LH2 complex with rhodopin content of 85% was isolated. Using a test for the thermal stability of LH2 complexes (DPA and with incorporated rhodopin), it was established that carotenoids of the early stages of biosynthesis (≤1 molecules per complex) did not interfere with rhodopin incorporation. It was found that when the LH2 complex with incorporated rhodopin was irradiated with light at the wavelength of 502 nm, BChl850 was photobleached at a rate close to that in the control LH2 complex. This indicates that rhodopin, after being incorporated into the DPA LH2 complex, is capable of generating singlet oxygen under illumination. Previously obtained data on heterogeneity of the carotenoid composition in DPA LH2 complexes (variation in the number of individual carotenoids molecules per complex in the general population) and our earlier suggestion about the structural role of carotenoids, namely, their ability to stabilize the LH2 complexes, were confirmed. Based on analysis of our results, as well as of the literature data, the interaction of singlet oxygen and carotenoids is discussed. [ABSTRACT FROM AUTHOR]

The interaction of singlet oxygen with bacteriochlorophyll (BChl) in the membranes, LH2 light-harvesting complexes and pigment extracts from Allochromatium (Alc.) vinosum strain MSU (previously Alc. minutissimum) was studied. The ability of carotenoids illuminated with light within their absorption region to generate singlet oxygen, which oxidizes BChl, was confirmed; BChl itself was not active in this process and acted a target for singlet oxygen. A sharp decrease in the amount of oxidized BChl in the LH2 complex under illumination in the carotenoid region was noted only at the carotenoid/complex ratio of about 0.1, i.e., when there is less than 1 carotenoid molecule per 10 LH2 complexes. It is assumed that carotenoids from the early stages of biosynthesis (neurosporene, ζ-carotene) are more active in the generation of singlet oxygen under illumination than more mature carotenoids. BChl photooxidation in LH2 complexes occurred only in the presence of oxygen. A decrease in oxygen concentration by 7 or more times resulted in a slowdown of the process or its complete stop. In model systems (pigment extract of the LH2 complex retaining the BChl/carotenoids ratio), BChl under illumination becomes both the main generator of singlet oxygen and a target for the action of the latter. Under these conditions, carotenoids do not protect BChl from oxidation. The ability of BChl850 in the LH2 complex to be oxidized by singlet oxygen suggests that it can be used as a natural sensor for this agent. [ABSTRACT FROM AUTHOR]

The effect of the qualitative and quantitative content of carotenoids on the assembly of light-harvesting complexes isolated from the heat-resistant purple sulfur bacterium Thermochromatium (T.) tepidum was studied. To decrease the content of carotenoids in the photosynthetic membranes and complexes, an inhibitor of carotenonegesis, diphenylamine (DPA), was used. The minimum carotenoid content of 40% was observed in T. tepidum cells grown in the presence of 120 μM DPA. At higher DPA concentrations, the growth of the culture stopped. Photosynthetic membranes and LH2 and LH1-RC complexes were isolated from DPA-treated cells; carotenoid content of these structures was 45 and 55%, respectively. Their spectral characteristics in the near IR region did not depend on the amount of carotenoids in the complexes and were similar to those of the native wild-type complexes. Simultaneously with a general decrease in the content of carotenoids, carotenoids from earlier stages of biosynthesis, such as neurosporene, ζ-carotene, and their hydroxy derivatives, appeared in the DPA-treated complexes. Emergence in DPA-treated complexes of carotenoids with a shorter chain of conjugated double bonds than in the control resulted in an increase in the efficiency of energy transfer from carotenoids to bacteriochlorophyll. [ABSTRACT FROM AUTHOR]

Regardless of cultivation conditions, two types of LH2 complexes are always assembled in photosynthetic membranes of Thiorhodospira (T.) sibirica: short-wave B800−830 and long-wave B800−850. The present work studied the effect of qualitative and quantitative content of carotenoids on assembling of these complexes. Diphenylamine (DPA), an inhibitor of carotenoid biosynthesis, was used to decrease the content of carotenoids in photosynthetic membranes and the complexes. In the cells of T. sibirica strain A12T, inhibition of carotenoid biosynthesis ranged from 40 to 50%. Complete suppression of carotenoid biosynthesis was achieved only once, which resulted in absorption spectrum changes of the DPA-LH2 complexes and lower stability of their structure; they were often destroyed during the isolation. The use of DPA with the cells of T. sibirica strain Kir-3 was more efficient. From inhibited cells of this strain, B800−850 and B800−830 complexes were isolated, in which the carotenoid content decreased to 80, 50, and 10%. Their spectral characteristics in the near infrared region were independent of the carotenoid quantity in the complexes and were similar to those of native complexes of the wild-type. At higher DPA concentration in the cultivation medium, accompanied by a decrease of the overall carotenoid content, such carotenoids of the earlier stages of biosynthesis as neurosporene, ζ-carotene, and their hydroxy derivatives, were found in the complexes. The efficiency of energy transfer from carotenoids to bacteriochlorophyll in all LH2 complexes was not shown to depend on the composition and quantity of carotenoids in the samples and was ~40%. Since the near-infrared spectrum of the LH2-complex of the B800−850 type was similar to that of the B800−850 complexes from Rhodospirillum molishianum, with 8 pairs of α/β-heterodimers forming the basis of their structure, it is possible to assume that the structure of the studied B800−850 complex from T. sibirica also consists of 8 pairs of α/β-heterodimers. A single carotenoid molecule is known to occur per the α/β-heterodimer; therefore, on average less than one carotenoid molecule (8 × 0.1 = 0.8) occurs per an LH2 complex from T.sibirica with 10% carotenoid content. This implies that the population of LH2 complexes contains both carotenoidless LH2 complexes and the complexes containing one or more carotenoid molecules. Obviously, LH2 complexes of both types are assembled regardless of the quantity of carotenoids per a complex, and carotenoids are not the required component for their correct assembly in vivo. [ABSTRACT FROM AUTHOR]

The possibility of singlet oxygen generation in five species of photosynthetic bacteria under irradiation with light was studied using the Singlet Oxygen Sensor Green (SOSG) dye that, when singlet oxygen is bound, is transformed into a fluorescent endoperoxide form. Irradiation was carried out using light of different spectral composition: red (absorption of bacteriochlorophyll, BChl) and white (absorption of BChl and carotenoids). No singlet oxygen generation was detected under red light irradiation, which indicated that BChl was not involved in this process. Under white light, as the irradiation time increased, an increase in SOSG fluorescence was recorded in the membranes of four bacterial species: Allochromatium vinosum MSU, Rhodobacter sphaeroides G1C, Rba. blasticus K-1 and Rhodopseudomonas faecalis. In Rba. sphaeroides, no increase in SOSG fluorescence was observed. It is assumed that in the white light this process occurs due to the pigments that absorb in the blue-green region of the spectrum, i.e. carotenoids with 8‒11 conjugated double bonds (neurosporene, spheroidene, lycopene, and rhodopin). Similar to Trolox, SOSG may slow down the bleaching process of BChl850 in the membranes of Alc. vinosum strain MSU and may therefore effectively perform the role of a singlet oxygen trap. [ABSTRACT FROM AUTHOR]

The effect of singlet oxygen was studied in the system of Allochromatium (Alc.) vinosum MSU membranes, rose bengal, and light (547−600nm) with and without a quencher. In the system without a quencher, bleaching of the BChl850 band of the LH2 light-harvesting complex was observed and an absorption peak emerged at 698 nm, which belonged to the oxidized product, 3-acetyl-chlorophyll. The efficiency of five quenchers of singlet oxygen for neutralization of the effect of singlet oxygen on bacteriochlorophyll in the light-harvesting complexes of the membranes Alc. vinosum MSU was studied: sodium ascorbate, histidine, imidazole, trolox, and sodium azide. All of them, with the exception of sodium azide, neutralized successively the effect of singlet oxygen. [ABSTRACT FROM AUTHOR]

The effect of light with different spectral composition: white, red and blue-green (the first one is absorbed by all the pigments of the cell, and the second and the third ones are absorbed by bacteriochlorophyll and carotenoids, respectively) on culture growth, carotenoid synthesis, and assembly of the light-harvesting complexes was studied for the purple sulfur bacteria Allochromatium (Alc.) minutissimum MSU and Alc. vinosum ATCC 17899. The working hypothesis on the growth of bacteria under blue-green illumination (absorbed by carotenoids) resulting in the inhibition of cell growth was tested. When equalizing the light by luxes, the intensity of illumination for each luminous flux was 1800 lx (white and red light, 4 W/m2; bluegreen light, 0.4 W/m2). The growth of the cells was recorded in white and red light, while in blue-green light an insignificant increase was observed only for Alc. vinosum at the end of the experiment (7-9 days). Regardless of the spectral composition of the light the B800-850 type LH2 complex was always assembled in Alc. minutissimum membranes, and two short-wave LH2 complexes of В800-820 and В800-840 type were assembled in the membranes of Alc. vinosum. Upon smoothing and increasing the luminous flux up to 6 W/m2 for every illumination mode, both cultures grew with approximately equal rates in blue-green light. In the membranes of Alc. minutissimum and Alc. vinosum the same types of LH2 complexes were assembled as in the case of 1800 lx illumination. It was found that blue-green light did not inhibit cell growth. At illumination of the cells collected at the end of the experiment with blue-green light for 6 h, no photooxidation of BChl850 was registered. However, in the membranes from the cells oxygen-saturated at isolation, ~50% of BChl850 was oxidized after 30 minutes of illumination. In the course of cell growth, oxygen is probably completely consumed and anaerobic conditions develop inside the cell. Under these conditions, formation of reactive oxygen species, BChl photooxidation and inhibition of the cell growth become impossible. [ABSTRACT FROM AUTHOR]

Incorporation of spirilloxanthin into carotenoidless LH2 and LH1-RC complexes from a purple sulfur bacterium Allochromatium (Alc.) minutissimum was studied. Carotenoidless cells of Alc. minutissimum were obtained using diphenylamine, a carotenoid biosynthesis inhibitor. In the course of incorporation of the carotenoid mixture, the composition of which corresponded to that of Alc. minutissimum control photosynthetic membranes, no selective incorporation of spirilloxanthin into the LH1-RC complex was detected. It is assumed that in vivo carotenoids are not incorporated into the LH2 and LH1-RC complexes from a common pool. Pure spirilloxanthin destroys both the LH2 and LH1-RC complexes. Within the concentration range of spirilloxanthin in the incorporated mixture from 27% to 52%, it was found to be incorporated into the LH2 and LH1-RC complexes with the efficiency of 13% and 33%, respectively. The possible existence of different sites of assembly for the LH2 and LH1-RC complexes is discussed, as well as of two fractions of LH2 complexes, in one of which rhodopin may be integrated, and in the other (minor) one, spirilloxanthin. [ABSTRACT FROM AUTHOR]

The system of the green microalgal order Protosiphonales ( Chlorophyceae, Chlorophyta) constructed based on morphological, ecological, and molecular genetic data is considered. Subdivision of the order (=macroclade Stephanosphaerinia) into four groups is proposed, with three probably corresponding to the families Stephanosphaeraceae, Chlorococcaceae, and Protosiphonaceae and the fourth having an indefinite status. The diagnostic characteristics of the families, genera, and species are discussed, as well as the independent rank of some taxa. Morphological descriptions of a number of species are supplemented. Based on polyphasic approach, strain ACSSI 23 is proposed as a candidate for a new genus and species. [ABSTRACT FROM AUTHOR]

B820 subunits from a purple sulfur bacterium Ectothiorhodospira haloalkaliphila strain ATCC 51935 were obtained by treatment of carotenoid free LH1-RC complexes of this bacterium with ß-octylglucopyranoside (ß-OG). The same complexes with 100% carotenoid content were unable to dißsociate to B820 subunits, but disintegrated to monomeric bacteriochlorophyll (BChl) regardless of their carotenoid composition. The degree of dissociation of the LH1-RC complexes with an intermediate content of carotenoids (the B820 formation) was directly dependent on the quantity of carotenoids in the samples. The resulting B820 subunits did not contain carotenoids. B820 subunits easily aggregated to form a complex with an absorption peak at 880 nm at decreased ß-OG concentration. Analysis of the spectra of the LH1-RC complexes isolated from the cells with different levels of carotenogenesis inhibition led to the conclusion of the heterogeneity of the samples with a predominance of them in (a) the fraction with 100% of carotenoids and (b) the fraction of carotenoid-free complexes. [ABSTRACT FROM AUTHOR]

Effect of illumination intensity and inhibition of carotenoid biosynthesis on assemblage of different spectral types of LH2 complexes in a purple sulfur bacterium Allochromatium (Alc.) vinosum ATCC 17899 was studied. Under illumination of 1200 and 500 lx, the complexes B800-850 and B800-840 and B800-820 were assembled. While rhodopine was the major carotenoid in all spectral types of the LH2 complex, a certain increase in the content of carotenoids with higher numbers of conjugated double bonds (anhydrorhodovibrin and didehydrorhodopin) was observed in the B800-820 complex. At 1200 lx, the cells grew slowly at diphenylamine (DPA) concentrations not exceeding 53 μM, while at illumination intensity decreased to 500 lx they could grow at 71 μM DPA (DPA cells). Independent on illumination level, the inhibitor is supposed to impair the functioning of phytoene synthetase (resulting in a decrease in the total carotenoid content) and of phytoene desaturase, which results in formation of neurosporene hydroxy derivatives and ζ-carotene. In the cells grown at 500 lx, small amounts of spheroidene and OH-spheroidene were detected. These carotenoids were originally found under conditions of carotenoid synthesis inhibition in bacteria with spirilloxanthin as the major carotenoid. Carotenoid content in the LH2 complexes isolated from the DPA cells was ~15% of the control (without inhibition) for the B800-850 and ~20% of the control for the B800-820 and B800-840 DPA complexes. Compared to the DPA pigment-containing membranes, the DPA complexes were enriched with carotenoids due to disintegration of some carotenoidless complexes in the course of isolation. These results support the supposition that some of the B800-820, B800-840, and B800-850 complexes may be assembled in the cells of Alc. vinosum ATCC 17899 without carotenoids. Comparison of the characteristics obtained for Alc. vinosum ATCC 17899 and the literature data on strain D of the same bacteria shows that they belong to two different strains, rather than to one as was previously supposed. [ABSTRACT FROM AUTHOR]

Up-to-date methods for isolation of cyanobacteria from soil samples, removal of accompanying microflora, obtaining axenic strains, and conditions and media for subsequent cultivation are reviewed. Characterization of soil as a specific habitat for cyanobacteria is provided. Comparative analysis of pH and elemental composition of the liquid phase of most soil types with the media for cultivating cyanobacteria is carried out. The functional role of the major components required for the cultivation of cyanobacteria is described. The problems associated with isolation, purification, and cultivation of soil cyanobacteria, as well as the relevant solutions, are discussed. [ABSTRACT FROM AUTHOR]

An alkaliphilic nonsulfur purple bacterial (NPB) strain “Green” was isolated from sediments of the littoral zone of the soda lake (mineralization 22 g/l, pH 9.5) in the Barguzin River valley (Eastern Siberia). The cells of the new isolate are ovoid or polymorphic at latter stages. The photosynthetic membrane structures are of vesicular type. Bacteriochlorophyll a and carotenoids of both spheroidene and spirilloxanthin type are the photosynthetic pigments. Two light-harvesting systems (LH1 and LH2) are present. The new isolate is a photoheterotroph and a facultative aerobe. It grows well in the dark on organic substrates; anaerobic phototrophic growth is poor. The isolate is alkaliphilic with pH optimum of 8.5–9.5. The most abundant cell growth occurred at 5–40 g/l NaCl (optimum at 10 g/l) and 30 °C. The DNA G+C base content was 69.9 mol %. Analysis of 16S rRNA gene sequences revealed a 10% difference with the most closely related NPB ( Rhodobacter species). Rubrimonas cliftoensis, a bacteriochlorophyll a-containing bacterium, is the closest relative (93.3% similarity). It is proposed that strain “Green” should be placed in the new genus and new species Rubribacterium polymorphum gen. nov., sp. nov. GenBank accession number: 16S rRNA-EU857676. [ABSTRACT FROM AUTHOR]

A novel strain, SHET, of aerobic bacteriochlorophyll a-containing bacteria was isolated from the surface layer of bottom sediments from the soda lake Shuluutai-Ekhe-Torom (Chita oblast, Eastern Siberia, Russia). The lake water has a total mineralization of 30 g/l and a pH of 9.2. The cells of strain SHET are cocci or short rods, which reproduce by uniform division. The cells are motile by means of flagella. The cell wall structure is of the gram-negative type. Sparse intracytoplasmic membrane vesicles are located close to the cell wall. The new isolate is an obligate aerobe and facultative alkaliphile which grows in a pH range of 7.5–9.5 (with an optimum at pH 8.5–9.0). The best growth of strain SHET occurred at 2.0 g/l NaCl and 23–28°C. Photosynthetic pigments are represented by bacteriochlorophyll a, with the maximum absorption at 865 nm in the in vivo spectrum, and carotenoids (spirilloxanthin derivatives). Analysis of the 16S rRNA gene sequences demonstrated that strain SHET is closely related to Roseococcus thiosulfatophilus of the α-1 subclass of Proteobacteria (98.6 % similarity). The DNA G+C base content is 69.1 mol %. Unlike Rsc. thiosulfatophilus, strain SHET grows well on sugars and glycerol and is not capable of utilizing thiosulfate as an energy source. The new isolate is a facultative alkaliphile and reduces nitrates to nitrites. On the basis of its phenotypic and genetic characteristics, strain SHET was described as a new species of the genus Roseococcus, Rsc. suduntuyensis sp. nov. [ABSTRACT FROM AUTHOR]

We investigated the formation of the B800-850 complex in cells of the bacterium Rhodopseudomonas palustris AB illuminated by red and blue light under anaerobic growth conditions. Under red illumination, the B800-850 complex was assembled with a reduced absorption band at 850 nm. The results of re-electrophoresis of the B800-850 complex and oxidation in the presence of potassium iridate suggest its heterogeneity. It may be a mixture of two complexes (B800 and B800-850). The B800-850 complex lacks the capacity for conformational transitions if assembled under blue illumination. Accordingly, the light-harvesting complex assembled in the blue light contains polypeptides that are not synthesized under normal conditions or at increased or decreased light intensities. The mechanism of regulation of the synthesis of the polypeptides of light-harvesting the B800-850 complex and its dependence on the spectral composition of the light is discussed. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

A new nonmotile purple sulfur bacterium (strain M9) was isolated from the steppe soda lake Lake Dzun Uldziin Nur (pH 9.4; mineralization, 3.3%) situated in southeastern Mongolia. Individual cells appear as vibrios 0.3–0.5 × 0.7–1 μm in size. The dividing cells often do not separate from each other, forming an almost closed ring. The internal photosynthetic membranes are represented by concentric lamellae lining the cell wall. Photosynthetic pigments are bacteriochlorophyll a and carotenoids of the spirilloxanthin series. The main carotenoid (>96%) is spirilloxanthin. Two typical light-harvesting complexes (LH1 and LH2) are present in the membranes in a 1 : 1 ratio. The bacterium is an anaerobe and facultative photoorganoheterotroph. Photolithoautotrophic growth on sulfide is scarce. Thiosulfate is utilized as an electron donor only in the presence of organic matter. Globules of elemental sulfur are formed as an intermediary product of sulfide and thiosulfate oxidation and are deposited outside the cells. The end product of oxidation is sulfate. In the presence of sulfide and carbonates, acetate, lactate, malate, pyruvate, propionate, succinate, and fumarate are used as additional sources of carbon in anoxygenic photosynthesis. Vitamins are not required. The bacterium is an alkaliphile, the pH optimum is at 8.3–9.1, the pH range is 7.6–10.1. The optimum NaCl concentration in the medium is 1 to 7%; the range is 0.5 to 0.9%. The optimum carbonate content in the medium is 2%; the range is 1 to 10%. The best growth occurs at 30–35°C. The DNA G+C content is 57.5 mol %. According to the results of analysis of the 16S rRNA gene sequences, the new isolate M9 belongs to the phylogenetic cluster containing representatives of the family Ectothiorhodospiraceae within the class “Gammaproteobacteria.” In this class, the new isolate forms a new branch, which occupies an intermediate position between the representatives of the genera Ectothiorhodospira and Thiorhodospira. Based on the phenotypic and genetic characteristics, the new purple sulfur bacterium was assigned to a new species of a new genus of the family Ectothiorhodospiraceae, Ectothiorhodosinus mongolicum gen. nov., sp. nov. [ABSTRACT FROM AUTHOR]