The physicochemical properties, species composition, and vertical distribution of microorganisms in the water column, shoreline microbial mat, and small shoreline mud volcanoes of the stratified soda Lake Doroninskoe were investigated in September 2007. The lake is located in the Transbaikal region, in the permafrost zone (51°25′N; 112°28′E). The maximal depth of the contemporary lake is about 6 m, the pH value of the water is 9.72, and the water mineralization in the near-bottom horizon is 32.3 g l−1. In summer, the surface oxygen-containing horizon of the water column becomes demineralized to 26.5 g l−1; at a depth of 3.5–4.0 m, an abrupt transition occurs to the aerobic zone containing hydrosulfide (up to 12.56 g l−1). Hydrosulfide was also detected in trace quantities in the upper water horizons. The density stratification of the water column usually ensures stable anaerobic conditions until the freezing period (November and December). The primary production of oxygenic phototrophs reached 176–230 μg l−1. High rates of dark CO2 assimilation (61–240 μg l−1) were detected in the chemocline. Within this zone, an alkaliphilic species of sulfur-oxidizing bacteria of the genus Thioalkalivibrio was detected (104 cells ml−1). Lithoheterotrophic bacteria Halomonas spp., as well as bacteriochlorophyll a-containing aerobic anoxygenic phototrophic bacteria (AAP) Roseinatronobacter sp. capable of thiosulfate oxidation, were isolated from samples collected from the aerobic zone (0–3 m). The water transparency in September was extremely low; therefore, no visible clusters of anoxygenic phototrophic bacteria (APBs) were detected at the boundary of the hydrosulfide layer. However, purple sulfur bacteria which, according to the results of the 16S rRNA gene analysis, belong to the species Thioalkalicoccus limnaeus, Ectothiorhodospira variabilis, “Ect. magna,” and Ect. shaposhnikovii, were isolated from samples of deep silt sediments. Ect. variabilis and Ect. shaposhnikovii were the major APB species in the shoreline algo-bacterial mat. The halotolerant bacterium Ect. shaposhnikovii, purple nonsulfur bacteria of the genus Rhodobacter, and AAP of Roseococcus sp. were isolated from the samples collected from mud volcanoes. All these species are alkaliphiles, moderate halophiles, or halotolerant microorganisms.
В сентябре 2007 г. исследовали физико-химические характеристики, видовой состав и вертикальное распределение микроорганизмов в воде, в прибрежном микробном мате и в прибрежных грязевых вулканчиках стратифицированного содового озера Доронинское. Озеро находится в Забайкальском крае в зоне многолетней мерзлоты (N 51°25; E 112°28). Максимальная глубина современного озера около 6 м, рН воды 9.72, минерализация воды у дна 32.3 г/л. В летний период верхняя кислородсодержащая зона водной толщи распреснена до 26.5 г/л и простирается до 3.54.0 м, после чего резко переходит в анаэробную зону, содержащую гидросульфид (до 12.56 г/л). В следовых количествах гидросульфид обнаружен также в верхних водных горизонтах. Плотностное расслоение водной толщи обычно обеспечивает стабильность анаэробных условий вплоть до периода ледостава (ноябрь декабрь). Первичная продукция оксигенных фототрофов достигала 176230 мкг/л. Зарегистрирован высокий уровень темновой фиксации углекислоты (61240 мкг/л) в зоне хемоклина. В этой зоне обнаружен алкалофильный вид сероокисляющих бактерий Thioalkalivibrio sp. в количестве свыше 104 кл/мл. Из аэробной зоны (03 м) выделены в культуру литогетеротрофные Halomonas spp., а также аэробные бактериохлорофилл-а-содержащие бактерии (АБХ) Roseinatronobacter sp., способные к окислению тиосульфата. Прозрачность водоема в сентябре была крайне низка, поэтому на границе сероводородного слоя не было отмечено заметного скопления аноксигенных фототрофных бактерий (АФБ). Однако из проб глубинного ила удалось выделить пурпурные серобактерии, идентифицированные по данным анализа нуклеотидов гена 16S рРНК, как Thioalkalicoccus limnaeus, Ectothiorhodospira variabilis, “Ect. magna” и Ect. shaposhnikovii. В прибрежном альгобактериальном мате основными видами АФБ были Ect. variabilis и Ect. shaposhnikovii. Из проб грязевых вулканчиков изолированы галотолерантные Ect. shaposhnikovii, несерные пурпурные бактерии рода Rhodobacter и АБХ Roseococcus sp. Все перечисленные виды являются алкалофилами, умеренными галофилами или галотолерантами.
Bioenergetics of the aerobic bacteriochlorophyll a-containing (BCl a) bacterium (ABC bacterium) Roseinatronobacter thiooxidans is a combination of photosynthesis, oxygen respiration, and oxidation of sulfur compounds under alkaliphilic conditions. The photosynthetic activity of Rna. thiooxidans cells was established by the photoinhibition of cell respiration and reversible photobleaching discoloration of the BCl a of reaction centers (RC), connected by the chain of electron transfer with cytochrome c 551 oxidation. The species under study, like many purple bacteria and some of the known ABC bacteria, possesses a light-harvesting pigment-protein (LHI) complex with the average number of 30 molecules of antenna BCl a per one photosynthetic RC. Under microaerobic growth conditions, the cells contained bc 1 complex and two terminal oxidases: cbb 3-cytochrome oxidase and the alternative cytochrome oxidase of the a 3 type. Besides, Rna. thiooxidans was shown to have several different soluble low- and high-potential cytochromes c, probably associated with the ability of utilizing sulfur compounds as additional electron donors.
Bioenergetics of the aerobic bacteriochlorophyll a-containing (BCl a) bacterium (ABC bacterium) Roseinatronobacter thiooxidans is a combination of photosynthesis, oxygen respiration, and oxidation of sulfur compounds under alkaliphilic conditions. The photosynthetic activity of Rna. thiooxidans cells was established by the photoinhibition of cell respiration and reversible photobleaching discoloration of the BCl a of reaction centers (RC), connected by the chain of electron transfer with cytochrome c551 oxidation. The species under study, like many purple bacteria and some of the known ABC bacteria, possesses a light-harvesting pigment-protein (LHI) complex with the average number of 30 molecules of antenna BCl a per one photosynthetic RC. Under microaerobic growth conditions, the cells contained bc1 complex and two terminal oxidases: cbb3-cytochrome oxidase and the alternative cytochrome oxidase of the a3 type. Besides, Rna. thiooxidans was shown to have several different soluble low- and high-potential cytochromes c, probably associated with the ability of utilizing sulfur compounds as additional electron donors.
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.
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.
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.
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).