Vasil A. Gaisin

Filamentous anoxygenic phototrophic bacteria from cyanobacterial mats of Alla hot springs (Barguzin Valley, Russia)

Alkaline hydrotherms of the Baikal rift zone are unique systems to study the diversity of thermophilic bacteria. In this study, we present data on the phototrophic bacterial community of cyanobacterial mats from the alkaline Alla hot spring. Using a clonal analysis approach, this study evaluated the species diversity, the proportion of oxygenic and anoxygenic phototrophs and their distribution between various areas of the spring. Novel group-specific PCR primers were designed and applied to detect representatives of the Chloroflexus and Roseiflexus genera in mat samples. For the first time, the presence of Roseiflexus-like bacteria was detected in the Baikal rift zone.

Rhodobaculum claviforme gen. nov., sp. nov., a new alkaliphilic nonsulfur purple bacterium

Two alkaliphilic strains of nonsulfur purple bacteria (NPB), B7-4 and B8-2, were isolated from moderately saline alkaline steppe lakes in southeast Siberia with pH values above 9.0. The isolates were motile, polymorphous cells (from short rods to long spindly cells) 1–2.5 × 2.5–7 μm. Intracellular membranes of vesicular type were mostly located at the cell periphery. The microorganisms contained bacteriochlorophyll a and carotenoids of the spheroidene and spirilloxanthin series. The photosynthetic apparatus was represented by LH2 and LH1 light-harvesting complexes. In the presence of organic compounds, the strains grew aerobically in the dark or anaerobically in the light. Capacity for photo- and chemoautotrophic growth was not detected. The cbbL gene encoding RuBisCO was not revealed. Optimal growth of both strains occurred at 2% NaCl (range from 0.5 to 4%), pH 8.0–8.8 (range from 7.5 to 9.7), and 25–35°C. The DNA G+C content was 67.6–69.8 mol %. Pairwise comparison of the nucleotides of the 16S rRNA genes revealed that strains B7-4 and B8-2 belonged to the same species (99.9% homology) and were most closely related to the aerobic alkaliphilic bacteriochlorophyll a-containing anoxygenic phototrophic bacterium (APB) Roseibacula alcaliphilum DeT (95.2%) and to NPB strains Rhodobaca barguzinensis VKM B-2406T (94.2%) and Rbc. bogoriensis LBB1T (93.9%). The isolates were closely related to the NPB Rhodobacter veldkampii DSM 11550T (94.8%) and to aerobic bacteriochlorophyll a-containing bacteria Roseinatronobacter monicus ROS 35T and Roseicitreum antarcticul ZS2-28T (93.5 and 93.9%, respectively). New strains were described as a new NPB genus and species of the family Rhodobacteriaceae, Rhodobaculum claviforme gen. nov., sp. nov., with B7-4T (VKM B-2708, LMG 28126) as the type strain.

Candidatus ‘Chloroploca asiatica’ gen. nov., sp. nov., a new mesophilic filamentous anoxygenic phototrophic bacterium

Five phylogenetically similar monocultures of mesophilic filamentous anoxygenic phototrophic bacteria (FAPB) were isolated from microbial mats of low-mineral (5–28 g/L) alkaline lakes in Buryat Republic, Transbaikalia and Mongolia, as well as from biofilms of an alkaline sulfide spring (3 g/L) of the Umhei hydrothermal system (Buryat Republic). New isolates were characterized by short trichomes (15–30 μm long and ∼1 μm in diameter), straight, curved, or wavy, surrounded by a thin iron-sorbing mucous sheath. Gliding motion of the trichomes was not observed. The trichomes formed bunches consisting of several filaments. Trichomes multiply by the separation of short fragments or single cells from the parental trichome. The cells in the filaments were elongated; they contained chlorosomes, gas vesicles, poly-β-hydroxybutyrate granules, and small polyphosphate inclusions. Bacteria contained bacteriochlorophylls c and a and γ-carotene. Absorption maxima of the pigments in the cells were observed at 462, (shoulder at 515), 742, 805, and 863 nm. The organisms were strict anaerobes capable of photoautotrophic growth with sulfide as an electron donor. Elemental sulfur emerged into the medium as a result of sulfide photooxidation. The organisms were tolerant to sulfide (up to 8 mM). Best growth occurred at pH 8.0, 3–15 g/L NaCl, and 1–5 g/L sodium bicarbonate. According to phylogenetic analysis, the 16S rRNA gene sequences of the FAPB isolates formed a separate cluster most closely related to the species cluster of the family Oscillochloridaceae, suborder Chloroflexinae, order Chloroflexales, class Chloroflexi. The differences with the closest 16S rRNA gene sequences of the known FAPB were 9–10%. The formal description of a new taxon, Candidatus’ Chloroploca asiatica’ gen. nov., sp. nov., is provided.

Anoxygenic phototrophic bacteria from microbial communities of Goryachinsk thermal spring (Baikal Area, Russia)

Species composition of anoxygenic phototrophic bacteria in microbial mats of the Goryachinsk thermal spring was investigated along the temperature gradient. The spring belonging to nitrogenous alkaline hydrotherms is located at the shore of Lake Baikal 188 km north-east from Ulan-Ude. The water is of the sulfate-sodium type, contains trace amounts of sulfide, and salinity does not exceed 0.64 g/L, pH 9.5. The temperature at the outlet of the spring may reach 54°C. The cultures of filamentous anoxygenic phototrophic bacteria, nonsulfur and sulfur purple bacteria, and aerobic anoxygenic phototrophic bacteria were identified using the pufLM molecular marker. The fmoA marker was used for identification of green sulfur bacteria. Filamentous cyanobacteria predominated in the mats, with anoxygenic phototrophs comprising a minor component of the phototrophic communities. Thermophilic bacteria Chloroflexus aurantiacus were detected in the samples from both the thermophilic and mesophilic mats. Cultures of nonsulfur purple bacteria similar to Blastochloris sulfoviridis and Rhodomicrobium vannielii were isolated from the mats developed at high (50.6–49.4°C) and low temperatures (45–20°C). Purple sulfur bacteria Allochromatium sp. and Thiocapsa sp., as well as green sulfur bacteria Chlorobium sp., were revealed in low-temperature mats. Truly thermophilic purple and green sulfur bacteria were not found in the spring. Anoxygenic phototrophic bacteria found in the spring were typical of the sulfur communities, for which the sulfur cycle is mandatory. The presence of aerobic bacteriochlorophyll a-containing bacteria identified as Agrobacterium (Rhizobium) tumifaciens in the mesophilic (20°C) mat is of interest.

Characterization of a New Strain of a Purple Nonsulfur Bacterium from a Thermal Spring

A new budding purple nonsulfur bacterium of the genus Rhodobacter (strain Ku-2) was isolated from a mat of a moderately thermal spring (Baikal rift zone, Buryatia, Russia). The bacterium had lamellar photosynthetic membranes, which are typical of only one Rhodobacter species, Rba. blasticus. The cells contained spheroidene carotenoids and bacteriochlorophyll a (Bchl a). In vivo absorption spectrum of the cells, with the major maximum at 863 nm and an additional peak at 887 nm, is characteristic of the pigment-protein complexes of Bchl a-containing membranes. The previously described Rba. blasticus strains do not exhibit the 887-nm maximum. The new isolate was photoheterotrophic, with optimal growth occurring at 35°C, 3 g/L NaCl, and pH 7–8. The DNA G+C content was 64.4 mol %. The similarity between the 16S rRNA gene sequences of strain Ku-2 and the Rba. blasticus type strain was 98.7%. The PufM amino acid sequences of strain Ku-2 and the earlier studied Rba. blasticus type strain were 89.5 % identical. Thus, strain Ku-2 belongs to the genus Rhodobacter and is phylogenetically close to Rba. blasticus.

Benthic phototrophic community from Kiran soda lake, south-eastern Siberia

Phototrophic bacterial mats from Kiran soda lake (south-eastern Siberia) were studied using integrated approach including analysis of the ion composition of water, pigments composition, bacterial diversity and the vertical distribution of phototrophic microorganisms in the mats. Bacterial diversity was investigated using microscopic examination, 16S rRNA gene Illumina sequencing and culturing methods. The mats were formed as a result of decomposition of sedimented planktonic microorganisms, among which cyanobacteria of the genus Arthrospira predominated. Cyanobacteria were the largest part of phototrophs in the mats, but anoxygenic phototrophs were significant fraction. The prevailing species of the anoxygenic phototrophic bacteria are typical for soda lakes. The mats harbored aerobic anoxygenic phototrophic bacteria, purple sulfur and non-sulfur bacteria, as well as new filamentous phototrophic Chloroflexi. New strains of Thiocapsa sp. Kir-1, Ectothiorhodospira sp. Kir-2 and Kir-4, Thiorhodospira sp. Kir-3 and novel phototrophic Chloroflexi bacterium Kir15-3F were isolated and identified.

Chloroflexus islandicus sp. nov., a thermophilic filamentous anoxygenic phototrophic bacterium from geyser Strokkur (Iceland).

A novel, thermophilic filamentous anoxygenic phototrophic bacterium, strain isl-2T, was isolated from the Strokkur Geyser, Iceland. Strain isl-2T formed unbranched multicellular filaments with gliding motility. The cells formed no spores and stained Gram-negative. The existence of pili was described in a species of the genus Chloroflexus for the first time, to our knowledge. Optimal growth occurred at a pH range of 7.5-7.7 and at a temperature of 55 °C. Strain isl-2T grew photoheterotrophically under anaerobic conditions in the light and chemoheterotrophically under aerobic conditions in the dark. The major cellular fatty acids were C18 : 1ω9, C16 : 0, C18 : 0 and C18 : 0-OH. The major quinone was menaquinone-10. The photosynthetic pigments were bacteriochlorophylls c and a as well as β- and γ-carotenes. The results of phylogenetic analysis of the 16S rRNA gene sequences placed strain isl-2T into the genus Chloroflexus of the phylum Chloroflexi with Chloroflexus aggregans DSM 9485T as the closest relative (97.0 % identity). The whole-genome sequence of isl-2T was determined. Average nucleotide identity values obtained for isl-2T in comparison to available genomic sequences of other strains of members of the genus Chloroflexus were 81.4 % or less and digital DNA-DNA hybridisation values 22.8 % or less. The results of additional phylogenetic analysis of the PufLM and BchG amino acid sequences supported the separate position of the isl-2T phylotype from the phylotypes of other members of the genus Chloroflexus. On the basis of physiological and phylogenetic data as well as genomic data, it was suggested that isl-2T represents a novel species within the genus Chloroflexus, with the proposed name Chloroflexus islandicus sp. nov. The type strain of the species is isl-2T (=VKM B-2978T,=DSM 29225T,=JCM 30533T).

Draft Genome Sequence of Chloroflexus sp. Strain isl-2, a Thermophilic Filamentous Anoxygenic Phototrophic Bacterium Isolated from the Strokkur Geyser, Iceland.

ABSTRACT We report here the draft genome sequence of the thermophilic filamentous anoxygenic phototrophic bacterium Chloroflexus sp. strain isl-2, which was isolated from the Strokkur geyser, Iceland, and contains 5,222,563 bp with a G+C content of 59.65%. The annotated genome sequence offers the genetic basis for understanding the strains ecological role as a phototrophic bacterium within the bacterial community.

Sharp water column stratification with an extremely dense microbial population in a small meromictic lake, Trekhtzvetnoe: Sharp water column stratification

Located on the shore of Kandalaksha Bay (the White Sea, Russia) and previously separated from it, Trekhtzvetnoe Lake (average depth 3.5 m) is one of the shallowest meromictic lakes known. Despite its shallowness, it features completely developed water column stratification with high-density microbial chemocline community (bacterial plate) and high rates of major biogeochemical processes. A sharp halocline stabilizes the stratification. Chlorobium phaeovibrioides dominated the bacterial plate, which reached a density of 2 × 108 cell ml-1 and almost completely intercepts H2 S diffusion from the anoxic monimolimnion. The resulting anoxygenic photosynthesis rate reached 240 μmol C l-1 day-1 , exceeding the oxygenic photosynthesis rate in the mixolimnion. The rates of other processes are also high, reaching 4.5 μmol CH4 l-1 day-1 for methane oxidation and 35 μmol S l-1 day-1 for sulfate reduction. Metagenomic analysis demonstrated that the Chl. phaeovibrioides population in the bacterial plate layer had nearly clonal homogeneity, although some fraction of these cells harbour a plasmid. The Chlorobium population was associated with bacteriophages that share homology with CRISPR spacers in the host. These features make the ecosystem of the Trekhtzvetnoe Lake a valuable model for studying regulation and evolution processes in natural high-density microbial systems.