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Том 85, № 5 (2016)

Experimental Articles

Formation of a subunit form of the core light-harvesting complex from sulfur purple bacteria Ectothiorhodospira haloalkaliphila with different carotenoid composition

Solov’ev A., Ashikhmin A., Moskalenko A.

Аннотация

B820 subunits from a purple sulfur bacterium Ectothiorhodospira haloalkaliphila strain ATCC 51935T 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.

Microbiology. 2016;85(5):515-522
pages 515-522 views

Biofilm of Aureobasidium pullulans var. pullulans on winter wheat kernels and its effect on other microorganisms

Wachowska U., Głowacka K., Mikołajczyk W., Kucharska K.

Аннотация

Winter wheat, grown under greenhouse conditions, was protected four times with a cell suspension of Aureobasidium pullulans var. pullulans during the growing season. After harvest, the distribution and survival rates of the studied biocontrol agent were analyzed under a scanning electron microscope. The abundance of filamentous fungi, yeasts, pseudomonads and Azotobacter bacteria was determined by inoculation onto selective agar media. A. pullulans produced mostly unicellular chlamydospores on the surface and in the brush of kernels. Multicellular blastospore conglomerates secreted extracellular polymeric substances (EPS), and their biofilms were found in the brush and crease of kernels. The application of a cell suspension of A. pullulans with the density of 104 CFU to winter wheat spikes, repeated four times, inhibited the growth of pseudomonads, Azotobacter bacteria and filamentous fungi.

Microbiology. 2016;85(5):523-530
pages 523-530 views

Microbial diversity associated with Tricholoma matsutake fruiting bodies

Li Q., Chen C., Penttinen P., Xiong C., Zheng L., Huang W.

Аннотация

Endophytes play an important role in the growth and development of the host. However, the study of endophytes is mostly focused on plants and animals, and reports on microorganisms associated with fungus are relatively rare. We studied the microorganisms associated with Tricholoma matsutake fruiting bodies picked from three main T. matsutake-producing areas in Sichuan, China, by both culture-dependent and culture- independent methods. Altogether 13 fungus, 15 yeast and 14 bacterial strains were isolated from the T. matsutake fruiting bodies. The most abundant cultivable fungus, yeast and bacteria isolates were assigned as Fusarium solanis, Cryptococcus sp. and Pseudomonas sp., respectively. Terminal-restriction fragment length polymorphism analysis (T-RFLP) showed that the bacteria in T. matsutake were abundant and diverse. Betaand gamma-proteobacteria, Acidobacteria, Bacteroidetes and Sphingobacterium were found in samples from all collecting sites. Among these bacteria, we may find some strains that can promote the growth of T. matsutake.

Microbiology. 2016;85(5):531-539
pages 531-539 views

Isolation, identification and characterization of Pediococcus pentosaceus LB44 and Weissella confusa LM85 for the presence of bacteriocin-like inhibitory substances (BLIS)

Kaur R., Tiwari S.

Аннотация

Lactic acid bacteria (LAB) were isolated from different soil samples such as dairy, garden, rhizospheric soil and sediments of lake. Thirty nine strains of bacteria were isolated using direct spreading method but only one showed the characteristics of LAB. Whereas, using accumulation with incubation method, fifty seven strains were found to be LAB out of sixty eight strains of bacteria isolated. Therefore, accumulation with incubation method was found to be more effective and faster. On the basis of higher growth and antimicrobial activity, two potential isolates LB44 and LM85 were selected for the further characterization. Strain LB44 and LM85 were identified using 16S rDNA amplification and sequencing as Pediococcus pentosaceus LB44 and Weissella confusa LM85, respectively. Cell free supernatant (CFS) of these strains showed antimicrobial activity, stability at higher temperatures, acidic pH and in the presence of organic solvents, detergents and surfactants. Antimicrobial activity was not affected by treatment with catalase and lipase but reduced after treatment with proteinase K reveals the presence of bacteriocin-like inhibitory substance in CFS. CFS inhibited the growth of other Gram-positive and Gram-negative bacteria such as Micrococcus luteus, Lactobacillus delbrueckii, L. curvatus, L. plantarum, L. acidophilus, Enterobacter cloaceae, Lactococcus lactis subsp. cremoris, L. lactis subsp. lactis, Bacillus subtilis, B. licheniformis, Salmonella typhi and Escherichia coli.

Microbiology. 2016;85(5):540-547
pages 540-547 views

Emended description of Methylovorus glucosotrophus govorukhina and trotsenko 1991

Doronina N., Kaparullina E., Trotsenko Y.

Аннотация

Phylogenetic analysis based on comparison of the 16S rRNA gene sequences in combination with comparative analysis of physiological, biochemical, and chemotaxonomic characteristics and DNA–DNA hybridization revealed that “Methylobacillus fructoseoxidans” 34 (VKM B-1609 = DSM 5897) and Methylovorus glucosotrophus 6B1T (ATCC 49758T = DSM 6874T = VKM B-1745T = NCIMB 13222T) belong to the same Methylovorus species. Extended description of the limited facultative methylotroph Methylovorus glucosotrophus is proposed, which includes the fructose-utilizing strain 34. Emended description of Methylovorus glucosotrophus is provided.

Microbiology. 2016;85(5):548-552
pages 548-552 views

Structure and seasonal trophodynamics of picophytoplankton in sevastopol bay and adjacent waters (the Black Sea)

Mukhanov V., Rylkova O., Churilova T., Sakhon E., Pimenov N.

Аннотация

Abundance and seasonal trophodynamics (specific growth rate, daily production, and grazing mortality) of the major picophytoplankton components, Synechococcus cyanobacteria (Syn) and picoeukaryotes (Pico-E), were studied at three stations in Sevastopol Bay and adjacent coastal waters (the Black Sea) in 2014 by flow cytometry and the dilution method. Pico-E abundance was shown to increase along the nutrient and pollution gradient from the coastal waters outside the bay (annual average of 7.3 ± 5.4 × 103 cells mL–1) to the eastern corner of the bay (28.7 ± 11.4 × 103 cells mL–1), while no relation was found between the water pollution status and Syn abundance (9.9 ± 8.7 × 103 cells mL–1; at all the stations, n = 27). Matter flows through the communities (daily production for Syn and Pico-E 0–16.6 and 0–19.3 µg C L–1 day–1, respectively; grazing mortality for Syn and PicoE 0–3.6 and 0–21.2 µg C L–1 day–1, respectively) were comparable to or even exceeded their biomass stocks (>0.05–6.8 and 0.9–26.5 µg C L–1 for Syn and PicoE, respectively), indicating high biomass turnover rates. The highest flow-to-stock ratio (up to 6 for Syn) and a significant imbalance between daily production (P) and grazing mortality (G) were observed in the most polluted and eutrophic waters of the bay in spring (Pico-E: P/G < 1) and late summer (Syn: P/G > 1). Black River inflow to the bay was hypothesized to be among the mechanisms maintaining this pronounced and long-term imbalance in the open system without any negative consequences for the picophytoplankton assemblages.

Microbiology. 2016;85(5):553-561
pages 553-561 views

Microbial community associated with Thioploca sp. sheaths in the area of the posolsk bank methane seep, southern baikal

Chernitsyna S., Khal’zov I., Khanaeva T., Morozov I., Klimenkov I., Pimenov N., Zemskaya T.

Аннотация

Bacterial mats formed by a colorless sulfur bacterium Thioploca sp. in the area of the Posolsk Bank cold methane seep (southern Baikal) were studied using electron microscopy and phylogenetic analysis. Morphologically the bacteria were identified as Thioploca ingrica. Confocal microscopy of DAPI-stained samples revealed numerous rod-shaped, filamentous, and spiral microorganisms in the sheaths, as well as inside and between the trichomes. Transmission electron microscopy revealed nonvacuolated bacteria and small cells without cell envelopes within the sheath. Bacteria with pronounced intracytoplasmic membranes characteristic of type I methanotrophs were observed at the outer side of the sheath. Based on analysis of the 16S rRNA gene sequences, the following phyla were identified in the sheath community: Bacteroidetes, Nitrospira, Chloroflexi, Planctomycetes, Verrucomicrobia, γ-, and δ-Proteobacteria, Euryarchaeota, Crenarchaeota, and Thaumarchaeota, as well as anammox bacteria. A hypothetical scheme of matter flows in the Lake Baikal bacterial mats was proposed based on the data on metabolism of the cultured homologues.

Microbiology. 2016;85(5):562-569
pages 562-569 views

Succession processes in the anoxygenic phototrophic bacterial community in Lake Kislo-Sladkoe (Kandalaksha Bay, White Sea)

Lunina O., Savvichev A., Krasnova E., Kokryatskaya N., Veslopolova E., Kuznetsov B., Gorlenko V.

Аннотация

The community of anoxygenic phototrophic bacteria (APB) in the water column of Lake Kislo- Sladkoe (Kandalaksha Bay, White Sea), which has recently become separated from the sea, was investigated in March?April 2012, March?April 2013, and in September 2013. The lake, which was previously considered meromictic, was in fact mixed and was strongly affected by the sea. In winter the lake is sometimes washed off with seawater, and this together with the seasonal cycles of succession processes determines the succession of the community. The consequences of the mixing in autumn 2011 could be observed in the APB community as late as autumn 2013. Green-colored green sulfur bacteria (GSB) usually predominated in the chemocline. In winter 2013 stagnation resulted in turbidity of water under the ice, which was responsible for both predominance of the brown GSB forms and the changes ratio of the species of purple sulfur bacteria (PSB) in anoxic water layers. Production of anoxygenic photosynthesis in the lake was at least 240 mg C m-2 day-1 in September and 0–20 mg C m–2 day–1 in March—April, which corresponded to 40 and 69%, respectively, of oxygenic photosynthesis. Okenone-containing purple sulfur bacteria, strain TcakPS12, were isolated in 2012 from lake water. The ells of this strain form filaments of not separated cells. Strain TcakPS12 exhibited 98% similarity with the type strains of Thiocapsa pendens DSM 236 and Thiocapsa bogorovii BBS, as well as with the strains AmPS10 and TcyrPS10, which were isolated from Lake Kislo-Sladkoe in 2010.

Microbiology. 2016;85(5):570-582
pages 570-582 views

Microbial community composition and methanotroph diversity of a subarctic wetland in Russia

Danilova O., Belova S., Gagarinova I., Dedysh S.

Аннотация

This study assessed the microbial diversity, activity, and composition of methane-oxidizing communities of a subarctic wetland in Russia with mosaic cover of Sphagnum mosses and lichens of the genera Cladonia and Cetraria. Potential methane-oxidizing activity of peat sampled from lichen-dominated wetland sites was higher than that in the sites dominated by Sphagnum mosses. In peat from lichen-dominated sites, major bacterial groups identified by high-throughput sequencing of the 16S rRNA genes were the Acidobacteria (35.4–41.2% of total 16S rRNA gene reads), Alphaproteobacteria (19.1–24.2%), Gammaproteobacteria (7.9–11.1%), Actinobacteria (5.5–13.2%), Planctomycetes (7.2–9.5%), and Verrucomicrobia (5.1–9.5%). The distinctive feature of this community was high proportion of Subdivision 2 Acidobacteria, which are not characteristic for boreal Sphagnum peat bogs. Methanotrophic community composition was determined by molecular analysis of the pmoA gene encoding particulate methane monooxygenase. Most (~80%) of all pmoA gene fragments revealed in peat from lichen-dominated sites belonged to the phylogenetic lineage represented by a microaerobic spiral-shaped methanotroph, “Candidatus Methylospira mobilis”. Members of the genus Methylocystis, which are typical inhabitants of boreal Sphagnum peat bogs, represented only a minor group of indigenous methanotrophs. The specific feature of a methanotrophic community in peat from lichen-dominated sites was the presence of uncultivated USCα (Upland Soil Cluster alpha) methanotrophs, which are typical for acidic upland soils showing atmospheric methane oxidation. The methanotrophic community composition in lichen-dominated sites of a tundra wetland, therefore, was markedly different from that in boreal Sphagnum peat bogs.

Microbiology. 2016;85(5):583-591
pages 583-591 views

Phylogenetic and functional prokaryotic diversity in the Hoito-Gol mesothermal mineral spring (Eastern Sayan Mountains, Buryat Republic)

Kashkak E., Bel’kova N., Danilova E., Dagurova O., Namsaraev B., Gorlenko V.

Аннотация

High-throughput sequencing was used for comparative analysis of microbial communities of the water and mat from the Hoito-Gol mesothermal mineral sulfide spring (Eastern Sayan Mountains, Buryat Republic). Activity of microbial communities was determined. While both spring biotopes were dominated by members of three bacterial phyla—Proteobacteria, Bacteroidetes, and Firmicutes—they differed drastically in the composition of predominant phylotypes (at the genus level). In the water, the organisms widespread in aquatic environments were predominant, mostly aerobic chemoorganotrophs of the genera Acinetobacter, Pedobacter, and Flavobacterium. In the microbial mat, the organisms actively involved in the sulfur cycle predominated, including sulfur-reducing bacteria Sulfurospirillum, sulfate-reducing deltaproteobacteria, sulfuroxidizing chemoautotrophic bacteria, anoxygenic phototrophic bacteria of the phyla Chloroflexi and Chlorobi, as well as purple bacteria belonging to the α-, ß-, and γ-Proteobacteria. Microbial mats of the spring exhibited higher phylogenetic diversity compared to high-temperature mats containing photosynthetic microorganisms.

Microbiology. 2016;85(5):592-603
pages 592-603 views

Diversity and physiological and biochemical properties of heterotrophic bacteria isolated from Lake Baikal neuston

Galach’yants A., Bel’kova N., Sukhanova E., Romanovskaya V., Gladka G., Bedoshvili E., Parfenova V.

Аннотация

For heterotrophic microorganisms (44 strains) isolated from the surface film of Lake Baikal, identification was carried out and their physiological and biochemical characteristics were determined. Compared to the water column, diversity of cultured heterotrophs was low, indicating formation of stable microbial communities at the air–water interface. Heterotrophic bacteria isolated from the surface microlayer exhibited the enzymatic activity comparable to that for strains from other biofilm associations. Deinococcus ficus strain NA202 was the most active component of the community, capable of utilization of the broadest spectrum of mono- and disaccharides, sugars, and amino acids. This strain possessed the highest diversity of extracellular enzymes and was the most resistant to UV radiation. The physiological and biochemical properties of this strain may be responsible for its adaptation to survival in extreme conditions of the surface microlayer. These results improve our understanding of occurrence of UV-resistant strains in freshwater ecosystems.

Microbiology. 2016;85(5):604-613
pages 604-613 views

Modeling of dissemination of microbial cells and phages from the sites of permafrost thawing

Skladnev D., Mulyukin A., Filippova S., Kulikov E., Letarova M., Yuzbasheva E., Karnysheva E., Brushkov A., Gal’chenko V.

Аннотация

A method is proposed for integral assessment of the propagation of microbial cells and phage particles during seasonal thawing of relic ice wedge layers. The results of field and laboratory investigation carried out in the upper part of permafrost exposure at Mamontova Gora (Yakutiya, Russia) are presented. Suspensions of yeast, bacteria, and two coliphages were introduced as biomarkers directly on the surface of thawing ice and in the meltwater flow. Microorganisms and phages were shown (a) to possess particular parameters of dissemination in the meltwater flow and (b) were able to move 132 m in 25–35 min with the stream water flow.

Microbiology. 2016;85(5):614-619
pages 614-619 views

Relations between bacterioplankton, heterotrophic nanoflagellates, and virioplankton in the littoral zone of a large plain reservoir: Impact of bird colonies

Rumyantseva E., Kosolapova N., Kosolapov D.

Аннотация

Interactions of the main components of microbial planktonic food web (bacteria, heterotrophic nanoflagellates, and viruses) were studied in a protected overgrown littoral zone of the Rybinsk Reservoir (Upper Volga). The effect of colonial bird settlements (the Laridae family) on these processes was determined. The following systems exhibited significant negative correlations: “heterotrophic nanoflagellates–large rod-shaped bacteria” (“predator–prey”), “viruses-bacteriophages–bacterial products” (“parasite–host”) and “heterotrophic nanoflagellates–viruses-bacteriophages”. Relations between biotic factors controlling bacterial development were more pronounced outside the zone affected by colonial bird settlements. Near the bird colony the role of viruses in mortality of planktonic bacteria increased. Reproduction of bacterial cells accelerated in response to the increase in feeding activity of heterotrophic nanoflagellates. Virusesbacteriophages and heterotrophic nanoflagellates probably eliminate different targets until medium-sized cells become predominant in the bacterial community. Then heterotrophic nanoflagellates consume bacterial cells infected with viruses.

Microbiology. 2016;85(5):620-628
pages 620-628 views

Role of the microbial community in formation of speleothem (moonmilk) in the Snezhnaya carst cave (abkhazia)

Kondratyeva L., Polevskaya O., Litvinenko Z., Golubeva E., Konovalova N.

Аннотация

The results of investigation of speleothem (moonmilk) from the Snezhnaya cave (West Caucasus, Abkhazia) are reported. The structure of microbial complexes from moonmilk was investigated by quantitative PCR; strains of culturable microorganisms were isolated and their preferred temperature and carbon sources were determined. Among eubacteria, ~34% belonged to the iron bacteria (Rhodoferax). Most bacterial strains were shown to be facultative psychrophiles with the maximum growth rate at 4°C. The microstructure and elemental composition of moonmilk were investigated using a scanning electron microscope (EVO-40HV, Carl Zeiss) and silicon drift X-ray detector X-MAX 80mm2. The visually plastic and homogeneous mass of moonmilk was shown to be highly heterogeneous, containing various microstructures. The elemental composition of some nanostructures depended on the structure of bacterial biofilms. Some loci of the biofilm were found to contain up to 46% (wt/wt) of iron oxides. High calcium content, up to 61.5% (wt/wt), was found only in cubic crystalline structures which were not involved in microbiological processes.

Microbiology. 2016;85(5):629-637
pages 629-637 views

Short Communications

Catechol 1,2-dioxygenase induced in Rhodococcus opacus strain 1CP cultured in the presence of 3-hydroxybenzoate

Subbotina N., Kolomytseva M., Baskunov B., Golovlev L.
Microbiology. 2016;85(5):638-641
pages 638-641 views
pages 642-645 views
pages 646-648 views

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