The structure of phototrophic communities of soda lakes of the southeastern Transbaikal Region

The structure of benthic phototrophic communities of 24 soda lakes of the southeastern Transbaikal Region was studied. The physicochemical properties of the lakes were determined. The results of enumeration of anoxygenic phototrophic bacteria (APB) belonging to various groups are presented. The influence of salinity on the structure of APB communities was investigated. The APB reaction to environmental conditions was determined. Massive development of phototrophic microorganisms in the form of mats and films was observed in the majority of the investigated lakes. The APB communities were characterized by a wide diversity and evenness of species composition. Purple sulfur bacteria of the families Ectothiorhodospiraceae and Chromatiaceae were predominant. Purple nonsulfur bacteria of the family Rhodobacteraceae, green filamentous bacteria Oscillochloris sp., and heliobacteria were also detected. According to preliminary data, no less than 15 species of APB occur in the studied lakes. Among them, three novel genera and four species have already been described. Identification of other isolates is still in progress. The lakes make an almost continuous series of fresh, brackish, and saline water bodies, varying in their degree of mineralization. It was demonstrated that the structure of APB communities was unaffected by changes in salinity from 5 to 40 g/l. At salt concentrations of lower than 5 g/l, the level of water mineralization became a limiting factor. Experiments with the isolated cultures showed that the APB were obligately dependent on the presence of carbonate ions in the medium. They were haloalkalitolerant or haloalkaliphilic. Thus, they are well adapted to the conditions of soda lakes with a low of moderate mineralization. It was demonstrated that soda lakes of the southeastern Transbaikal Region represent a special type of habitat which harbors a peculiar autochthonous microflora and differs from both highly mineralized soda lakes and shallow saline water bodies of the sea origin.     

The effect of the pH level on the communities of anoxygenic phototrophic bacteria of soda lakes of the southeastern Transbaikal Region

The effect of pH on the structure of the communities of anoxygenic phototrophic bacteria (APB) was studied under laboratory conditions. Samples of natural APB communities were inoculated into media that differed in pH values, which were 7, 9.5, or 10.5. The structure of the APB communities in the obtained enrichment cultures at all pH values depended also on the mineralization levels of the media, which were the same as in the lakes from which samples were taken. The same dependence of the community structure on salinity was observed as in the case of the natural communities that had been described previously. APB were most diverse in the enrichment cultures grown at pH 9.5. The shift of the pH to either neutral or extremely alkaline values restricted the species diversity within the APB community, resulting in marked predominance of the most adapted forms. It was shown that the status of Ectothiorhodospira species within the community could serve not only as an indicator of salinity but also as an indicator of pH in soda lakes with a water mineralization of higher than 5 g/l. The statuses of various APB groups in the community as dependent on pH and salinity are discussed, as well as possible changes in these statuses due to changes in the water level and other environmental parameters in the studied lakes.     

Communities of anoxygenic phototrophic bacteria in the saline soda lakes of the Kulunda steppe (Altai Krai)

The saline soda lakes of the Kulunda steppe (Altai krai) are small and shallow; they are characterized by a wide range of salinity and alkalinity, as well as by the extreme instability of their water and chemical regimes. Accumulations of anoxygenic phototrophic bacteria (APB) visible to the unaided eye were noted only in several lakes with high rates of sulfate reduction in their bottom sediments. However, enumeration of APB cells by inoculation revealed their presence in all 17 lakes. APB cell numbers varied from 10³ to 10⁹ CFU cm^?3. In the APB communities of all lakes, purple sulfur bacteria of the family Ectothiorhodospi- raceae were predominant. In 14 out of the 17 lakes, purple nonsulfur bacteria of the family Rhodobacteraceae were also detected (10³–10⁷ CFU cm^?3). Purple sulfur bacteria of the family Chromatiaceae were less abundant: Halochromatium sp. (10⁴–10⁷ CFU cm^?3) were found in six lakes, while Thiocapsa sp. (10⁴ CFU cm³) were detected in one lake. On the whole, the APB communities of the soda lakes of the Kulunda steppe were characterized by the low diversity and evenness of their species compositions, as well as by the pronounced dominance of the members of the family Ectothiorhodospiraceae. There was no correlation between the structures of the APB communities and alkalinity. However, the dependence of the species composition of APB (mainly ectothiorhodospiras) on water mineralization was revealed. High mineralization (above 200 g l^?1) was a limiting factor that affected the APB communities on the whole, restricting the APB species diversity to extremely halophilic bacteria of the genus Halorhodospira.     

The effect of the pH level on the communities of anoxygenic phototrophic bacteria of soda lakes of the southeastern Transbaikal region

The effect of pH on the structure of the communities of anoxygenic-phototrophic-bacteria (APB) was studied under laboratory conditions. Samples of natural APB communities were inoculated into media that differed in pH values, which were 7, 9.5, or 10.5. The structure of the APB communities in the obtained enrichment cultures depended on the pH values and on the mineralization levels of the media, which were the same as in the lakes from which samples were taken. The same dependence of the community structure on salinity was observed as in the case of the natural communities that had been described previously. APB were most diverse in the enrichment cultures grown at pH 9.5. The shift of the pH to either neutral or extremely alkaline values affected the species diversity within the APB community, resulting in marked predominance of the most adapted forms. It was shown that the status of Ectothiorhodospira within the community could serve as an indicator of both salinity and pH in soda lakes with a water mineralization of higher than 5 g/l. The statuses of various APB groups in the community as dependent on pH and salinity are discussed, as well as possible changes in these statuses due to changes in the water level and other environmental parameters in the studied lakes.     

Primary production of organic matter and phototrophic communities in the soda lakes of the Kulunda steppe (Altai krai)

The rates of photosynthesis and dark CO₂ fixation were determined in 12 soda lakes of the Kulunda steppe. Characterization of the phototrophic communities was given, and the cell numbers of anoxygenic phototrophic bacteria (APB) were determined. The photosynthetic production in different lakes was substantially different, constituting from 0.01 to 1.32 g C m⁻² day⁻¹. The main part of carbon dioxide was assimilated in the process of oxygenic photosynthesis. Anoxygenic photosynthesis was recorded only in 5 of the 12 lakes studied. Its values varied between 0.06 and 0.42 g C m⁻² day⁻¹, constituting from 8 to 34% of the total photosynthetic activity. Anoxygenic photosynthesis was revealed in the lakes where the number of APB reached 10⁷–10⁹ CFU cm⁻³. Dark CO₂ fixation constituted 0.01–0.15 g C m⁻² day⁻¹. Positive correlation was observed between the primary production value and water alkalinity. No relationship between productivity and water mineralization was revealed in the 30–200 g l⁻¹ range, whereas an increase in salinity above 200 g l⁻¹ suppressed the photosynthetic activity. The mechanisms of influence of the environmental factors on the rate of photosynthesis are discussed.     

Salinity as a determinant of the structure of biological communities in salt lakes

The paper considers the extent to which salinity determines the structure of biological communities (composition and species richness and diversity) in saline lakes, i.e. inland bodies of water with salinities in excess of 3 g l-1. It also considers the extent to which oxygen, ionic composition, pH, hydrological patterns (degree of permanence and impermanence of water), geographical position, palaeoclimatic events, chance, human intervention, and biological interactions especially predation determine biological communities in salt lakes. It suggests that salinity is less significant as a determinant of community structure in salt lakes than has been assumed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Biological fractionation of stable carbon isotopes at the aerobic/anaerobic water interface of meromictic water bodies

Mass-spectrometric investigation of carbon isotope composition (δ¹³C) was carried out for suspended organic matter and dissolved mineral compounds for the water column of some meromictic water bodies differing in salinity and trophic state. As a rule, a more pronounced carbon isotope fractionation (resulting from the metabolism of phytoplankton and anoxygenic phototrophic bacteria) was revealed in the zones of enhanced oxygenic and anoxygenic photosynthesis. Carbon isotope fractionation at the border between oxidized and reduced waters depends both on the activity of microbial communities and on the dominant species of phototrophic microorganisms. Analysis of the distribution profiles of the isotopic composition of suspended organic matter and dissolved mineral carbon revealed active mineralization of the organic matter newly formed via anoxygenic photosynthesis in the monimolimnion by microbial communities, resulting in the release of isotopically light carbon dioxide. Mineral carbon in the anaerobic zones of highly productive meromictic water bodies is therefore enriched with the light ¹²C isotope.     

High abundances of aerobic anoxygenic phototrophs in saline steppe lakes

We studied the distribution of anoxygenic phototrophs in 23 steppe lakes in the Transbaikal region (Russia), in Uzbekistan (Central Asia) and in the Crimean peninsula (Ukraine). The lakes varied in their mineral content and composition (salinities from 0.2 to 300 g L(-1) ). The Transbaikal lakes were alkaline (pH>9), with high amounts of soda. The Uzbek and Crimean lakes were more pH neutral, frequently with high amounts of sulfates. The presence of anoxygenic phototrophs was registered by infrared epifluorescence microscopy, infrared fluorometry and pigment analyses. In mostly shallow, fully oxic lakes, the anoxygenic phototrophs represented 7-65% of the total prokaryotes, with the maxima observed in Transbaikal soda lakes Gorbunka (32%), Khilganta (65%), Zanday (58%) and Zun-Kholvo (46%). Some of the lakes contained over 1 μg bacteriochlorophyll L(-1) . In contrast, only small amounts of anoxygenic phototrophs were present in highly mineralized lakes (>100 g total salts L(-1) ); Borzinskoe, Tsagan-Nur (Transbaikal), Staroe (Crimea) and in the residual part of the south-west Aral Sea (Uzbekistan). The oxic environment and the specific diurnal changes of bacteriochlorophyll concentration observed suggest that the phototrophic community was mostly composed of aerobic anoxygenic phototrophs. The high abundances and bacteriochlorophyll concentrations point to an important role of aerobic anoxygenic phototrophs in the habitats studied.     

Taxonomic and ecological characterization of cyanobacteria from some brackish and saline lakes of Southern Transbaikal Region

The species composition of cyanobacteria from 12 lakes of Southern Transbaikal Region was studied. In the studied lakes, a total of 28 species of cyanobacteria were detected, including 10 species previously not retrieved from the lakes of Southern Transbaikal Region. The morphological and ecophysiological characteristics, as well as the phylogenetic position, of pure cyanobacterial cultures were determined. According to the results of 16S rRNA sequencing, the Cya 1, Cya 2, and Cya 10 cultures were identified as representatives of the genus Phormidium; the Cya 5 and Cya 6 cultures were assigned to the genus Nodularia. On the basis of its morphological properties, the Cya 4 culture was identified as Pseudanabaena frigida. The studied microorganisms were moderate alkaliphiles and could grow within a broad salinity range (0–100 g/l NaCl).     

Structure and formation properties of the haloalkaliphilic community of Lake Khilganta

The structural features of a cyanobacterial mat from Lake Khilganta (Southeastern Transbaikal Region) developing at different values of salinity and pH were determined based on our long-term investigation of the natural community, as well as results obtained during experimentation with its laboratory analogue. At water mineralization of 40–50 g/l, Microcoleus chthonoplastes and Phormidium molle play a key role in the formation of the cyanobacterial mat. As water mineralization increases, the diversity of cyanobacteria in the natural mat increases as well, reaches its maximum at 80 g/l NaCl, and decreases at 100 g/l. In the laboratory community, Nodularia sp. prevailed. It was able to form matlike structures within a broad pH range and at a salinity of up to 50 g/l NaCl. As the water mineralization level increased up to 100 g/l or higher, a replacement of the dominant complexes occurred both in the laboratory and natural communities: cyanobacterial species were substituted with green algae.     

Prokaryotic communities of the north-eastern Mongolian soda lakes

We have studied the activity and composition of several geochemically significant physiological groups of bacteria in more than twenty alkaline salt lakes of the north-east Mongolia steppe with water salinity from 3 to 390 g l^?1 and pH values ranging from 9.0 to 10.6. Active and diverse microbial communities have been found in most of the lakes. The methanotrophic bacteria were represented by the Type I members. Among the culturable forms of sulfur-oxidizing bacteria obligately chemolithoautotrophic and haloalkaliphilic representatives of the genera Thioalkalimicrobium and Thioalkalivibrio were detected in the sediments at high numbers (up to 10⁶ cells ml^?1). The largest population of anaerobic phototrophic bacteria was represented by purple sulfur bacteria of the Ectothiorhodospiraceae family. Salinity was the key factor in determining the activity and the composition of the microbial communities. The most diverse and active prokaryotic populations, including aerobic and anaerobic phototrophic, methanogenic, methanotrophic, sulfur-oxidizing, sulfate-reducing and nitrifying bacteria, were found in lakes with salinity less than 60 g l^?1. In hypersaline lakes with a salinity >100 g l^?1, the sulfur cycle remained active due to the activity of extremely halotolerant and alkaliphilic sulfur bacteria, while other important functional groups responsible for nitrification and methane oxidation processes were not detected. Overall, the prokaryotic communities of the Mongolian alkaline salt lakes represent an interesting new example of a diverse community of haloalkaliphilic bacteria well adopted to a broad salinity range.     

Diversity of diazotrophs in the sediments of saline and soda lakes analyzed with the use of the nifH gene as a molecular marker

Phylogenetic analysis of the nifH genes, encoding the Fe protein of the nitrogenase enzymatic complex, was carried out for pure cultures of anoxygenic phototrophic bacteria of diverse origin, as well as for heterotrophic alkaliphilic sulfate reducers isolated from saline and soda lakes. Topology of the nitrogenase tree correlated with that of the 16S rRNA gene tree to a considerable degree, which made it possible to use the nifH gene as a molecular marker for investigation of diazotrophic bacterial communities in sediments of hyper saline and soda lakes. Although diazotrophs were revealed in all environmental samples, their phylogenetic diversity was relatively low. Sulfate-reducing deltaproteobacteria and photo- and chemotrophic gammaproteobacteria were predominant in integrated samples. Analysis of the upper sediment layers revealed predominance of phototrophic diazotrophs of various phyla, including purple sulfur and nonsulfur proteobacteria, green nonsulfur bacteria, heliobacteria, and cyanobacteria. Some phylotypes could not be identified, probably indicating the presence of bacterial groups which have not yet been studied by conventional microbiological techniques.     

Diversity of extremophilic purple phototrophic bacteria in Soap Lake, a Central Washington (USA) Soda Lake

Culture-based and culture-independent methods were used to explore the diversity of phototrophic purple bacteria in Soap Lake, a small meromictic soda lake in the western USA. Among soda lakes, Soap Lake is unusual because it consists of distinct upper and lower water bodies of vastly different salinities, and its deep waters contain up to 175 mM sulfide. From Soap Lake water new alkaliphilic purple sulfur bacteria of the families Chromatiaceae and Ectothiorhodospiraceae were cultured, and one purple non-sulfur bacterium was isolated. Comparative sequence analysis of pufM, a gene that encodes a key photosynthetic reaction centre protein universally found in purple bacteria, was used to measure the diversity of purple bacteria in Soap Lake. Denaturing gradient gel electrophoresis and subsequent phylogenetic analyses of pufMs amplified from Soap Lake water revealed that a significant diversity of purple bacteria inhabit this soda lake. Although close relatives of several of the pufM phylotypes obtained from cultured species could also be detected in Soap Lake water, several other more divergent pufM phylotypes were also detected. It is possible that Soap Lake purple bacteria are major contributors of organic matter into the ecosystem of this lake, especially in its extensive anoxic and sulfidic deep waters.     

The phototrophic community found in Lake Khilganta (an alkaline saline lake located in the southeastern Transbaikal region)

The structure of the phototrophic community found in Lake Khilganta (the Agin-Buryat Autonomous Area), a shallow saline soda lake (depth, 35-45 cm; water mineralization, 45 g/l; alkalinity, 30 mg-equiv/l; pH 9.5) has been studied. The bottom of the lake is covered with a 10- to 15-mm microbial mat, whose basis is formed by the filamentous cyanobacterium Microcoleus chthonoplastes. The mat exhibits pronounced layering and contains a significant amount of minerals. Six zones, which have characteristic colors and consistencies and are composed of intermittent layers, have been identified along the vertical profile. Live phototrophic bacteria have been found in the three upper zones. The bulk of the cyanobacteria is concentrated in the upper zone. In the lower zones, the development of purple bacteria has been observed. The diurnal dynamics of the vertical distribution of phototrophic microorganisms, which results from variations in the physicochemical environmental parameters, is described. Ectothiorhodospira sp. are dominant among the anoxyphotobacteria present. Their number, determined according to the inoculation method, is 10(6)-10(7) cells/ml. The purple bacteria of the genera Allochromatium, Thiocapsa, and Rhodovulum are also present. Experiments with isolated pure cultures have shown that the anoxygenic photosynthetic bacteria of Lake Khilganta are halotolerant and alkalitolerant or alkaliphilic. In liquid enrichment cultures, at pH 9.5, the ratio of anoxyphotobacteria species is close to that observed in the lake. When the pH is increased to 10.4, it is Ectothiorhodospira, which is the most adapted to life under increased mineralization and alkalinity, that predominantly develops. Photosynthetic activity has been observed in the three upper mat zones and constitutes, on average, 1.5 g C/(m2 h); the share of anoxygenic photosynthesis accounts for 75-95% of the total productivity. The main role in sulfide oxidation belongs to the phototrophic anoxyphotobacteria and cyanobacteria. In terms of the physicochemical conditions and structure of the phototrophic community, Lake Khilganta is similar to shallow saline water bodies of marine origin. The main differences consist in the increased alkalinity and in the consequent prevalence of alkaliphilic and alkalitolerant microorganisms and in the absence of representatives of the neutrophilic group of green sulfur bacteria.     

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.

     

MARINE‐DERIVED DINOFLAGELLATES IN ANTARCTIC SALINE LAKES: COMMUNITY COMPOSITION AND ANNUAL DYNAMICS1*[link]

The saline lakes of the Vestfold Hills in Antarctica offer a remarkable natural laboratory where the adaptation of planktonic protists to a range of evolving physiochemical conditions can be investigated. This study illustrates how an ancestral marine community has undergone radical simplification leaving a small number of well‐adapted species. Our objective was to investigate the species composition and annual dynamics of dinoflagellate communities in three saline Antarctic lakes. We observed that dinoflagellates occur year‐round despite extremely low PAR during the southern winter, which suggests significant mixotrophic or heterotrophic activity. Only a small number of dominant dinoflagellate species were found in each lake, in contrast to the species‐rich Southern Ocean from which the lake communities are believed to be derived. We verified that the lake species were representatives of the marine polar dinoflagellate community, and not freshwater species. Polarella glacialis Montresor, Procaccini et Stoecker, a bipolar marine species, was for the first time described in a lake habitat and was an important phototrophic component in the higher salinity lakes. In the brackish lakes, we found a new sibling species to the brackish‐water species Scrippsiella hangoei (J. Schiller) J. Larsen, previously observed only in the Baltic Sea.     

Potential activity of methane and ammonia oxidation by methanotropic communities from soda lakes of the southern Transbaikal

Radioisotopic measurements of the methane consumption by mud samples taken from nine Southern Transbaikal soda lakes (pH 9.5-10.6) showed an intense oxidation of methane in the muds of lakes Khuzhirta, Bumalai Nur, Gorbunka, and Suduntuiskii Torom, with the maximum oxidation rate in the mud of lakes Khuzhirta (33.2 nmol/(ml day)). The incorporation rate of the radioactive label from 14CH4 into 14CO2 was higher than into acid-stable metabolites. Optimum pH values for methane oxidation in water samples were 7-8, whereas mud samples exhibited two peaks of methane oxidation activity (at pH 8.15-9.4 and 5.8-7.0). The majority of samples could oxidize ammonium to nitrites; the oxidation was inhibited by methane. The PCR amplification analysis of samples revealed the presence of genes encoding soluble and particulate methane monooxygenase and methanol dehydrogenase. Three alkaliphilic methanotrophic bacteria of morphotype I were isolated from mud samples in pure cultures, one of which, B5, was able to oxidize ammonium to nitrites at pH 7-11. The data obtained suggest that methanotrophs are widely spread in the soda lakes of Southern Transbaikal, where they actively oxidize methane and ammonium.     

Opposing patterns of zooplankton diversity and functioning along a natural stress gradient: when the going gets tough,the tough get going

Salinity represents a major structuring factor in aquatic habitats which strongly affects species richness. We studied the relationships among species richness, density and phylogenetic diversity of zooplankton communities along a natural salinity gradient in astatic soda pans in the Carpathian Basin (Hungary, Austria and Serbia). Diversity and density showed opposing trends along the salinity gradient. The most saline habitats had communities of one or two species only, with maximum densities well above 1000 ind l^?1. Similarity of communities increased with salinity, with most of the highly saline communities being dominated by one highly tolerant calanoid copepod, Arctodiaptomus spinosus, which was at the same time the only soda‐water specialist. Salinity obviously constrained species composition and resulted in communities of low complexity, where few tolerant species ensure high biomass production in the absence of antagonistic interactions. The pattern suggests that environmental stress may result in highly constrained systems which exhibit high rates of functioning due to these key species, in spite of the very limited species pool.     

Diversity, activity, and abundance of sulfate-reducing bacteria in saline and hypersaline soda lakes

Soda lakes are naturally occurring highly alkaline and saline environments. Although the sulfur cycle is one of the most active element cycles in these lakes, little is known about the sulfate-reducing bacteria (SRB). In this study we investigated the diversity, activity, and abundance of SRB in sediment samples and enrichment cultures from a range of (hyper)saline soda lakes of the Kulunda Steppe in southeastern Siberia in Russia. For this purpose, a polyphasic approach was used, including denaturing gradient gel electrophoresis of dsr gene fragments, sulfate reduction rate measurements, serial dilutions, and quantitative real-time PCR (qPCR). Comparative sequence analysis revealed the presence of several novel clusters of SRB, mostly affiliated with members of the order Desulfovibrionales and family Desulfobacteraceae. We detected sulfate reducers and observed substantial sulfate reducing rates (between 12 and 423 micromol/dm(3) day(-1)) for most lakes, even at a salinity of 475 g/liter. Enrichments were obtained at salt saturating conditions (4 M Na(+)), using H(2) or volatile fatty acids as electron donors, and an extremely halophilic SRB, strain ASO3-1, was isolated. Furthermore, a high dsr gene copy number of 10(8) cells per ml was detected in a hypersaline lake by qPCR. Our results indicate the presence of diverse and active SRB communities in these extreme ecosystems.