Phytoplankton is the basis of the food web and an indicator of environmental change in aquatic ecosystems. Phytoplankton assessment uses microscopy, which estimates the composition, absolute abundance and biomass of taxa, and metabarcoding, which estimates the composition, high richness and diversity, and relative abundance of taxa. Problems remain with the consistency of results from these two methods and the quantification of metabarcoding. Using 18S rRNA metabarcoding and microscopy we compared the relative or absolute abundance and biomass of phytoplankton taxa (class or genus/species) in the south basin of Lake Baikal in spring over 3 years. Absolute abundance/biomass of phytoplankton taxa estimated by metabarcoding was obtained by combining relative abundances of amplicon sequence variants (ASV produced by error-correcting method) derived from the V8-V9 region of 18S rRNA gene amplicon sequencing (primers were used that accurately represented the mean relative abundance of different microalgae) with total or class-specific abundance/biomass of phytoplankton estimated by light microscopy. Many Spearman correlations were found between relative (non- or clr-transformed) or absolute abundances/biomasses of the same phytoplankton classes or genus/species. Correlation coefficients were higher between absolute values than between relative values. Correlations were found between relative or absolute abundance/biomass, estimated by both methods, of the classes Bacillariophyceae, Coscinodiscophyceae, Mediophyceae, Chrysophyceae, Cryptophyceae, and Chlorophyceae, but not Dinophyceae and Trebouxiophyceae. Correlations were found between relative or absolute abundance/biomass of dominant species and ASVs of diatoms (Ulnaria, Aulacoseira, Stephanodiscus), Chrysophyceae (Dinobryon), and Cryptophyceae (Cryptomonas). Thus, the consistency of the dynamics of the relative or absolute abundance/biomass of phytoplankton taxa estimated by the two methods was revealed. Absolute abundances/biomasses of taxa estimated by metabarcoding in combination with microscopy improve the accuracy of metabarcoding-based ecological assessment.

Water bodies located in the north-east of Yakutia during the period of ice cover were studied. High-throughput sequencing of 16S rRNA gene libraries revealed a high taxonomic diversity of microbial communities in the studied ecotopes. The majority of the under-ice communities was represented by Proteobacteria, Actinobacteriota, Cyanobacteria, Verrucomicrobiota, Bacteroidota, and Planctomycetota. The difference in community diversity between deep-water lakes Labynkyr, Vodorazdelnoye, and Ulu, the source of the Labynkyr River, and shallow-water lakes Myamichi and Mertvoye, the mouth of the Labynkyr River, is shown.

Metabarcoding using high throughput sequencing of amplicons of the 18S rRNA gene is one of the widely used methods for assessing the diversity of microeukaryotes in various ecosystems. We investigated the effectiveness of the V4 and V8-V9 regions of the 18S rRNA gene by comparing the results of metabarcoding microeukaryotic communities using the DADA2 (ASV), USEARCH-UNOISE3 (ZOTU), and USEARCH-UPARSE (OTU with 97% similarity) algorithms. Both regions showed similar levels of genetic variability and taxa identification accuracy. Richness for DADA2 datasets of both regions was lower than for UNOISE3 and UPARSE datasets, which is due to more accurate error correction in amplicons. Microeukaryotic communities (autotrophs and heterotrophs) structure identified using both regions showed a significant relationship with phytoplankton (autotrophs) communities structure based on microscopy in a seasonal freshwater sample series. The strongest relationship was found between the phytoplankton species and V8-V9 ASVs produced by DADA2.

Microorganisms exhibit seasonal succession governed by physicochemical factors and interspecies interactions, yet drivers of this process in different environments remain to be determined. We used high-throughput sequencing of 16S rRNA and 18S rRNA genes to study seasonal dynamics of bacterial and microeukaryotic communities at pelagic site of Lake Baikal from spring (under-ice, mixing) to autumn (direct stratification). The microbial community was subdivided into distinctive coherent clusters of operational taxonomic units (OTUs). Individual OTUs were consistently replaced during different seasonal events. The coherent clusters change their contribution to the microbial community depending on season. Changes of temperature, concentrations of silicon, and nitrates are the key factors affected the structure of microbial communities. Functional prediction revealed that some bacterial or eukaryotic taxa that switched with seasons had similar functional properties, which demonstrate their functional redundancy. We have also detected specific functional properties in different coherent clusters of bacteria or microeukaryotes, which can indicate their ability to adapt to seasonal changes of environment. Our results revealed a relationship between seasonal succession, coherency, and functional features of freshwater bacteria and microeukaryotes.

Phytoplankton and bacterioplankton play a key role in carbon cycling of aquatic ecosystems. In this study, we found that co-occurrence patterns between different types of phytoplankton, bacterioplankton, and environmental parameters in Lake Baikal during spring were different over the course of three consecutive years. The composition of phytoplankton and bacterial communities was investigated using microscopy and 16S rRNA gene pyrosequencing, respectively. Non-metric multidimensional scaling (NMDS) revealed a relationship between the structure of phytoplankton and bacterial communities and temperature, location, and sampling year. Associations of bacteria with diatoms, green microalgae, chrysophyte, and cryptophyte were identified using microscopy. Cluster analysis revealed similar correlation patterns between phytoplankton abundance, number of attached bacteria, ratio of bacteria per phytoplankton cell and environmental parameters. Positive and negative correlations between different species of phytoplankton, heterotrophic bacteria and environmental parameters may indicate mutualistic or competitive relationships between microorganisms and their preferences to the environment.

Algal-bacterial associations were studied in unialgal xenic cultures of Synedra acus subsp. radians, Asterionella formosa and Fragillaria crotonensis planktonic diatoms from Lake Baikal, using epifluorescent and scanning electron microscopy. It was found that rod- and ovoid-shaped bacteria colonized cell walls of diatoms. Cloning and sequencing of fragments of 16S rRNA gene in diatom cultures revealed members of Gammaproteobacteria (Pseudomonas sp.), Betaproteobacteria (Janthinobacterium sp., Hydrogenophaga sp., Methylophilus sp.), Bacteroidetes (Flavobacterium sp., Pedobacter sp.), and Acinobacteria (Nocardioides sp.).

The pelagic zone of Lake Baikal is an ecological niche where phytoplankton bloom causes increasing microbial abundance in spring which plays a key role in carbon turnover in the freshwater lake. Co-occurrence patterns revealed among different microbes can be applied to predict interactions between the microbes and environmental conditions in the ecosystem. We used 454 pyrosequencing of 16S rRNA and 18S rRNA genes to study bacterial and microbial eukaryotic communities and their co-occurrence patterns at the pelagic zone of Lake Baikal during a spring phytoplankton bloom. We found that microbes within one domain mostly correlated positively with each other and are highly interconnected. The highly connected taxa in co-occurrence networks were operational taxonomic units (OTUs) of Actinobacteria, Bacteroidetes, Alphaproteobacteria, and autotrophic and unclassified Eukaryota which might be analogous to microbial keystone taxa. Constrained correspondence analysis revealed the relationships of bacterial and microbial eukaryotic communities with geographical location.

Pyrosequencing of amplicons V3-V4 of the 16S rRNA gene of 30 samples from the photic layer across the three basins of Lake Baikal has revealed 44 958 sequences of the domain Bacteria. The most representative phylums among all classified sequences in the samples were Actinobacteria, Bacteroidetes, Verrucomicrobia, Proteobacteria, Acidobacteria, and Cyanobacteria. As much as 38.5% of the total number of sequences has been determined to the level of genus. The taxonomic composition of bacterial communities was similar regardless of some differences in the composition and abundance of spring phytoplankton. Closely related or identical sequences of bacteria inhabiting the epilimnion of other lakes worldwide have been discovered for most sequences. This confirms the similarity of microbial communities in freshwater ecosystems.

Background: Interest in studies concerning the effect of organic carbon sources on the growth of diatoms is largely aimed at subsequent physiological changes occurring in their cells. There are no data on structural changes in the cytoplasm and their relationship with changes in the composition of fatty acids in the course of mixotrophic culturing of freshwater diatoms. To elucidate the role of lipids in the growth of diatom cells in autotrophic and mixotrophic cultures, it is necessary to obtain information on the distribution of fatty acids among intracellular compartments and on possible ultrastructural changes in the cells.Results: In this study, the results demonstrated that Synedra acus cells cultured in the presence of 80 mM glycerol contained lipid bodies of increased size, while cells from cultures supplemented with 40 mM glucose accumulated polysaccharide (chrysolaminarin) granules. An increase in the relative amounts of palmitic and stearic acids was revealed at the exponential growth phase of S. acus in the medium with 80 mM glycerol, which was indicative of the accumulation of fatty acids contained in triacylglycerols.Conclusions: Synedra acus subsp. radians have an ability to proliferate in the presence of high concentrations of organic substances and their transport into cells is evidence for its high adaptation potential, which, along with other factors, accounts for the dominance of this diatom in the spring-summer plankton of the oligotrophic Lake Baikal.

Production of axenic cultures of diatoms is a highly relevant task, as these widespread organisms are objects of research in different fields of biology, from ecology to whole‐genome studies, and promising as producers of various organic molecules, including biologically active substances. However, culturing of diatoms isolated from natural communities is accompanied by the development of associated microorganisms. We propose a procedure for establishing an axenic culture of the freshwater diatom Synedra asus subsp. radians (Kütz.) Skabitsch. from Lake Baikal. Such a culture has been produced and maintained by subculturing in our laboratory. The procedure involves filtration (to remove free bacteria), detergent treatment (to remove microorganisms associated with diatom frustules), selection of effective antibiotics and their concentrations nontoxic for diatoms (by the disk diffusion method), cell treatment with the selected antibiotic (ciprofloxacin), repeated filtration, and monoclonal culturing of diatom cells. Axenity of the culture was verified microscopically (after DAPI staining) and by molecular biological methods.