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Pelagic microbes have adopted distinct strategies to inhabit the pelagial of lakes and oceans and can be broadly categorized in two groups: free‐living, specialized oligotrophs and patch‐associated generalists or copiotrophs. In this review, we aim to identify genomic traits that enable pelagic freshwater microbes to thrive in their habitat. To do so, we discuss the main genetic differences of pelagic marine and freshwater microbes that are both dominated by specialized oligotrophs and the difference to freshwater sediment microbes, where copiotrophs are more prevalent. We phylogenomically analysed a collection of >7700 metagenome‐assembled genomes, classified habitat preferences on different taxonomic levels, and compared the metabolic traits of pelagic freshwater, marine, and freshwater sediment microbes. Metabolic differences are mainly associated with transport functions, environmental information processing, components of the electron transport chain, osmoregulation and the isoelectric point of proteins. Several lineages with known habitat transitions (Nitrososphaeria, SAR11, Methylophilaceae, Synechococcales, Flavobacteriaceae, Planctomycetota) and the underlying mechanisms in this process are discussed in this review. Additionally, the distribution, ecology and genomic make‐up of the most abundant freshwater prokaryotes are described in details in separate chapters for Actinobacteriota, Bacteroidota, Burkholderiales, Verrucomicrobiota, Chloroflexota, and 'Ca. Patescibacteria'. [ABSTRACT FROM AUTHOR]

Summary: Diplonemids are considered marine protists and have been reported among the most abundant and diverse eukaryotes in the world oceans. Recently we detected the presence of freshwater diplonemids in Japanese deep freshwater lakes. However, their distribution and abundances in freshwater ecosystems remain unknown. We assessed abundance and diversity of diplonemids from several geographically distant deep freshwater lakes of the world by amplicon‐sequencing, shotgun metagenomics and catalysed reporter deposition‐fluorescent in situ hybridization (CARD‐FISH). We found diplonemids in all the studied lakes, albeit with low abundances and diversity. We assembled long 18S rRNA sequences from freshwater diplonemids and showed that they form a new lineage distinct from the diverse marine clades. Freshwater diplonemids are a sister‐group to a marine clade, which are mainly isolates from coastal and bay areas, suggesting a recent habitat transition from marine to freshwater habitats. Images of CARD‐FISH targeted freshwater diplonemids suggest they feed on bacteria. Our analyses of 18S rRNA sequences retrieved from single‐cell genomes of marine diplonemids show they encode multiple rRNA copies that may be very divergent from each other, suggesting that marine diplonemid abundance and diversity both have been overestimated. These results have wider implications on assessing eukaryotic abundances in natural habitats by using amplicon‐sequencing alone. [ABSTRACT FROM AUTHOR]

Abstract: Different bacterioplankton species represent different food quality resources for heterotrophic nanoflagellate (HNF) communities, potentially affecting HNF growth, community dynamics and carbon flow to higher trophic levels. However, our knowledge of such dynamics is still very limited. Here, we describe the results of 11 experiments with natural HNF communities from distinct seasonal phases in two freshwater habitats. The HNF communities were released from predation pressure of zooplankton and incubated with 16 distinct ecologically relevant prey bacterial strains from important Betaproteobacteria genera (Limnohabitans, Polynucleobacter, and Methylopumilus) and one Actinobacteria strain from the Luna 2 cluster. We observed remarkable prey‐ and season‐specific variability in community HNF growth parameters, i.e., doubling time, volumetric gross growth efficiency (GGE), and length of lag phase. All strains, except for the actinobacterium, supported rapid HNF population growth with an average doubling time of 10 h and GGE of 29%. Our analysis revealed that 59% of the variability in flagellate GGE data was explained by the length of lag phase after prey amendments. This indicates a considerable “adaptation time,” during which the predator communities undergo compositional shifts toward flagellate bacterivores best adapted to grow on the offered prey. Importantly, the rapid HNF growth detected on various bacteria tightly corresponds to doubling times reported for fast growing bacterioplankton groups. We propose a conceptual model explaining the tight linkages between rapid bacterial community shifts and succeeding HNF community shifts, which optimize prey utilization rates and carbon flow from various bacteria to the microbial food chain. [ABSTRACT FROM AUTHOR]

While mechanisms of different carbon dioxide (CO2) assimilation pathways in chemolithoautotrohic prokaryotes are well understood for many isolates under laboratory conditions, the ecological significance of diverse CO2 fixation strategies in the environment is mostly unexplored. Six stratified freshwater lakes were chosen to study the distribution and diversity of the Calvin-Benson-Bassham (CBB) cycle, the reductive tricarboxylic acid (rTCA) cycle, and the recently discovered archaeal 3-hydroxypropionate/4-hydroxybutyrate (HP/HB) pathway. Eleven primer sets were used to amplify and sequence genes coding for selected key enzymes in the three pathways. Whereas the CBB pathway with different forms of RubisCO (IA, IC and II) was ubiquitous and related to diverse bacterial taxa, encompassing a wide range of potential physiologies, the rTCA cycle in Epsilonproteobacteria and Chloribi was exclusively detected in anoxic water layers. Nitrifiying Nitrosospira and Thaumarchaeota, using the rTCA and HP/HB cycle respectively, are important residents in the aphotic and (micro-)oxic zone of deep lakes. Both taxa were of minor importance in surface waters and in smaller lakes characterized by an anoxic hypolimnion. Overall, this study provides a first insight on how different CO2 fixation strategies and chemical gradients in lakes are associated to the distribution of chemoautotrophic prokaryotes with different functional traits. [ABSTRACT FROM AUTHOR]

Trap fluid of aquatic carnivorous plants of the genus Utricularia hosts specific microbiomes consisting of commensal pro- and eukaryotes of largely unknown ecology. We examined the characteristics and dynamics of bacteria and the three dominant eukaryotes, i.e. the algae-bearing ciliate Tetrahymena utriculariae (Ciliophora), a green flagellate Euglena agilis (Euglenophyta), and the alga Scenedesmus alternans (Chlorophyta), associated with the traps of Utricularia reflexa. Our study focused on ecological traits and life strategies of the highly abundant ciliate whose biomass by far exceeds that of other eukaryotes and bacteria independent of the trap age. The ciliate was the only bacterivore in the traps, driving rapid turnover of bacterial standing stock. However, given the large size of the ciliate and the cell-specific uptake rates of bacteria we estimated that bacterivory alone would likely be insufficient to support its apparent rapid growth in traps. We suggest that mixotrophy based on algal symbionts contributes significantly to the diet and survival strategy of the ciliate in the extreme (anaerobic, low pH) trap-fluid environment. We propose a revised concept of major microbial interactions in the trap fluid where ciliate bacterivory plays a central role in regeneration of nutrients bound in rapidly growing bacterial biomass. [ABSTRACT FROM AUTHOR]

The ecological relevance and factors shaping dynamics of Limnohabitans sp. have been largely studied by fluorescence in situ hybridization with a 16S rRNA probe targeting the R-BT group (lineages LimBCDE), but not lineage LimA. Consequently, ecology and distribution of LimA remained unknown. We developed a double hybridization strategy using a novel 23S rRNA probe specifically targeting LimA and LimE that in combination with the existing R-BT probe can discriminate LimA populations. This technique was applied for more than 1000 samples from 46 freshwater sites including long-term data sets from oligo-mesotrophic Lake Zurich, CH and meso-eutrophic Římov reservoir, CZ. LimA was ubiquitously distributed and highly abundant. Observed ecological preferences of LimA in Lake Zurich were in general similar to already reported for Limnohabitans with highest numbers in surface waters during growing seasons. Three times higher densities of LimA were detected in Římov reservoir, where they were significantly more abundant at the riverine zone especially after flood events that introduced fresh terrestrial DOM (dissolved organic matter). Moreover, statistical analyses of biological and physicochemical parameters obtained from small dynamic water bodies confirmed a correspondence between LimA and allochthonous DOM, in opposite to R-BT that was more related to algal primary production. [ABSTRACT FROM AUTHOR]

We studied the seasonal growth potential of opportunistic bacterial populations in Lake Zurich ( Switzerland) by a series of grazer-free dilution culture assays. Pronounced shifts in the composition of the bacterial assemblages were observed within one doubling of total cell numbers, from initially abundant A ctinobacteria to other fast-growing microbial lineages. Small populations with growth potentials far above community average were detected throughout the year with striking seasonal differences in their respective taxonomic affiliations. Members of C ytophaga- F lavobacteria ( CF) were disproportionally proliferating only during phytoplankton blooms in spring and summer, while B eta- and G ammaproteobacteria showed superior growth at all other occasions. Growth rates of A lphaproteobacteria and esp. S phingomonadaceae were significantly correlated to water temperatures and were far above community average in summer. Within the genus F lavobacterium, two species-like populations showed a tendency for fast growth in most experiments, while four others were exclusively proliferating either during a spring or during a summer phytoplankton bloom. Their high growth potentials but low in situ abundances hint at a tight control by bacterivorous grazers and at a consequently accelerated carbon flux to higher trophic levels. [ABSTRACT FROM AUTHOR]

While typical freshwater and marine bacteria have been found to co-occur in brackish habitats, it is unknown if they are active members of the local bacterial assemblages or if their presence is the result of passive transport only. We followed the seasonal dynamics of typical freshwater bacteria (R-BT lineage of Betaproteobacteria; Ac1 Actinobacteria; LD12 Alphaproteobacteria) and of marine SAR11 Alphaproteobacteria in the brackish water of the Gulf of Gdańsk (southern Baltic Sea), and we assessed their incorporation of thymidine and leucine at three distinct environmental conditions. The temporal development of bacteria was driven not only by local conditions but also by phenomena resulting from the dynamic hydrology of the site. Both temperature and salinity were important factors influencing bacterial community composition, as reflected by the clear distinction of three assemblages in spring and summer and during a period of enhanced freshwater influx. During spring, high proportions of R-BT Betaproteobacteria and Ac1 Actinobacteria incorporated the radiolabeled tracers, and all three freshwater lineages were most active during the subsequent phase of low salinity. The summer period was characterized by highest abundances of Ac1 Actinobacteria and of both alphaproteobacterial lineages. All studied freshwater lineages were active members of the brackish water communities at specific environmental conditions, including LD12 Alphaproteobacteria, which have so far been considered to thrive exclusively in freshwater habitats. By contrast, the presence of the typical marine SAR11 bacteria seemed to result from passive inflow with more saline waters from the Baltic proper. [ABSTRACT FROM AUTHOR]

Summary The vernal successions of phytoplankton, heterotrophic nanoflagellates (HNF) and viruses in temperate lakes result in alternating dominance of top-down and bottom-up factors on the bacterial community. This may lead to asynchronous blooms of bacteria with different life strategies and affect the channelling of particular components of the dissolved organic matter (DOM) through microbial food webs. We followed the dynamics of several bacterial populations and of other components of the microbial food web throughout the spring phytoplankton bloom period in a pre-alpine lake, and we assessed bacterial uptake patterns of two constituents of the labile DOM pool ( N-acetyl-glucosamine [NAG] and leucine). There was a clear genotypic shift within the bacterial assemblage, from fast growing Cytophaga-Flavobacteria (CF) affiliated with Fluviicola and from Betaproteobacteria (BET) of the Limnohabitans cluster to more grazing resistant AcI Actinobacteria (ACT) and to filamentous morphotypes. This was paralleled by successive blooms of viruses and HNF. We also noted the transient rise of other CF (related to Cyclobacteriaceae and Sphingobacteriaceae) that are not detected by fluorescence in situ hybridization with the general CF probe. Both, the average uptake rates of leucine and the fractions of leucine incorporating bacteria were approximately five to sixfold higher than of NAG. However, the composition of the NAG-active community was much more prone to genotypic successions, in particular of bacteria with different life strategies: While 'opportunistically' growing BET and CF dominated NAG uptake in the initial period ruled by bottom-up factors, ACT constituted the major fraction of NAG active cells during the subsequent phase of high predation pressure. This indicates that some ACT could profit from a substrate that might in parts have originated from the grazing of protists on their bacterial competitors. [ABSTRACT FROM AUTHOR]

We investigated the diversity of planktonic Betaproteobacteria and the seasonal population changes of betaproteobacterial taxa in an oligo-mesotrophic lake (Piburger See, Austria). Focus was put on the vertical distribution of the investigated populations and on differences between their respective cell fractions with apparent amino acid incorporation. On average, 66% of betaproteobacterial cells and 73% of their diversity could be attributed to four clades within three lineages that were further analysed by fluorescence in situ hybridization. The numbers of bacteria from the R-BT subclade of the beta I lineage and from the PnecB subgroup of the beta II lineage were rather constant throughout the water column. In contrast, members of another subgroup of beta II (PnecC) and bacteria related to Methylophilus (beta IV) were particularly numerous in the oxygen-depleted zone. In general, only moderate seasonal changes in abundance were observed in the upper water layers, whereas there was a clear relationship between decreasing oxygen levels and the rise of bacteria from the PnecC and beta IV clades in deeper strata. On average, almost 80% of beta I bacteria, but < 15% of cells from the beta IV clade, showed amino acid incorporation. Our results suggest that the studied populations occupy distinct vertical and ecophysiological niches in Piburger See. [ABSTRACT FROM AUTHOR]

The pelagic zone of large, deep freshwater lakes features pronounced horizontal and vertical gradients of physicochemical parameters, which in turn might allow for a nonuniform occurrence of specifically adapted bacterial taxa. We, therefore, studied the spatial distribution patterns of different heterotrophic bacteria, picocyanobacteria, and the dominant primary producer, the filamentous cyanobacterium Planktothrix rubescens, in a large, canyon-shaped, prealpine lake (Lake Zurich, Switzerland), in six vertical profiles along a 21.7-km longitudinal transect. Highest total densities and proportions of cells with high nucleic acid content were in the warm epilimnion and the hypoxic zone of the hypolimnion. P. rubescens formed a dense layer in the metalimnion throughout the lake, whereas picocyanobacteria populated the water layers above. The epilimnion was mainly inhabited by ultramicrobacteria related to the LD12-lineage of Alphaproteobacteria and to Actinobacteria; the latter group preferred the shallow regions. Cytophaga-Flavobacteria, in particular a population related to Fluviicola sp. were more frequent in and below the layer of maximal P. rubescens abundances. Betaproteobacteria, on the other hand, were highly abundant in the epi- and hypolimnion, but not in the P. rubescens layer. Four betaproteobacterial subpopulations with contrasting longitudinal and/or vertical habitat preferences were distinguished: putatively methylotrophic bacteria of the LD28 lineage (beta IV) preferentially inhabited the hypolimnion, Polynucleobacter acidiphobus was found throughout the epilimnion, Limnohabitans (R-BT065) more in the shallow regions of the lake, and Polynucleobacter necessarius ssp. asymbioticus only in hypoxic waters. Our results stress the potential importance of spatial niche differentiation in freshwater bacterioplankton. [ABSTRACT FROM AUTHOR]

We explored the seasonal and vertical patterns of cell numbers, biomass, and amino acid incorporation of three major phylogenetic groups of the bacterioplankton in an oligomesotrophic alpine lake. Throughout the year the numerically dominant bacteria in the oxygenated epilimnion were small Actinobacteria. The timing and vertical location of their spring peak coincided with that of the heterotrophic flagellates (HNF). A second maximum of the relative proportions of Actinobacteria occurred during an autumn bloom of the diatom Asterionella formosa. Betaproteobacteria were negatively related to HNF abundances and formed maximal numbers and biomass during the summer stratification in the increasingly oxygen-depleted hypolimnion. Members of the Cytophaga-Flavobacteria subclade (CF) of Bacteroidetes preferably inhabited the completely anoxic zone of the hypolimnion. The numerically dominant Actinobacteria contributed only little to total bacterial biomass, whereas Betaproteobacteria and the comparatively rare (albeit large) CF represented the bulk of the bacterial carbon pool. Amino acid incorporation was mainly observed in Actinobacteria and Betaproteobacteria. Actinobacteria showed a clear preference for amino acids other than leucine. These bacteria represented the most active biomass (with respect to amino acid uptake), whereas the opposite was the case for CF. The contrasting vertical and seasonal patterns suggest that the three phylogenetic groups to some extent represent different functional compartments of the bacterioplankton. [ABSTRACT FROM AUTHOR]