The lion's share of bacteria in various environments cannot be cloned in the laboratory and thus cannot be sequenced using existing technologies. A major goal of single-cell genomics is to complement gene-centric metagenomic data with whole-genome assemblies of uncultivated organisms. Assembly of single-cell data is challenging because of highly non-uniform read coverage as well as elevated levels of sequencing errors and chimeric reads. We describe SPAdes, a new assembler for both single-cell and standard (multicell) assembly, and demonstrate that it improves on the recently released E+V-SC assembler (specialized for single-cell data) and on popular assemblers Velvet and SoapDeNovo (for multicell data). SPAdes generates single-cell assemblies, providing information about genomes of uncultivatable bacteria that vastly exceeds what may be obtained via traditional metagenomics studies. SPAdes is available online ( http://bioinf.spbau.ru/spades ). It is distributed as open source software.

SPAdes, a new genome assembly algorithm and its applications to single-cell sequencing ( 7th Annual SFAF Meeting, 2012)

Microbial communities are essential components of aquatic ecosystems through their contribution to food web dynamics and biogeochemical processes. Aquatic microbial diversity is immense and a general challenge is to understand how metabolism and interactions of single organisms shape microbial community dynamics and ecosystem-scale biogeochemical transformations. Metagenomic approaches have developed rapidly, and proven to be powerful in linking microbial community dynamics to biogeochemical processes. In this review, we provide an overview of metagenomic approaches, followed by a discussion on some recent insights they have provided, including those in this special issue. These include the discovery of new taxa and metabolisms in aquatic microbiomes, insights into community assembly and functional ecology as well as evolutionary processes shaping microbial genomes and microbiomes, and the influence of human activities on aquatic microbiomes. Given that metagenomics can now be considered a mature technology where data generation and descriptive analyses are relatively routine and informative, we then discuss metagenomic-enabled research avenues to further link microbial dynamics to biogeochemical processes. These include the integration of metagenomics into well-designed ecological experiments, the use of metagenomics to inform and validate metabolic and biogeochemical models, and the pressing need for ecologically relevant model organisms and simple microbial systems to better interpret the taxonomic and functional information integrated in metagenomes. These research avenues will contribute to a more mechanistic and predictive understanding of links between microbial dynamics and biogeochemical cycles. Owing to rapid climate change and human impacts on aquatic ecosystems, the urgency of such an understanding has never been greater.

/pdf/linking-metagenomics-to-aquatic-microbial-ecology-and-1st6q2tcsw.pdf

Linking metagenomics to aquatic microbial ecology and biogeochemical cycles

Microcystis is a cyanobacterium that forms toxic blooms in freshwater ecosystems around the world. Biological variation among taxa within the genus is apparent through genetic and phenotypic differences between strains and via the spatial and temporal distribution of strains in the environment, and this fine-scale diversity exerts strong influence over bloom toxicity. Yet we do not know how varying traits of Microcystis strains govern their environmental distribution, the tradeoffs and links between these traits, or how they are encoded at the genomic level. Here we synthesize current knowledge on the importance of diversity within Microcystis and on the genes and traits that likely underpin ecological differentiation of taxa. We briefly review spatial and environmental patterns of Microcystis diversity in the field and genetic evidence for cohesive groups within Microcystis. We then compile data on strain-level diversity regarding growth responses to environmental conditions and explore evidence for variation of community interactions across Microcystis strains. Potential links and tradeoffs between traits are identified and discussed. The resulting picture, while incomplete, highlights key knowledge gaps that need to be filled to enable new models for predicting strain-level dynamics, which influence the development, toxicity, and cosmopolitan nature of Microcystis blooms. This article is protected by copyright. All rights reserved.

https://sfamjournals.onlinelibrary.wiley.com/doi/pdf/10.1111/1462-2920.15615

The genetic and ecophysiological diversity of Microcystis.

Cyanobacterial blooms have substantial direct and indirect negative impacts on freshwater ecosystems including releasing toxins, blocking light needed by other organisms, and depleting oxygen. There is growing concern over the potential for climate change to promote cyanobacterial blooms, as the positive effects of increasing lake surface temperature on cyanobacterial growth are well documented in the literature; however, there is increasing evidence that cyanobacterial blooms are also being initiated and persisting in relatively cold‐water temperatures (< 15°C), including ice‐covered conditions. In this work, we provide evidence of freshwater cold‐water cyanobacterial blooms, review abiotic drivers and physiological adaptations leading to these blooms, offer a typology of these lesser‐studied cold‐water cyanobacterial blooms, and discuss their occurrence under changing climate conditions.

https://openpub.fmach.it/bitstream/10449/79017/1/2023%20LOL%20Obertegger.pdf

Blooms also like it cold

The amazing morphological diversity of phytoplankton has to be considered an evolutionarily driven compendium of strategies to cope with the strong variability and unpredictability of the pelagic environment. Phytoplankton collects unicellular and colonial photosynthetic organisms adapted to live in apparent suspension in turbulent water masses. Turbulence represents a key driver of phytoplankton dynamics in all aquatic ecosystems and phytoplankton morphological variability is the evolutionary response of this group of photosynthetic organisms to the temporal and spatial scales of variability of turbulence. This paper reviews the existing literature on the effects exerted by turbulence on phytoplankton populations and is aimed at showing how deeply turbulence contributes to the shape and size structure of phytoplankton assemblages. Our aim is to explore how turbulence governs phytoplankton access to resources and, at the same time, how the shape and size structure of phytoplankton represent the evolutionary way in which this group of organisms has optimised its survival in the highly dynamic aquatic environment. The paper is intended to serve as an homage to the (phytoplankton) ecologist Colin S. Reynolds. His life-long work highlighted how profoundly the ecology of phytoplankton depends on the physical constraints governing the movements of the water masses in which phytoplankton evolved and lives.

https://link.springer.com/content/pdf/10.1007/s10750-020-04217-x.pdf

Life in suspension and its impact on phytoplankton morphology: an homage to Colin S. Reynolds

Bacteria play key roles in the function and diversity of aquatic systems, but aside from study of specific bloom systems, little is known about the diversity or biogeography of bacteria associated with harmful cyanobacterial blooms (cyanoHABs). CyanoHAB species are known to shape bacterial community composition and to rely on functions provided by the associated bacteria, leading to the hypothesized cyanoHAB interactome, a coevolved community of synergistic and interacting bacteria species, each necessary for the success of the others. Here, we surveyed the microbiome associated with Microcystis aeruginosa during blooms in 12 lakes spanning four continents as an initial test of the hypothesized Microcystis interactome. We predicted that microbiome composition and functional potential would be similar across blooms globally. Our results, as revealed by 16S rRNA sequence similarity, indicate that M. aeruginosa is cosmopolitan in lakes across a 280° longitudinal and 90° latitudinal gradient. The microbiome communities were represented by a wide range of operational taxonomic units and relative abundances. Highly abundant taxa were more related and shared across most sites and did not vary with geographic distance, thus, like Microcystis, revealing no evidence for dispersal limitation. High phylogenetic relatedness, both within and across lakes, indicates that microbiome bacteria with similar functional potential were associated with all blooms. While Microcystis and the microbiome bacteria shared many genes, whole-community metagenomic analysis revealed a suite of biochemical pathways that could be considered complementary. Our results demonstrate a high degree of similarity across global Microcystis blooms, thereby providing initial support for the hypothesized Microcystis interactome.

https://aslopubs.onlinelibrary.wiley.com/doi/epdf/10.1002/lno.11361

The global Microcystis interactome

Global warming and eutrophication contribute to the worldwide increase in cyanobacterial blooms, and the level of cyanobacterial biomass is strongly associated with rises in methane emissions from surface lake waters. Hence, methane metabolizing microorganisms may be important for modulating carbon flow in cyanobacterial blooms. Here, we surveyed methanogenic and methanotrophic communities associated with floating Microcystis aggregates in ten lakes spanning four continents, through sequencing of 16S rRNA and functional marker genes. Methanogenic archaea (mainly Methanoregula and Methanosaeta) were detectable in five of the ten lakes and constituted the majority (~50 - 90%) of the archaeal community in these lakes. Three of the ten lakes contained relatively more abundant methanotrophs than the other seven lakes, with the methanotrophic genera Methyloparacoccus, Crenothrix, and an uncultured species related to Methylobacter dominating and nearly exclusively found in each of those three lakes. These three are among the five lakes in which methanogens were observed. OTU richness and abundance of methanotrophs were strongly positively correlated with those of methanogens, suggesting that their activities may be coupled. These Microcystis-aggregate-associated methanotrophs may be responsible for a hitherto overlooked sink for methane in surface freshwaters, and their co-occurrence with methanogens sheds light on the methane cycle in cyanobacterial aggregates.

Global co-occurrence of methanogenic archaea and methanotrophic bacteria in Microcystis aggregates.

Rnf is a membrane protein complex that has been shown to be important in energy conservation. Here, Desulfovibrio alaskensis G20 and Rnf mutants of G20 were grown with different electron donor and acceptor combinations to determine the importance of Rnf in energy conservation and the type of ion gradient generated. The addition of the protonophore TCS strongly inhibited lactate-sulfate dependent growth whereas the sodium ionophore ETH2120 had no effect, indicating a role for the proton gradient during growth. Mutants in rnfA and rnfD were more sensitive to the protonophore at 5 µM than the parental strain, suggesting the importance of Rnf in the generation of a proton gradient. The electrical potential (ΔΨ), ΔpH and proton motive force were lower in the rnfA mutant than in the parental strain of D.alaskensis G20. These results provide evidence that the Rnf complex in D. alaskensis functions as a primary proton pump whose activity is important for growth.

The role of Rnf in ion gradient formation in Desulfovibrio alaskensis.

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The global Microcystis interactome

Global co-occurrence of methanogenic archaea and methanotrophic bacteria in Microcystis aggregates.

The role of Rnf in ion gradient formation in Desulfovibrio alaskensis.