Technical challenges in metatranscriptomic studies applied to the bacterial communities of freshwater ecosystems.

Metatranscriptome analysis relates to the transcriptome of microbial communities directly sampled in the environment. Accessing the mRNA pool in natural bacterial communities presents some technical challenges such as the RNA extraction, rRNA depletion, and the choice of the high-throughput sequencing technique. The lack of technical details in scientific articles is a major problem to correctly obtained mRNA from a microbial community and thus the corresponding sequencing data. In our study, we present the methodological procedure that was developed in order to access to the metatranscriptome of the microbial communities during two cyanobacterial blooms successively occurring in a freshwater eutrophic lake. Each procedure step was detailed and discussed with regard to the choices and difficulties encountered and to the recent literature. Finally, the two major limits for metatranscriptomic approaches targeting bacterial communities from natural environments were (i) the removal of rRNA in order to increase the putative mRNA reads number after sequencing, and (ii) for most of the bacterial communities living in natural environments, the lack of reference genomes in databases that leads to the non-assignation of numerous reads. Once these challenges overcome, we managed to access putative mRNA of dominant species, i.e. cyanobacteria (from 6 to 72 % of mRNA assigned), and of the surrounding bacteria (from 1 to 5 % of mRNA assigned).

Influence of local and global environmental parameters on the composition of cyanobacterial mats in a tropical lagoon.

Cyanobacteria-dominated microbial mat communities thrive widely and year round in coral reefs and tropical lagoons, with periodic massive development of benthic blooms. We studied the diversity and spatiotemporal variation of the cyanobacterial dominance in mats of the shallow lagoon of La Réunion Island in the Indian Ocean by means of denaturing gradient gel electrophoresis and cloning-sequencing approaches targeting the 16S rRNA gene, combined with macromorphological and micromorphological characterization of corresponding phenotypes. The mat-forming cyanobacteria were highly diversified with at least 67 distinct operational taxonomic units identified in the lagoon, encompassing the entire morphological spectrum of the phylum Cyanobacteria, but with striking dominance of Oscillatoriales and Nostocales. It appeared also that selective pressures acting at different geographical scales have an influence on the structure and composition of these mats dominated by cyanobacteria. First, large changes were observed in their diversity and composition in relation to local changes occurring in their environment. Second, from the data obtained on the richness and composition of the mats and from the comparison with similar studies in the world, tropical mats seem to display wider cyanobacterial richness than in temperate and cold areas. Moreover, these tropical mats share more species with mats in other tropical regions than with those in temperate and cold climatic regions, suggesting that marine cyanobacteria in biofilms and mats display a biogeographic structure.

Planctomycetes in lakes: poor or strong competitors for phosphorus?

Experiments were conducted with water samples from two perialpine lakes with differing eutrophication status in order to examine the effects of inorganic-nutrient amendments (nitrogen as NO3(-) or NH4(+) and phosphorus as PO4(3-)) on the dynamics, structure, and composition of Planctomycetes and to test the hypothesis that the community structure of Planctomycetes members and that of the other bacteria (without Planctomycetes, here referred to as bacteria-wP, the most represented groups within the community) would be similarly impacted by nutrient additions. Initial samples were characterized by high total nitrogen-to-total phosphorus ratios (range, 39 to 55), suggesting P rather than N was the limiting nutrient for microbial communities. Consistent with this, P additions stimulated phytoplankton growth and affected the community structure of bacteria-wP but, surprisingly, not that of Planctomycetes. N additions did not significantly affect the community structures of bacteria-wP and Planctomycetes or the Planctomycetes phylotype composition. The estimated generation time of Planctomycetes was 123 h. These findings could suggest that the generally well-accepted statement that bacteria (as a whole) are superior to phytoplankton in the ability to obtain phosphorus under P limitation might actually not hold for Planctomycetes. Planctomycetes might be poor competitors for P that do not respond quickly to the nutrient supply, which may help explain why their abundance is low in aquatic systems. The alternative view that Planctomycetes could be strong competitors for P (storing it) is also discussed. Our findings highlight the need for further studies examining Planctomycetes-phosphorus relationships in aquatic ecosystems.

The phyto-bacterioplankton couple in a shallow freshwater ecosystem: Who leads the dance?

Bloom-forming phytoplankton dynamics are still unpredictable, even though it is known that several abiotic factors, such as nutrient availability and temperature, are key factors for bloom development. We investigated whether biotic factors, i.e. the bacterioplankton composition (via 16SrDNA metabarcoding), were correlated with phytoplankton dynamics, through a weekly monitoring of a shallow lake known to host recurrent cyanobacterial blooms. We detected concomitant changes in both bacterial and phytoplankton community biomass and diversity. During the bloom event, a significant decrease in phytoplankton diversity, was detected, with a first co-dominance of Ceratium, Microcystis and Aphanizomenon, followed by a co-dominance of the two cyanobacterial genera. In the same time, we observed a decrease of the particle-associated (PA) bacterial richness and the emergence of a specific bacterial consortium that was potentially better adapted to the new nutritional niche. Unexpectedly, changes in PA bacterial communities occurred just before the development the emergence of the phytoplanktonic bloom and the associated modification of the phytoplanktonic community composition, suggesting that changes in environmental conditions leading to the bloom, were first sensed by the bacterial PA community. This last was quite stable throughout the bloom event, even though there were changes in the blooming species, suggesting that the association between cyanobacterial species and bacterial communities may not be as tight as previously described for monospecific blooming communities. Finally, the dynamics of the free-living (FL) bacterial communities displayed a different trajectory from those of the PA and phytoplankton communities. This FL communities can be viewed as a reservoir for bacterial recruitment for the PA fraction. Altogether, these data also highlight s that the spatial organization within these different microenvironments in the water column is a relevant factor in the structuring of these communities.

Assessment of cyanotoxins in water and fish in an African freshwater lagoon (Lagoon Aghien, Ivory Coast) and the application of WHO guidelines.

In comparison with northern countries, limited data are available on the occurrence and potential toxicity of cyanobacterial blooms in lakes and ponds in sub-Saharan countries. With the aim of enhancing our knowledge on cyanobacteria and their toxins in Africa, we performed a 17-month monitoring of a freshwater ecosystem, Lagoon Aghien (Ivory Coast), which is used for multiple practices by riverine populations and for drinking water production in Abidjan city. The richness and diversity of the cyanobacterial community were high and displayed few variations during the entire survey. The monthly average abundances ranged from 4.1 × 104 to 1.8 × 105 cell mL-1, with higher abundances recorded during the dry seasons. Among the five cyanotoxin families analyzed (anatoxin-a, cylindrospermopsin, homoanatoxin, microcystins, saxitoxin), only microcystins (MC) were detected with concentrations ranging from 0 to 0.364 µg L-1 in phytoplankton cells, from 32 to 1092 µg fresh weight (FW) kg-1 in fish intestines, and from 33 to 383 µg FW kg-1 in fish livers. Even if the MC concentrations in water and fish are low, usually below the thresholds defined in WHO guidelines, these data raise the issue of the relevance of these WHO guidelines for sub-Saharan Africa, where local populations are exposed throughout the year to these toxins in multiple ways.

Fish gut-associated bacterial communities in a tropical lagoon (Aghien lagoon, Ivory Coast).

Aghien lagoon (Ivory Coast) is a eutrophic freshwater lagoon that harbors high biomasses of phytoplankton. Despite Increasing interest in fish gut microbiomes diversity and functions, little data is currently available regarding wild species from tropical west African lakes. Here, gut-associated bacterial communities are investigated in four fish species that are consumed by locale populations, namely the Cichlidae Hemichromis fasciatus, Tilapia guineensis and Sarotherodon melanotheron, and the Claroteidae Chrysichthys nigrodigitatus. Species-related differences are identified, that can be attributed to host phylogeny and diet. Important variations throughout the year are observed in T. guineensis and C. nigrodigitatus. This result emphasized the importance of time-series sampling and comparison with environmental variables even in tropical regions, that are not often conducted in wild populations. Effects of environmental factors (anthropogenic or not) on the microbiota and potential outcomes for fish health and populations sustainability need to be further explored. Interestingly, fish appear as major reservoirs of bacterial diversity, suggesting that they could contribute to the overall stability and resilience of bacterial communities present in the Aghien lagoon.

Characterization of Potential Threats from Cyanobacterial Toxins in Lake Victoria Embayments and during Water Treatment.

Africa's water needs are often supported by eutrophic water bodies dominated by cyanobacteria posing health threats to riparian populations from cyanotoxins, and Lake Victoria is no exception. In two embayments of the lake (Murchison Bay and Napoleon Gulf), cyanobacterial surveys were conducted to characterize the dynamics of cyanotoxins in lake water and water treatment plants. Forty-six cyanobacterial taxa were recorded, and out of these, fourteen were considered potentially toxigenic (i.e., from the genera Dolichospermum, Microcystis, Oscillatoria, Pseudanabaena and Raphidiopsis). A higher concentration (ranging from 5 to 10 µg MC-LR equiv. L−1) of microcystins (MC) was detected in Murchison Bay compared to Napoleon Gulf, with a declining gradient from the inshore (max. 15 µg MC-LR equiv. L−1) to the open lake. In Murchison Bay, an increase in Microcystis sp. biovolume and MC was observed over the last two decades. Despite high cell densities of toxigenic Microcystis and high MC concentrations, the water treatment plant in Murchison Bay efficiently removed the cyanobacterial biomass, intracellular and dissolved MC to below the lifetime guideline value for exposure via drinking water (<1.0 µg MC-LR equiv. L−1). Thus, the potential health threats stem from the consumption of untreated water and recreational activities along the shores of the lake embayments. MC concentrations were predicted from Microcystis cell numbers regulated by environmental factors, such as solar radiation, wind speed in the N−S direction and turbidity. Thus, an early warning through microscopical counting of Microcystis cell numbers is proposed to better manage health risks from toxigenic cyanobacteria in Lake Victoria.

Impacts of nutrient loading and fish grazing on the phytoplankton community and cyanotoxin production in a shallow tropical lake: Results from mesocosm experiments.

Given the increasing eutrophication of water bodies in Africa due to increasing anthropogenic pressures, data are needed to better understand the responses of phytoplankton communities to these changes in tropical lakes. These ecosystems are used by local human populations for multiple purposes, including fish and drinking water production, potentially exposing these populations to health threats if, for example, an increase in toxic cyanobacterial blooms is associated with increasing eutrophication. To test the short-term response of the phytoplankton community to the addition of nutrients (phosphorus and nitrogen, alone or in combination) and Nile tilapia, we developed an in situ mesocosm experiment in a freshwater lagoon located near Abidjan (Ivory Coast). We found that phytoplankton growth (estimated by chlorophyll-a quantification) was highly stimulated when both nitrogen and phosphorus were added, while there was no clear evidence for such colimitation by these two nutrients when considering their concentrations in the lagoon. Phytoplankton growth was accompanied by significant changes in the diversity and composition of this community and did not lead to an increase in the proportions of cyanobacteria. However, the addition of fish to some mesocosms resulted in a drastic decrease in phytoplankton biomass and a dominance of chlorophytes in this community. Finally, these experiments showed that the addition of nitrogen, alone or combined with phosphorus, stimulated microcystin production by cyanobacteria. In addition, no evidence of microcystin accumulation in the fish was found. Taken together, these data allow us to discuss strategies for controlling cyanobacterial blooms in this tropical ecosystem.

Spatiotemporal changes in the structure and composition of a less-abundant bacterial phylum (Planctomycetes) in two perialpine lakes.

We used fingerprinting and cloning-sequencing to study the spatiotemporal dynamics and diversity of Planctomycetes in two perialpine lakes with contrasting environmental conditions. Planctomycetes, which are less-abundant bacteria in freshwater ecosystems, appeared to be structured in the same way as the entire bacterial community in these ecosystems. They were more diversified and displayed fewer temporal variations in the hypolimnia than in the epilimnia. Like the more-abundant bacterial groups in aquatic systems, Planctomycetes communities seem to be composed of a very small number of abundant and widespread operational taxonomic units (OTUs) and a large number of OTUs that are present at low abundance. This indicates that the concept of "abundant or core" and "rare" bacterial phylotypes could also be applied to less-abundant freshwater bacterial phyla. The richness and diversity of Planctomycetes were mainly driven by pH and were similar in both of the lakes studied, whereas the composition of the Planctomycetes community seemed to be determined by a combination of factors including temperature, pH, and nutrients. The relative abundances of the dominant OTUs varied over time and were differently associated with abiotic factors. Our findings demonstrate that less-abundant bacterial phyla, such as Planctomycetes, can display strong spatial and seasonal variations linked to environmental conditions and suggest that their functional role in the lakes studied might be attributable mainly to a small number of phylotypes and vary over space and time in the water column.

Insights into the cyanosphere: capturing the respective metabolisms of cyanobacteria and chemotrophic bacteria in natural conditions?

Specific interactions have been highlighted between cyanobacteria and chemotrophic bacteria within the cyanosphere, suggesting that nutrients recycling could be optimized by cyanobacteria/bacteria exchanges. In order to determine the respective metabolic roles of the cyanobacterial and bacterial consortia (microbiome), a day-night metatranscriptomic analysis was performed on Dolichospermum sp. (N2 -fixer) and Microcystis sp. (non N2 -fixer) natural blooms occurring successively within a French peri-urban lake. The taxonomical and functional analysis of the metatranscriptoms have highlighted specific association of bacteria within the cyanosphere, driven by the cyanobacteria identity, without strongly modifying the functional composition of the microbiomes, suggesting functional redundancy within the cyanosphere. Moreover, the functional composition of these active communities was driven by the living mode. During the two successive bloom events, it appeared that NH4 + (newly fixed and/or allochthonous) was preferentially transformed into amino acids for the both the microbiome and the cyanobacteria, while phosphate metabolism was enhanced, suggesting that due to a high cellular growth, P limitation might take place within the cyanosphere consortium.

Assessment of some key indicators of the ecological status of an African freshwater lagoon (Lagoon Aghien, Ivory Coast).

The supply of drinking water is a vital challenge for the people who live on the African continent, as this continent is experiencing strong demographic growth and therefore increasing water demands. To meet these needs, surface water resources are becoming increasingly mobilized because underground resources are not always available or have already been overexploited. This situation is the case in the region of Abidjan in the Ivory Coast, where the drinking water deficit is a growing problem and it is therefore necessary to mobilize new water resources to ensure the supply of drinking water. Among the potential resources, local managers have identified a freshwater lagoon, Lagoon Aghien, That is in close proximity to the city of Abidjan. With the aim of enhancing knowledge on the ecological functioning of the lagoon and contributing to the assessment of its ability to provide drinking water, several physical and chemical parameters of the water and the phytoplankton community of the lagoon were monitored for 17 months (December 2016-April 2018) at six sampling stations. Our findings show that the lagoon is eutrophic, as evidenced by the high concentrations of total phosphorus (>140 µg L-1), nitrogen (1.36 mg L-1) and average chlorophyll-a (26 to 167 µg L-1) concentrations. The phytoplankton community in the lagoon is dominated by genera typical of eutrophic environments including mixotrophic genera such as Peridinium and by cyanobacteria such as Cylindrospermopsis/Raphidiopsis, Microcystis and Dolichospermum that can potentially produce cyanotoxins. The two rainfall peaks that occur in June and October appeared to be major events in terms of nutrient flows entering the lagoon, and the dynamics of these flows are complex. Significant differences were also found in the nutrient concentrations and to a lesser extent in the phytoplankton communities among the different stations, especially during the rainfall peaks. Overall, these results reveal that the quality of the lagoon's water is already severely degraded, and this degradation could increase in future years due to increasing urbanization in the watershed. These results therefore raise questions about the potential use of the lagoon as a source of drinking water if measures are not taken very quickly to protect this lagoon from increasing eutrophication and other pollution sources.

A review of the socioecological causes and consequences of cyanobacterial blooms in Lake Victoria.

Africa is experiencing high annual population growth in its major river basins. This growth has resulted in significant land use change and pollution pressures on the freshwater ecosystems. Among them, the Lake Victoria basin, with more than 42 million people, is a unique and vital resource that provides food and drinking water in East Africa. However, Lake Victoria (LV) has experienced a progressive eutrophication and substantial changes in the fish community leading to recurrent proliferation of water hyacinth and cyanobacteria. Based on an extensive literature review, we show that cyanobacterial biomasses and microcystin concentrations are higher in the bays and gulfs (B&Gs) than in the open lake (OL), with Microcystis and Dolichospermum as the dominant genera. These differences between the B&Gs and the OL are due to differences in their hydrological conditions and in the origins, type and quantities of nutrients. Using data from the literature, we describe the multiple ways in which the human population growth in the LV watershed is connected to the increasing occurrence of cyanobacterial blooms in the OL and B&Gs. We also described the consequences of cyanobacterial blooms on food resources and fishing and on direct water use and water supply of local populations, with their potential consequences on the human health. Finally, we discuss the actions that have been taken for the protection of LV. Although many projects have been implemented in the past years in order to improve the management of waste waters or to reduce deforestation and erosion, the huge challenge of the reduction of cyanobacterial blooms in LV by the control of eutrophication seems far from being achieved.

Can participatory approaches strengthen the monitoring of cyanobacterial blooms in developing countries? Results from a pilot study conducted in the Lagoon Aghien (Ivory Coast).

Monitoring of cyanobacteria in freshwater ecosystems is a complex task, which is time consuming and expensive due to the chaotic population dynamics and highly heterogeneous distribution of cyanobacteria populations in water bodies. The financial cost constitutes a strong limitation for the implementation of long-term monitoring programs in developing countries, particularly in Africa. The work presented here was performed in the framework of an international project addressing the sustainable monitoring and management of surface water resources used for the production of drinking water in three African countries. We tested the potential of a citizen approach for monitoring cyanobacterial blooms, which are a growing threat to the drinking water supply. This pilot study was designed, implemented and evaluated in close interaction with the Pasteur Institute of the Ivory Coast and with the populations of three villages located on the shoreline of a freshwater lagoon located near Abidjan city. Based on the use of a smartphone application, the citizens of the three villages were invited to report water color changes, as these changes could reflect cyanobacteria proliferations. A two-year experimentation period has shown that it is possible to mobilize the local populations to monitor cyanobacterial blooms. The data collected by citizens were consistent with the data obtained by a classical monitoring of cyanobacteria performed over seven months, but it appeared that new approaches were needed to validate the citizen data. This participatory approach also provided great improvements to the understanding and awareness of local populations regarding water quality and cyanobacterial bloom issues. Finally, we discuss some of the difficulties and limitations of our participatory monitoring approach that should be considered by further implementations. Despite these difficulties, our work suggests that citizen monitoring is a promising approach that may complement the classical approach to sustainable monitoring of cyanobacteria in developing countries.

Comment on "Meta-analysis reveals declines in terrestrial but increases in freshwater insect abundances".

Van Klink et al (Reports, 24 April 2020, p. 417) argue for a more nuanced view of insect decline, and of human responsibility for this decline, than previously suggested. However, shortcomings in data selection and methodology raise questions about their conclusions on trends and drivers. We call for more rigorous methodology to be applied in meta-analyses of ecological data.

Toxic benthic freshwater cyanobacterial proliferations: Challenges and solutions for enhancing knowledge and improving monitoring and mitigation.

1. This review summarises knowledge on the ecology, toxin production, and impacts of toxic freshwater benthic cyanobacterial proliferations. It documents monitoring, management, and sampling strategies, and explores mitigation options. 2. Toxic proliferations of freshwater benthic cyanobacteria (taxa that grow attached to substrates) occur in streams, rivers, lakes, and thermal and meltwater ponds, and have been reported in 19 countries. Anatoxin- and microcystin-containing mats are most commonly reported (eight and 10 countries, respectively). 3. Studies exploring factors that promote toxic benthic cyanobacterial proliferations are limited to a few species and habitats. There is a hierarchy of importance in environmental and biological factors that regulate proliferations with variables such as flow (rivers), fine sediment deposition, nutrients, associated microbes, and grazing identified as key drivers. Regulating factors differ among colonisation, expansion, and dispersal phases. 4. New -omics-based approaches are providing novel insights into the physiological attributes of benthic cyanobacteria and the role of associated microorganisms in facilitating their proliferation. 5. Proliferations are commonly comprised of both toxic and non-toxic strains, and the relative proportion of these is the key factor contributing to the overall toxin content of each mat. 6. While these events are becoming more commonly reported globally, we currently lack standardised approaches to detect, monitor, and manage this emerging health issue. To solve these critical gaps, global collaborations are needed to facilitate the rapid transfer of knowledge and promote the development of standardised techniques that can be applied to diverse habitats and species, and ultimately lead to improved management.

Metagenomic approach studying the taxonomic and functional diversity of the bacterial community in a mesotrophic lake (Lac du Bourget--France).

The main goals of this work were to identify the metabolic pathways of the bacterial community in a lacustrine ecosystem and to establish links between taxonomic composition and the relative abundances of these metabolic pathways. For this purpose, we analysed a 16S rRNA gene library obtained by gene amplification together with a sequence library of both insert ends on c. 7700 fosmids. Whatever the library used, Actinobacteria was the most abundant bacterial group, followed by Proteobacteria and Bacteroidetes. Specific aquatic clades such as acI and acIV (Actinobacteria) or LD12 and GOBB-C201 (Alphaproteobacteria) were found in both libraries. From comparative analysis of metagenomic libraries, the metagenome of this lake was characterized by overrepresentation of genes involved in the degradation of xenobiotics mainly associated with Alphaproteobacteria. Actinobacteria were mainly related to metabolic pathways involved in nucleotide metabolism, cofactors, vitamins, energy, replication and repair. Betaproteobacteria appeared to be characterized by the presence of numerous genes implicated in environmental information processing (membrane transport and signal transduction) whereas glycan and carbohydrate metabolism pathways were overrepresented in Bacteroidetes. These results prompted us to propose hypotheses on the ecological role of these bacterial classes in lacustrine ecosystems.

Diversity, spatial distribution and activity of fungi in freshwater ecosystems.

High-throughput sequencing has given new insights into aquatic fungal community ecology over the last 10 years. Based on 18S ribosomal RNA gene sequences publicly available, we investigated fungal richness and taxonomic composition among 25 lakes and four rivers. We used a single pipeline to process the reads from raw data to the taxonomic affiliation. In addition, we studied, for a subset of lakes, the active fraction of fungi through the 18S rRNA transcripts level. These results revealed a high diversity of fungi that can be captured by 18S rRNA primers. The most OTU-rich groups were Dikarya (47%), represented by putative filamentous fungi more diverse and abundant in freshwater habitats than previous studies have suggested, followed by Cryptomycota (17.6%) and Chytridiomycota (15.4%). The active fraction of the community showed the same dominant groups as those observed at the 18S rRNA genes level. On average 13.25% of the fungal OTUs were active. The small number of OTUs shared among aquatic ecosystems may result from the low abundances of those microorganisms and/or they constitute allochthonous fungi coming from other habitats (e.g., sediment or catchment areas). The richness estimates suggest that fungi have been overlooked and undersampled in freshwater ecosystems, especially rivers, though they play key roles in ecosystem functioning as saprophytes and parasites.

Linking phytoplankton pigment composition and optical properties: A framework for developing remote-sensing metrics for monitoring cyanobacteria.

This study has been performed in the framework of a research program aiming to develop a low-cost aerial sensor for the monitoring of cyanobacteria in freshwater ecosystems that could be used for early detection. Several empirical and mechanistic remote-sensing tools have been already developed and tested at large scales and have proven useful in monitoring cyanobacterial blooms. However, the effectiveness of these tools for early detection is hard to assess because such work requires the detection of low concentrations of characteristic pigments amid complex ecosystems exhibiting several confounding factors (turbidity, blooms of other species, etc.). We developed a framework for performing high-throughput measurements of the absorbance and reflectance of small volumes (∼ = 20 mL) of controlled mixtures of phytoplankton species and studied the potential of this framework to validate remote-sensing proxies of cyanobacteria concentration. The absorption and reflectance spectra of single and multiple cultures carried a specific signal that allowed for the quantitative analysis of culture mixes. This specific signal was shown to be related to known pigment absorbance spectra. The concentrations of chlorophyll-a and -b, phycocyanin and phycoerythrin could be obtained from direct absorbance measurements and were correlated with the concentration obtained after pigment extraction (R2 ≥ 0.96 for all pigments). A systematic test of every possible two-band and three-band normalized difference between optical indices was then performed, and the coincidental correlation with chlorophyll-b (absent in cyanobacteria) was used as an indicator of non-specificity. Two-band indices were shown to suffer from non-specificity issues and could not yield strong and specific relationships with phycocyanin or phycoerythrin (maximum R2 < 0.5). On the other hand, the three-band modified normalized difference indices yielded strong specific relationships (R2 > 0.8).

Highly plastic genome of Microcystis aeruginosa PCC 7806, a ubiquitous toxic freshwater cyanobacterium.

BACKGROUND: The colonial cyanobacterium Microcystis proliferates in a wide range of freshwater ecosystems and is exposed to changing environmental factors during its life cycle. Microcystis blooms are often toxic, potentially fatal to animals and humans, and may cause environmental problems. There has been little investigation of the genomics of these cyanobacteria. RESULTS: Deciphering the 5,172,804 bp sequence of Microcystis aeruginosa PCC 7806 has revealed the high plasticity of its genome: 11.7% DNA repeats containing more than 1,000 bases, 6.8% putative transposases and 21 putative restriction enzymes. Compared to the genomes of other cyanobacterial lineages, strain PCC 7806 contains a large number of atypical genes that may have been acquired by lateral transfers. Metabolic pathways, such as fermentation and a methionine salvage pathway, have been identified, as have genes for programmed cell death that may be related to the rapid disappearance of Microcystis blooms in nature. Analysis of the PCC 7806 genome also reveals striking novel biosynthetic features that might help to elucidate the ecological impact of secondary metabolites and lead to the discovery of novel metabolites for new biotechnological applications. M. aeruginosa and other large cyanobacterial genomes exhibit a rapid loss of synteny in contrast to other microbial genomes. CONCLUSION: Microcystis aeruginosa PCC 7806 appears to have adopted an evolutionary strategy relying on unusual genome plasticity to adapt to eutrophic freshwater ecosystems, a property shared by another strain of M. aeruginosa (NIES-843). Comparisons of the genomes of PCC 7806 and other cyanobacterial strains indicate that a similar strategy may have also been used by the marine strain Crocosphaera watsonii WH8501 to adapt to other ecological niches, such as oligotrophic open oceans.

Competition between microcystin- and non-microcystin-producing Planktothrix agardhii (cyanobacteria) strains under different environmental conditions.

The factors that control the production of microcystins (hepatotoxins) during cyanobacterial blooms, and the function of these metabolites remain largely unknown. In an attempt to provide answers to these questions, we compared the fitness of microcystin (MC)-producing and non-MC-producing Planktothrix agardhii strains under various experimental conditions. More specifically, we investigated the effects of temperature, light intensity and nitrate concentrations on several MC-producing and non-MC-producing strains in monoculture and competition experiments. In the monoculture experiments, no significant difference in cell growth rates was found for any of the environmental conditions tested. On the other hand, at the end of the competition experiments, we found that when the environmental conditions limited cell growth, MC-producing strains were clearly winning out over the non-MC-producing ones. This suggested that, under growth-limiting conditions, the benefits of producing MC outweigh the cost. Moreover, the reverse was found under non-growth-limiting conditions, suggesting that under environmental conditions that favour cyanobacterial growth, the cost of MC production must outweigh its benefits. These findings suggest that environmental factors may have an indirect effect on the MC production rate, and on the selection of MC-producing and non-MC-producing strains, via their direct impact on both the cell growth rate and the cell densities in the cultures. Several hypotheses have been advanced concerning the possible function of MCs, but none of them seems to be supported by our data.