The filamentous cyanobacterium Anabaena sp. PCC 7120 expresses during the differentiation of heterocysts a short peptide PatS and a protein HetN, both containing an RGSGR pentapeptide essential for activity. Both act on the master regulator HetR to guide heterocyst pattern formation by controlling the binding of HetR to DNA and its turnover. A third small protein, PatX, with an RG(S/T)GR motif is present in all HetR-containing cyanobacteria. In nitrogen-depleted medium, inactivation of patX does not produce a discernible change in phenotype, but its overexpression blocks heterocyst formation. Mutational analysis revealed that PatX is not required for normal intercellular signaling, but it nonetheless is required when PatS is absent to prevent rapid ectopic differentiation. Deprivation of all three negative regulators – PatS, PatX, and HetN – resulted in synchronous differentiation. However, in nitrogen-containing medium, such deprivation leads to extensive fragmentation, cell lysis, and aberrant differentiation, while either PatX or PatS as the sole HetR regulator can establish and maintain a semiregular heterocyst pattern. These results suggest that tight control over HetR by PatS and PatX is needed to sustain vegetative growth and regulated development. The mutational analysis has been interpreted in light of the opposing roles of negative regulators of HetR and the positive regulator HetL. Keywords: cyanobacteria; heterocyst, regulation of differentiation

Cyanobacteria are highly diverse, widely distributed photosynthetic bacteria inhabiting various environments ranging from deserts to the cryosphere. Throughout this range of niches, they have to cope with various stresses and kinds of deprivation which threaten their growth and viability. In order to adapt to these stresses and survive, they have developed several global adaptive responses which modulate the patterns of gene expression and the cellular functions at work. Sigma factors, two-component systems, transcriptional regulators and small regulatory RNAs acting either separately or collectively, for example, induce appropriate cyanobacterial stress responses. The aim of this review is to summarize our current knowledge about the diversity of the sensors and regulators involved in the perception and transduction of light, oxidative and thermal stresses and nutrient starvation responses. The studies discussed here point to the fact that various stresses affecting the photosynthetic capacity are transduced by common mechanisms.

Hepatotoxic microcystins (MCs) are the most widespread class of cyanotoxins and the one that has most often been implicated in cyanobacterial toxicosis. One of the main challenges in studying and monitoring MCs is the great structural diversity within the class. The full chemical structure of the first MC was elucidated in the early 1980s and since then the number of reported structural analogues has grown steadily and continues to do so, thanks largely to advances in analytical methodology. The structures of some of these analogues have been definitively elucidated after chemical isolation using a combination of techniques including nuclear magnetic resonance, amino acid analysis and tandem mass spectrometry (MS/MS). Others have only been tentatively identified using liquid chromatography-MS/MS without chemical isolation. An understanding of the structural diversity of MCs, the genetic and environmental controls for this diversity and the impact of structure on toxicity are all essential to the ongoing study of MCs across several scientific disciplines. However, because of the diversity of MCs and the range of approaches that have been taken for characterizing them, comprehensive information on the state of knowledge in each of these areas can be challenging to gather. We have conducted an in-depth review of the literature surrounding the identification and toxicity of known MCs and present here a concise review of these topics. At present, at least 269 MCs have been reported. Among these, about 20% (54 of 269) appear to be the result of chemical or biochemical transformations of MCs that can occur in the environment or during sample handling and extraction of cyanobacterial, including oxidation products, methyl esters, or post-biosynthetic metabolites. The toxicity of many MCs has also been studied using a range of different approaches and a great deal of variability can be observed between reported toxicities, even for the same congener. This review will help clarify the current state of knowledge on the structural diversity of MCs as a class and the impacts of structure on toxicity, as well as to identify gaps in knowledge that should be addressed in future research.

During blooms, cyanobacteria produce diverse modified peptides. Among these are the microginins, which inhibit zinc-containing metalloproteases. Ten microginins, microginins KR767 (1), KR801(2), KR835 (3), KR785 (4), KR604 (5), KR638 (6), KR781 (7), KR815 (8), FR3 (9), and FR4 (10) were isolated from the extract of a bloom material of Microcystis sp. (IL-405) collected from the Kishon Reservoir, Israel in the fall of 2009. The structures of the pure compounds were elucidated using 1D and 2D NMR techniques and high-resolution mass spectrometry. The absolute configuration of the chiral centers of the amino acids were determined by Marfey’s and advance Marfey’s methods and by comparison of ¹H and ¹³C NMR chemical shifts of the Ahda derivatives with those of known microginins. These microginins differ in sequence and absolute configuration of the chiral centers of the Ahda moieties and by N-methylation of Ahda amine group and extent of chlorination of Ahda terminal methyl group. The compounds were evaluated for inhibition of the zinc metalloprotease aminopeptidase M and exhibited low- to sub-nanomolar IC₅₀ values.

Cyanobacteria constitute pioneer colonizer of specific environments whom settlement in new biotopes precedes the establishment of composite microbial consortia. Some heterotrophic bacteria constitute cyanobacterial partners that are considered as their cyanosphere, being potentially involved in mutualistic relationships through exchange and recycling of key nutrients, and sharing of common goods. Several non-axenic cyanobacterial strains have been recently isolated along with their associated cyanosphere from the thermal mud of Balaruc-les-Bains (France) and the biofilms of the retention basin where they develop. The community structure and relationships among members of the isolated cyanobacterial strains were characterized using a metagenomic approach combined with taxonomic and microscopic description of the microbial consortia. Results provide insights into the potential role and metabolic capabilities of microorganisms of thermal mud-associated cyanobacterial biofilms. Thus, the physical proximity, host-specificity and complimentary functions advocate for their complementarity between cyanobacteria and their associated microbiota. Besides these findings, our results also highlight the great influence of the reference protein database chosen when performing functional annotation of the metagenomes from organisms of the cyanosphere and the difficulty of selecting one unique database that appropriately cover both autotroph and heterotroph metabolic specificities.

Abstract: The genus Acaryochloris is unique among phototrophic organisms due to the dominance of chlorophyll d in its photosynthetic reaction centres and light-harvesting proteins. This allows Acaryochloris to capture light energy for photosynthesis over an extended spectrum of up to ~760 nm in the near infra-red (NIR) spectrum. Acaryochloris sp. has been reported in a variety of ecological niches, ranging from polar to tropical shallow aquatic sites. Here, we report a new Acarychloris strain isolated from an NIR-enriched stratified microbial layer 4-6 mm under the surface of stromatolite mats located in the Hamelin Pool of Shark Bay, Western Australia. Pigment analysis, by traditional spectrometry/fluorometry, flow cytometry and spectral confocal microscopy identify unique patterns in pigment distribution that likely reflect niche adaption. For example, unlike the original A. marina species (type strain MBIC11017), this new strain, Acarychloris LARK001, shows little change in the chlorophyll d/a ratio in response to changes in light wavelength, displays a different Fv/Fm response and lacks detectable levels of phycocyanin. Indeed, 16S rRNA analysis supports the identity of the A. marina LARK001 strain as distinct from the A. marina HICR111A strain first isolated from Heron Island, previously found on the Great Barrier Reef, under coral rubble on the reef flat. Taken together, A. marina LARK001 is a new cyanobacterial strain adapted to the stromatolite matts in Shark Bay.

Cyanobacteria biomasses are sources of secondary metabolites and nutritious ingredients such as vitamins, essential amino acids, and unsaturated fatty acids. Biochemical composition, presence of cyanotoxins and contaminants are major concerns to be addressed on such edible biomasses. Macrocolonies of a filamentous diazotrophic Nostoc species known as Llayta are found at Andean wetlands and consumed since pre-Columbian times in South America. Its biochemical composition has been previously conducted to assess their nutritious quality and cyanotoxicity. Macrocolonies of filamentous cyanobacteria are niches for colonization by diverse microorganisms; however, the Llayta microcolonies cyanosphere is unknown. Based on a culture-independent approach, we report the identification of members of the resilient microflora associated with Llayta trichomes after Gentamicin treatments. We have also reconstructed the genomes of the Llayta macrocolony-forming Nostoc sp. cyanobacterium (6,781,030 bp; GC content of 41.2%) and the genomes of five dominant bacteria genera (Mesorhizobium, Microvirga, Paracoccus, Aquimonas, and Blastomonas). The detection of genes and genes clusters involved in primary and secondary metabolism is described. Our results provide new insights on the metabolic capabilities and biotechnological potential of the Andean Nostoc cyanobacterium, and the ecological role and adaptive strategies of microorganisms living under extreme environmental conditions at the Andean wetlands.

During a survey in 2015 an impressive assemblage of organisms were found in a hypersaline pond of the Messolonghi saltworks. The salinity ranged between 50 and 180 ppt and the organisms recorded fell in the categories of Cyanobacteria (17 species), Chlorophytes (4 species), Diatoms (23 species), Dinoflagellates (1 species), Protozoa (40 species), Rotifers (8 species), Copepods (1 species), Artemia sp., one nematode and Alternaria sp. (Fungi). Fabrea salina was the most prominent protist in all samples and salinities. This ciliate has the potential to be a live-food candidate for marine fish larvae. Asteromonas gracilis proved a sturdy microalga performing excellently in a broad spectrum of culture salinities ies. Most of the specimens were identified only to the genus level and, based on their morphology, as there are no relevant records in Greece, there is a possibility for some of them to be either new species or strikingly different strains of certain species recorded elsewhere.

Cyanobacteria are an ancient lineage of slow-growing photosynthetic bacteria and a prolific source of natural products with diverse chemical structures and potent biological activities and toxicities. The chemical identification of these compounds remains a major bottleneck. Strategies that can prioritize the most prolific strains and novel compounds are of great interest. Here, we combine chemical analysis and genomics to investigate the chemodiversity of secondary metabolites based on their pattern of distribution within some cyanobacteria. Planktothrix being a cyanobacterial genus known to form blooms worldwide and to produce a broad spectrum of toxins and other bioactive compounds, we applied this combined approach on four closely related strains of Planktothrix. The chemical diversity of the metabolites produced by the four strains was evaluated using an untargeted metabolomics strategy with high-resolution LC-MS. Metabolite profiles were correlated with the potential of metabolite production identified by genomics for the different strains. Although, the Planktothrix strains present a global similarity in term biosynthetic cluster gene for microcystin, aeruginosin and prenylagaramide for example, we found remarkable strain-specific chemo-diversity. Only few of the chemical features were common to the four studied strains. Additionally, the MS/MS data were analyzed using Global Natural Products Social Molecular Networking (GNPS) to identify molecular families of the same biosynthetic origin. In conclusion, we present an efficient integrative strategy for elucidating the chemical diversity of a given genus and link the data obtained from analytical chemistry to biosynthetic genes of cyanobacteria.

A strain of the filamentous non N-fixing cyanobacterium Phormidium sp. isolated from the Messolonghi (W. Greece) saltworks, was cultured in the laboratory at 6 different combinations of salinity (20-40-60 ppt) and illumination (low-2000 lux and high-8000 lux). At salinities of 60 and 40 ppt and in high illumination (XL-8000 lux) the growth rate (μmax) presented the highest values (0.491 and 0.401 respectively) compared to the corresponding at 20 ppt (0.203). In general and at all salinities, the higher illumination (XL) gave the highest growth rates and shorter dublication time (tg) in comparison to the lower illumination (L). On the contrary, phycocyanin, phycoerythrin and allophycocyanin production was extremely increased in the lower illumination (L) in all salinities, from ~14fold at 40 and 60 ppt to 269fold at 20 ppt of those corresponding to higher illumination (XL). Similar analogies were also recorded for the other two billiproteins. Chlorophyll-a content was also higher in lower illumination at all salinities in contrast to total carotenoids that did not exhibit such a pattern. The high growth rate and high phycocyanin content along with the rapid sedimentation of its cultured biomass can set this marine Phormidium species as a promising canditate for mass culture.

Environmental genetics-related modern methods are shown as important indicators of various cyanotoxins syntheses, and their knowledge and use are critically analyzed. Microcystins and other cyanotoxins loads and syntheses are related to different drivers, like various chemical elements and compounds (especially nutrients, such as nitrogen and phosphorus, and their ratio), then to the light, conductivity, temperature, and other climatical and hydrological factors, to which spatial and geographical features (such as the surface of the water bodies) have to be added. The biotic relationships include different specific and supraspecific, uni- and bilateral links between the cyanobacteria, and subsequently their synthesized toxins, and protozoans (or protoctists), chromists, macrophytes, different systematical and ecological groups of zooplankton, and others. The importance of, but also the gaps in, the knowledge and the scarcity of studies involving ectocrines mediated interactions between different groups of algae and plants are highlighted. The paper ends with an interesting classification of lakes' trophicity, illustrated with conceptual diagrams, based on possible scenarios of cyanobacteria behavior.

Background: The Balaruc-les-Bains’ thermal mud was found to be colonized predominantly by microorganisms, with cyanobacteria constituting the primary organism in the microbial biofilm observed on the mud surface. The success of cyanobacteria in colonizing this specific ecological niche can be explained in part by their taxa-specific adaptation capacities, and also the diversity of bioactive natural products that they synthesize. This array of components has physiological and ecological properties that may be exploited for various applications.

Cyanobacterial species (blue-green algae) constitute the major part of the phytoplanktonic biomass during the summer in freshwater ponds. The aim of the research work was to study the biodiversity of cyanobacteria among 20 different freshwater ponds of the Pudukkottai district of Tamil Nadu, India. The morphological identification of cyanobacterial species was carried out using a trinocular microscope. The results showed that the maximum number of cyanobacterial species belonged to Oscillatoriaceae, Nostocaceae, Microcystaceae, Scenedesmaceae, and Desmidiaceae families. Among 25 different families of Cyanobacteria about 42 distinct species were identified. These results showed that the freshwater ponds of the Pudukkottai district have an abundance of cyanobacteria species.

Llayta is a dietary supplement used by rural communities in Perú and northern Chile since pre-Columbian days. Llayta is the biomass of colonies of a Nostoc cyanobacterium grown in wetlands of the Andean highlands, harvested, sun-dried and sold as an ingredient for human consumption. The biomass has a substantial content of essential amino acids (58% of total amino acids) and polyunsaturated fatty acids (33% total fatty acids). This ancestral practice is being loss and the causes were investigated by an ethnographic approach to register the social representations of Llayta, to document how this Andean feeding practice is perceived and how much the community knows about Llayta. Only 37% of the participants (mostly adults) have had a direct experience with Llayta; other participants (mostly children) did not have any knowledge about it. These social responses reflect anthropological and cultural tensions associated to lack of knowledge on Andean algae, sites where to find Llayta, where it is commercialized, how it is cooked and on its nutritional benefits. The loss of this ancestral feeding practice, mostly on northern Chile, is probably associated to cultural changes, migration of the rural communities, and a very limited access to the available information. We propose that Llayta consumption can be revitalized by developing appropriate educational strategies and investigating potential new food derivatives based on the biomass from the isolated Llayta cyanobacterium.

Control of nutrients, mainly nitrogen (N) and phosphorus (P), plays a significant role in preventing cyanobacterial blooms (harmful algal blooms (HABs)). This study aimed at evaluating changes in the risk of the occurrence of cyanobacterial blooms and advancing the understanding of how N and P affect the growth of cyanobacteria in a eutrophic lake, Lake Vombsjön, in southern Sweden. Statistical analysis was used to demonstrate the pattern of cyanobacterial blooms, that the highest content present in September and the later that algal blooms occur, the more likely it is a cyanobacterial bloom as cyanobacteria became dominating in October and November (90%). Two hypothesises tested in Lake Vombsjön confirmed namely that a high total phosphorus (TP) level correlates with an abundance of cyanobacteria and that low N:P ratio (total nitrogen/total phosphorus < 20) favours the growth of cyanobacteria. To control the growth of cyanobacteria in Lake Vombsjön, the TP level should be kept below 20 µg/L and the N:P ratio be maintained at a level of over 20. The two species Planktothrix agardhii, and Pseudanabaena spp. should be carefully monitored especially in late autumn. Future work should consider any high degree of leakage from the sediment of the dissolved phosphorus available there.

Photosynthesis is one the most important biological processes on earth, producing life-giving oxygen and is the basis for a large variety of plant products. Measurable properties of photosynthesis provide information about its biophysical state and, in turn, the physiological conditions of a photoautotrophic organism. For instance, chlorophyll fluorescence of an intact photosystem is not linear as in the case of a single fluorescent dye in solution, but shows temporal changes related to the quantum yield of the photosystem. Commercial photosystem analyzers already use the fluorescence kinetics characteristics of photosystems to infer the viability of organisms under investigation. Here, we provide a novel approach based on an optical Fabry-Pérot microcavity or that enables the readout of photosynthetic properties and activity for an individual cyanobacterium. This approach offers a completely new dimension of information, which would normally be lost due to averaging in ensemble measurements obtained from a large population of bacteria.

Cyanobacterial macrocolonies known as Llayta are found at Andean wetlands and consumed since pre-Columbian times in South America. Macrocolonies of filamentous cyanobacteria are niches for colonization by other microorganisms; however, the microbiome of edible Llayta has not been explored. Based on a culture-independent approach, we report the presence, identification and metagenomic genome reconstruction of Cyanocohniella sp. LLY associated to Llayta trichomes. The assembled genome of strain LLY is now available for further inquiries, and may be instrumental for taxonomic advances on this genus. All known members of the Cyanocohniella genus have been isolated from salty European habitats. A biogeographic gap for the Cyanocohniella genus is partially filled by the existence of strain LLY at Andes Mountains wetlands in South America as a new habitat. This is the first genome available for members of this genus. Genes involved in primary and secondary metabolism are described providing new insights on the putative metabolic capabilities of Cyanocohniella sp. LLY. The reconstructed genome of strain LLY is now available and instrumental for further inquiries and taxonomic advances on the genus Cyanocohniella.

Cyanobacteria are prokaryotes whose taxonomy follows the same rules of a code (the International Botanical Nomenclature Code, IBNC) built for eukaryotic photosynthetic organisms. Hence, names of cyanobacteria follow the same rules and are assigned to biological entities (species) that should correspond to eukaryotic species. The main difficulty in the current situation is that the species concept in eukaryotes is based theoretically mainly on the biological species concept, that is centered on genetic exchange through sexual reproduction or lack of them. However, as shown, this difference is important from a theoretical point of view, but also in eukaryotes, the boundaries between different species are very rarely checked experimentally by direct observation of sexual barriers and hybridization events. The main concept for species delimitation is hence that related to morphology and, more recently and always in relation to morphology, DNA sequences. The introduction of distances obtained from matrixes of aligned sequences in the framework of a barcoding project provides a quantitative interpretation of species delimitation in relation to genetic distance that can be used both in eukaryotes and prokaryotes. However, the introduction of quantitative criteria needs the definition of distance thresholds to identify the boundaries between different species and, for doing that, it is necessary to test the distance thresholds in models of traditionally defined and recognized species. An alternative approach may be the comparison of the molecular distance (quantitative approach) to data about the capability of strains/species to exchange genetic information. Unfortunately data about this last question is still scarce. The adoption of molecular criteria to check species boundaries based on morphological characters has proved particularly challenging in cyanobacteria: a known example is provided. In conclusion, the only possible approach appears to be the association of molecular data to the increase of available data about the cell structure and the variation thereof in different physiological situations, particularly at the ultrastructural level. A further necessity is the check of the typus for a large number of cyanobacteria species, often based on old basionyms. In many of these cases the typus is often a drawing and more rarely a herbarium specimen or a microscope slide. In many cases an epitypification or a neotypification appears to be necessary.

The purpose of this work is to define optimal growth conditions for batch culture of the cyanobacterium Arthrospira maxima and the microalgae Chlorella vulgaris, Isochrysis galbana and Nannochloropsis gaditana. Thus, we study the effect of three variables on algae growth: i.e., inoculum:culture medium ratio, light:darkness photoperiod and type of culture medium, including both synthetic media and wastewaters. The results showed that the initial inoculum volume did not affect the amount of biomass at the end of the growth (14 days), whereas an excess (18 h) or defect (6 h) in the number of hours of light is determinant for its development. The contribution of nutrients from different culture media modified the growth of the different species. A. maxima was favoured in seawater enriched with Guillard's F/2 as well as C. vulgaris and N. gaditana but in fresh water medium. I. galbana had the greatest growth in the marine environment enriched with Walne’s media. Nitrate was the limiting growth reagent at the end of the exponential phase of growth for C. vulgaris and N. gaditana, while iron was for A. maxima and I. galbana. All species demonstrated their capability to grow in effluents from a wastewater treatment plant and they efficiently consume nitrogen, especially the three microalgae species.

Cyanobacterial mass developments in eutrophic ponds and lakes are a major concern for lake management, as many cyanobacteria produce a huge variety of toxic secondary metabolites, e.g. microcystins. The aim of this research was to culture a strain of the cyanobacterium Microcystis sp strain BM25, to observe its biomass production and to isolate and purify protease inhibitors from this cyanobacterial biomass. Different secondary metabolites were isolated following a standard bioassay-guideline. Isolation was performed, with an enzymatic protease assay as bioassay. High performance liquid chromatography was used to identify different fractions of secondary metabolite from the strain BM25. Moreover, protease homogenates were isolated from Daphnia magna in order to test the inhibitors against naturally occurring major digestive proteases trypsin and chymotrypsin. It was measured that 60% MeOH and the 80% MeOH C18-SPE fraction inhibits chymotrypsin activity 98% (6 nmol pNA min^-1 mg^-1) and 99 % (4 nmol pNA min^-1 mg^-1), respectively. In contrast, trypsin activity was not inhibited by methanolic extracts of this cyanobacterium strain.

Cyanobacteria are photosynthetic microorganisms that colonize diverse environments worldwide, ranging from ocean to freshwaters, soils, and extreme environments. Their adaptation capacities and the diversity of natural products (molecules, metabolites, or compounds) that they synthesize support the cyanobacterial success for the colonization of their respective ecological niches. Although cyanobacteria are well-known for their toxin production and their relative deleterious consequences, they also produce a large variety of molecules that exhibit beneficial properties with high potential for various fields of application (e.g., synthetic analog of the dolastatin 10 used against Hodgkin lymphoma). The present review specially focuses on the beneficial activities of cyanobacterial molecules described so far. Based on an analysis of 670 papers, it appears that more than 90 genera of cyanobacteria have been found to produce compounds with potential beneficial activities, most of them belonging to the orders Oscillatoriales, Nostocales Chroococcales, and Synechococcales. The rest of the cyanobacterial orders (i.e., Pleurocapsales, Chroococcidiopsales, and Gloeobacterales) remain poorly explored in terms of their molecular diversity and relative bioactivity. The diverse cyanobacterial molecules presenting beneficial bioactivities belong to 10 different chemical classes (alkaloids, depsipeptides, lipopeptides, macrolides/lactones, peptides, terpenes, polysaccharides, lipids, polyketides, and others) that exhibit 14 major kinds of bioactivity. However, no direct relation between the chemical class and the bioactivity of these molecules has been demonstrated. We further selected and specifically described 50 molecule families according to their specific bioactivities and their potential uses in pharmacology, cosmetology, agriculture, or other specific fields of interest. This up-to-date review takes advantage of the recent progresses in genome sequencing and biosynthetic pathway elucidation, and presents new perspectives for the rational discovery of new cyanobacterial metabolites with beneficial bioactivity.

Modern day civilization is dependent on energy generation by fossil fuels. But the major drawback of using fossil fuels is environmental pollution. Microalgae are potential candidate for production of various products of interest, such as proteins, mini food, pigments and triglycerides that can be converted into biofuels. Lignocellulosic feedstocks are the most abundantly available raw material of plants that can serve as a promising feedstock for cultivating bacteria, fungi, yeasts and microalgae to produce biofuels and other value-added products. Owing to the abundant availability of these low/no cost substrates, can be utilized as feedstocks for cultivating microalgae to generate biogas/biodiesel. Likewise, there is much room to exploit defatted algal biomass to be used as animal/fish feed and oil producing/accumulating genes knowledge in future to produce high and good quality biodiesel and biogas.

Cyanobacteria are microorganisms able to adapt to a wide variety of environmental conditions and abiotic stresses. They produce a very large number of metabolites that can participate in the adaptation of cyanobacteria to a large range of resources such as light, temperature, or nutrient. The metabolites variation is one way to understand the physiological status and adaptation of cells. In this study, we aim to understand how the diversity and the dynamics of the whole metabolome is dependent of the growth phases and under control of abiotic factors (e.g. light intensity and temperature). The cyanobacteria Aliinostoc sp. PMC 882.14 was selected for its large number of biosynthetic gene clusters. Metabolomes were analyzed by using mass spectrometry (qTOF-MS/MS) combined with untargeted analysis to investigate the metabolite dynamics. Significant variations were characterized between exponential and stationary phases, whatever the culture conditions (“control”, “higher light”, or “higher temperature”). ”Higher light” and “higher temperature” favored the synthesis of metabolites belonging to the same molecular families. Among highly regulated metabolites, we observe the presence of mycosporine-like amino acids (MAAs), and various variants of somamides, microginins, and microviridins. Through Aliinostoc sp. PMC 882.14, this study shows the importance of knowing the physiological state of cyanobacteria for comparative global metabolomics and questions the regulation processes involve into metabolite families production. Our results also open up new perspectives in the context of the production of targeted bioactive metabolites.

The presence of harmful algal blooms (HABs) and cyanotoxins in drinking water sources poses a great threat to human health. The current study employed molecular techniques to determine the occurrence of non-toxic and toxic cyanobacteria species in the Limpopo River basin based on the phylogenetic analyses of 16S rRNA gene. The bottom sediments samples were collected from selected rivers: Limpopo, Crocodile, Mokolo, Mogalakwena, Nzhelele, Lephalale, Sand Rivers (South Africa); Notwane (Botswana), Shashe River and Mzingwane River (Zimbabwe). The physical-chemical analysis of the bottom sediments showed the availability of nutrients, nitrates and phosphates, in excess of 0.5 mg/l for most of river sediments, alkaline pH and salinity in excess of 500 mg/l. The FlowCam showed the dominant cyanobacteria species identified from the samples were Microcystis species, followed by Cylindrospermopsis raciborskii, Phormidium and Planktothrix species and this was confirmed by molecular techniques. Nevertheless, two samples showed the amplification of cylindrospermopsin polyketide synthetase gene (S3 and S9) while two samples showed amplification for microcystin/nodularin synthetase gene (S8 and S13). Thus these findings may imply the presence of toxic cyanobacteria species in the river sediments. The presence of cyanobacteria may be hazardous to human because rural communities and farmers who abstract water from Limpopo river catchment for human consumption, livestock and wildlife watering and irrigation.

Cyanobacteria are extensively studied and cultured because they can produce many value-added substances among which are pigments, mainly the phycobiliproteins phycocyanin (PC), phycoerythrin (PE), allophycocyanin (APC) and chlorophyll-a and carotenoids as well. As numerous cyanobacterial species await optimization for maximizing pigment production, we examined here two local marine species, Phormidium sp. and Cyanothece sp. batch cultured under 18-19.5 oC, at 40 ppt salinity with Walne’s nutrient medium, using white LED light of low (2000 lux) and high (8000 lux) intensity and additionally blue, green and red LED light. Significant differences were found among the intensities and colors of light used. Maximum growth was induced by high white light in both species (2.15 g dw/L in Phormidium and 1.47 g/L in Cyanothece). Next to them was green light (1.25 g/L) in Cyanothece and low white and green (1.26 – 1.33 g/L) in Phormidium. Green light maximized phycocyanin content in Phormidium (0.45 mg/mL), while phycoerythrin was maximized (0.17 mg/mL) by blue light and allophycocyanin by all colors (~0.80 mg/mL). All colors maximized phycocyanin in Cyanothece (~0.32 mg/mL) while phycoerythrin and allophycocyanin were maximized under green light (~0.138 and 0.38 mg/mL respectively). In Phormidium maximization of chlorophyll-a (9.3 μg/mL) was induced by green light while total carotenoids and b-carotene (3.05 and 0.89 μg/mL respectively) by high white light. In Cyanothece both white light intensities along with green light maximized chlorophyll-a content (~9 μg/mL) while high white light and green maximized total carotenoids (2.6-3.0 μg/mL).

Microcystins (MCs) are cyanobacterial toxins and potent inhibitors of protein phosphatases 1 (PP1) and 2A (PP2A), which are involved in plant cytoskeleton (microtubules and F-actin) organization. Therefore, studies on the toxicity of cyanobacterial products on plant cells have so far being focused on MCs. In this study, we investigated the effects of extracts from 16 (4 MC-producing and 12 non-MC-producing) cyanobacterial strains from several habitats, on various enzymes (PP1, trypsin, elastase), on the plant cytoskeleton and H2O2 levels in Oryza sativa (rice) root cells. Seedling roots were treated for various time periods (1, 12 and 24h) with aqueous cyanobacterial extracts and underwent either immunostaining for α-tubulin or staining of F-actin with fluorescent phalloidin. DCF-DA staining was performed for H2O2 imaging. The enzyme assays confirmed the bioactivity of the extracts of not only MC-rich (MC+), but also MC-devoid (MC-) extracts, which induced major time-dependent alterations on both components of the plant cytoskeleton. These findings suggest that a broad spectrum of bioactive cyanobacterial compounds, apart from MCs or other known cyanotoxins (such as cylindrospermopsin), can affect plants by disrupting the cytoskeleton.

Artificial groundwater recharge is commonly used for drinking water supply. The resulting water quality is highly dependent on the raw water quality. In many cases, pre-treatment is required. Pre-treatment improves the drinking water quality, although how and to what extent it affects the subsequent pond water quality and infiltration process, is still unknown. We evaluated two treatment systems by applying different pre-treatment methods for raw water from a eutrophic and temperate lake. An artificial recharge pond was divided into two parts, where one received raw water, only filtered through a micro-screen with 500 µm pores (control treatment), while the other part received pre-treated lake water using chemical flocculation with polyaluminium chloride (PACl) combined with sand filtration, i.e. continuous contact filtration (contact filter treatment). Water quality such as cyanobacterial biomass, microcystin-LR as well as organic matter and nutrients were measured in both treatment processes. We found cyanobacterial biomass and microcystin-LR level after the contact filter treatment was significantly different from the control treatment and also significantly different in the pond water. In addition, with contact filter treatment, total phosphorus (TP) and organic matter removal were significantly improved in the end water, TP was reduced by 96 % (< 20 µg/L) and the total organic carbon (TOC) was reduced by 66 % instead of 55 % (TOC content around 2.1 mg/L instead of 3.0 mg/L). This full-scale onsite experiment demonstrated effective pre-treatment would benefit a more stable water quality system, with less variance and lower cyanotoxin risk. In a broader drinking water management perspective, the presented method is promising to reduce cyanotoxin risk, as well as TP and TOC, which are all predicted to increase with global warming and extreme weather.

Man-made shallow fishponds in the Czech Republic have been facing a high eutrophication since 1950s. Anthropogenic eutrophication and feeding of fish have strongly affected the physico-chemical properties of water and its aquatic community composition leading to harmful algal bloom formation. In our current study, we have characterised the phytoplankton community across three hypertrophic ponds to assess the phytoplankton dynamics during the vegetation season. We microscopically identified and quantified 29 cyanobacterial taxa comprised of non-toxigenic and toxigenic species. Further, a detailed cyanopeptides (CNPs) profiling was performed using molecular networking analysis of liquid chromatography tandem mass spectrometry (LC–MS/MS) data coupled with dereplication strategy. This MS networking approach coupled with dereplication on online global natural product social networking (GNPS) web platform led us to putatively identify forty CNPs: fourteen anabaenopeptins, ten microcystins, five cyanopeptolins, six microginins, two cyanobactins, a dipeptide radiosumin, a cyclooctapeptide planktocyclin and epidolastatin12. We have applied the binary logistic regression to estimate the CNPs producer by correlating the GNPS data with the species abundance. Usage of The combination of molecular networking and dereplication on online global natural product social networking (GNPS) web platform has proved as a valuable approach for rapid and simultaneous detection of high number of peptides, and rapidly assessing the risk for harmful bloom.

Cyanobacteria produce a wide range of structurally diverse cyanotoxins and bioactive cyanopeptides in freshwater, marine, and terrestrial ecosystems. The health significance of these metabolites, which include genotoxic- and neurotoxic agents, is confirmed by continued associations between the occurrence of animal and human acute toxic events and, in the long term, by associations between cyanobacteria and neurodegenerative diseases. One of the implicated mechanisms includes a misincorporation of cyanobacterial non-proteogenic amino acids leading to mistranslation and protein misfolding. A better understanding of the interaction between the cyanopeptide metabolism and the nervous system will be crucial to target or to prevent pathogenic response.

Cyanobacteria are ubiquitous phototrophic bacteria that inhabit diverse environments across the planet. They dominate many eutrophic lakes impacted by excess nitrogen (N) and phosphorus (P) forming dense accumulations of biomass known as cyanobacterial harmful algal blooms or cyanoHABs. Their dominance in eutrophic lakes is attributed to a variety of unique adaptations including N and P concentrating mechanisms, N fixation, colony formation that inhibits predation, vertical movement via gas vesicles, and the production of toxic or otherwise bioactive molecules. While some of these molecules have been explored for their medicinal benefits, others are potent toxins harmful to humans, animals, and other wildlife known as cyanotoxins. In humans these cyanotoxins affect various tissues, including the liver, central and peripheral nervous system, kidneys, and reproductive organs among others. They induce acute effects at low doses in the parts-per-billion range and some are tumor promoters linked to chronic diseases such as liver and colorectal cancer. The occurrence of cyanoHABs and cyanotoxins in lakes presents challenges for maintaining safe recreational aquatic environments and the production of potable drinking water. CyanoHABs are a growing problem in the North American (Laurentian) Great Lakes basin. This review summarizes information on the occurrence of cyanoHABs in the Great Lakes, toxicological effects of cyanotoxins, and appropriate numerical limits on cyanotoxins in finished drinking water.

The transition from the Peptide/RNA world to the Protein/RNA world in the hydrothermal vent environment was a major event in the history of life. The advent of proteins utterly changed the conditions of emerging life, representing a watershed in its development. During subsequent translation various protein enzymes emerged driving protocells into a more complex and interconnected system. With their astonishing versatility, the protein enzymes catalyzed crucial biochemical reactions within protocells into more complex biomolecules in diverse metabolic pathways, whereas structural proteins provided strength and permeability in the cell membrane. Four major events followed after availability of various kinds of protein molecules during prebiotic synthesis. These are: (1) the modification of the phospholipid membrane into the plasma membrane; (2) the origin of primitive cytoplasm; (3) the beginnings of the virus world; and (4) the advent of DNA. The first innovation mediated by proteins was the improvement of the cell membrane. The phospholipid membrane was initially evolved in a vent environment from the gradual modification of a fatty acid membrane via an intermediate phosphatidate acid by non-enzymatic reactions. The phospholipid is then synthesized from phosphatidate acid by a series of enzymes. To make the phospholipid membrane more permeable, various protein molecules interacted with the cell membrane. Proteins not only stabilized the wall membrane, but also acted as pumps, preventing some molecules from the protocells from crossing the membrane barriers, while permitting other selected molecules and ions to enter and leave the protocell. The second modification led by proteins is the gradual conversion of the interior of the protocell from a water-like medium into a gel-like cytoplasm, which became the storehouse of a wide range of biomolecules including amino acids, proteins, nucleic acids, ribosomes, as well as salt and water. The third innovation utilizing the newly synthesized proteins was the emergence of the ancient virus world. In the milieu of different kinds of mRNAs in the prebiotic soup, jelly-roll capsid genes originated de novo within genomes of nonviral mRNAs by overprinting. These fragile capsid genes were possibly coated by proteins on the mineral substrate for stability and durability, transforming them into ancient viral particles. These protein coats were random and were not encoded by encased genes. Some protocells might have engulfed these viral particles, when the capsid genes utilized the ribosomes of the host to translate into the appropriate capsid proteins. These capsid proteins then coated the viral genes to make new copies of primordial viruses inside the protocell. Since then, viruses became capsid-encoding organisms. These primordial mRNA viruses parasitized RNA-based protocells, manipulating them to make new copies of themselves. This was the beginning of a relentless war between viruses and their protocellular hosts. The next stage in viral evolution was the emergence of a primitive retrovirus (pre-retrovirus) with a new kind of replicative strategy in a sense that it could turn its RNA into DNA using its own reverse transcriptase enzyme. This is the beginning of the Retro world that facilitated the transition from RNA to DNA genomes. The infection of RNA protocells with pre-retroviruses progressively transferred the RNA genome to a viral DNA genome by retro-transcription. The advent of DNA by the pre-retrovirus marks the fourth innovation, when a number of enzymes had already developed and were utilized by pre-retroviruses. With continued infection, DNA viruses slowly transferred not only their core replication enzymes, such as helicase, primase, and DNA polymerase, to RNA protocells, but also to their DNAs as well. Thus, began the DNA world, when DNA replaced RNA as the major genome of the protocells. With the advent of DNA, replication of information was entirely dissociated from its expression. Because DNA is much more stable than mRNA with more storage capacity, it is a superb archive for information systems in the form of base sequences. DNA progressively took over the replicative storage function of mRNA, leaving the latter for protein synthesis. The new protocell with the DNA genome will diversify into large populations of DNA protocells that will outcompete populations of RNA protocells. Genetic information began to flow from DNA to mRNA to protein in a two-step process involving transcription and translation. In the biological stage, DNA replication was central to the binary fission of the first cell, orchestrated by the duplication of genomes and then the division of the parent cell into two identical daughter cells. It was carried out by a set of enzymes that formed a Z-ring at the site of replication. With the onset of binary fission, the population of primitive cells grew rapidly in the hydrothermal vent environment, undergoing Darwinian evolution and diversification. These primordial hyperthermophiles, presumably the first life, obtained food and energy directly from the vent environment. However, such a situation was self-limiting, so the early cells evolved their own mechanisms for generating metabolic energy and synthesizing the molecules necessary for their reproduction. The earliest fossil record (≥ 3.5 Ga) of biotic activity is preserved in the Archean hydrothermal and sedimentary rocks of the Nuvvuagittuq Craton of Canada, the Isua Craton of Greenland, the Pilbara Craton of Australia, the Kaapvaal Craton of South Africa, and the Singhbhum Craton of India in the form of the carbonaceous remains of microbial cells, cellular microfossils, and stromatolites. These microscopic fossils provide crucial evidence of the origin and early evolution of prokaryotic cells, beginning with hyperthermophiles. Molecular phylogenetic analysis suggests that both domains of life ¬– Bacteria and Archaea probably split from the last universal common ancestor (LUCA), a hyperthermophilic organism. In the younger sequences of these Archean cratons, two kinds of photosynthetic bacteria, anoxygenic green sulfur bacteria, and oxygenic cyanobacteria, appeared in quick succession from the thermophilic ancestor, indicating a shift of niche from a benthic to a planktonic, with reduced thermotolerance. The development of anoxygenic and oxygenic photosynthesis would have allowed life to escape the hydrothermal setting and invade a newly evolved habitat—broad continental shelves to tap solar energy. Cyanobacteria invaded the global ocean, turned it into blue and green, produced oxygen for the first time, and left their signatures in the carbonates and stromatolites.

The emergence of proteins in the prebiotic world was a watershed event at the origin of life. With their astonishing versatility, the protein enzymes catalyzed crucial biochemical reactions within protocells into more complex biomolecules in diverse metabolic pathways, whereas structural proteins provided strength and permeability in the cell membrane. Five major biochemical innovations followed in succession after availability of various kinds of protein molecules during decoding and translation of mRNAs. These are: (1) the modification of the phospholipid membrane into the plasma membrane; (2) the origin of primitive cytoplasm; (3) primitive gene regulation; (4) the beginnings of the virus world; and (5) the advent of DNA. The creative role of viruses during prebiotic synthesis led to the origin of the DNA world, when DNA replaced mRNA as the major genome of the protocells. With the advent of DNA, replication of information was entirely dissociated from its expression. Because DNA is much more stable than mRNA with more storage capacity, it is a superb archive for information systems in the form of base sequences. DNA progressively took over the replicative storage function of mRNA, leaving the latter for protein synthesis. Genetic information began to flow from DNA to mRNA to protein in a two-step process involving transcription and translation. In the biological stage, DNA replication was central to the binary fission of the first cell, orchestrated by the duplication of genomes and then the division of the parent cell into two identical daughter cells. With the onset of binary fission, the population of primitive cells grew rapidly in the hydrothermal vent environment, undergoing Darwinian evolution and diversification by mutation. The habitat of the earliest fossil record (≥ 3.5 Ga) from the Archean sedimentary rocks of Canada, Greenland, Australia, South Africa, and India offers a new window on the early radiation of microbial life. The development of anoxygenic and then oxygenic photosynthesis from early hyperthermophiles would have allowed life to escape the hydrothermal setting to the mesophilic global ocean.