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Significance To adapt to the daily light–dark cycle, diurnal rhythms are used by the photosynthetic cyanobacteria Prochlorococcus and Synechococcus, which are the most abundant photosynthetic organisms on earth. Field studies revealed that cyanobacterial virus (cyanophage) populations in the oceans showed transcriptional rhythms. To explore the underlying mechanism, we used cyanophage laboratory cultures to find that some showed adsorption rhythms and all showed transcriptional rhythms. We discovered that the cyanophage transcriptional rhythm is partially caused by the photosynthetic activity of host cells, explaining transcriptional rhythms of field cyanophage populations. Our study shows that cultured viruses have diurnal infection rhythms which are critical for understanding how light–dark cycles shape the interaction of cyanophages and their hosts in the oceans., As an adaptation to the daily light–dark (diel) cycle, cyanobacteria exhibit diurnal rhythms of gene expression and cell cycle. The light–dark cycle also affects the life cycle of viruses (cyanophages) that infect the unicellular picocyanobacteria Prochlorococcus and Synechococcus, which are the major primary producers in the oceans. For example, the adsorption of some cyanophages to the host cells depends on light, and the burst sizes of cyanophages are positively correlated to the length of light exposure during infection. Recent metatranscriptomic studies revealed transcriptional rhythms of field cyanophage populations. However, the underlying mechanism remains to be determined, as cyanophage laboratory cultures have not been shown to exhibit diurnal transcriptional rhythms. Here, we studied variation in infection patterns and gene expression of Prochlorococcus phages in laboratory culture conditions as a function of light. We found three distinct diel-dependent life history traits in dark conditions (diel traits): no adsorption (cyanophage P-HM2), adsorption but no replication (cyanophage P-SSM2), and replication (cyanophage P-SSP7). Under light–dark cycles, each cyanophage exhibited rhythmic transcript abundance, and cyanophages P-HM2 and P-SSM2 also exhibited rhythmic adsorption patterns. Finally, we show evidence to link the diurnal transcriptional rhythm of cyanophages to the photosynthetic activity of the host, thus providing a mechanistic explanation for the field observations of cyanophage transcriptional rhythms. Our study identifies that cultured viruses can exhibit diurnal rhythms during infection, which might impact cyanophage population-level dynamics in the oceans.

Background Cyanobacteria are ideal model organisms to exploit photosynthetically derived electrons or fixed carbon for the biotechnological synthesis of high value compounds and energy carriers. Much effort is spent on the rational design of heterologous pathways to produce value-added chemicals. Much less focus is drawn on the basic physiological responses and potentials of phototrophs to deal with natural or artificial electron and carbon sinks. However, an understanding of how electron sinks influence or regulate cellular physiology is essential for the efficient application of phototrophic organisms in an industrial setting, i.e., to achieve high productivities and product yields. Results The physiological responses of the cyanobacterium Synechocystis sp. PCC 6803 to electron sink variation were investigated in a systematic and quantitative manner. A variation in electron demand was achieved by providing two N sources with different degrees of reduction. By additionally varying light and CO2 availabilities, steady state conditions with strongly differing source–sink ratios were established. Balancing absorbed photons and electrons used for different metabolic processes revealed physiological responses to sink/source ratio variation. Surprisingly, an additional electron sink under light and thus energy limitation was found not to hamper growth, but was compensated by improved photosynthetic efficiency and activity. In the absence of carbon and light limitation, an increase in electron demand even stimulated carbon assimilation and growth. Conclusion The metabolism of Synechocystis sp. PCC 6803 is highly flexible regarding the compensation of additional electron demands. Under light limitation, photosynthesis obviously does not necessarily run at its maximal capacity, possibly for the sake of robustness. Increased electron demands can even boost photosynthetic activity and growth. Electronic supplementary material The online version of this article (10.1186/s13068-019-1378-y) contains supplementary material, which is available to authorized users.

Cyanobacteria, also known as blue-green algae, are known to be a rich source of secondary metabolites with diverse chemical structures and biological activities. Over 1100 natural constituents of cyanobacterial origin have been reported in the literature. These include various natural biologically active constituents with antimicrobial, antiviral, immunosuppressive, and anticancer activities. Terrestrial cyanobacteria of genus Fischerella (Family Fischerellaceae), belonging to order Stigonematales have recently received enormous attention from the researchers due to the presence of vast array bioactive compounds. Fischerella species are filamentous cyanobacteria with creeping thallus and sheath in filaments. Several species within the genus are known to produce interesting bioactive constituents. Perhaps the best characterized are F. ambigua and F. muscicola, which have been reported to produce antibacterial, antifungal and antialgal isonitrile containing indole alkaloids, such as hapalindoles, ambiguine isonitriles, fischerindoles, and wetwitindolinones. Keeping in mind these resources, a detailed literature survey on a cultured, fresh water and terrestrial cyanobacterium Fischerella ambigua (Nӓg.) Gom. has been carried out. The current review describes the chemical constituents isolated from different extracts of Fischerella ambigua, as well as their biological activities.

The aim and novelty of this paper are found in assessing the influence of inhibitors and antibiotics on intact cell MALDI-TOF mass spectra of the cyanobacterium Synechococcus sp. UPOC S4 and to check the impact on reliability of identification. Defining the limits of this method is important for its use in biology and applied science. The compounds included inhibitors of respiration, glycolysis, citrate cycle, and proteosynthesis. They were used at 1–10 μM concentrations and different periods of up to 3 weeks. Cells were also grown without inhibitors in a microgravity because of expected strong effects. Mass spectra were evaluated using controls and interpreted in terms of differential peaks and their assignment to protein sequences by mass. Antibiotics, azide, and bromopyruvate had the greatest impact. The spectral patterns were markedly altered after a prolonged incubation at higher concentrations, which precluded identification in the database of reference spectra. The incubation in microgravity showed a similar effect. These differences were evident in dendrograms constructed from the spectral data. Enzyme inhibitors affected the spectra to a smaller extent. This study shows that only a long-term presence of antibiotics and strong metabolic inhibitors in the medium at 10−5 M concentrations hinders the correct identification of cyanobacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF).

A novel symmetrical macrocyclic phthalate hexaester (1) and a known macrocyclic phthalate tetraester (2) were isolated during a natural product-exploring program on the cyanobacterium Moorea producens. Their structures were elucidated based on spectroscopic data, including nuclear magnetic resonance and high-resolution mass spectra. In the antibacterial activity test, compounds 1 and 2 showed no bioactivity at the concentrations tested.

Cryoturbated peat circles (pH 4) in the Eastern European Tundra harbor up to 2 mM pore water nitrate and emit the greenhouse gas N2O like heavily fertilized agricultural soils in temperate regions. The main process yielding N2O under oxygen limited conditions is denitrification, which is the sequential reduction of nitrate/nitrite to N2O and/or N2. N2O reduction to N2is impaired by pH < 6 in classical model denitrifiers and many environments. Key microbes of peat circles are important but largely unknown catalysts forC- andN-cycling associated N2O fluxes. Thus, we hypothesized that the peat circle community includes hitherto unknown taxa and is essentially unable to efficiently perform complete denitrification, i.e., reduce N2O, due to a lowin situpH. 16S rRNA analysis indicated a diverse active community primarily composed of the bacterial class-level taxa Alphaproteobacteria, Acidimicrobiia, Acidobacteria, Verrucomicrobiae, and Bacteroidia, as well as archaeal Nitrososphaeria. Euryarchaeota were not detected.13C2- and12C2-acetate supplemented anoxic microcosms with endogenous nitrate and acetylene at anin situnear pH of 4 were used to assess acetate dependent carbon flow, denitrification and N2O production. Initial nitrate and acetate were consumed within 6 and 11 days, respectively, and primarily converted to CO2and N2, suggesting complete acetate fueled denitrification at acidic pH. Stable isotope probing coupled to 16S rRNA analysis via Illumina MiSeq amplicon sequencing identified acetate consuming key players of the familyBurkholderiaceaeduring complete denitrification correlating withRhodanobacterspp. The archaeal community consisted primarily of ammonia-oxidizing Archaea of Nitrososphaeraceae, and was stable during the incubation. The collective data indicate that peat circles (i) host acid-tolerant denitrifiers capable of complete denitrification at pH 4–5.5, (ii) other parameters like carbon availability rather than pH are possible reasons for high N2O emissionsin situ, and (iii)Burkholderiaceaeare responsive key acetate assimilators co-occurring withRhodanobactersp. during denitrification, suggesting both organisms being associated with acid-tolerant denitrification in peat circles.

Programmed cell death (PCD) in unicellular organisms is in some instances an altruistic trait. When the beneficiaries are clones or close kin, kin selection theory may be used to explain the evolution of the trait, and when the trait evolves in groups of distantly related individuals, group or multilevel selection theory is invoked. In mixed microbial communities, the benefits are also available to unrelated taxa. But the evolutionary ecology of PCD in communities is poorly understood. Few hypotheses have been offered concerning the community role of PCD despite its far-reaching effects. The hypothesis we consider here is that PCD is a black queen. The Black Queen Hypothesis (BQH) outlines how public goods arising from a leaky function are exploited by other taxa in the community. Black Queen (BQ) traits are essential for community survival, but only some members bear the cost of possessing them, while others lose the trait In addition, BQ traits have been defined in terms of adaptive gene loss, and it is unknown whether this has occurred for PCD. Our conclusion is that PCD fulfils the two most important criteria of a BQ (leakiness and costliness), but that more empirical data are needed for assessing the remaining two criteria. In addition, we hold that for viewing PCD as a BQ, the original BQH needs to include social traits. Thus, despite some empirical and conceptual shortcomings, the BQH provides a helpful avenue for investigating PCD in microbial communities. ispartof: MOLECULAR ECOLOGY vol:30 issue:5 pages:1110-1119 ispartof: location:England status: published

Made available in DSpace on 2021-06-25T10:32:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Due to the public and environmental health impact of cyanotoxins, investigations have been focused on finding environmental friendly algaecides from aquatic plants. The present study had the objective to evaluate the population control and physiological response of Microcystis aeruginosa (Kützing) Kützing (strain BCCUSP232) exposed to Pistia stratiotes L. extracts. Aqueous and ethanolic extracts of P. stratiotes at different concentrations (10, 25, and 50 mg L−1) were submitted to M. aeruginosa and reduced significantly (p

Three carbamidocyclophanes, A, F and V, and carbamidocylindrofridin A were isolated from the cultured freshwater cyanobacterium Cylindrospermum stagnale, collected in the Canary Islands. The chemical structures of these compounds were elucidated through NMR, HRMS and ECD spectroscopy. The absolute configuration of carbamidocyclophane A was confirmed using X-ray-diffraction. All compounds showed apoptotic capacity against the SK-MEL-1, SK-MEL-28 and SK-MEL-31 tumour cells. Carbamidocylindrofridin A had the highest anti-tumour potential, with an IC50 of 1 ± 0.3 μM in the SK-MEL-1 cell line, This work was supported by the Fundación de la Universidad Autónoma de Madrid (FUAM)

Sustainable production of biofuels and biochemicals has been broadly accepted as a solution to lower carbon dioxide emissions. Besides being used as lubricants or detergents, oleochemicals are also attractive biofuels as they are compatible with existing transport infrastructures. Cyanobacteria are autotrophic prokaryotes possessing photosynthetic abilities with mature genetic manipulation systems. Through the introduction of exogenous or the modification of intrinsic metabolic pathways, cyanobacteria have been engineered to produce various bio-chemicals and biofuels over the past decade. In this review, we specifically summarize recent progress on photosynthetic production of fatty acids, fatty alcohols, fatty alk(a/e)nes, and fatty acid esters by genetically engineered cyanobacteria. We also summarize recent reports on fatty acid and lipid metabolisms of cyanobacteria and provide perspectives for economic cyanobacterial oleochemical production in the future.

Background Spirulina platensis, an edible cyanobacterium, is considered as a valuable and natural resource of novel anticancer agents. This study aimed to investigate the anticancer potential of major bioactive metabolites from Spirulina platensis on hepatocellular carcinoma cells. The total phenolic and alkaloid content of S. platensis were determined using spectrophotometric procedures and thin-layer chromatography. ‍‍Cellular viability of HepG2 cancer cells and normal fibroblasts was evaluated using MTT assay after 24 h treatment with 0.02–2 mg/ml of alkaloids, phenolic compounds, aqueous, and methanol extracts from Spirulina platensis. Results Total phenolic and total alkaloid compounds were 150.5 ± 1.18 mg gallic acid equivalents/mg extract and 11.4 ± 0.05 mg atropine equivalents/mg extract, respectively. All tested extracts and compounds demonstrated the inhibitory effect on the viability of HepG2 cells in a dose-dependent manner without cytotoxicity on normal cells. The most potent anticancer activity was induced by alkaloids (2 ± 0.001 mg/ml) with 80% reduction in cell viability and an IC50 of 0.53 ± 0.08 mg/ml. IC50 values of the aqueous extract, the methanolic extract, and phenolic compounds were 1.7 ± 0.14, 1.28 ± 0.22, and 0.86 ± 0.14 mg/ml, respectively. Conclusions This is the first report to demonstrate anticancer effects of alkaloids and phenolic compounds of Spirulina platensis in relation to liver cancer.

Background Based on the peak expression times during infection, early, middle, and late genes have been characterized in viruses (cyanophages) that infect the unicellular cyanobacterium Prochlorococcus. Laboratory experiments show that some cyanophages can only replicate in the light and thus exhibit diurnal infection rhythms under light-dark cycles. Field evidence also suggests synchronized infection of Prochlorococcus by cyanophages in the oceans, which should result in progressive expression of cyanophage early, middle, and late genes. However, distinct temporal expression patterns have not been observed in cyanophage field populations. Results In this study, we reanalyzed a previous metatranscriptomic dataset collected in the North Pacific Subtropical Gyre. In this dataset, it was previously shown that aggregate transcripts from cyanophage scaffolds display diurnal transcriptional rhythms with transcript abundances decreasing at night. By mapping metatranscriptomic reads to individual viral genes, we identified periodically expressed genes from putative viruses infecting the cyanobacteria Prochlorococcus and Synechococcus, heterotrophic bacteria, and algae. Of the 41 cyanophage genes, 35 were from cyanomyoviruses. We grouped the periodically expressed cyanomyovirus genes into early, middle, and late genes based on the conserved temporal expression patterns of their orthologs in cyanomyovirus laboratory cultures. We found that the peak expression times of late genes in cyanophage field populations were significantly later than those of early and middle genes, which were similar to the temporal expression patterns of synchronized cyanophage laboratory cultures. Conclusions The significantly later peak expression times of late genes in cyanomyovirus field populations suggest that cyanophage infection of Prochlorococcus is synchronized in the North Pacific Subtropical Gyre. The night-time peak expression of late genes also suggests synchronized lysis of Prochlorococcus at night, which might result in synchronized release of dissolved organic matter to the marine food web.

A filamentous cyanobacterium, Trichormus sp. IMU26, capable of rapid growth and high phycobiliprotein (PBP) production under N2-fixing conditions was isolated. The strain has been maintained in N-free BG-11 medium for more than 20 months upon isolation. Nitrogen supply resulted in higher soluble protein and saccharide content but a lower growth rate and PBP production in Trichormus sp. IMU26. Short-term N-P deprivation induced PBP production with no clear change in growth while growth and PBP content decreased in the longer incubation period. Induction of PBP production in N-P-deprived cells was characterized by FTIR analysis and change in carotenoid-pigment interactions. Rapid induction of zeaxanthin and ?-carotene production and slight reduction of echinenone and canthaxanthin levels might be associated with increased PBP levels in short-term N-P deprivation of Trichormus sp. IMU26. Overall, long-term diazotrophic growth of N2-fixing cyanobacteria may increase the PBP yield. © 2020, Springer Nature B.V. TAGEM-12/AR-GE/13 This study was supported by a grant from the Turkish Ministry of Agriculture and Forestry General Directorate of Agricultural Research (TAGEM-12/AR-GE/13).

Some cyanobacteria use light outside the visible spectrum for oxygenic photosynthesis. The far-red light (FRL) region is made accessible through a complex acclimation process that involves the formation of new phycobilisomes and photosystems containing chlorophyll f. Diverse cyanobacteria ranging from unicellular to branched-filamentous forms show this response. These organisms have been isolated from shaded environments such as microbial mats, soil, rock, and stromatolites. However, the full spread of chlorophyll f-containing species in nature is still unknown. Currently, discovering new chlorophyll f cyanobacteria involves lengthy incubation times under selective far-red light. We have used a marker gene to detect chlorophyll f organisms in environmental samples and metagenomic data. This marker, apcE2, encodes a phycobilisome linker associated with FRL-photosynthesis. By focusing on a far-red motif within the sequence, degenerate PCR and BLAST searches can effectively discriminate against the normal chlorophyll a-associated apcE. Even short recovered sequences carry enough information for phylogenetic placement. Markers of chlorophyll f photosynthesis were found in metagenomic datasets from diverse environments around the globe, including cyanobacterial symbionts, hypersaline lakes, corals, and the Arctic/Antarctic regions. This additional information enabled higher phylogenetic resolution supporting the hypothesis that vertical descent, as opposed to horizontal gene transfer, is largely responsible for this phenotype’s distribution.

Thermosynechococcus is a genus of thermophilic unicellular cyanobacteria that are dominant in microbial mats at about 50-65°C in alkaline hot springs of eastern Asia. We used PacBio SMRT Sequencing to sequence the complete genome of a novel strain of thermophilic cyanobacterium, Thermosynechococcus sp. CL-1, isolated from the Chin-Lun hot spring (pH 9.3, 62°C) in Taiwan. Genome-scale phylogenetic analysis and average nucleotide identity (ANI) results suggested that CL-1 is a new species in the genus Thermosynechococcus. Comparative genome analysis revealed divergent genome structures of Thermosynechococcus strains. In addition, the distinct genetic differences between CL-1 and the other Thermosynechococcus strains are related to photosynthesis, transporters, signal transduction, the chaperone/usher system, nitric oxide protection, antibiotic resistance, prokaryotic immunity systems, and other physiological processes. This study suggests that Thermosynechococcus strains have actively acquired many putative horizontally transferred genes from other bacteria that enabled them to adapt to different ecological niches and stressful conditions in hot springs.

Microorganisms may play an important role in the binding of sediments and the formation of sedimentary structures, by means of the formation of biofilms and microbial mats. In this work, filamentous cyanobacteria from three different environments were compared: a biofilm from a lacustrine environment with intervals of flooding/desiccation, a biofilm from a continental saline environment, and a microbial mat from a thermal spring. The optical identification of the cyanobacteria allowed to establish that in the biofilms there is a dominance of the Order Oscillatoriales, while in the microbial mat both cyanobacteria of the Orders Nostocales and Oscillatoriales dominated. Two rock samples isolated from the thermal spring which genesis was possibly influenced by the activity of cyanobacteria are described and classified. One of them is a travertine/microbial framestone with stromatolitic and thrombolytic texture. The second one is classified as a mudstone/microbial boundstone. Finally, a comparison between the sedimentary structures identified in those rocks with similar structures in the formations La Luna, Paja and Tetuán, deposited during regressive phases of the Colombian Cretaceous epyric sea, and microbial mat features previously described is performed. Based on morphological resemblance, fibrillar networks identified locally in those formations are interpreted as possible biolaminations originated from the activity of cyanobacteria. RESUMEN Los microorganismos pueden jugar un papel importante en la cohesión de los sedimentos y la formación de estructuras sedimentarias, a través de la formación de biopelículas y tapetes microbianos. En el presente trabajo se compararon las cianobacterias filamentosas de tres ambientes diferentes: una biopelícula de un ambiente lacustre con intervalos de inundación/desecación, una biopelícula de un ambiente continental salino, y un tapete microbiano aislado de una fuente termal. La identificación óptica de las cianobacterias permitió establecer que en las biopelículas predomina el Orden Oscillatoriales, mientras que en el tapete microbiano predominan a la vez cianobacterias de los Ordenes Nostocales y Oscillatoriales. Se describen y clasifican dos muestras de roca tomadas de la fuente termal cuya génesis posiblemente estuvo influenciada por la actividad de cianobacterias. Una de ellas es un travertino/ "framestone microbiana con textura estromatolítica y trombolítica. La segunda se clasificó como una lodolita/ "boundstone" microbiana. Finalmente, se realiza una comparación entre las estructuras sedimentarias identificadas en dichas rocas con estructuras similares en las formaciones La Luna, Tetuán y Paja, depositadas durante fases regresivas del mar epicontinental de Colombia durante el Cretácico, y las características de tapetes microbianos descritas previamente. Con base en las similitudes morfológicas, las redes fibrilares identificadas de manera localizada en dichas formaciones se interpretan como posibles bioláminas originadas por la actividad de cianobacterias.

Arthrospira platensis is one of the most important cultured microalgal species in the world. Arthrospira complete dry biomass (ACDB) has been reported as an interesting feedstock for many industries, including biodiesel production. The A. platensis by-product of biodiesel production (lipid-free biomass, LFB) is a source of proteins, functional molecules, and carbohydrates, and can also be reused in several applications. The current study investigated the efficiency of ACDB and LFB in bioremediation of dye (Ismate violet 2R, IV2R) from textile effluents. In addition, the potential of ACDB and LFB loaded by IV2R as a feed for Rotifer, Brachionus plicatilis, was examined. The surface of the adsorbents was characterized by SEM, FTIR, and Raman analysis to understand the adsorption mechanism. The batch sorption method was examined as a function of adsorbent dose (0.02–0.01 g L−1), solution initial concentration (10–100 mg L−1), pH (2–10), and contact time (15–180 min). The kinetic studies and adsorption isotherm models (Freundlich, Langmuir, Tempkin, and Halsey) were used to describe the interaction between dye and adsorbents. The results concluded that the adsorption process increased with increasing ACDB and LFB dose, contact time (120 min), initial IV2R concentration (10 mg L−1), and acidity pH (2 and 6, respectively). For the elimination of industrial textile wastewater, the ACDB and LFB sorbents have good elimination ability of a dye solution by 75.7% and 61.11%, respectively. The kinetic interaction between dye and adsorbents fitted well to Langmuir, Freundlish, and Halsey models for LFB, and Langmuir for ACDB at optimum conditions with R2 &gt, 0.9. In addition, based on the bioassay study, the ACDB and LFB loaded by IV2R up to 0.02 g L−1 may be used as feed for the marine Rotifer B. plicatilis.

Formation of the multi-subunit oxygen-evolving photosystem II (PSII) complex involves a number of auxiliary protein factors. In this study we compared the localization and possible function of two homologous PSII assembly factors, Psb28-1 and Psb28-2, from the cyanobacterium Synechocystis sp. PCC 6803. We demonstrate that FLAG-tagged Psb28-2 is present in both the monomeric PSII core complex and a PSII core complex lacking the inner antenna CP43 (RC47), whereas Psb28-1 preferentially binds to RC47. When cells are exposed to increased irradiance, both tagged Psb28 proteins additionally associate with oligomeric forms of PSII and with PSII-PSI supercomplexes composed of trimeric photosystem I (PSI) and two PSII monomers as deduced from electron microscopy. The presence of the Psb27 accessory protein in these complexes suggests the involvement of PSI in PSII biogenesis, possibly by photoprotecting PSII through energy spillover. Under standard culture conditions, the distribution of PSII complexes is similar in the wild type and in each of the single psb28 null mutants except for loss of RC47 in the absence of Psb28-1. In comparison with the wild type, growth of mutants lacking Psb28-1 and Psb27, but not Psb28-2, was retarded under high-light conditions and, especially, intermittent high-light/dark conditions, emphasizing the physiological importance of PSII assembly factors for light acclimation.

Cylindrospermopsin (CYN)-producing cyanobacterial blooms such as Raphidiopsis, Aphanizomenon, Anabaena, Umezakia, and Lyngbya spp. are occurring more commonly and frequently worldwide. CYN is an environmentally stable extracellular toxin, which inhibits protein synthesis, and, therefore, can potentially affect a wide variety of aquatic biota. Submerged and floating macrophytes, as primary producers in oligotrophic habitats, are at risk of exposure and information on the effects of CYN exposure at environmentally relevant concentrations is limited. In the present study, we investigated CYN uptake in the floating macrophyte Lemna minor with exposure to reported environmental concentrations. The effects were evaluated in terms of bioaccumulation, relative plant growth, and number of fronds per day. Variations in the concentrations and ratios of the chlorophylls as stress markers and carotenoids as markers of oxidative stress defense were measured. With exposure to 25 &mu, g/L, L. minor could remove 43% of CYN within 24 h but CYN was not bioaccumulated. Generally, the pigment concentrations were elevated with exposure to 0.025, 0.25, and 2.5 &mu, g/L CYN after 24 h, but normalized quickly thereafter. Changes in relative plant growth were observed with exposure to 0.25 and 2.5 &mu, g/L CYN. Adverse effects were seen with these environmentally realistic concentrations within 24 h, however, L. minor successfully recovered within the next 48&ndash, 96 h.

Although microcystins (MCs) are the most commonly studied cyanotoxins, their significance to the producing organisms remains unclear. MCs are known as endotoxins, but they can be found in the surrounding environment due to cell lysis, designated as extracellular MCs. In the present study, the interactions between MC producing and the non-producing strains of Microcystis aeruginosa, PCC 7806 and PCC 7005, respectively, and a green alga, Desmodesmus subspicatus, were studied to better understand the probable ecological importance of MCs at the collapse phase of cyanobacterial blooms. We applied a dialysis co-cultivation system where M. aeruginosa was grown inside dialysis tubing for one month. Then, D. subspicatus was added to the culture system on the outside of the membrane. Consequently, the growth of D. subspicatus and MC contents were measured over a 14-day co-exposure period. The results showed that Microcystis negatively affected the green alga as the growth of D. subspicatus was significantly inhibited in co-cultivation with both the MC-producing and -deficient strains. However, the inhibitory effect of the MC-producing strain was greater and observed earlier compared to the MC-deficient strain. Thus, MCs might be considered as an assistant factor that, in combination with other secondary metabolites of Microcystis, reinforce the ability to outcompete co-existing species.

Microcystins (MCs) are hepatotoxic and potentially carcinogenic cyanotoxins. They exhibit high structural variability, with nearly 250 variants described to date. This variability can result in incomplete detection of MC variants during lake surveys due to the frequent use of targeted analytical methods and a lack of standards available for identification and quantitation. In this study, Lake Uluabat in Turkey was sampled during the summer of 2015. Phylogenetic analysis of the environmental mcyA sequences suggested Microcystis spp. were the major MC contributors. A combination of liquid chromatography-tandem mass spectrometry (LC-MS/MS), liquid chromatography with UV detection and mass spectrometry (LC-UV-MS), and a novel liquid chromatography-high resolution mass spectrometry (LC-HRMS) method, together with thiol and periodate reactivity, revealed more than 36 MC variants in the lake samples and a strain of M. aeruginosa (AQUAMEB-24) isolated from Lake Uluabat. Only MCs containing arginine at position-4 were detected in the culture, while MC-LA, -LY, -LW and -LF were also detected in the lake samples, suggesting the presence of other MC producers in the lake. The previously unreported MCs MC-(H2)YR (dihydrotyrosine at position-2) (17), [epoxyAdda(5)]MC-LR, [DMAdda(5)]MC-RR (1) and [Mser(7)]MC-RR (8) were detected in the culture and/or field samples. This study is a good example of how commonly used targeted LC-MS methods can underestimate the diversity of MCs in freshwater lakes and cyanobacteria cultures and how untargeted LC-MS methods can be used to comprehensively assess MC diversity present in a new system.

In plants, chlorophyll (Chl) a and b are interconvertible by the action of three enzymes—chlorophyllide a oxygenase, Chl b reductase (CBR) and 7-hydroxymethyl chlorophyll a reductase (HCAR). These reactions are collectively referred to as the Chl cycle. In plants, this cyclic pathway ubiquitously exists and plays essential roles in acclimation to different light conditions at various developmental stages. By contrast, only a limited number of cyanobacteria species produce Chl b, and these include Prochlorococcus, Prochloron, Prochlorothrix and Acaryochloris. In this study, we investigated a possible existence of the Chl cycle in Chl b synthesizing cyanobacteria by testing in vitro enzymatic activities of CBR and HCAR homologs from Prochlorothrix hollandica and Acaryochloris RCC1774. All of these proteins show respective CBR and HCAR activity in vitro, indicating that both cyanobacteria possess the potential to complete the Chl cycle. It is also found that CBR and HCAR orthologs are distributed only in the Chl b-containing cyanobacteria that habitat shallow seas or freshwater, where light conditions change dynamically, whereas they are not found in Prochlorococcus species that usually habitat environments with fixed lighting. Taken together, our results implicate a possibility that the Chl cycle functions for light acclimation in Chl b-containing cyanobacteria.

This review covers the structure, synthesis, biological activity, biosynthesis and distribution of cyanobacterial alkylresorcinols., Covering: up to 2019 Alkylresorcinols are amphiphilic metabolites, well-known for their diverse biological activities, produced by both prokaryotes and eukaryotes. A few classes of alkylresorcinol scaffolds have been reported from the photoautotrophic cyanobacteria, ranging from the relatively simple hierridins to the more intricate cylindrocyclophanes. Recently, it has emerged that cyanobacteria employ two different biosynthetic pathways to produce unique alkylresorcinol scaffolds. However, these convergent pathways intersect by sharing biosynthetic elements which lead to common structural motifs. To obtain a broader view of the biochemical diversity of these compounds in cyanobacteria, we comprehensively cover the isolation, structure, biological activity and biosynthesis of their mono- and dialkylresorcinols. Moreover, we provide an overview of the diversity and distribution of alkylresorcinol-generating biosynthetic gene clusters in this phylum and highlight opportunities for discovery of novel alkylresorcinol scaffolds. Because some of these molecules have inspired notable syntheses, different approaches used to build these molecules in the laboratory are showcased.

Marine biofouling has severe economic impacts and cyanobacteria play a significant role as early surface colonizers. Despite this fact, cyanobacterial biofilm formation studies in controlled hydrodynamic conditions are scarce. In this work, computational fluid dynamics was used to determine the shear rate field on coupons that were placed inside the wells of agitated 12-well microtiter plates. Biofilm formation by three different cyanobacterial strains was assessed at two different shear rates (4 and 40 s-1 ) which can be found in natural ecosystems and using different surfaces (glass and perspex). Biofilm formation was higher under low shear conditions, and differences obtained between surfaces were not always statistically significant. The hydrodynamic effect was more noticeable during the biofilm maturation phase rather than during initial cell adhesion and optical coherence tomography showed that different shear rates can affect biofilm architecture. This study is particularly relevant given the cosmopolitan distribution of these cyanobacterial strains and the biofouling potential of these organisms. This work was financially supported by project UID/EQU/ 00511/2019 – Laboratory for Process Engineering, Environment, Biotechnology and Energy – LEPABE funded by national funds through FCT/MCTES (PIDDAC) and by the CVMAR+i – Industrial Innovation and Marine Biotechnology Valorisation, funded by INTERREG V A Espanha Portugal (POCTEP) [0302_CVMAR_I_1_P]. The authors also acknowledge support from the EU COST Actions iPROMEDAI (TD1305) and ENBA (CA15216), and M.J.R. acknowledges a PhD grant from FCT (SFRH/BD/140080/2018).

Cyanobacteria may live in the water column and in the benthos of aquatic environments, or be symbionts of other organisms, as in the case of Phormidium-like cyanobacteria, known to influence the ecology of freshwater and marine ecosystems. A strain of Phormidium-like cyanobacteria has been recently isolated as a free-living epiphyte of leaves of Posidonia oceanica (L.) Delile in the Mediterranean sea and its biology and ecology is herein investigated. It was identified as Halomicronema metazoicum, previously known uniquely as a symbiont of marine sponges. We cultivated it in a range of light irradiances, temperatures and salinities, to establish the most suitable conditions for the production of allelopatic and toxic compounds. The bioactivity of its spent culture medium was measured by means of standard toxicity tests performed on two model organisms. Our results indicate that at least two bioactive compounds are produced, at low and high irradiance levels and at two temperatures. The main compounds influencing the survival of model organisms are produced at the highest temperature and high or intermediate irradiance levels. The present research contributes to the understanding of critical toxigenic relationships among cyanobacteria and invertebrates, possibly influencing the ecology of such a complex environment as P. oceanica. Future isolation, identification and production of bioactive compounds will permit their exploitation for biotechnologies in the field of ecological conservation and medical applications.

Combination of biotic and abiotic factors influences the effects of naturally occurring or anthropogenic chemicals on photosynthetic microorganisms in the aquatic environment. Nonetheless, the combined effects of physical stressors and species-species interaction on chemicals’ toxicity are still poorly understood. The present study examines the responses of the green alga Chlamydomonas reinhardtii and the cyanobacterium Synechocystis sp. alone and in mixtures to copper exposure under increasing visible light intensities. Cell growth, chlorophyll bleaching, oxidative stress and membrane permeability were determined by flow cytometry in both mono- and multi-species tests. The results revealed that species-species interactions influenced copper toxicity under different light regimes at 4 h and 48 h - exposure. For a given light condition, monocultures of Synechocystis sp. were more sensitive to copper than those of C. reinhardtii. In long-term incubation C. reinhardtii sensitivity to copper diminished in presence of Synechocystis sp. under low-intensity light, however it was enhanced under high-intensity light. The present results revealed the complex interplay between visible light intensity variations, species-species interaction and copper effects to phytoplankton in long- term exposure.

The N2 fixing cyanobacterium Trichodesmium is a critically important organism in oligotrophic marine ecosystems, supplying "new" nitrogen (N) to the otherwise N-poor tropical and subtropical regions where it occurs. Low concentrations of phosphorus (P) in these regions can constrain Trichodesmium distribution and N2 fixation rates. Physiological characterization of a single species in a mixed community can be challenging, and 'omic approaches are increasingly important tools for tracking nutritional physiology in a taxon-specific manner. As such, studies examining the dynamics of gene and protein markers of physiology (e.g., nutrient stress) are critical for the application and interpretation of such 'omic data in situ. Here we leveraged combined transcriptomics, proteomics, and enzyme activity assays to track the physiological response of Trichodesmium erythraeum IMS101 to P deficiency and subsequent P re-supply over 72 h of sampling. P deficiency resulted in differential gene expression, protein abundance, and enzyme activity that highlighted a synchronous shift in P physiology with increases in the transcripts and corresponding proteins for hydrolyzing organic phosphorus, taking up phosphate with higher affinity, and modulating intracellular P demand. After P deficiency was alleviated, gene expression of these biomarkers was reduced to replete levels within 4 h of P amendment. A number of these gene biomarkers were adjacent to putative pho boxes and their expression patterns were similar to a sphR response regulator. Protein products of the P deficiency biomarkers were slow to decline, with 84% of the original P deficient protein set still significantly differentially expressed after 72 h. Alkaline phosphatase activity tracked with proteins for this enzyme. With the rapid turnover time of transcripts, they appear to be good biomarkers of a P stress phenotype, whereas proteins, with a slower turnover time, may better reflect cellular activities. These results highlight the importance of validating and pairing transcriptome and proteome data that can be applied to physiological studies of key species in situ.

Publisher's version (útgefin grein)., Marine sponges and cyanobacteria have a long history of co-evolution, with documented genome adaptations in cyanobionts. Both organisms are known to produce a wide variety of natural compounds, with only scarce information about novel natural compounds produced by cyanobionts. In the present study, we aimed to address their toxicological potential, isolating cyanobacteria (n = 12) from different sponge species from the coast of Portugal (mainland, Azores, and Madeira Islands). After large-scale growth, we obtained both organic and aqueous extracts to perform a series of ecologically-relevant bioassays. In the acute toxicity assay, using nauplii of Artemia salina, only organic extracts showed lethality, especially in picocyanobacterial strains. In the bioassay with Paracentrotus lividus, both organic and aqueous extracts produced embryogenic toxicity (respectively 58% and 36%), pointing to the presence of compounds that interfere with growth factors on cells. No development of pluteus larvae was observed for the organic extract of the strain Chroococcales 6MA13ti, indicating the presence of compounds that affect skeleton formation. In the hemolytic assay, none of the extracts induced red blood cells lysis. Organic extracts, especially from picoplanktonic strains, proved to be the most promising for future bioassay-guided fractionation and compounds isolation. This approach allows us to classify the compounds extracted from the cyanobacteria into effect categories and bioactivity profiles., Funding: This work was financed by UID/Multi/04423/2013 and by the Structured Program of R&D&I INNOVMAR—Innovation and Sustainability in the Management and Exploitation of Marine Resources (reference NORTE-01-0145-FEDER-000035, Research Line NOVELMAR), funded by the Northern Regional Operational Program (NORTE2020) through the European Regional Development Fund (ERDF).and by the grants PTDC/MAR/099642/2008, PhD grants SFRH/BD/73033/2010 and the Fellowship grant BI/PTDC/MAR/099642/2008/2011-030.

Cyanobacteria are prokaryotic phototrophs capable of achieving high cellular densities with minimal inputs. These prokaryotic organisms can grow using sunlight as energy source and carbon dioxide as carbon source what makes them promising candidates as microbial cell factories for the production of numerous compounds such as chemicals, fuels, or biocatalysts. In this study, we have successfully designed and constructed using synthetic biology approach two recombinant strains of Synechococcus elongatus PCC7942 for heterologous expression of the industrially relevant Bacillus subtilis CotA laccase. One of the strains (PCC7942-NSI-CotA) was constructed through integration of the laccase gene into neutral site I of the cyanobacterial genome whilst the other (PCC7942-NSII-CotA) targeted neutral site II of the genome. Of the two strains the one with CotA laccase integrated in neutral site II (PCC7942-NSII-CotA) was superior in terms of growth rate and enzymatic activity toward typical laccase substrates: ABTS [2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonate)] and syringaldazine. That may suggest that two of the traditionally used neutral sites of S. elongatus PCC7942 are not equally suitable for the expression of certain transgenes. The PCC7942-NSII-CotA produced protein was capable of decolourising three classes of dyes namely: anthraquinonic-, azo-, and indigoid-type over 7 days of incubation making the strain a potentially useful microbial cell factory for the production of broad-spectrum biodegradation agent. Interestingly, presence of additional synthetic redox mediator ABTS had no effect on the degradation of these dyes.

Lower-energy photons do the work, too Plants and cyanobacteria use chlorophyll-rich photosystem complexes to convert light energy into chemical energy. Some organisms have developed adaptations to take advantage of longer-wavelength photons. Nürnberg et al. studied photosystem complexes from cyanobacteria grown in the presence of far-red light. The authors identified the primary donor chlorophyll as one of a few chlorophyll molecules in the far-red light–adapted enzymes that were chemically altered to shift their absorption spectrum. Kinetic measurements demonstrated that far-red light is capable of directly driving water oxidation, despite having less energy than the red light used by most photosynthetic organisms. Science , this issue p. 1210

Insight into how environmental change determines the production and distribution of cyanobacterial toxins is necessary for risk assessment. Management guidelines currently focus on hepatotoxins (microcystins). Increasing attention is given to other classes, such as neurotoxins (e.g., anatoxin-a) and cytotoxins (e.g., cylindrospermopsin) due to their potency. Most studies examine the relationship between individual toxin variants and environmental factors, such as nutrients, temperature and light. In summer 2015, we collected samples across Europe to investigate the effect of nutrient and temperature gradients on the variability of toxin production at a continental scale. Direct and indirect effects of temperature were the main drivers of the spatial distribution in the toxins produced by the cyanobacterial community, the toxin concentrations and toxin quota. Generalized linear models showed that a Toxin Diversity Index (TDI) increased with latitude, while it decreased with water stability. Increases in TDI were explained through a significant increase in toxin variants such as MC-YR, anatoxin and cylindrospermopsin, accompanied by a decreasing presence of MC-LR. While global warming continues, the direct and indirect effects of increased lake temperatures will drive changes in the distribution of cyanobacterial toxins in Europe, potentially promoting selection of a few highly toxic species or strains. © 2018 by the authors. Licensee MDPI, Basel, Switzerland., The authors acknowledge COST Action ES 1105 “CYANOCOST—Cyanobacterial blooms and toxins in water resources: Occurrence impacts and management” and COST Action ES 1201 “NETLAKE—Networking Lake Observatories in Europe” for contributing to this study through networking and knowledge sharing with European experts in the field. Evanthia Mantzouki was supported by a grant from the Swiss State Secretariat for Education, Research and Innovation (SERI) to Bas Ibelings and by supplementary funding from the University of Geneva. We thank Clare Ahnlund, Ena Suarez and Irene Gallego for helping out with the Swiss survey. We thank Wendy Beekman and Els J. Faassen for the nutrient and toxin analysis.

Cosmopolitan Gram-negative cyanobacteria may affect human and animal health by contaminating terrestrial, marine and freshwater environments with toxins, such as lipopolysaccharide (LPS). The cyanobacterial genus Scytonema (S) produces several toxins, but to our knowledge the bioactivity of genus Scytonema LPS has not been investigated. We recently reported that cyanobacterium Oscillatoria sp. LPS elicited classical and alternative activation of rat microglia in vitro [1]. Thus, we hypothesized that treatment of brain microglia in vitro with either cyanobacteria S. javanicum or S. ocellatum LPS might stimulate classical and alternative activation with concomitant release of superoxide anion (O2−), matrix metalloproteinase-9 (MMP-9) and cytokines and chemokines. Microglia were isolated from neonatal rats and treated in vitro with either S. javanicum LPS, S. ocellatum LPS, or E. coli LPS (positive control) in a concentration-dependent manner for 18 hours at 35.9 °C. We observed that treatment of microglia with either E. coli LPS, S. javanicum or S. ocellatum LPS generated statistically significant and concentration-dependent O2−, MMP-9 and pro-inflammatory cytokines IL-6 and TNF-α, pro-inflammatory chemokines MIP-2/CXCL-2, CINC-1/CXCL-1 and MIP-1α/CCL3, and the anti-inflammatory cytokine IL-10. Thus, our results provide experimental support for our working hypothesis because both S. javanicum and S. ocellatum LPS elicited classical and alternative activation of microglia and concomitant release of O2-, MMP-9 and cytokines and chemokines in a concentration-dependent manner. To our knowledge this is the first report on the toxicity of cyanobacteria S. javanicum and S. ocellatum LPS to microglia, an immune cell type involved in neuroinflammation and neurotoxicity in the central nervous system.&nbsp

Swinholides are 42-carbon ring polyketides with a 2-fold axis of symmetry. They are potent cytotoxins that disrupt the actin cytoskeleton. Swinholides were discovered from the marine sponge Theonella sp. and were long suspected to be produced by symbiotic bacteria. Misakinolide, a structural variant of swinholide, was recently demonstrated to be the product of a symbiotic heterotrophic proteobacterium. Here, we report the production of swinholide A by an axenic strain of the terrestrial cyanobacterium Nostoc sp. strain UHCC 0450. We located the 85-kb trans -AT polyketide synthase (PKS) swinholide biosynthesis gene cluster from a draft genome of Nostoc sp. UHCC 0450. The swinholide and misakinolide biosynthesis gene clusters share an almost identical order of catalytic domains, with 85% nucleotide sequence identity, and they group together in phylogenetic analysis. Our results resolve speculation around the true producer of swinholides and demonstrate that bacteria belonging to two distantly related phyla both produce structural variants of the same natural product. In addition, we described a biosynthesis cluster from Anabaena sp. strain UHCC 0451 for the synthesis of the cytotoxic and antifungal scytophycin. All of these biosynthesis gene clusters were closely related to each other and created a group of cytotoxic macrolide compounds produced by trans -AT PKSs of cyanobacteria and proteobacteria. IMPORTANCE Many of the drugs in use today originate from natural products. New candidate compounds for drug development are needed due to increased drug resistance. An increased knowledge of the biosynthesis of bioactive compounds can be used to aid chemical synthesis to produce novel drugs. Here, we show that a terrestrial axenic culture of Nostoc cyanobacterium produces swinholides, which have been previously found only from marine sponge or samples related to them. Swinholides are polyketides with a 2-fold axis of symmetry, and they are potent cytotoxins that disrupt the actin cytoskeleton. We describe the biosynthesis gene clusters of swinholide from Nostoc cyanobacteria, as well as the related cytotoxic and antifungal scytophycin from Anabaena cyanobacteria, and we study the evolution of their trans -AT polyketide synthases. Interestingly, swinholide is closely related to misakinolide produced by a symbiotic heterotrophic proteobacterium, demonstrating that bacteria belonging to two distantly related phyla and different habitats can produce similar natural products.

Allelopathic compounds produced by cyanobacteria may play important roles in the dynamics of several biological systems. The main goal of this work was to investigate reciprocal allelopathic effects between species of two relevant groups of marine cyanobacteria: The picocyanobacterium Synechococcus sp. and the filamentous species Nodularia spumigena. Our experimental design consisted of cell-free filtrates and co-cultures. We demonstrated that Synechococcus sp. had a strong inhibitory effect on N. spumigena, and surprisingly, there was no reciprocal effect from the filamentous cyanobacteria. We detected this effect both in co-cultures and cell-free filtrate bioassays. These effects depended on light conditions in the culture of the allelopathic species. This allelopathic effect against N. spumigena triggered physiological responses leading to reduced chlorophyll a (Chl a) and carotenoid (Car) content, cell shape distortions and, often, cell lysis. Surprisingly, no evidence was found of allelopathic effects of our strain of N. spumigena (a well-known allelopathic species) against Synechococcus sp. These results support the fact that allelopathic interactions between Synechococcus sp. and N. spumigena may be a factor influencing the formation of massive bloom of the former organisms in many aquatic ecosystems, like the Baltic Sea, where the two species constitute a relevant fraction of phytoplankton biomass. © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. This study was supported by BMN grants, Poland, No. 538-G245-B568-17 and FCT Project UID/Multi/04 423/ 2013.

Cyanobacteria are found globally due to their adaptation to various environments. The occurrence of cyanobacterial blooms is not a new phenomenon. The bloom-forming and toxin-producing species have been a persistent nuisance all over the world over the last decades. Evidence suggests that this trend might be attributed to a complex interplay of direct and indirect anthropogenic influences. To control cyanobacterial blooms, various strategies, including physical, chemical, and biological methods have been proposed. Nevertheless, the use of those strategies is usually not effective. The isolation of natural compounds from many aquatic and terrestrial plants and seaweeds has become an alternative approach for controlling harmful algae in aquatic systems. Seaweeds have received attention from scientists because of their bioactive compounds with antibacterial, antifungal, anti-microalgae, and antioxidant properties. The undesirable effects of cyanobacteria proliferations and potential control methods are here reviewed, focusing on the use of potent bioactive compounds, isolated from seaweeds, against microalgae and cyanobacteria growth. © 2018 by the authors. Acknowledgments: This work was supported by FCT Project UID/Multi/04423/2013,by the Structured Program of R&D&I INNOVMAR—Innovation and Sustainability in the Management and Exploitation of Marine of R&D&I INNOVMAR—Innovation and Sustainability in the Management and Exploitation of Marine Resources Resources (reference NORTE-01-0145-FEDER-000035, Research Line NOVELMAR), funded by the Northern Regional Operational Program (NORTE2020) through the European Regional Development Fund (ERDF) and bCyV Mthaer +pIro(0je3c0t2 _CCVVMMaAr+RI_ I(_013_0P2_)CfuVnMdeAdRb_yI_t1h_ePp) rofugnradmedI nbteyrrtehge VpAroEgrsapman hIna—terProegrt uVg aAl ( PEOspCaTnEhPa)—20P1o4r-t2u0g2a0l.

The production and release of allelopathic compounds is an important adaptation by which some species of cyanobacteria can achieve a competitive advantage over other primary producers. In the present study we tested the allelopathic activity of the picocyanobacterium Synechococcus sp. against the following coexisting unicellular eukaryote microalgae: Porphyridium purpureum, Stichococcus bacillaris, Prymnesium parvum and Nitzschia dissipata. With these species, we covered a wide range of taxonomic groups. We demonstrated that both the addition of Synechococcus sp. cell-free filtrate and coculture inhibited the growth, chlorophyll content and photosynthetic rate of P. purpureum and S. bacillaris. Conversely, P. parvum, a well-known mixotroph, was positively affected by both Synechococcus sp. treatments. In contrast, N. dissipata was not affected by either the picocyanobacterial filtrate or coculture. These results suggest that the negative allelopathic effect is related to a reduction in the photosynthetic rate, and that Synechococcus sp. allelopathy should be taken into account in the interactions between picocyanobacteria and eukaryote competitors coexisting in a planktonic system. © CSIRO. This study was supported by grants from BMN, Poland (number 538-G245-B568-17) and FCT Project UID/Multi/04423/2013.

Previous studies have demonstrated the modulation of glutathione transferases (GSTs) induced by microcystin (MC) alone or in combination with other cyanobacterial secondary metabolites in bivalves. However, interspecies information about which and how GST isoforms are affected by these secondary metabolites is still scarce, especially considering the dynamic process involving their uptake and elimination routes. In this context, the role of GSTs gene expression changes in response to a toxic Microcystis aeruginosa extract were examined for Mytilus galloprovincialis and Ruditapes philippinarum during exposure and recovery phases. The expression levels of sigma 1, sigma 2, pi and mu-class GST genes were analyzed in the hepatopancreas of both bivalve species during cyanobacteria extract exposure (24 h) and post-exposure (24 and 72 h). Only a significant induction of sigma 1-class GST expression was observed for R. philippinarum upon 24-hour exposure of both bivalve species to Microcystis extract. During the recovery phase, GST transcriptional changes for M. galloprovincialis were characterized by an early induction (24 h) of sigma 1 and sigma 2 transcripts. On the other hand, GST transcriptional changes for R. philippinarum during post-exposure phase were characterized by an early induction (24 h) of sigma 1 and mu transcripts and a later induction (72 h) of the four analyzed GST transcripts. Such differences reflect variable GST response mechanisms to cope with MC-producing cyanobacterial blooms exposure between these two bivalve species, revealing a higher sensitivity of R. philippinarum to Microcystis-induced stress than M. galloprovincialis. The results also suggest a much higher level of activity of the GST detoxification system during the recovery phase compared to the period of the stress exposure for both bivalve species. © 2018, Springer Science+Business Media, LLC, part of Springer Nature. Acknowledgements This research was partially supported by the European Regional Development Fund (ERDF) through the COMPETE—Operational Competitiveness Programme and national funds through FCT (Foundation for Science and Technology), under the project PTDC/MAR-EST/4614/2012 (FCOMP-01-0124-FEDER-029452) and UID/Multi/04423/2013 and by the project ALERTOX-NET-EAPA_317/2016 of the Interreg Atlantic Area Program. José Carlos Martins is supported by a research contract financed by national funds through FCT, I.P., within the scope of the project DL57/2016/ CP1331/CT0002.

Cylindrospermopsin (CYN) is a toxic secondary metabolite produced by filamentous cyanobacteria which could work as an allelopathic substance, although its ecological role in cyanobacterial-algal assemblages is mostly unclear. The competition between the CYN-producing cyanobacterium Chrysosporum (Aphanizomenon) ovalisporum, and the benthic green alga Chlorococcum sp. was investigated in mixed cultures, and the effects of CYN-containing cyanobacterial crude extract on Chlorococcum sp. were tested by treatments with crude extracts containing total cell debris, and with cell debris free crude extracts, modelling the collapse of a cyanobacterial water bloom. The growth inhibition of Chlorococcum sp. increased with the increasing ratio of the cyanobacterium in mixed cultures (inhibition ranged from 26% to 87% compared to control). Interestingly, inhibition of the cyanobacterium growth also occurred in mixed cultures, and it was more pronounced than it was expected. The inhibitory effects of cyanobacterial crude extracts on Chlorococcum cultures were concentration-dependent. The presence of C. ovalisporum in mixed cultures did not cause significant differences in nutrient content compared to Chlorococcum control culture, so the growth inhibition of the green alga could be linked to the presence of CYN and/or other bioactive compounds.

Toxicity-data of two carbamate insecticides, carbaryl and carbofuran, and three fungicides, ziram, zineb and mancozeb with rice-field N2-fixing cyanobacterium Cylindrospermum sp., obtained by in vitro growth and at soil-water interface, were analyzed by the probit method. Growth enhancing concentration, no-observed effective concentration, minimum inhibitory concentration, the highest permissive concentration and lethal concentration100 (LC100) were determined experimentally. The LC50 values of carbaryl, carbofuran, ziram, zineb and mancozeb in N2-fixing liquid medium were 56.2, 588.8, 0.07, 4.2 and 3.4 μg/mL, respectively, whereas the corresponding LC100 values were 100.0, 1500.0, 0.17, 25.0 and 9.0 μg/mL, respectively. The LC50 values of these pesticides in succession in N2-fixing agar medium were 44.7, 239.9, 0.07, 1.8 and 2.3 μg/mL, respectively, whereas the corresponding LC100 values were 100.0, 600.0, 0.17, 10.0 and 7.0 μg/mL, respectively. Similar results with nitrate supplemented liquid and agar media indicated that nitrate supplementation had toxicity reducing effect. The LC50 and LC100 values of toxicity in the N2-fixing liquid medium at soil-water interface were 91.2 and 200.0 μg/mL for carbaryl, 2 317 and 6 000 μg/mL for carbofuran, 0.15 and 0.50 μg/mL for ziram, 16.4 and 50.0 μg/mL for zineb, and 7.2 and 25.0 μg/mL for mancozeb, respectively. Each LC100 value at soil-water interface with a pesticide was significantly higher than its corresponding LC100 value at liquid/agar media. It can be concluded that, under the N2-fixing conditions, the cyanobacterium tolerated higher levels of each pesticide at soil-water interface.

Oxygenic photosynthesis crucially depends on proteins that possess Fe (2+) or Fe/S complexes as co-factors or prosthetic groups. Here, we show that the small regulatory RNA (sRNA) IsaR1 (Iron-Stress-Activated RNA 1) plays a pivotal role in acclimation to low-iron conditions. The IsaR1 regulon consists of more than 15 direct targets, including Fe (2+)-containing proteins involved in photosynthetic electron transfer, detoxification of anion radicals, citrate cycle, and tetrapyrrole biogenesis. IsaR1 is essential for maintaining physiological levels of Fe/S cluster biogenesis proteins during iron deprivation. Consequently, IsaR1 affects the acclimation of the photosynthetic apparatus to iron starvation at three levels: (1) directly, via posttranscriptional repression of gene expression; (2) indirectly, via suppression of pigment; and (3) Fe/S cluster biosynthesis. Homologs of IsaR1 are widely conserved throughout the cyanobacterial phylum. We conclude that IsaR1 is a critically important riboregulator. These findings provide a new perspective for understanding the regulation of iron homeostasis in photosynthetic organisms. German Federal Ministry of Education and Research [0316165] DFG [HE 2544/9-1] Academy of Finland [253269, 271832, 273870] Portuguese Fundacao para a Ciencia e a Tecnologia [IF/00881/2013, UID/Multi/04326/2013-CCMAR] European Commission FP7 Marie Curie Initial Training Network "Photo.COMM'' [317184] info:eu-repo/semantics/publishedVersion

We have found a novel group of plants that appear to obligately accumulate salt (sodium chloride) although the level in their environment is low. The group is taxonomically diverse including angiosperms (Gunnera spp.), gymnosperms (Cycas) and a fern (Azolla species) but all of them form a symbiosis with a cyanobacterium (blue-green alga) that carries out nitrogen fixation, a rare occurrence in plants. The functional basis for the high salt contents is not clear and, despite the high internal salt levels, the plants are unable to grow in high salt environments., The majority of plant species are glycophytes and are not salt-tolerant and maintain low sodium levels within their tissues; if. high tissue sodium concentrations do occur, it is in response to elevated environmental salt levels. Here we report an apparently novel and taxonomically diverse grouping of plants that continuously maintain high tissue sodium contents and share the rare feature of possessing symbiotic cyanobacteria. Leaves of Gunnera magellanica in Tierra del Fuego always had sodium contents (dry weight basis) of around 4.26 g kg−1, about 20 times greater than measured in other higher plants in the community (0.29 g kg−1). Potassium and chloride levels were also elevated. This was not a response to soil sodium and chloride levels as these were low at all sites. High sodium contents were also confirmed in G. magellanica from several other sites in Tierra del Fuego, in plants taken to, and cultivated in Madrid for 2 years at low soil salt conditions, and also in other free living or cultivated species of Gunnera from the UK and New Zealand. Gunnera species are the only angiosperms that possess cyanobacterial symbionts so we analysed other plants that have this rather rare symbiosis, all being glycophytes. Samples of Azolla, a floating aquatic fern, from Europe and New Zealand all had even higher sodium levels than Gunnera. Roots of the gymnosperm Cycas revoluta had lower sodium contents (2.52 ± 0.34 g kg−1) but still higher than the non-symbiotic glycophytes. The overaccumulation of salt even when it is at low levels in the environment appears to be linked to the possession of a cyanobacterial symbiosis although the actual functional basis is unclear.

Allelopathy of picocyanobacterium Synechococcus sp. may be involved in the formation of massive blooms and their subsequent global expansion in many aquatic ecosystems. However, the effect of the allelopathic activity of this species on filamentous cyanobacteria remains unknown. In this study, we tested the allelopathic activity of Synechococcus sp. on growth, pigment content and chlorophyll fluorescence of filamentous cyanobacteria Aphanizomenon flos-aquae, Nostoc sp., Phormidium sp. and Rivularia sp. by single and repeated addition of cell-free filtrate. Negative effects against Nostoc sp. and Phormidium sp. were amplified by repeated filtrate additions compared with single filtrate addition. A maximum reduction in growth and a maximum quantum yield of photosystem II (PSII) photochemistry (Fv/Fm) relative to controls were observed for Phormidium sp. However, the addition of picocyanobacterial filtrate stimulated the growth and Fv/Fm of A. flos-aquae. Synechococcus sp. filtrate had no allelopathic effects on the growth and Fv/Fm of Rivularia sp. Moreover, filtrates caused significant decreases in the chlorophyll a (Chl a) contents of Phormidium sp. and Rivularia sp. cells. The addition of filtrate also resulted in increased cell carotenoid (Car) content in A. flos-aquae and Nostoc sp. These results showed for the first time that picocyanobacterium Synechococcus sp. negatively and positively affected coexisting filamentous cyanobacteria. © 2017 Elsevier The authors would like to thank the anonymous reviewers and Editor for their valuable comments and suggestions to improve the quality of the paper. This study was supported by BMN grants, Poland, no. 538-G245-B568-17 .

Blooms of harmful cyanobacteria that synthesize cyanotoxins are increasing worldwide. Agronomic plants can uptake these cyanotoxins and given that plants are ultimately ingested by humans, this represents a public health problem. In this research, parsley and coriander grown in soil and watered through 7 days with crude extracts containing microcystins (MCs) or cylindrospermopsin (CYN) in 0.1–1 μg mL−1 concentration range were evaluated concerning their biomass, biochemical parameters and uptake of cyanotoxins. Although biomass, chlorophylls (a and b), carotenoids and glutathione-S-transferase of parsley and coriander exposed to the crude extracts containing MC or CYN had shown variations, these values were not statistically significantly different. Protein synthesis is not inhibited in coriander exposed to MC or CYN and in parsley exposed to MC. Also, glutathione reductase (GR) and glutathione peroxidase (GPx) in parsley and coriander was not affected by exposure to MC, and in coriander, the CYN did not induce statistically significant differences in these two antioxidative enzymes. Only parsley showed statistically significant increase in protein content exposed to 0.5 μg CYN mL−1 (3.981 ± 0.099 mg g−1 FW) compared to control (2.484 ± 0.145 mg g−1 FW), statistically significant decrease in GR exposed to 0.1 μg CYN mL−1 (0.684 ± 0.117 nmol min−1 mg−1 protein) compared to control (1.30 ± 0.06 nmol min−1 mg−1 protein) and statistically significant increase in GPx exposed to 1 μg CYN mL−1 (0.054 ± 0.026 nmol min−1 mg−1 protein) compared to 0.5 μg CYN mL−1 (0.003 ± 0.001 nmol min−1 mg−1 protein). These changes may be due to the induction of defensive mechanisms by plants by the presence of toxic compounds in the soil or probably to a low generation of reactive oxygen species. Furthermore, the parsley and coriander leaves and stems after 10 days of exposure did not accumulate microcystins or cylindrospermopsin. © 2016, Springer-Verlag Berlin Heidelberg. European Social Funding (FSE) under the Human Potential Operational Program (POPH) of National Strategic Reference Board (QREN) supports the fellowship SFRH/BPD/44459/2008 to Ana L. Pereira. This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT—Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the program PT2020. Thanks are given to Benedita Monteiro and Catarina Santos for the growth, extraction and quantification of microcystins from M. aeruginosa and cylindrospermopsin from A. ovalisporum respectively.

A phycocyanin-deletion mutant of Synechocystis (cyanobacteria) was generated upon replacement of the CPC-operon with a kanamycin resistance cassette. The Δcpc transformant strains (Δcpc) exhibited a green phenotype, compared to the blue-green of the wild type (WT), lacked the distinct phycocyanin absorbance at 625nm, and had a lower Chl per cell content and a lower PSI/PSII reaction center ratio compared to the WT. Molecular and genetic analyses showed replacement of all WT copies of the Synechocystis DNA with the transgenic version, thereby achieving genomic DNA homoplasmy. Biochemical analyses showed the absence of the phycocyanin α- and β-subunits, and the overexpression of the kanamycin resistance NPTI protein in the Δcpc. Physiological analyses revealed a higher, by a factor of about 2, intensity for the saturation of photosynthesis in the Δcpc compared to the WT. Under limiting intensities of illumination, growth of the Δcpc was slower than that of the WT. This difference in the rate of cell duplication diminished gradually as growth irradiance increased. Identical rates of cell duplication of about 13h for both WT and Δcpc were observed at about 800μmolphotonsm−2s−1 or greater. Culture productivity analyses under simulated bright sunlight and high cell-density conditions showed that biomass accumulation by the Δcpc was 1.57-times greater than that achieved by the WT. Thus, the work provides first-time direct evidence of the applicability of the Truncated Light-harvesting Antenna (TLA)-concept in cyanobacteria, entailing substantial improvements in the photosynthetic efficiency and productivity of mass cultures upon minimizing the phycobilisome light-harvesting antenna size.

Objective To explore the ability of Synechocystis sp. PCC6803 in transforming 6-deoxypseudoanisatin. Methods The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocystis sp. PCC6803 under continuous white light at 30°C for 5 days. The crude converted product was detected using thin-layer chromatography (TLC) and further analyzed using high-performance liquid chromatography (HPLC) as well as HPLC with electron spray ionization mass spectrometry (HPLC-ESI-MS). Results TLC results showed that 6-deoxypseudoanisatin was converted into a less polar product. HPLC and MS data indicated that the retention time of the converted product increased in comparison with the standard of 6-deoxypseudoanisatin. Conclusion Thus, the study appears to demonstrate that Synechocystis sp. PCC6803 can transform 6-deoxypseudoanisatin. The polarity of the converted product is less than that of 6-deoxypseudoanisatin.

Macroscopic gelatinous colonies of freshwater cyanobacterium Aphanothece sacrum, a luxury ingredient for Japanese cuisine, were found to contain a new oxylipin-derived macrolide, sacrolide A (1), as an antimicrobial component. The configuration of two chiral centers in 1 was determined by a combination of chiral anisotropy analysis and conformational analysis of different ring-opened derivatives. Compound 1 inhibited the growth of some species of Gram-positive bacteria, yeast Saccharomyces cerevisiae and fungus Penicillium chrysogenum, and was also cytotoxic to 3Y1 rat fibroblasts. Concern about potential food intoxication caused by accidental massive ingestion of A. sacrum was dispelled by the absence of 1 in commercial products. A manual procedure for degrading 1 in raw colonies was also developed, enabling a convenient on-site detoxification at restaurants or for personal consumption.

Effects of chemical fertilizers (urea, super phosphate and potash) on toxicities of two carbamate insecticides, carbaryl and carbofuran, individually to the N2-fixing cyanobacterium, Cylindrospermum sp. were studied in vitro at partially lethal levels (below highest permissive concentrations) of each insecticide. The average number of vegetative cells between two polar heterocysts was 16.3 in control cultures, while the mean value of filament length increased in the presence of chemical fertilizers, individually. Urea at the 10 ppm level was growth stimulatory and at the 50 ppm level it was growth inhibitory in control cultures, while at 100 ppm it was antagonistic, i.e. toxicity-enhancing along with carbaryl, individually to the cyanobacterium, antagonism was recorded. Urea at 50 ppm had toxicity reducing effect with carbaryl or carbofuran. At 100 and 250 ppm carbofuran levels, 50 ppm urea only had a progressive growth enhancing effect, which was marked well at 250 ppm carbofuran level, a situation of synergism. Super phosphate at the 10 ppm level only was growth promoting in control cultures, but it was antagonistic at its higher levels (50 and 100 ppm) along with both insecticides, individually. Potash (100, 200, 300 and 400 ppm) reduced toxicity due to carbaryl 20 and carbofuran 250 ppm levels, but potash was antagonistic at the other insecticide levels. The data clearly showed that the chemical fertilizers used were antagonistic with both the insecticides during toxicity to Cylindrospermum sp.

Eukaryotes co-opted photosynthetic carbon fixation from prokaryotes by engulfing a cyanobacterium and stably integrating it as a photosynthetic organelle (plastid) in a process known as primary endosymbiosis. The sheer complexity of interactions between a plastid and the surrounding cell that started to evolve over 1 billion years ago, make it challenging to reconstruct intermediate steps in organelle evolution by studying extant plastids. Recently, the photosynthetic amoeba Paulinella chromatophora was identified as a much sought-after intermediate stage in the evolution of a photosynthetic organelle. This article reviews the current knowledge on this unique organism. In particular it describes how the interplay of reductive genome evolution, gene transfers, and trafficking of host-encoded proteins into the cyanobacterial endosymbiont contributed to transform the symbiont into a nascent photosynthetic organelle. Together with recent results from various other endosymbiotic associations a picture emerges that lets the targeting of host-encoded proteins into bacterial endosymbionts appear as an early step in the establishment of an endosymbiotic relationship that enables the host to gain control over the endosymbiont.

Proteome-wide Amino aCid and Elemental composition (PACE) analysis is a novel and informative way of interrogating the proteome. The PACE approach consists of in silico decomposition of proteins detected and quantified in a proteomics experiment into 20 amino acids and five elements (C, H, N, O and S), with protein abundances converted to relative abundances of amino acids and elements. The method is robust and very sensitive; it provides statistically reliable differentiation between very similar proteomes. In addition, PACE provides novel insights into proteome-wide metabolic processes, occurring, e.g., during cell starvation. For instance, both Escherichia coli and Synechocystis down-regulate sulfur-rich proteins upon sulfur deprivation, but E. coli preferentially down-regulates cysteine-rich proteins while Synechocystis mainly down-regulates methionine-rich proteins. Due to its relative simplicity, flexibility, generality and wide applicability, PACE analysis has the potential of becoming a standard analytical tool in proteomics.

Spirulina, now named Arthrospira, is a microscopic and filamentous cyanobacterium that has a long history of use as a safe food lacking toxicity. It is commercially produced in large outdoor ponds under controlled conditions. The aim of this review article is to summarize available recent information concerning human clinical potential and applications of Spirulina, as well as clinical data related to the safety and side effects of Spirulina. Potential health benefits of Spirulina are mainly due to its chemical composition, which includes proteins (the highest protein content of any natural food, 55%-70%), carbohydrates, essential amino acids, minerals (especially iron), essential fatty acids, vitamins, and pigments. In this respect, three major bioactive components of Spirulina, the protein phycocyanin (a biliprotein pigment), sulfated polysaccharides and gamma linolenic acid seem to play significant role in imparting improved human body functions. Furthermore, new experimental evidence supports the immunomodulation and antiviral effects of Spirulina supplementation. According to the Dietary Supplements Information Expert Committee of United States Pharmacopeial Convention the available clinical evidence does not indicate a serious risk to health or other public health concerns for Spirulina. However, a few cases of severe side-effects have been reported.