Blue-green algae (cyanobacteria): prospects and perspectives

Summary Photosynthetic, prokaryotic blue-green algae (cyanobacteria) occur in a wide range of natural habitats of diverse ionic composition and as such, represent an important source of biological material for biosolar energy conversion programs using saline water. The gasvacuolate, filamentous Spirulina is grown in seminatural culture in Lake Texcoco, Mexico, as a major source of single-cell protein for animal nutrition. Pilot-scale trials in other areas of the world have also demonstrated the suitability of blue-green algae, including Spirulina, for growth under brackish conditions. The carbohydrate accumulation profiles of blue-green algae differ in isolates from freshwater, marine and hypersaline habitats, with a trend towards sucrose or trehalose accumulation in stenohaline freshwater strains grown in media containing NaCl, while euryhaline and marine forms frequently accumulate glucosylglycerol. Many halotolerant isolates from hypersaline habitats accumulate glycinebetaine in response to osmotic stress. This knowledge may provide scope for future improvement in the N₂ fixation rates of blue-green algae in saline media, using betaine-accumulating N₂-fixing strains in preference to other, saltsensitive isolates.     

Accessory polypeptides in phycobilisomes of red algae and cyanobacteria

Phycobilisomes of red algae and cyanobacteria contain small amounts of nonpigmented polypeptides in addition to the major constituent biliprotein pigments. The localization of these polypeptides is analyzed by gel electrophoresis of phycobilisome fragments obtained by selective dissociation and subsequent separation. Five groups of biliprotein aggregates are determined, belonging to the 6, 11, 16, 18 and 23 S categories. Accessory nonpigmented high molecular weight proteins (80,000 MW) are exclusively bound to phycobilisome core fractions and thylakoids, thus apparently serving as links between the phycobilisomes and the photosynthetic units of the thylakoids. In contrast, smaller nonpigmented accessory polypeptides of 20,000 to 60,000 MW are preferably found in the peripheral biliprotein stacks. They may either form a compatible link between the phycobilisome core and periphery or bind and co-polymerize with hexameric biliproteins in the peripheral stacks to enhance or effect binding of the aggregates. Furthermore, they may determine the arrangement and composition of the phycobilisomes during development and chromatic adaptation.Abbreviations PE phycoerythrin - PEC phycoerythrocyanin - PC phycocyanin - APC allophycocyanin     

Physiological characteristics of cyanobacteria in pulp and paper waste-treatment systems

An investigation was made into the physiological characteristics of the extensive cyanobacterial communities in pulp and paper secondary waste-treatment systems, including the capacity of isolates to biodegrade organic contaminants in these systems. Although pulp and paper waste-treatment systems were found to be severely light-limited, photosynthesis-irradiance curves indicated that shade-adapted cyanobacterial communities could fix conspicuous amounts of inorganic carbon via photosynthesis. Of 21 cyanobacterial strains isolated from pulp and paper waste-treatment systems located in 4 countries, all except one were capable of glucose uptake in the light, and 19 also showed uptake in the dark. In the 6 species tested, glucose and acetate addition stimulated growth in low light in all except one species. Aphanocapsa rivularis grew equally well on glucose and acetate in the dark but Pseudanabaena sp. grew well on acetate, and minimally on glucose. Growth stimulation by glucose and acetate in these two strains was greater in low than in high light. Pseudanabaena sp. and Phormidium animale both accumulated 2,4-dichlorophenol and 3-chlorobenzoate but showed minimal mineralization to CO₂. None of the four species tested could accumulate or degrade phenol or dichloroacetate. It is concluded that, depending on the light conditions, cyanobacteria contribute organic carbon in photosynthesis, and/or remove small organic molecules during mixotrophic and heterotrophic growth but are not important degraders of contaminants in these waste treatment systems.     

Changes in the community structure and activity of betaproteobacterial ammonia-oxidizing sediment bacteria along a freshwater-marine gradient

To determine whether the distribution of estuarine ammonia-oxidizing bacteria (AOB) was influenced by salinity, the community structure of betaproteobacterial ammonia oxidizers (AOB) was characterized along a salinity gradient in sediments of the Ythan estuary, on the east coast of Scotland, UK, by denaturant gradient gel electrophoresis (DGGE), cloning and sequencing of 16S rRNA gene fragments. Ammonia-oxidizing bacteria communities at sampling sites with strongest marine influence were dominated by Nitrosospira cluster 1-like sequences and those with strongest freshwater influence were dominated by Nitrosomonas oligotropha-like sequences. Nitrosomonas sp. Nm143 was the prevailing sequence type in communities at intermediate brackish sites. Diversity indices of AOB communities were similar at marine- and freshwater-influenced sites and did not indicate lower species diversity at intermediate brackish sites. The presence of sequences highly similar to the halophilic Nitrosomonas marina and the freshwater strain Nitrosomonas oligotropha at identical sampling sites indicates that AOB communities in the estuary are adapted to a range of salinities, while individual strains may be active at different salinities. Ammonia-oxidizing bacteria communities that were dominated by Nitrosospira cluster 1 sequence types, for which no cultured representative exists, were subjected to stable isotope probing (SIP) with 13C-HCO3-, to label the nucleic acids of active autotrophic nitrifiers. Analysis of 13C-associated 16S rRNA gene fragments, following CsCl density centrifugation, by cloning and DGGE indicated sequences highly similar to the AOB Nitrosomonas sp. Nm143 and Nitrosomonas cryotolerans and to the nitrite oxidizer Nitrospira marina. No sequence with similarity to the Nitrosospira cluster 1 clade was recovered during SIP analysis. The potential role of Nitrosospira cluster 1 in autotrophic ammonia oxidation therefore remains uncertain.     

Molecular and phylogenetic characterization of potentially toxic cyanobacteria in Tunisian freshwaters

This study presents a genetic characterization of 27 potentially toxic cyanobacterial strains isolated from seven reservoirs located in the north and centre of Tunisia. These strains belonged mainly to Microcystis aeruginosa, Cylindrospermopsis raciborskii and Planktothrix agardhii species. Their toxicological potential was evaluated by molecular biology tools, which showed that none of the isolated strains carried segments of the gene cluster responsible for the production of cylindrospermopsin and saxitoxin. The majority of Microcystis isolates were able to synthesize microcystin, since they presented the six characteristic segments of the microcystin synthetase mcy cluster (mcyA, -B, -C, -D, -E and -G). This was further confirmed by MALDI-TOF analysis that showed the presence of eight microcystin variants, including microcystin-LR. The taxonomic identification of the strains was assessed based on the variability of the 16S rRNA gene sequences. Furthermore, the 16S-23S rRNA ITS sequences of Microcystis isolates and rpoC1 sequences of Cylindrospermopsis strains were also used in the phylogenetic analysis.     

滨海盐渍化土壤中蓝细菌多样性及分布

【目的】蓝细菌在贫瘠土壤的固氮、固碳中发挥着重要作用,然而土壤盐渍化对蓝细菌多样性及群落结构的影响还不清楚。本研究以莱州湾南岸及黄河口海水入侵盐渍化土壤为例,研究蓝细菌的多样性、群落结构及丰度沿盐度梯度的分布情况。【方法】利用自动核糖体间隔基因分析(ARISA)技术与群落相似性分析(ANOSIM)探究蓝细菌群落的差异与空间分布格局;通过16S rRNA基因克隆文库、测序与系统进化分析解析了3个典型盐度梯度样品中蓝细菌的群落组成;实时定量PCR测定蓝细菌16S rRNA拷贝数(丰度);BEST多元分析探寻影响蓝细菌分布的主要环境因子。【结果】蓝细菌在莱州湾南岸盐土中分布广泛,其群落结构在低(0.63%?1.27%)、中(1.55%?2.00%)、高盐度(2.39%?5.11%)样品组之间差异显著(P=0.03),而在不同含水量样品组间则不显著(P=0.09)。总体来看,群落结构主要受土壤盐度与含水量这两个因子联合控制(P=0.02)。低盐样品中蓝细菌多样性和丰富度最低,并以Halomicronema和Acaryochloris为优势类群。Leptolyngbya在中、高盐土中占优势。Arthrospira与Geitlerinema仅在低盐土中检测到,而Oscillatoria则仅出现在高盐土中。随盐度升高蓝细菌丰度呈下降趋势,低盐土样中蓝细菌丰度(2.14×105 copies/g干土)显著高于中盐土(1.25×105 copies/g干土)(P<0.05);高盐度土为1.20×105 copies/g干土。【结论】盐渍化程度是调控莱州湾南岸滨海土壤中蓝细菌群落结构与丰度的最重要环境因子,可能对滨海土壤微生物碳氮循环产生重要影响。     

LexA-binding sequences in Gram-positive and cyanobacteria are closely related

The lexA gene of the cyanobacterium Anabaena sp. strain PCC7120 has been cloned by PCR amplification with primers designed after TBLASTN analysis of its genome sequence using the Escherichia coli LexA sequence as a probe. After over-expression in E. coli and subsequent purification, footprinting experiments demonstrated that the Anabaena LexA protein binds to the sequence TAGTACTAATGTTCTA, which is found upstream of its own coding gene. Directed mutagenesis and sequence comparison of promoters of other Anabaena genes, as well as those of several cyanobacteria, allowed us to define the motif RGTACNNNDGTWCB as the LexA box in this bacterial phylum. Substitution of a single nucleotide in this motif present in the Anabena lexA promoter is sufficient to enable it to bind the Bacillus subtilis LexA protein. These data indicate that Cyanobacteria and Gram-positive bacteria are phylogenetically closely related.Communicated by R. Devoret     

Development of a biofertilizer based on filamentous nitrogen-fixing cyanobacteria for rice crops in Chile

The purpose of this study was to develop a biofertilizer based on filamentous nitrogen-fixing cyanobacteria selected from rice fields and to generate a technological package compatible with its use for the rice crop in Chile. Thirty-four Chilean rice fields, located between Maule and BioBío regions, were sampled during the 1998/1999 and 1999/2000 growing seasons. A total of 9 species and 3 varieties of cyanobacteria were found, and the nitrogen fixation rate under laboratory conditions was determined for 6 of them. Only 4 were used for the small-scale production of a biofertilizer, which was assayed in field trials. To check the efficiency of the biofertilizer during the rice crop, the nitrogen fixation rates in soil samples were estimated. Additionally, the biofertilizer application efficiency was tested in combination with nitrogen synthetic fertilizer, in rates that were previously established in field trials. Biofertilization allowed a decrease of up to 50% in the use of nitrogen synthetic fertilizer (50 kg N ha⁻¹), resulting in the same grain yield (7.4 t ha⁻¹) and quality in relation to the fertilized control. The use of biofertilizers based on local strains of cyanobacteria shows promise to increase nitrogen use efficiency in rice.     

Biotic communities of freshwater marshes and mangroves in relation to saltwater incursions: implications for wetland regulation

An ecosystem-level study was conducted in the Guandu wetlands insubtropical coastal Taiwan to examine how salinity influences the abundance,diversity, and structure of biotic communities. We surveyed eight permanentstudy sites, spanning freshwater marshes, to the gate on the dyke, andmesohaline mangroves representing a gradient of the extent of saltwaterincursions. Analyses of abiotic variables showed that salinity was the primarydetermining factor for discriminating habitat types in the wetlands, butcommunities differed in their sensitivity to salinity. The composition of plantand insect communities was most affected by the salinity gradient, suggestingthe utility of these communities for ecological monitoring of saltwaterincursions. However, spatial changes in communities at higher trophic levels,including macrobenthos, mollusks, fish, and birds, could not be explained simplyby the salinity gradient. Instead, changes in these communities were morerelevant to the composition of other biotic communities. Our results show thatspecies richness and diversity of plant communities were higher in the marshesthan in the mangroves. Nevertheless, insect communities censused in themangroves had higher diversity, despite lower abundance and species richness.Macrobenthos surveyed in the mangroves showed higher biomass and number of taxa.Mollusks and fish were also more abundant at sites near the gate compared to themarsh sites. This suggests that maintaining a tidal flux by means of gateregulation is necessary for conserving the spatial heterogeneity andbiodiversity of coastal wetlands.     

Diversity and bioactivities of nostocacean cyanobacteria isolated from paddy soil in Vietnam

Nostocacean cyanobacteria are one of the important components of paddy fields due to their ability to fix atmospheric nitrogen and supply phytohormones for crop growth. In this study, 13 Nostoc strains isolated from paddy soils in Vietnam were classified using a polyphasic approach. The results showed a high diversity of the isolated strains that represented seven morphotypes corresponding to five genotypes, with 16S rRNA gene sequence similarity values ranging between 94.97–99.78% compared to the available sequences from GenBank. Bioassay assessment revealed that 11 out of 13 strains possessed antibacterial activities, three of which exhibited cytotoxic activities on MCF7 and HCT116 cells with an IC₅₀ ranging from 47.8 μg mL^?1 to 232.0 μg mL^?1. Interestingly, strains with identical 16S rRNA gene sequences displayed different antibacterial and cytotoxic activity profiles.     

Biotransformation and toxicity of aniline and aniline derivatives in cyanobacteria

Agmenellum quadruplicatum strain PR-6 and Oscillatoria sp. strain JCM grown photoautotrophically in the presence of aniline metabolized the aromatic amine to formanilide, acetanilide and p-aminophenol. The metabolites were isolated by either thin-layer, gas-liquid or high pressure liquid chromatography and identified by comparison of their chromatographic, ultraviolet absorbance and mass spectral properties with those of authentic compounds. The toxicity of aniline derivatives towards Agmenellum quadruplicatum strain PR-6 indicated that the cyanobacterium was extremely sensitive to o-, m- and p-aminophenols, and phenylhydroxylamine.Abbreviations TLC thin layer chromatography - HPLC high pressure liquid chromatography - GC/MS gas chromatography/mass spectrometry - m/e mass to charge ratio     

Stimulation of nitrogenase activity and photosynthesis in some cyanobacteria by glyoxylate

Abstract The effect og glyoxylate on nitrogenase activity (C₂H₂ reduction) and photosynthesis (H¹⁴CO₃ fixation and O₂ evolution) was in vestigated in the three heterocystous cyanobacteria Anabaena cylindrica, A. variabiltis and N. muscorum. Glyoxylate had virtually no effect on the rate of dark respiration and was unable to sustain photoheterotrophic growth, though some slight stimulation (= 30%) of photorophic growth was noted. A considerable stimulation of both nitrogenase and photosynthetic activities was observed in presence of glyoxylate. In the light the stimulation increased with time up to about 15-25 h after adding optimal concentrations of 4–6 mM glyoxylate. Placing glyoxylate treated samples in the dark or adding DCMU (30 μM) in the light, showed that glyoxylate initially supported significantly higher nitrogenase activity than did samples in absence of glyoxylate. However, after a prolonged incubation in the dark or in presence of DCMU glyoxylate is unable to relieve the adverse effects of such conditions. The stimulation of the nitrogenase activity was even more pronounced when the glyoxylate was added to cells preincubated in the dark (“carbon starved”) than for cells kept constantly in light. The results suggest that glyoxylate, or a metabolite, may act as an inhibitor of cyanobacterial photorespiration and this hypothesis is discussed.     

Inducible defence against a ciliate grazer Pseudomicrothorax dubius,in two strains of Phormidium (cyanobacteria)

Experiments were done with two strain of filamentous, mat-forming Phormidium and their ciliate grazer Pseudomicrothorax dubius, to explain why the ciliates remain hungry in an apparent surplus of food, except for the first 24 hours after feeding. Under grazing pressure, both strains of cyanobacteria showed statistically significant increases in the number of filaments terminating in an empty sheath, compared to the control. Direct observations revealed that the mechanism behind this effect was active withdrawal of the trichomes inside the sheaths when disturbed by grazers. As P. dubius is unable to ingest trichomes with such endings, we conclude that cyanobacteria are not limited to chemical means of defence against grazers but can also defend themselves by means of movement and changes in filament morphology. This is apparently the first report on behavioural defence observed in cyanobacteria.     

Oxidative and hydrolytic transformations of hydroxylamine compounds in cyanobacteria

Ammonia-incubated cyanobacteria liberated H₂O₂, accumulated hydroxylamine compounds and nitrite and catalyzed dismutation of hydroxylamine as well as oxidations of ammonia, glutamine, and oximes. Ethyl acetohydroximate-adapted Phormidium released excess H₂O₂ and phototrophically metabolized the oxime via hydrolysis and dismutation to nitrite and ammonia, which were consumed by nitrite reductase and glutamine synthetase. Added ammonia stimulated H₂O₂ production and oxime metabolism via glutamate dehydrogenase pathway.Abbreviations EAH Ethyl acetohydroximate - GDH glutamate dehydrogenase - GS glutamine synthetase - NiR nitrite reductase     

Excretion of fermentation products in dark and anaerobically incubated cyanobacteria

An arbitrarily chosen selection of 37 cyanobacterial strains of the Oldenburg culture collection were tested for their ability of fermentation and secretion of fermentation products. In all examined strains at least one fermentation product could be detected. For the most part fermentation products were only shed in traces. Thus, for a large part of the investigated strains fermentation does not seem to be a sufficient metabolism to survive dark and anaerobic periods. Only five strains secreted remarkable amounts of products. Glycollate was mostly found in combination with formate and/or traces of oxalate. Lactate, ethanol and acetate were found in combination or single. Most of those strains sheding high amounts of glycollate and formate, did not show a remarkable lactate, ethanol or acetate excretion; those excreting high amounts of lactate, ethanol or acetate produced only minor volumes of glycollate and formate. It was not possible to find similar fermentation patterns by comparing fermentation of species belonging to the same family. Organisms fermenting or not fermenting could be found among marine, brackish and freshwater cyanobacteria. Fermentation, therefore seems to be a unique, and likely old capability among cyanobacteria, which was partly lost during evolution.     

Isolation and characterization of plasma membranes from cyanobacteria

The two-phase partition system in comparison to the traditional methods used thus far (density gradients) for the isolation of the plasma membrane from cyanobacteria is described. The advantages of the two-phase system are: A short-time preparation of 3–4 h compared to 16–48 h required for the density gradient method; a purer fraction, resulting from separation according to membrane surface charge and hydrophobicity, not specific density; and, ease of scaling-up for obtaining large quantities.
Also, the different biological activities attributed to this membrane to date are summarized. Findings on the typical plasma membrane ATPase (P-type ATPase) as well as the nutrient transporters and the corresponding proteins are included. As for the electron transport chain, one may conclude that this membrane contains a complete system (similar to that of the mitochondrion), portraying apparently F-type (F₀F₁) ATPase activity.