Tetrazolium reduction and nitrogenase activity in heterocystous blue-green algae

Summary Heterocysts reduce triphenyl tetrazolium chloride (TTC) faster than vegetative cells apparently because the absence of the O₂-evolving photosystem II and the high electron transport activity in these cells. Although the rate of TTC reduction in vegetative cells is increased by the continuous removal of O₂ evolved in photosynthesis, it has not been possible to obtain rates of TTC reduction comparable with those in heterocysts probably because of the continued competition for electrons between TTC and O₂. The use of nitro-blue tetrazolium chloride (NBT) as a redox indicator has revealed the presence in filaments under aerobic conditions of a gradient of electron transport activity with strongest reducing power in the heterocysts, proheterocysts and vegetative cells next to heterocysts, and with gradually diminishing activity midway between two heterocysts. This pattern is indistinct in filaments grown under micro-aerophilic conditions. The strong electron transport activity in vegetative cells adjacent to heterocysts appears to promote reducing conditions in the heterocysts. Both, red-formazan formation in the heterocysts and blue-formazan deposition in vegetative cells greatly inhibit nitrogenase activity, and this was adversely affected also by the detachment of heterocysts from vegetative cells. The findings are consistent with the idea that the association of heterocysts with vegetative cells in essential for nitrogen fixation to occur in heterocystous blue-green algae.     

Nitrogen fixation (acetylene reduction) associated with communities of heterocystous and non-heterocystous blue-green algae in mangrove forests of Sinai

Summary High rates of nitrogen fixation (acetylene reduction) are associated with communities of heterocystous and non-heterocystous blue-green algae, which are widespread and abundant in the coastal mangrove forests of the Sinai Peninsula.Heterocystous forms, particularly representatives of the Rivulariaceae, grow in aerobic environments, where nitrogenase activity may be limited by the availability of nutrients such as Fe and PO₄–P. Desiccated communities of Scytonema sp. reduce acetylene within ten minutes of wetting by tidal sea water. Communities dominated by the non-heterocystous Hydrocoleus sp., Hyella balani, Lyngbya aestuarii, Phormidium sp. and Schizothrix sp., occur in close contact with anaerobic sediments and reduce acetylene in the dark as well as in the light.Nitrogen fixation in all these communities is light dependant and may be supplemented by an alternative source of reductant in the dark. The indications are that nitrogen fixation by these communities of blue-green algae, makes a significant contribution to the overall nitrogen input of the mangrove ecosystem.     

Nitrogenase activity of blue-green algae on seasonally flooded soils in Bangladesh

Blue-green algal communities formed an extensive cover on soils at five deepwater rice-growing locations in Bangladesh during the month before the arrival of floodwater. The principal taxa wereAnabaena, Cylindrospermum, Lyngbya, Microcoleus, Nostoc, Prophyrosiphon notarisii, Scytonema mirabile andTolypothrix byssoidea. One of two locations studied after the floodwaters had receded also had an extensive cover, mainlyScytonema mirabile. Nitrogenase activity assayed at mid-day was from one to three orders of magnitude higher per unit area of community than bare soil. Nostoc showed higher activity than Tolypothrix, whether expressed per unit area or biomass. Whole field estimates of N₂ fixed by blue-green algal communities during the pre-flood period ranged from 1.0 to 10.2 kg N ha^–1. Much of this is probably not recycled until floodwaters cover the fields. However N₂ fixed after floodwaters have receded is probably recycled rapidly due to ploughing.     

Nitrogenase activity,amino acid pool patterns and amination in blue-green algae

Summary The free amino acid pools in the nitrogen-fixing blue-green algae Anabaena cylindrica, A. flos-aquae and Westiellopsis prolifica contain a variety of amino acids with aspartic acid, glutamic acid and the amide glutamine being present in much higher concentrations than the others. This pattern is characteristic of that found in organisms having glutamine synthetage/glutamate synthetase [glutamine amide-2-oxoglutarate amino transferase (oxido-reductase)] as an important pathway of ammonia incorporation. Under nitrogen-starved conditions the level of acetylene reduction (nitrogen fixation) and the glutamine pool both increase but the free ammonia pool decreases, suggesting that ammonia rather than glutamine regulates nitrogen fixation.Glutamine synthetase has been demonstrated in Anabaena cylindrica using the -glutamyl transferase assay and also using a biosynthetic assay in which P_i release from ATP during glutamine synthesis was measured. The enzyme (-glutamyl transferase assay) is present in nitrogen-fixing cultures and activity is higher in aerobic than in microaerophilic cultures. Ammonium-grown cultures have lowest levels of all and activity in the presence of nitrate-nitrogen (150 mg nitrogen 1^-1) is lower than in aerobic cultures growing on elemental nitrogen. Ammonium-nitrogen and nitrate-nitrogen have no effect on glutamine synthetase in vitro. Glutamate synthetase also operates in nitrogen-fixing cultures of Anabaena cylindrica.     

Ca requirement for aerobic nitrogen fixation by heterocystous blue-green algae

The requirement of Ca²⁺ for growth and nitrogen fixation has been investigated in two strains of heterocystous blue-green algae (Anabaena sp. and Anabaena ATCC 33047). With combined nitrogen (nitrate or ammonium) or with N₂ under microaerobic conditions, Ca²⁺ was not required for growth, at least in concentrations greater than traces. In contrast, Ca²⁺ was required as a macronutrient for growth and nitrogen fixation with air as the nitrogen source. Addition of Ca²⁺ to an aerobic culture without Ca²⁺ promoted, after a lag of several hours, development of nitrogenase activity and cell growth. Provision of air to a microaerobic culture in the absence of Ca²⁺ promoted a drastic drop in nitrogenase activity, which rapidly recovered its initial level upon restoration of microaerobic conditions. Development of nitrogenase activity in response to either Ca²⁺ or low oxygen tension was dependent on de novo protein synthesis. The role of Ca²⁺ seems to be related to protection of nitrogenase from inactivation, by conferring heterocysts resistance to oxygen.     

Nitrogenase activity in the blue-green alga Plectonema boryanum strain 594

Summary The non-heterocystous filamentous blue-green alga, Plectonema boryanum strain 594 reduces acetylene to ethylene, incorporates ¹⁵N₂ into cell protoplasm, and grows readily in medium free of combined nitrogen, when incubated in a gas phase without added oxygen. Cells grown in the presence of 50 mg/l of ammonium-nitrogen do not reduce acetylene, and a concentration of 0.015 atm. CO in the gas phase inhibits nitrogenase activity completely but inhibits ¹⁴CO₂ incorporation by only 28%. Nitrogenase activity is inhibited after 2 h treatment with 3×10^-5 M DCMU and is inhibited completely in air.     

Nitrogenase activity in the non-heterocystous cyanobacterium Oscillatoria sp. grown under alternating light-dark cycles

The non-heterocystous cyanobacterium Oscillatoria sp. strain 23 fixes nitrogen under aerobic conditions. If nitrate-grown cultures were transferred to a medium free of combined nitrogen, nitrogenase was induced within about 1 day. The acetylene reduction showed a diurnal variation under conditions of continuous light. Maximum rates of acetylene reduction steadily increased during 8 successive days. When grown under alternating light-dark cycles, Oscillatoria sp. fixes nitrogen preferably in the dark period. For dark periods longer than 8 h, nitrogenase activity is only present during the dark period. For dark periods of 8 h and less, however, nitrogenase activity appears before the beginning of the dark period. This is most pronounced in cultures grown in a 20 h light – 4 h dark cycle. In that case, nitrogenase activity appears 3–4 h before the beginning of the dark period. According to the light-dark regime applied, nitrogenase activity was observed during 8–11 h. Oscillatoria sp. grown under 16 h light and 8 h dark cycle, also induced nitrogenase at the usual point of time, when suddenly transferred to conditions of continuous light. The activity appeared exactly at the point of time where the dark period used to begin. No nitrogenase activity was observed when chloramphenicol was added to the cultures 3 h before the onset of the dark period. This observation indicated that for each cycle, de novo nitrogenase synthesis is necessary.     

Phosphorus availability and nitrogenase activity in aquatic blue-green algae

Metabolically active phosphorus-starved cultures of blue-green algae assimilate ³²P rapidly in the light and in the dark. The uptake of phosphorus results in a rapid (within 15 min) stimulation in acetylene reduction by Anabaena cylindrica, A. flosaquae, Anabacnopsis circuiaris and Chlorogloea fritschii, with a response being obtained to less than 5 μg/1 of phosphorus. Uptake of phosphorus also causes a rapid increase in respiration in the dark but not in photo respiration, and the size of the cellular ATP pool and the ¹⁴CO₂ fixation rate both increase more slowly. The metabolism of phosphorus-sufficient cells, which assimilate phosphorus more slowly, shows little response when phosphorus is provided. Excess phosphorus is stored in the vegetative cells of blue-green algae as polyphosphate bodies which may form within 60 min of adding phosphorus to phosphorusstarved cells and which serve as a source of phosphorus for the algae when exogenous phosphorus is limiting. Preliminary results from Scottish waters suggest that urban effluents are important sources of available-phosphorus for algal growth and that the levels entering fresh waters from agricultural land are, per unit volume, lower. In both types of water the levels of available-phosphorus are rather similar to the levels of orthophosphate-phosphorus present. Most detergents tested serve as a source of phosphorus for nitrogen-fixing blue-green algae and cause a rapid stimulation in reduction when added to phosphorus-starved cultures. Of the detergents assayed, the biological types were richest in available phosphorus. The addition of detergents may result in a rapid increase in number of polyphosphate bodies present in the algae. Detergents in general also contain an inhibitor of algal metabolism. Whether a stimu-lation or an inhibition occurs depends on the quantities of detergent added and on whether or not the alga is phosphorus-deficient.     

Fine control of citrate synthase activity in blue-green algae

Summary The citrate synthases of four blue-green algae, two unicellular (Aphanocapsa spp.) and two filamentous (Nostoc sp., Phormidium sp.) were inhibited by -ketoglutarate but not by NADH. This control of citrate synthase activity reflects the lack of -ketoglutarate dehydrogenase in blue-green algae and the strictly biosynthetic role played by the glutamate branch of the tricarboxylic acid cycle. The citrate synthases were also inhibited by ATP and the enzyme of one of the unicellular organisms was also sensitive to inhibition by NADPH. These effectors may function in regulating the flow of fixed carbon into lipids rather than the glutamate family of amino acids.Contribution No. 1649 from the University of Miami, Rosenstiel School of Marine and Atmospheric Science, 10 Rickenbacker Causeway, Miami, Florida 33149, U.S.A.     

Mesosomes in blue-green algae

Summary Mesosome-like, unit-membrane structures are clearly defined in the blue-green algae, Spirulina and three strains of Synechococcus, after osmium or potassium permanganate fixation and observation with the electron microscope. The membranous structures are distinct from the photosynthetic membranes and, in the case of Spirulina, are frequently observed in cells and can occur in large volume within the cell.     

Immunostimulating activity of the lipopolysaccharides of blue-green algae]

The whole cells of blue-gree algae and lipopolysaccharides isolated from these cells were shown to stimulate the production of macro-(mainly) and microglobulin antibodies in rabbits. The macro- and microphage indices in rabbits increased significantly after the injection of LPS isolated from blue-green algae 24--48 hours before infecting the animals with a virulent Y. pseudotuberculosis strain. Besides, the inhibiting action of this strain on the migration of phagocytes to the site of infection was abolished immediately after the injection. The use of the indirect hemagglutination test allowed to prove the absence of close antigenic interrelations between blue-green algae and the following organisms: Spirulina platensis, Microcystis aeruginosa, Phormidium africanum and P. uncinatum.     

Catalase activity in thermal blue-green algae in relation to temperature

Biologia Plantarum - The author has studied the relation of the catalase activity in thermal blue-green algae to temperature. Experimental data were taken from the first phase of the reaction, so...     

Hydrocarbons in green and blue-green algae

Liquid column chromatography and thin-layer chromatography were used to determine the total content of hydrocarbons and gas chromatography was used to evaluate composition of hydrocarbons in green algae (Chlorella kessleri, C. vulgaris, Chlorella sp.,Scenedesmus acutus, S. acuminatus, S. obliquus) and the blue-green alga (Spirulina platensis) cultivated under autotrophic or heterotrophic conditions. InC. kessleri cultivated under heterotrophic conditions the content of hydrocarbons was found to be about 10^-2 % (per dry mass), whereas under autotrophic conditions it was about 10^-3 % (per dry mass). The highest content of hydrocarbons was detected in species of the genusScenedesmus cultivated autotrophically (10^-1 %). Heptadecane and hexacosane were found as major alkanes, 1-heptadecene was detected among alkenes.     

Morphology-habitat associations in blue-green algae

Morphological and habitat features have been recorded on punchcards for three genera in the Oscillatoriaceae and two in the Nostocaceae. As a result it is concluded that generic divisions in the Oscillatoriaceae are arbitrary and artificial, but that in the Nostocaceae, Cylindrospermum represents a grouping of distinct forms to a greater degree than does the amorphous genus Anabaena.

Some morphological comparisons between the two groups are attempted and detectable associations between morphological and habitat features are suggested : together these may contribute to an eventual separation of taxa on a more logical basis than at present.     

Ammonia assimilation in blue-green algae

Summary The occurrence of alanine dehydrogenase (AlaDH), glutamate dehydrogenase (GDH), and 2-ketoglutarate: glutamine amidotransferase (GGAT), has been surveyed in a number of blue-green algae. Among nine unicellular strains grown with nitrate, and belonging to five of the major typological groups, AlaDH was present in seven, and GDH in all eight that were assayed. In ten filamentous strains grown with nitrate, and belonging to the three nonheterocyst-forming and four heterocyst-forming groups, AlaDH was present in six, but both AlaDH and GDH were present in only one strain. In those strains which could be grown with N₂ as sole nitrogen source, levels of GDH were generally lower, and AlaDH higher in cells fixing N₂ than in those growing with nitrate. GGAT was undetectable in N₂-grown cells. Two unicellular and three filamentous strains were tested for their ability to use L-alanine, L-glutamate, L-glutamine, and L-asparagine as sole sources of nitrogen. Of these, L-asparagine was utilized most effectively. There was little difference in levels of GDH in cells grown with nitrate or with L-asparagine, while the levels of AlaDH were slightly lower in cells grown with L-asparagine.