Fixation of Elemental Nitrogen by Marine Blue-green Algae

Three blue-green algae, Calothrix scopulorum, Nostoc entophytum,and Oscillatoria brevis, isolated from the upper littoral andsupralittoral fringe of the sea-shore were obtained in pureculture and tested for fixation of elemental nitrogen. Appreciablefixation by Calothrix and Nostoc was detected, a proportionof the total nitrogen fixed being liberated into the culturemedium. There was no evidence of fixation by Oscillatoria. Thisappears to be the first evidence that blue-green algae isolatedin pure culture from marine habitats fix nitrogen.     

Resistance to blue-green algal toxins by Bosmina longirostris

Bosmina longirostris was resistant to strains of Microcysrtsaeruginosa and Anabaena flosaquae previously reported to havelethal toxic effects on cladocerans. These blue-green algaewere of poor nutritional value to B.longirostris; survivorshipwas increased when fed these algae but reproduction was negligible.Results of feeding selectivity experiments showed that B.longirostrisdid not avoid consuming these blue-green algae, indicating thatthe mechanism for resistance must be post-ingestion. These resultssuggest that, unlike the other cladocerans tested, B.longirostriscould potentially coexist with toxic blue-green algal blooms.     

Acetylene reduction by nitrogen-fixing blue-green algae

Summary Known nitrogen-fixing species of blue-green algae are capable of reducing acetylene to ethylene, but acetylene is not reduced by Anacystis nidulans, which does not fix nitrogen. Cycad root nodules which contain blue-green algae as endophytes reduce acetylene. Acetylene reduction is inhibited by carbon monoxide. Nitrate or ammonium-nitrogen has no immediate effect on algae reducing acetylene, but algae grown on nitrate-nitrogen gradually lose their capacity to reduce acetylene. Nitrate-nitrogen also inhibits heterocyst formation in these algae and there is a fairly direct correlation between the abundance of heterocysts in a particular sample and its capacity to reduce acetylene. Aphanizomenon flosaquae reduces acetylene and fixes nitrogen in unialgal culture and there is strong presumptive evidence that these reductions are carried out by the alga rather than by associated bacteria. The molar ratios of ethylene: ammonia produced vary within the range 1.4–1.8.     

Algal nitrogen fixation in the tropics

Summary a)Nitrogen fixation in rice fields. Nitrogen-fixing blue-green algae grow abundantly in tropical regions and are particularly common in paddy fields. Their possible role in the nitrogen accumulation of soil has been studied. The most vigorous nitrogen-fixing blue-green algae have been assessed for use as green manure in rice fields and favorable effects have been reported in India and other countries. b)Nitrogen fixation by algae in water. The planktonic blue-green algae occur abundantly at certain time of the year in sea water and lake water, and some of them are known to be nitrogen fixers. Certain Japanese species of blue-green algae can withstand high temperatures including ten nitrogen-fixing species from hot-spring waters. c)Nitrogen fixation by symbiotic blue-green algae. Certain species of blue-green algae form associations with other organisms such as fungi, liverworts, ferns and seed plants. The relationship between these two organisms is on one occasion commensal and on others symbiotic. Certain symbiotic blue-green algae are provided with the ability to fix the atmospheric nitrogen.     

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.     

Quantitative relationship between two reaction centersin the photosynthetic system of blue-green algae

The quantitative relationship between reaction centers I andII was studied with blue-green algae Anabaena cylindrica, Anabaenavariabilis and Anacystis nidulans grown under different lightconditions. The number of reaction centers I was estimated fromthe P700 content and that of reaction centers II, from the O₂yield of repetitive short flashes. Supplementary determinationswere done with three other blue-green algae and one red alga.The maximum number of reaction centers II counted from the O₂yield of repetitive short flashes was markedly smaller thanthe total number of P700 in all algae tested when the algaewere grown under weak light; in the extreme case (Anabaena cylindrica),the ratio was only 0.258?0.015. This ratio became larger andclose to unity when the algae were grown under stronger light.Variation in the number of reaction centers in a single cellsuggested that reaction center I was a variable component. Ourresults indicate that the proportion of the two reaction centersmay markedly vary in blue-green algae depending on the growthconditions (Received November 13, 1978; )     

The occurrence of the hydrogenase in some blue-green algae

Several blue-green algae were surveyed for the occurrence of the hydrogenase which was assayed by the oxyhydrogen or Knallgas reaction in the intact organisms. In aerobically grown cultures, the reaction was detectable in Anabaena cylindrica, Nostoc muscorum and in two Anabaena variabilis species, whereas virtually no activity was observed in Anacystis nidulans and Cyanophora paradoxa. In these latter two algae, the reaction was, however, found after growth under molecular hydrogen for several days, which drastically increased the activity levels with all the algae tested. In the nitrogen fixing species, the activity of the Knallgas reaction was enhanced when all combined nitrogen was omitted from the media. H₂ and hydrogenase could not significantly support the CO₂-fixation in photoreduction experiments with all blue-green algae investigated here. Hydrogenase was assayed by the dithionite and methyl viologen dependent evolution of hydrogen and was found to be present with essentially the same specific activity levels in preparations of both heterocysts and vegetative cells from Anabaena cylindrica. Na₂S₂O₄ as well as H₂ supported the C₂H₂-reduction of the isolated heterocysts. The H₂-dependent C₂H₂-reduction did not require the presence of oxygen but was strictly light-dependent where H₂ served as an electron donor to photosystem I of these cells. It is concluded that hydrogen can be utilized by two different pathways in blue-green algae.Abbreviations Chl chlrophyll - CP creatine phosphate - CP kinase creatine phosphokinase - DCMU N-(3,4-dichlorophenyl)N,N-dimethylurea     

Nitrogen fixation and heterocysts in the blue-green alga Anabaena cylindrica

The site of nitrogen fixation in the blue-green alga Anabaenacylindrica Lemra (Fogg strain) was investigated. Less than 4%of the total nitrogen fixed during a relatively short period(5-15 min) was recovered in heterocysts. When estimated on thecellular nitrogen basis, vegetative cells can fix molecularnitrogen at the same rate as do heterocysts. There was no positivecorrelation between nitrogen fixation and heterocyst formation.Results do not support the hypothesis that the heterocyst isthe main site for nitrogen fixation in blue-green algae. ¹ This work was supported by grant (No. 38814) from the Ministryof Education. (Received July 23, 1971; )     

Cyanidium caldarium as a bridge alga between Cyanophyceae and Rhodophyceae: Evidence from immunodiffusion studies

The primer-independent phosphorylase isozyme, a₂, of Cyanidiumcaldarium was used for immunization of rabbits. The immune serumwas tested against pure a₂ isozymes from blue-green, red, andgreen algae. Double immunodiffusion in agar indicated that therewas structural similarity in the isozyme from Cyanidium caldarium,the blue-green algae, Oscillaloria princeps, Pleclonema nostocorumand the red alga, Rhodymenia pertusa. Complete fusion of theprecipitin lines was obtained with these algae. However, onlypartial fusion was observed with the a₂ isozymes from Chlorophyceaesuch as Chlorella pyrenoidosa and Spirogyra setiformis. Spurformation on the precipitin lines occurred when the isozymesfrom these algae were tested against the immuneserum. The results were interpreted as indicative of the possible transitionstatus of Cyanidium caldarium between prokaryotic blue-greenalgae and eukaryotic red algae. It would appear that the Chlorophyceaeevolved along different lines from Cyanophyceae than did theRhodophyceae. (Received November 25, 1975; )     

The effects of nutrient,pH and herbicide levels on algal growth

Growth inhibition of algae increased as herbicide concentrations increased, particularly with prometryn and fluometuron. However, picloram had no effect on algal growth while dinoseb inhibited only Lyngbya. There were no differences in growth rate of algae treated with different levels of potassium or phosphorus. High levels of calcium or magnesium increased growth rate of the algae tested. High levels of nitrogen or pH increased growth rates except when combined with prometryn or fluometuron.     

Biomass production,total protein,chlorophylls, lipids and fatty acids of freshwater green and blue-green algae under different nitrogen regimes

Two green algae (Chlorella vulgaris and Scenedesmus obliquus) and four blue-green algae (Anacystis nidulans, Microcystis aeruginosa, Oscillatoria rubescens and Spirulina platensis) were grown in 81 batch cultures at different nitrogen levels. In all the algae increasing N levels led to an increase in the biomass (from 8 to 450 mg/l), in protein content (from 8 to 54 %) and in chlorophyll. At low N levels, the green algae contained a high percentage of total lipids (45 % of the biomass). More than 70 % of these were neutral lipids such as triacylglycerols (containing mainly 16:0 and 18:1 fatty acids) and trace amounts of hydrocarbons. At high N levels, the percentage of total lipids dropped to about 20 % of the dry weight. In the latter case the predominant lipids were polar lipids containing polyunsaturated C₁₆ and C₁₈ fatty acids. The blue-green algae, however, did not show any significant changes in their fatty acid and lipid compositions, when the nitrogen concentrations in the nutrient medium were varied. Thus the green but not the blue-green algae can be manipulated in mass cultures to yield a biomass with desired fatty acid and lipid compositions. The data may indicate a hitherto unrecognized distinction between prokaryotic and eukaryotic organisms.     

Nitrogen fixation by the unicellular blue-green alga Aphanothece

Summary Frequent growth of unicellular blue-green alga Aphanothece sp. was observed in medium free of combined nitrogen. Its generation time was 12 h and more than 2 mg of nitrogen was fixed in 25 days. Its growth and nitrogen fixation were comparable to other heterocystous algae.     

Growth and Nitrogen Fixation by Westiellopsis prolifica Janet

The blue-green alga Westiellopsis prolifica Janet has been isolatedin unialgal bacteria-free culture. Evidence of the ability ofthis algs to fix elementary nitrogen was obtained by demonstrationof increase in total combined nitrogen in cultures by the micro-Kjeldahlmethod and also by the uptake of ¹⁵N as demonstrated by theuse of the mass spectrometer. The growth and nitrogen-fixingcapacity of the alga was studied in relation to temperatureand light intensity. Increase in dry weight was greatest at40° C but the optimum for nitrogen fixation was between30 and 35°C. A considerable proportion of the nitrogen fixedwas released in soluble organic form into the medium.     

Responses of a eutrophic pond community to separate and combined effects of N:P supply and planktivorous fish: a mesocosm experiment

We conducted an outdoor mesocosm experiment of factorial design consisting of three levels of nutrient supply (no nutrient addition and additions of nitrogen and phosphorus in ratios of 10:1 and 45:1) cross-classified with two levels of bluegill (Lepomis macrochirus) (presence and absence). Nutrient supply significantly affected total phosphorus (TP), total nitrogen (TN), TN: TP ratio, turbidity, Secchi depth, phytoplankton chlorophyll, filamentous blue-green algae, periphyton chlorophyll, Asplanchna and non-predatory rotifers. The presence of bluegill significantly increased TP, turbidity, diatoms, unicellular green algae, colonial blue-green algae, filamentous blue-green algae, periphyton chlorophyll, Asplanchna and non-predatory rotifers, and decreased Secchi depth, cladocerans, cyclopoid copepodids, copepod nauplii and chironomid tube densities. Nutrient supply and fish effects were not independent of each other as shown by significant nutrient × fish interaction effects for TP, Secchi depth, filamentous blue-green algae, periphyton chlorophyll, Asplanchna and non-predatory rotifers.     

Measurement of photorespiration in algae

The rates of true and apparent photosynthesis of two unicellular green algae, one diatom and four blue-green algae were measured in buffer at pH 8.0 at subsaturating concentrations of dissolved inorganic carbon (13-27 micromolar). Initial rates of depletion from the medium of inorganic carbon and ¹⁴C activity caused by the algae in a closed system were measured by gas chromatography and by liquid scintillation counting, respectively. The rate of photorespiration was calculated as the difference between the rates of apparent and true photosynthesis. The three eucaryotic algae and two blue-green algae had photorespiratory rates of 10 to 28% that of true photosynthesis at air levels of O₂. Reduction of the O₂ level to 2% caused a 52 to 91% reduction in photorespiratory rate. Two other blue-green algae displayed low photorespiratory rates, 2.4 to 6.2% that of true photosynthesis at air levels of O₂, and reduction of the O₂ concentration had no effect on these rates.     

Toxicity of Selenium to the Blue-green Algae, Anacystis nidulans and Anabaena variabilis

Selenate, selenite, selenomethionine, and selenopurine are toxicto Anacystis nidulans and Anabaena variabilis. These compoundsare less toxic in culture medium containing sulphate than insulphur-free medium, thus suggesting a protective role of sulphuragainst selenium toxicity Selenite is more toxic in agar plate cultures and less toxicin liquid cultures than selenate. Cells grown in the highestgrowth-permitting concentration of selenite in liquid mediumform a variable number of red granules No such granulation occursin cells treated with other seleno-compounds, indicating therebythat the mode of selenite action in blue-green algae differsfrom that of other selenium compounds     

UV-B treated algae exhibiting different responses as a food source for Daphnia magna

The response of green algae Scenedesmus quadricauda and Selenastrumcapricornutum to different levels of UV-B radiation was studied.Daphnia magna, the next step in the food web, was fed with UV-Btreated algae. UV-B radiation induced the synthesis of UV absorbingsubstances and photosynthetic pigments, enhanced the level ofrespiratory potential and suppressed growth of the treated algae.Biomass production in S. quadricauda was higher than in S. capricornutum,but the production of photosynthetic pigments and relative amountsof UV absorbing substances as well as respiratory potentialwere more pronounced in S. capricornutum. A short-term feedingexperiment with D. magna showed that in general, females ingestedhigher numbers of cells of algae treated with a high UV-B dose,but the biomass of ingested algae did not differ significantly.The exception was S. capricornutum treated with a high UV-Bdose; a lower biomass was ingested, probably reflecting thesmaller UV-treated cells.     

Effects of Acid Stress on Aerobic Decomposition of Algal and Aquatic Macrophyte Detritus: Direct Comparison in a Radiocarbon Assay

Radiolabeled phytoplankton and macrophyte lignocelluloses were incubated at pHs 4 and 7 in water from a naturally acidic freshwater wetland (Okefenokee Swamp; ambient pH, 3.8 to 4.2), a freshwater reservoir (L-Lake; pH 6.7 to 7.2), and a marine marsh (Sapelo Island; pH ~7.8). The data suggest that acidity is an important factor in explaining the lower decomposition rates of algae in Okefenokee Swamp water relative to L-Lake or Sapelo Island water. The decomposition of algal substrate was less sensitive to low pH (~5 to 35% inhibition) than was the decomposition of lignocellulose (~30 to 70% inhibition). These substrate-dependent differences were greater and more consistent in salt marsh than in L-lake incubations. In both freshwater sites, the extent to which decomposition was suppressed by acidity was greater for green algal substrate than for mixed diatom or blue-green algal (cyanobacteria) substrates. The use of different bases to adjust pH or incubation in a defined saltwater medium had no significant effect on substrate-dependent differences. Although pH differences with lignocellulose were larger in marine incubations, amendment of lakewater with marine bacteria or with calcium, known to stabilize exoenzymes in soils, did not magnify the sensitivity of decomposition to acid stress.     

Isolation and characterization of rapidly-growing,marine, nitrogen-fixing strains of blue-green algae

Five strains of heterocystous blue-green algae capable of high rates of growth and nitrogenase activity were isolated from shallow coastal environments. Growth of the organisms was characterized with respect to temperature, NaCl concentration in the medium, and nitrogen source. The temperature optima ranged from 35–42°C, and all but one of the strains displayed a requirement for added NaCl. The generation times under N₂-fixing conditions were 5.1–5.9 h, and were as low as 3.4 h for growth on NH₄Cl. Nitrogenase activity (C₂H₂ reduction) was high throughout the logarithmic growth phase of each strain. The maximum value observed for one strain was 65.5 nmoles C₂H₄ produced/mg protein x min, and the average values for the five strains ranged from 24.5–46.7 nmoles C₂H₄/mg protein x min. The organisms all belong to the genusAnabaena. The growth and nitrogenase activity of these strains are much higher than those of the heterocystous blue-green algae commonly used for investigation of nitrogen metabolism, and they thus should prove to be useful physiological tools. Their prevalence, as judged by the ease of their enrichment and isolation, in bay and estuarine environments suggests that they are important contributors of combined nitrogen.