Respiration in Blue-Green Algae

The low rate of endogenous respiration exhibited by the blue-green algae Anacystis nidulans and Phormidium luridum was not increased by the addition of respiratory substrates. However, endogenous respiration was inhibited by low concentrations of cyanide and by high carbon monoxide tensions. In addition, the uncouplers dinitrophenol and carbonyl cyanide p-trifluoromethoxyphenylhydrazone both stimulated the respiratory rate. The transition of cells from the aerobic steady state to anaerobiosis was accompanied by a decrease in the concentration of cellular nicotinamide adenine dinucleotide phosphate (NADP(+)) and adenosine triphosphate (ATP), whereas the concentration of nicotinamide adenine dinucleotide (NAD(+)) was unchanged. Concomitant with the metabolite decreases were stoichiometric increases io reduced NADP(+) (NADPH), adenosine diphosphate, and adenosine monophosphate. A decrease in ATP was also observed after the addition of uncouplers. These data are interpreted as evidence for the association of oxidative phosphorylation with the oxidation of NADP(+)-linked substrates in these algae. Membrane fragments isolated from the algal cells oxidized succinate, malate, ferrocytochrome c, ascorbate-tetramethyl-p-phenylenediamine, and reduced 2,6-dichlorophenol indophenol but did not oxidize NADPH or reduced NAD(+) in a cyanide-sensitive system. Oxidative phosphorylation has not yet been demonstrated in these fragments, but a dark ATP-P(i) exchange, distinct from the lighttriggered exchange associated with photosynthesis, is readily observed. This exchange was inhibited by phloridzin, Atabrine, and uncouplers in concentrations which suggest that the mechanism of oxidative phosphorylation in blue-green algae is different from that found in other bacteria and in mitochondria. These results led to the conclusion that the biochemical basis for obligate autotrophy in these organisms does not lie in the metabolic events associated with terminal electron transport and energy conservation.     

Photooxidative Death in Blue-Green Algae

When incubated in the light under 100% oxygen, wild-type blue-green algae (Anacystis nidulans, Synechococcus cedrorum) die out rapidly at temperatures of 4 to 15 C, and at 35 C (or at 26 C in the case of S. cedrorum) in the absence of CO(2). Photosynthesis is impaired in these cells long before they die. Blocking of photosystem II at high temperatures in the presence of CO(2) sensitizes the algae to photooxidative death. Photooxidative death and bleaching of photosynthetic pigments are separable phenomena. Photooxidative conditions were demonstrated in Israeli fish ponds using A. nidulans as the test organism during dense summer blooms, when dissolved CO(2) is low, and in winter, when water temperatures generally drop below 15 C. This finding suggests that photooxidative death may be responsible for the sudden decomposition of blue-green blooms in summer, and may be a factor in the absence of blue-green blooms in winter.     

Ultrastructure of Blue-Green Algae

Two freshwater blue-green algae, Tolypothrix tenuis and Fremyella diplosiphon, and an oscillatorialike marine alga, were found to possess structures on the photosynthetic lamellae which appear to correspond to the phycobilisomes of red algae. These homologous structures are important because they contain the phycobilins which are accessory pigments involved in photosynthesis. As in the red algae, the phycobilisomes were attached on the outer side of each lamellae, i.e., the side facing away from its own membrane pair. Although our study on Anacystis nidulans has not thus far revealed the presence of phycobilisomes, some observations were made on the structure of the polyhedral bodies. After negative staining, the polyhedral bodies were seen to be composed of regularly spaced subunits arranged in a crystalline array. Elongated segmented rods, which differed from the polyhedral bodies, were found in the nuclear region of apparently healthy Tolypothrix cells.     

钝顶螺旋藻（Spirulina platensis）藻胆体Langmuir-Blodgett膜

从钝顶螺旋藻中分离制备完整藻胆体 ,然后滴加于空气 水界面上 ,应用LB膜技术制备藻胆体LB膜。结果表明 ,藻胆体在空气 水界面上具有很好的成膜性能。将藻胆体LB单层膜转移到刚揭开的云母表面 ,喷一层金 ,然后用扫描隧道显微镜观察。结果表明 ,藻胆体在Langmuir Blodgett膜中的排列方式与其在体内类囊体膜表面的排列方式类似 ,一排排聚集在一起 ,然后排列成膜。藻胆体的“核”吸附在云母表面 ,而藻胆体的“杆”伸向外面。由于钝顶螺旋藻易于规模化培养 ,藻胆体容易批量制备 ,加之藻胆体具有的独特的光物理、光化学特性和良好的成膜性能 ,以及本身就是纳米量级的颗粒 (5 0 70nm) ,预示着藻胆体在纳米光电子器件中具有很好的应用前景。     

Cylindrical Phycobilisomes from a Blue-Green Alga, Anabaena variabilis

Cylindrical 52.5-nm-long phycobilisomes were observed in Anabaenavariabilis, differing from the generally accepted hemidiscoidalmorphology. The central part of such a phycobilisome has a network-likefine structure of slightly greater diameter (16 nm) than theconnected end parts of stacked-disc structure (12 nm in diameter).On the basis of this morphology, the molecular mass of thisphycobilisome was calculated to be 3.27?10⁶, about 60% of whichis accounted for by phycocyanin with the rest being due to allophycocyanin.Separately prepared 23 S allophycocyanin particles with a molecularmass of 1.13?10⁶ have the dimensions (16?23 nm) and network-likefine structure similar to the central part of phycobilisomes,while an aggregate form of phycocyanin (18 S) has a fine structureof stacked discs similar to the connecting end part of phycobilisomes,suggesting that the central part constitutes the core at whichthese phycobilisomes attach to the thylakoid membranes. (Received June 5, 1982; Accepted September 21, 1982)     

Lysis of Blue-Green Algae by Myxobacter

Enrichment from local fishponds led to the isolation of a bacterium capable of lysing many species of unicellular and filamentous blue-green algae, as well as certain bacteria. The isolate is an aflagellate, motile rod which moves in a gliding, flexuous manner; the organism is capable of digesting starch and agar, but not cellulose and gelatin. Its deoxyribonucleic acid base pair composition (per cent guanine plus cytosine approximately 70) shows a close resemblance to that of the fruiting myxobacteria. Algae in lawns on agar plates were lysed rapidly by the myxobacter, but only limited and slow lysis occurred in liquid media, and no lysis took place when liquid cultures were shaken. No diffusible lytic factors would be demonstrated. Continuous observation of the lytic process under a phase-contrast microscope suggested that a close contact between the polar tip of the myxobacter and the alga is necessary for lysis. The lytic action is limited to the vegetative cells of the algae, whereas heterocysts are not affected. The gas vacuoles of the algal host are the only remnant visible after completion of digestion by the myxobacter.     

Pyridine Nucleotide-Dependent Glucose Dehydrogenase Activity in Blue-Green Algae

Pyridine nucleotide-dependent glucose dehydrogenase activity (GPND) is described for the first time in cell-free extracts of certain blue-green algae. When glucose is added to these crude cell extracts, nicotinamide adenine dinucleotide phosphate is reduced at twice the rate as nicotinamide adenine dinucleotide; but evidence suggests that this activity is due to a single enzyme. The distribution and level of GPND in selected blue-green algae correlates with the heterotrophic potential of each species. In all blue-green algae where GPND was detected, O(2) uptake coupled to the GPND reaction was also observed. Both GPND and O(2) uptake apparently occur in the soluble fraction of the cell. An essential role for GPND in the heterotrophic metabolism of blue-green algae is postulated.     

Heterotrophic Growth of Blue-Green Algae in Dim Light

A unicellular blue-green alga, Agmenellum quadruplicatum, and a filamentous blue-green alga, Lyngbya lagerheimíi, were grown heterotrophically in dim light with glucose as major source of carbon and possibly energy. The dim-light conditions did not support autotrophic growth. The two blue-green algae appeared to have the same metabolic block, namely an incomplete tricarboxylic acid cycle, as has been found in other obligately phototrophic blue-green algae. Under dim-light conditions, glucose made a greater contribution to cell constituents (amino acids) of A. quadruplicatum and L. lagerheimii than under high-light conditions.     

Metabolism of delta-Aminolevulinic Acid in Red and Blue-Green Algae

δ-Aminolevulinic acid was incorporated in vivo into C-phycocyanin and B-phycoerythrin in two species of the Rhodophyta (Cyanidium caldarium, Porphyridium cruentum) and three species of the Cyanophyta (Anacystis nidulans, Plectonema boryanum, Phormidium luridum). Amino acid analysis of phycocyanin-¹⁴C from C. caldarium cells which had been incubated with δ-aminolevulinate-4-¹⁴C showed that 84% of the radioactivity incorporated was present in the phycocyanobilin chromophore and less than 16% of the radioactivity cochromatographed with amino acids. These results indicate that δ-aminolevulinate is utilized predominantly via the porphyrin pathway in C. caldarium. Conversely, analysis of phycocyanin-¹⁴C prepared from cells of A. nidulans, P. boryanum, and P. luridum which had been incubated with radiolabeled δ-aminolevulinate demonstrated that 85%, 81%, and 93%, respectively, of the radioactivity incorporated cochromatographed with amino acids. The ratio of incorporated radioactivity in amino acids and phycoerythrobilin was 40:60 in P. cruentum phycoerythrin obtained from cells which had been incubated with δ-aminolevulinate-4-¹⁴C. Succinate-2-3-¹⁴C appeared to be as good a carbon source of amino acids as did C₄ and C₅ of δ-aminolevulinate. These data demonstrate a major alternate route (other than the porphyrin pathway) of δ-aminolevulinate metabolism in red and blue-green algae. The factors responsible for the extent to which δ-aminolevulinate is utilized for synthesis of porphyrins and their derivatives and routes of δ-aminolevulinate catabolism in the organisms employed are discussed.     

Biochemical Basis of Obligate Autotrophy in Blue-Green Algae and Thiobacilli

Differential rates of incorporation of sugars, organic acids, and amino acids during autotrophic growth of several blue-green algae and thiobacilli have been determined. In obligate autotrophs (both blue-green algae and thiobacilli), exogenously furnished organic compounds make a very small contribution to cellular carbon; acetate, the most readily incorporated compound of those studied, contributes about 10% of newly synthesized cellular carbon. In Thiobacillus intermedius, a facultative chemoautotroph, acetate contributes over 40% of newly synthesized cellular carbon, and succinate and glutamate almost 90%. In the obligate autotrophs, carbon from pyruvate, acetate, and glutamate is incorporated into restricted groups of cellular amino acids, and the patterns of incorporation in all five organisms are essentially identical. These patterns suggest that the tricarboxylic acid cycle is blocked at the level of alpha-ketoglutarate oxidation. Enzymatic analyses confirmed the absence of alpha-ketoglutarate dehydrogenase in the obligate autotrophs, and also revealed that they lacked reduced nicotinamide adenine dinucleotide oxidase, and had extremely low levels of malic and succinic dehydrogenase. These enzymatic deficiencies were not manifested by the two facultative chemoautotrophs examined. On the basis of the data obtained, an interpretation of obligate autotrophy in both physiological and evolutionary terms has been developed.     

Glycolic Acid Oxidase Activity in Cell-free Preparations of Blue-Green Algae

Glycolic acid oxidase activity was detected in cell-free preparations of Anabaena flos-aquae and Oscillatoria sp. by the reduction of 2,6-dichlorophenolindophenol and by the formation of glyoxylate. Enzyme activity was localized in the 20,000 times gravity supernatant fraction, and optimal activity was obtained at pH 8.0. Activity was lost on storing the preparation at 4 C and could not be restored by addition of flavin mononucleotide.     

Interaction of Two Populations of the Blue-Green Algae in Co-Culture

Interaction of the populations of two species of blue-green algae Tolypothrix tenuis and Anabaena variabilis "1058", in mixed culture, was observed in three different ways: (1) The growth competition was studied in mixed-culture, where the two species grew in one batch of culture. (2) The effect of the filtrate, the extracellular products, from the culture of one species on the growth of the other species was studied. (3) In fitrated culture, two algal species were cultivated separately in either side a U-form container partitioned by a micro-pore membrane in the middle. The extracellular products were permeable through the membrane from one side to the other side. The influonce of the biologically active substances prosduced by the algae at different growth stages can be observed and estimated. The growth was measured by dry weight of biomass andchlorophyll-a content. The proportion of components of phycobiliprotein (e. g. ratio of phycocyanin to phycoerythrin) was estimated as the specific growth rate of two blue-greens in the mixed cultures. 1. The results obtained are summarized as follows. There were three types of the bioactivity of the extracellular products: (1) The lethal effects on each other caused the by the lethal agents of these two blue-greens were different. (2) The suppressive effects on growth by each other in one community were also found. (3) Effect on growth promotion exhibited only in the extracellular products of A. variabilis "1058". 2. The results indicate that there occur a direct competitive interaction of two populations. This Competition was controlled by the bio-active substances of the extracellular products which might regulate the structural composition and inturn, the succession of the comunity.     

Fatty Acid Composition of Unicellular Strains of Blue-Green Algae

The fatty acids of 34 strains of unicellular blue-green algae provisionally assigned to the genera Synechococcus, Aphanocapsa, Gloeocapsa, Microcystis, and Chlorogloea by Stanier et al. have been chemically characterized. The strains analyzed can be divided into a series of compositional groups based upon the highest degree of unsaturation of the major cellular fatty acids. Twenty strains fall into the group characterized by one trienoic fatty acid isomer (alpha-linolenic acid), and seven strains fall into a group characterized by another trienoic acid isomer (gamma-linolenic acid). These groups in many cases correlate well with groupings based upon other phenotypic characters of the strains, e.g., deoxyribonucleic acid base composition. The assignment of a strain to a compositional group is not altered when the strain is grown under a variety of different culture conditions. All strains contain glycolipids with the properties of mono- and digalactosyldiglycerides.