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.     

Photosynthetic and Respiratory Activity in Anacystis nidulans Adapted to Osmotic Stress

Anacystis nidulans cultures growing photoautotrophically in a minimal medium were exposed to different concentrations of NaCl, up to 0.4 molar. Initially, a marked decrease in photosynthetic activity took place, which was in direct relation to the salt concentration and which was not associated with a change in endogenous respiratory activity. After a period of exposure to salinity, a process of adaptation became apparent, being manifested in a partial reversal of the decline in photosynthesis and a marked increase in endogenous respiration.     

Phycobilisomes in Blue-Green Algae

Fifteen species of freshwater blue-green algae, including unicellular, filamentous, and colonial forms, were subjected to a variety of fixatives, fixation conditions, and stains for comparison of the preservation of phycobilisomes. Absorption spectra of the corresponding in vivo and released photosynthetic pigments, in 10 of the species that were maintained in culture, demonstrated the presence of phycocyanin in all 10 species and phycoerythrin in only 2 of them. Spectroscope and electron microscope evidence was obtained for localization of phycobiliproteins in phycobilisomes of Nostoc muscorum. Phycobilisomes were observed in all species examined in situ, strenghening the hypothesis that phycobilisomes are common to all phycobiliprotein-containing photosynthetic blue-green algae.     

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.     

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.     

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.     

Complemental Effect of Osmotic Stabilizers and Their Roles in Degradation Cell Walls of Blue-Green Algae

The compound osmotic stabilizers consisted of several salt solutions exerted a greater effect on the isolation of blue-green algae spheroplasts than a single salt solution. However, the effect of compound osmotic stabilizers on the spheroplast stability could be multiphasic. Some osmotic stabilizers, such as the solution of (NH4) 2C4H406, (NH4) 2SO4 and MgSO4, exerted degradation on cell walls of the blue-green alga; among which the (NH4) 2C4H406 solution (0.15 mol/L) had the greatest degradation resulting in formation of spheroplasts. The spheroplasts were sensitive to hypotonic condition but were less transparent.     

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.     

Chromatic Regulation of Photosystem Composition in the Photosynthetic System of Red and Blue-Green Algae

A chromatic adaptation in the photosynthetic quantum yield forthe light mainly absorbed by chlorophyll a (Chl a light) firstfound by Yocum (1951) was studied with one red and three blue-greenalgal strains. When the cells were grown under a weak Chl alight, the quantum yield in all the strains increased. Comparisonof photosystem (PS) compositions, including phycobilin (PBP)and Chl a antennae, reaction centers I and II, in the cellsgrown under the light mainly absorbed by PBP and Chl a revealedthat changes in quantum yield could be attributed to changesin the ratio of PS I/II; PS I/II becomes larger than 1 underPBP light but decreases to 1 in most cases under Chl a light.The change in the PS I/II ratio is due solely to the changesin the PS I population in the cell; PS II remains constant.These results are similar to the intensity-dependent responsein PS composition. A common hypothesis for both the chromatic and intensity-inducedregulation of PS composition was proposed based on the ideaof balance between the electron flow from H₂O to NADP drivenby PS I and II and the cyclic one driven by PS I. (Received May 16, 1985; Accepted September 4, 1985)     

Shear-oriented Microfibrils in the Mucilaginous Investments of Two Motile Oscillatoriacean Blue-Green Algae

Trichomes of two oscillatoriacean blue-green algae execute screw-like gliding motion, but the two organisms differ from each other with respect to the screw sense of motion. Electron microscopy of serial longitudinal sections reveals extracellular microfibrils which lie roughly parallel to stream-lines at the surface of each organism. The author proposes that the microfibrils are oriented by shear in a zone just external to the outer unit membrane-like component of the cell wall.     

Formation of Radioactive Citrulline during Photosynthetic C14O2-Fixation by Blue-Green Algae

Citrulline has been isolated and identified from extracts ofNostoc muscorum. All members of the Cyanophyceae hitherto investigatedshow a relatively large amount of the CO₂ fixed during photosynthesisin citrulline (ranging as high as 20 per cent. in Nostoc) whencompared to the trace amounts found in the Chlorophyceae. Nostocalso has the ability to fix C¹⁴ in citrulline during dark fixation,but at a rate slower than in light. As no free urea or arginine was found in Nostoc, it is likelythat citrulline is functioning in reactions other than thoseleading to arginine and urea synthesis. Other possible functions for citrulline are briefly discussed.     

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.     

An Exclusive Increase in the Concentration of ATP as a Result of Osmotic Stress in Escherichia coli B

The ATP level was exclusively increased among the nucleotides in Escherichia coli under osmotic stress, concomitant with a decrease of guanosine phosphates levels. The profile of ATP formation was different from that resulting from chemicals. E. coli contains a specific system to enhance the ATP level and it might be possible that the source of the increased ATP is guanosine phosphates.     

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.