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PubMed1993

An extended corona attached to metaphase kinetochores of the green alga Oedogonium.

Mitotic cells of the green alga Oedogonium were treated with the anti-microtubule agent oryzalin (1.0-0.1 microM) for 5 to 10 min. Within 5 min treatment of living cells, metaphase spindles became spherical with disorganized chromosomes, and anaphase spindles collapsed. At lower concentrations, the effects were slower, and partial recovery was observed about 10 to 20 min after the drug was washed out. Following breakdown of the spindle, considerable disorganized activity detected by time-lapse continued within the nucleus, isolated from the cytoplasm by its intact nuclear membrane. Under the electron microscope, spindle microtubules (MTs) were absent in oryzalin-treated cells. Paired metaphase kinetochores displayed an array of fine filamentous material extended, usually straight, about 3 microns into the nucleoplasm. In cells recovering from oryzalin treatment, MTs became associated with kinetochores in the usual manner. However, this filamentous array, the "extended corona" (EC), was almost undetectable, even when the MTs were short and poorly organized. The EC is appreciably larger by metaphase than the corona of prophase chromosomes and so it may assemble during early mitosis. Fine filaments interspersed with kinetochore MTs have been described in carefully fixed cells of this alga (M.J. Schibler, J.D. Pickett-Heaps, Eur. J. Cell Biol. 22, 687-698 (1980)). The EC apparently represents a less organized form of this material remaining after its scaffold of MTs has been removed. These fibers appear involved in MT capture upon spindle recovery from anti-MT drugs. They could function during prometaphase and even anaphase movement along spindle MTs.

Dinitrobenzenes↗

PubMed1978

Hydrogen metabolism in blue-green algae.

This manuscript reviews the literature on hydrogen metabolism in blue-green algae and reports some new data from this laboratory. H2-formation by intact cells is found to be catalyzed exclusively by nitrogenase. Its rate appears to be variable from strain to strain used byt is--in our hands--very small. Therefore, blue-green algae are presumably of limited value in projects of solar energy conversion to form molecular hydrogen. These organisms are also able to consume the gas in a reaction catalysed by hydrogenase. Hydrogen is mainly consumed in an oxygen dependent reaction, as in aerobic nitrogen fixing bacteria. It can also serve as an electron donor for nitrogen fixation under certain physiological conditions. In experiments with a cell-free preparation, hydrogenase is found to be membrane-bound. The enzyme is characterized with respect to its specifity towards electron donors and acceptors.

Aerobiosis↗

PubMedDate not supplied

[Structure and cell division of the blue-green alga Plectonema boryanum].

Our data on the structure of the blue-green alga Plectonema boryanum, as well as the analysis of the data of other authors, show that its cells resemble bacterial cells in the basic features of their fine structure (the cell wall, cytoplasmic membrane, nucleoid, membraneous formations, ribosomal apparatus). Besides the organelles typical of blue-green algae, organelles similar to mesosomes and formations looking like lysosomes were found in Pl. boryanum. Data obtained in the course of the formation of cell partitions evidently speak in favour of the many-sided activity of the lamellae of the photosynthetic apparatus (the parachromatophore) of Pl. boryanum.

Cell Membrane↗

PubMedDate not supplied

Purification and characterization of cytochrome c6 from the unicellular green alga Scenedesmus obliquus.

Purification of a soluble cytochrome c6 from the unicellular green alga Scenedesmus obliquus by a simple and rapid method is described. The purification procedure includes ammonium sulfate precipitation and non-denaturating PAGE. The N-terminal sequence of the first 20 amino acids was determined and shows 85% similarity and 75% identity to the sequence of cytochrome c6 from the green alga Monoraphidium braunii. The ferrocytochrome shows typical UV/VIS absorption peaks at 552.9, 521.9 and 415.7 nm. The apparent molecular mass was estimated to be 12 kDa by SDS-PAGE. EPR-spectroscopy at 20 K shows resonances indicative for two distinct low-spin heme forms.

Amino Acid Sequence↗

PubMed1976

Ion metabolism in a halophilic blue-green alga, Aphanothece halophytica.

The intracellular ion content of the halophilic blue-green alga, Aphanthece halophytica was studied as a function of age, external sodium and external potassium concentration. Intracellular Na+ was found to be about 0.38 millimoles/g dry mass. Intracellular K+ concentrations were as high as 1 M and varied directly with external salinity. Intracellular Ca++ and Mg++ were in the range previously reported for fresh water blue-green algae despite their extremely high extracellular concentrations. Average cell size is consistent at room temperature with two exceptions. When the outside K+ is lower than 6.5 mM the cells tend to be smaller with less intracellular K+ and high Ca++. In stationary phase cultrues the cells are larger with high intracellular Mg++ and low K+.

Calcium↗

PubMed1995

Detection of green algae (Chlorophyceae) for the diagnosis of drowning.

The plankton test (generally, diatom test) is one of the methods available to diagnose the cause of death of submerged bodies. The solubilization method using tissue solubilizer Soluene-350 was used in this study to detect not only diatoms but also green algae, based on the fact that the solubilizer does not digest the cell walls of green algae which are made from cellulose. Detection of green algae from organs of submerged cadavers is very informative to determine drowning in fresh water, and also in cases where only few diatoms are detected in the organs.

Acetone↗

PubMed2007

A clade uniting the green algae Mesostigma viride and Chlorokybus atmophyticus represents the deepest branch of the Streptophyta in chloroplast genome-based phylogenies.

BACKGROUND: The Viridiplantae comprise two major phyla: the Streptophyta, containing the charophycean green algae and all land plants, and the Chlorophyta, containing the remaining green algae. Despite recent progress in unravelling phylogenetic relationships among major green plant lineages, problematic nodes still remain in the green tree of life. One of the major issues concerns the scaly biflagellate Mesostigma viride, which is either regarded as representing the earliest divergence of the Streptophyta or a separate lineage that diverged before the Chlorophyta and Streptophyta. Phylogenies based on chloroplast and mitochondrial genomes support the latter view. Because some green plant lineages are not represented in these phylogenies, sparse taxon sampling has been suspected to yield misleading topologies. Here, we describe the complete chloroplast DNA (cpDNA) sequence of the early-diverging charophycean alga Chlorokybus atmophyticus and present chloroplast genome-based phylogenies with an expanded taxon sampling. RESULTS: The 152,254 bp Chlorokybus cpDNA closely resembles its Mesostigma homologue at the gene content and gene order levels. Using various methods of phylogenetic inference, we analyzed amino acid and nucleotide data sets that were derived from 45 protein-coding genes common to the cpDNAs of 37 green algal/land plant taxa and eight non-green algae. Unexpectedly, all best trees recovered a robust clade uniting Chlorokybus and Mesostigma. In protein trees, this clade was sister to all streptophytes and chlorophytes and this placement received moderate support. In contrast, gene trees provided unequivocal support to the notion that the Mesostigma + Chlorokybus clade represents the earliest-diverging branch of the Streptophyta. Independent analyses of structural data (gene content and/or gene order) and of subsets of amino acid data progressively enriched in slow-evolving sites led us to conclude that the latter topology reflects the true organismal relationships. CONCLUSION: In disclosing a sister relationship between the Mesostigmatales and Chlorokybales, our study resolves the long-standing debate about the nature of the unicellular flagellated ancestors of land plants and alters significantly our concepts regarding the evolution of streptophyte algae. Moreover, in predicting a richer chloroplast gene repertoire than previously inferred for the common ancestor of all streptophytes, our study has contributed to a better understanding of chloroplast genome evolution in the Viridiplantae.

Algal Proteins↗

PubMed1969

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.

Adenosine Triphosphate↗

PubMed2001

Identification of anatoxins in blue-green algae food supplements using liquid chromatography-tandem mass spectrometry.

Blue-green algae (cyanobacteria) in tablets and capsules, which are marketed as health food supplements, were investigated for the presence of neurotoxins related to anatoxin-a. These neurotoxins, which are nicotinic agonists, were investigated using isocratic micro-liquid chromatograph-tandem mass spectrometry (micro-LC-MS-MS). The investigated compounds were anatoxin-a and homoanatoxin-a, together with their degradation products, dihydroanatoxin-a, epoxyanatoxin-a, dihydrohomoanatoxin-a and epoxyhomoanatoxin-a which were synthesized from the parent toxins. The analytes were extracted with methanol followed by isocratic chromatography on a micro C18 reversed-phase column using acetonitrile-water, 50:50 (v/v), containing 20 mm acetic acid at 30 microl min(-1). The toxins were ionized in an ionspray (IS) interface operating in the positive ion mode, where the intact protonated molecules, [M + H]+, were generated at m/z 166, m/z 168, m/z 182, m/z 180, m/z 182 and m/z 196, for anatoxin-a, dihydroanatoxin-a, epoxyanatoxin-a, homoanatoxin-a, dihydrohomoanatoxin-a and epoxyhomoanatoxin-a, respectively. These served as precursor ions for collision-induced-dissociation (CID) and diagnostic product ions for these anatoxins were identified to carry out toxin confirmation by selected reaction monitoring (SRM) LC-MS-MS analysis. Dihydrohomoanatoxin-a and a novel isomer of epoxyanatoxin-a were identified in blue-green algae tablets. This finding suggests that a potential human health hazard could be associated with the consumption of these food supplements.

Chromatography, Liquid↗

PubMed1982

Sequence of the gene coding for the beta-subunit of dinitrogenase from the blue-green alga Anabaena.

The nitrogen fixation nif K gene of the blue-green alga Anabaena, which codes for the beta-subunit of dinitrogenase, has been subjected to sequence analysis. The nif K protein is predicted to be 512 amino acids long, to have a M(r) or 57,583, and to contain six cysteine residues. Three of these cysteines are within peptides homologous to FeS cluster-binding cysteinyl peptides from ferredoxins and from a high potential iron protein and, thus, may be ligands to which FeS clusters bind in dinitrogenase. The sequences surrounding the cysteine residues are 70% homologous to the corresponding cysteinyl tryptic peptides of the Azotobacter vinelandii dinitrogenase, although the positions of the cysteine residues are not always conserved between the two proteins. A 15-amino acid coding sequence precedes nif K on its transcript. Amino acid codon usage is highly asymmetric and parallels that found for the Anabaena dinitrogenase reductase gene (nif H). Putative promoter and ribosome binding site sequences were identified for nif K. These regulatory sequences are homologous to sequences preceding nif D; nif D codes for the alpha-subunit of dinitrogenase but is separated from nif K on the chromosome by 11,000 nucleotides. The nif K promoter also is virtually identical to a promoter-like sequence that immediately precedes the start of the transcript for the large subunit of ribulosebisphosphate carboxylase from maize chloroplasts. This homology appears to support the theory that chloroplasts evolved from blue-green algae.

Journal Article↗

PubMed2006

The complete chloroplast genome sequence of the chlorophycean green alga Scenedesmus obliquus reveals a compact gene organization and a biased distribution of genes on the two DNA strands.

BACKGROUND: The phylum Chlorophyta contains the majority of the green algae and is divided into four classes. While the basal position of the Prasinophyceae is well established, the divergence order of the Ulvophyceae, Trebouxiophyceae and Chlorophyceae (UTC) remains uncertain. The five complete chloroplast DNA (cpDNA) sequences currently available for representatives of these classes display considerable variability in overall structure, gene content, gene density, intron content and gene order. Among these genomes, that of the chlorophycean green alga Chlamydomonas reinhardtii has retained the least ancestral features. The two single-copy regions, which are separated from one another by the large inverted repeat (IR), have similar sizes, rather than unequal sizes, and differ radically in both gene contents and gene organizations relative to the single-copy regions of prasinophyte and ulvophyte cpDNAs. To gain insights into the various changes that underwent the chloroplast genome during the evolution of chlorophycean green algae, we have sequenced the cpDNA of Scenedesmus obliquus, a member of a distinct chlorophycean lineage. RESULTS: The 161,452 bp IR-containing genome of Scenedesmus features single-copy regions of similar sizes, encodes 96 genes, i.e. only two additional genes (infA and rpl12) relative to its Chlamydomonas homologue and contains seven group I and two group II introns. It is clearly more compact than the four UTC algal cpDNAs that have been examined so far, displays the lowest proportion of short repeats among these algae and shows a stronger bias in clustering of genes on the same DNA strand compared to Chlamydomonas cpDNA. Like the latter genome, Scenedesmus cpDNA displays only a few ancestral gene clusters. The two chlorophycean genomes share 11 gene clusters that are not found in previously sequenced trebouxiophyte and ulvophyte cpDNAs as well as a few genes that have an unusual structure; however, their single-copy regions differ considerably in gene content. CONCLUSION: Our results underscore the remarkable plasticity of the chlorophycean chloroplast genome. Owing to this plasticity, only a sketchy portrait could be drawn for the chloroplast genome of the last common ancestor of Scenedesmus and Chlamydomonas.

Base Sequence↗

PubMed1975

Flash kinetics and light intensity dependence of oxygen evolution in the blue-green alga Anacystis nidulans.

Patterns of oxygen evolution in flashing light for the glue-green alga Anacystis nidulans are compared with those for broken spinach chloroplasts and whole cells of the green alga Chlorella pyrenoidosa. The oscillations of oxygen yield with flash number that occur in both Anacystis and Chlorella, display a greater degree of damping than do those of isolated spinach chloroplasts. The increase in damping results from a two- to threefold increase in the fraction (alpha) of reaction centers "missed" by a flash. The increase in alpha cannot be explained by non-saturing flash intensities or by the dark reduction of the oxidized intermediates formed by the flash. Anaerobic conditions markedly increase alpha in Anacystis and Chlorella but have no effect on alpha in broken spinach chloroplasts. The results signify that the mechanism of charge separation and water oxidation involved in all three orgainsms is the same, but that the pool of secondary electron acceptors between Photosystem II and Photosystem I is more reduced in the dark, in the algal cells, than in the isolated spinach chloroplasts. Oxygen evolution in flashing light for Anacystis and Chlorella show light saturation curves for the oxygen yield of the third flash (Y3) that differ markedly from those of the steady-state flashes(YS). In experiments in which all flashes are uniformly attenuated, Y3 requires nearly twice as much light as YS to reach half-saturation. Under these conditions Y3 has a sigmoidal dependence on intensity, while that of YS is hyperbolic. These differences depend on the number of flashes attenuated. When any one of the first three flashes is attenuated, the variation of Y3 with intensity resembles that of YS. When two of the first three flashes are attenuated, Y3 is intermediate in shape between the two extremes. A quantitative interpretation of these results based on the model of Kok et al. (Kik, B., Forbush, B.and McGloin, M. (1970) Photochem. Photobiol. 14, 307-321) fits the experimental data.

Chlorella↗

PubMed1980

Oscillations of the NAD(P)H pool size and of the redox state of a cytochrome b during dark respiration of the blue-green alga, Anacystis nidulans.

Oscillations of the oxygen uptake rate of the blue-green alga (cyanobacterium) Anacystis nidulans were induced by light pulses. The pool size of NAD(P)H and the redox state of a cytochrome b showed oscillations of similar shape and frequency. Phase diagrams revealed that these three oscillations were presumably linked. The cytochrome b should be a part of the respiratory chain of this blue-green alga. The oscillations were inducible only in a limited physiological state of the alga.

Cyanobacteria↗

PubMed1990

The impact of aluminium on green algae isolated from two hydrochemically different headwater streams, Bavaria, Germany.

Two strains of green algae (Scenedesmus sp. and Chlorella sp.) have been isolated from two headwater streams differing in stream chemistry (Eger river: acidic, low alkalinity; Püttlach river: slightly alkaline, high alkalinity). In this study the growth response of these indigenous algae to increased concentrations of aluminium (Al) is investigated. A semi-continuous culture technique was used for Al-toxicity studies. Algal response was determined by calculating growth rates from turbidity and cell counts. Those Al-species which are well known to be toxic were estimated by equilibrium calculations using the WATEQ computer program (Truesdall & Jones, 1973). The pH-value of the culture media was usually pH=5, except for one of the test series. Tested concentrations ranged from c=4 to 220 micromol litre(-1). The isolated strains of green algae were highly sensitive to increased Al-concentrations. The strain isolated from the Püttlach river was more sensitive than the Eger river algae. A total growth inhibition occurs at Al-concentrations of c=4 micromol litre(-1) if the whole Al was added at once. If Al was added gradually into the growth media the response of the algae was delayed. This is due to Al-enrichment in cells. In our long term toxicity studies, growth inhibition occurs even at nearly neutral pH-conditions (pH=6.5) although Al toxicity is expected at pH-values less than pH=5.5. This new result confirms the need of long-term studies in continuous cultures under simulated natural conditions. This might be the only way to achieve valid conclusions about the fate and the toxicity of environmental pollutants.

Journal Article↗

PubMed1990

Reflection confocal laser scanning microscopy of eyespots in flagellated green algae.

The reflection properties of different types of eyespots in three unicellular, flagellated green algae (Tetraselmis chui, Chlamydomonas eugametos, Hafniomonas reticulata) were investigated using confocal laser scanning microscopy in the epireflection mode. The eyespots differed with respect to the number of eyespot lipid globule layers and surface appearance (concave/convex). A strong reflection signal was observed in all eyespots, and a detailed quantitative analysis by optical xy (horizontal) and xz (vertical) sectioning was performed. By applying both sectioning capabilities, multi- and single/double-layered eyespots as well as concave and convex eyespot surfaces could be distinguished using living, immobilized cells. Focusing of the reflected light was only observed in eyespots with concave surfaces. In xz series of multi-layered eyespots at reduced laser intensities (0.01%), the intensity profiles of the reflection revealed a series of alternating maxima and minima with increasing reflection intensities toward the cell surface. At very low laser intensities (0.001%), multi-layered eyespots exhibited about twice the reflection intensity at the presumptive photoreceptor site compared to single/double-layered eyespots. Our results provide the first experimental evidence to support the proposal that multi-layered eyespots act as interference reflectors in photoaxis of green algae.

Chlorophyta↗

PubMed1987

Blue-green algae toxicosis in five dairy cows.

Five Holstein cows developed a sudden clinical syndrome of ataxia, muscle tremors, recumbency, and bloody diarrhea. The pond where these cows obtained water contained a near pure culture of Microcystis aeruginosa, a toxic blue-green algae. All cows affected were treated with activated charcoal, procaine penicillin, glucose, and calcium and magnesium gluconate. All 5 cows were clinically normal ten days later. Many practicing veterinarians regard blue-green algae toxicosis as a rare syndrome that results in rapid death for consuming animals; however, this toxicosis may be common and not lethal. Because no diagnostic test is available for blue-green algae toxicosis, this condition rarely is diagnosed.

Animals↗

PubMed1998

Characterization of novel extrachromosomal DNA from giant-celled marine green algae.

Cloned HinfI fragments of the plasmid-like 2.2-kb DNA from the green alga Ernodesmis verticillata (Kützing) Borgesen hybridized solely to single or double bands within the 2.2-kb DNA in genomic Southern blots. Heterologous probes for nuclear and chloroplast genes hybridized only to high-molecular-weight (HMW) DNA. Thus, the low-molecular-weight (LMW) DNA is extrachromosomal and lacks extensive homology to nuclear or chloroplast genes. There was cross-hybridization to LMW DNA from other Caribbean isolates of E. verticillata, but not to that from a Pacific isolate. Under reduced stringency, cross-hybridization to LMW DNA from the related green alga Boergesenia forbesii (Harvey) Feldmann was also observed, suggesting that the LMW DNA may have a common origin and/or function in these algae. Six out of sixteen unique clones hybridized to discrete bands in northern blots, indicating that the LMW DNA may be actively transcribed in vivo. Four of the putatively transcribed clones have regions with significant deduced amino-acid sequence identity to psa and psb gene products, implying that the plasmid-like molecules might have originated from chloroplast DNA. Sequencing data also indicated a high G/C content, as well as the presence of frequent tandem and direct repeats in many of the cloned fragments. Sequencing and restriction analyses suggest that most of the cloned fragments are portions of different DNA molecules, providing evidence that the 2.2-kb extrachromosomal DNA in Ernodesmis is novel in that it represents a fairly heterogeneous population of molecules.

Amino Acid Sequence↗

PubMed1993

Inhibition of reverse transcriptase activity by extracts of cultured blue-green algae (cyanophyta).

Lipophilic and hydrophilic extracts of over 900 strains of cultured blue-green algae (cyanophyta) were examined in vitro for their ability to inhibit the reverse transcriptases (RT) of avian myeloblastosis virus (AMV) and human immunodeficiency virus, type 1 (HIV-1). Eighteen (2.0%) aqueous extracts showed activity against AMV and HIV RTs. The maximal level of RT inhibition achieved by some of the active extracts was equivalent to that measured for 3'-azido-2',3'-di-deoxythymidine (AZT) at 668 ng/ml. Examination of partially purified fractions prepared by C18 column chromatography demonstrated that the RT inhibition observed could not be attributed entirely to the degradation of transcript DNA, template RNA, or enzyme protein in the reaction mixture. Thus, these results indicate that cultured blue-green algae may represent a novel source of compounds that inhibit RT activity, including that of HIV-1.

Antiviral Agents↗