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Temperature shift-induced responses in lipids in the blue-green alga, Anabaena variabilis: the central role of diacylmonogalactosylglycerol in thermo-adaptation.

Changes in fatty acids and lipid molecular species after shift of growth temperature were studied in the blue-green alga, Anabaena variabilis. In the first 10 h after a temperature shift from 38 to 22 degrees C, lipid synthesis was markedly suppressed. During this period most of the palmitic acid of the diacylmonogalactosylglycerol was desaturated to palmitoleic acid. Thereafter lipid synthesis resumed, and, in the following hours, the relative contents of palmitic and palmitoleic acids were almost restored to original levels. On the other hand, the oleic and linoleic acids were almost restored to original levels. On the other hand, the oleic and inoleic acids were desaturated to alpha-linolenic acid in all the lipid classes. The desaturation reaction of the C18 acids were slower than those of the C16 acids. In the first 5 h after a growth-temperature shift from 22 to 38 degrees C, lipid synthesis was considerably stimulated. During this period, the relative content of palmitic acid increased and that of palmitoleic acid decreased in the diacylmonogalactosylglycerol, and in the following 20 h, they were restored. The oleic and linoleic acids increased with a concomitant decrease in alpha-linolenic acid in all the lipid classes. The decreases in unsaturation in the C16 and C18 acids were due to the stimulated synthesis of more saturated fatty acids. Among the major molecular species of the lipids a particular change was seen in 1-oleoyl-2-palmitoylmonogalactosyl-sn-glycerol. This species rapidly decreased after the downward temperature shift and rapidly increased after the upward temperature shift. From the viewpoint of thermo-adaptive regulation of membrane fluidity, the central role played by the diacylmonogalactosylglycerol in temperature acclimation is discussed.

Adaptation, Physiological↗

RNA synthesis and the accumulation of guanine nucleotides during growth shift down in the blue-green alga Anacystis nidulans.

Shift down of growth rate in the blue-green alga Anacystis nidulans by reduction of the incident light intensity produced a reduction in the rate of stable RNA accumulation which was correlated with increased concentrations of three phosphorylated compounds, two of which were identified as guanosine 5'-diphosphate-3'-diphosphate (ppGpp) and guanosine 5'-triphosphate3'-diphosphate (pppGpp). The step also causes a large but transient increase in the concentration of GTP. Stable RNA fails to accumulate for a considerable length of time after the concentrations of ppGpp and pppGpp have fallen, suggesting the involvement of another mechanism in the control of stable RNA accumulation.

Cyanobacteria↗

DNA sequence organization in the alga Euglena gracilis.

The sequence organization of nuclear DNA in the single-celled alga Euglena gracilis has been studied by a combination of techniques: (1) the comparison of the reassociation kinetics of DNA fragments 300, 2000 and 8100 nucleotides long; (2) the reassociation of 32P-labeled DNA fragments of various lengths with driver fragments 300 nucleotides long; (3) the hyperchromicity of DNA structures formed by the reassociation of repetitive sequences; (4) and the direct measurement of the size of the duplex regions of reassociated repetitive DNA resistant to S1 nuclease. The single copy DNA sequences are approximately 1500 nucleotide pairs long and are interspersed with repetitive DNA sequences. The repetitive DNA, consisting of both highly repetitive and middle repetitive sequences, consists of one fraction of nucleotide sequences (0.67) with an average size of 4900 nucleotide pairs and a second fraction (0.33) with an average size of 1000 nucleotide pairs, 34% of the DNA consists of foldback sequences which are present on 45% of the DNA 4000 nucleotides long.

Base Sequence↗

Purification and characterization of an agglutinin in the red alga Agardhiella tenera.

The red alga, Agardhiella tenera was found to contain a glycoprotein which agglutinates mouse leukemia cells, L5178Y but not L1210. It also agglutinates guinea pig and rabbit erythrocytes, and has weak activity against human A, B and O, mouse, horse and sheep erythrocytes and hamster and mouse lymphocytes. The agglutination was not inhibited by simple sugars. The major active component was purified and determined to be a beta-structure protein containing 2.7% glucose as sugar moiety. The molecular weight was estimated to be 12,000 by gel filtration and 13,000 by polyacrylamide gel electrophoresis, in the presence of sodium dodecylsulfate. Its isoelectric point was 6.1, and it contained high amounts of glycine, serine and threonine, but no half cystine or histidine. It had no subunit structure, and the C- and N-terminal amino acids were threonine and arginine, respectively.

ABO Blood-Group System↗

Antiferromagnetic exchange interaction in the two-iron-two-sulphur ferredoxin from the blue-green alga Spirulina maxima studied with a highly sensitive magnetic balance.

1. A highly sensitive magnetic balance of the Faraday type is described. 2. The magnetic susceptibility of the oxidized and reduced forms of the two-iron-two-sulphur ferredoxin from the blue-green alga Spirulina maxima has been measured over a wide temperature range. 3. The results can be interpreted within a simple model involving antiferromagnetically coupled iron atoms at the active site. The coupling, expressed as --J, is estimated to be 182 +/- 20/cm and 98 +5/-10 /cm for the oxidized and reduced forms, respectively.

Binding Sites↗

Electrophoretic study of histones in the unicellular alga Olisthodiscus luteus.

Basic proteins were prepared from isolated nuclei of the unicellular alga Olisthodiscus luteus. The ratio of DNA/RNA/basic protein for these nuclei was 1:0.17:1.1, respectively, and the amino acid composition of the basic protein was very similar to that of Euglena and calf histones. The Olisthodiscus basic proteins were separated into four major components by polyacrylamide gel electrophoresis in four gel systems: (1) low pH disc gels containing 2.5 M urea; (2) two-dimensional low pH-urea gels in which the second dimension contained 1% Triton X-100; (3) slab gels containing 0.1% sodium dodecyl sulfate (SDS); (4) two-dimensional gels combining systems (1) and (3). The presence of four rather than five major histone fractions was shown to be not merely the result of proteolytic degradation. Results from the urea-containing gels suggest that an H1-like histone is missing, while electrophoresis in the SDS-containing gels shows the presence of a component resembling H1. In view of recent reports documenting the presence of five major histones in lower eukaryotes as well as in higher organisms, the presence of only four histones in Olisthodiscus suggests that this primitive eukaryote is unusual in its histone complement.

Amino Acids↗

Depression of nitrate reductase in the presence of excess ammonium in a unicellular alga growing under conditions of phosphate limitation.

Chemostat cultures of the unicellular alga Cyanidium caldarium have shown that under conditions of phosphate limitation nitrate reductase is completely derepressed even in cells growing in a large excess of ammonium, but that it occurs mainly in a catalytically inactive form. It is hypothesized that phosphate limitation contributes to maintaining intracellular level of glutamine suitable to stimulate inactivation but not repression of nitrate reductase. It is not excluded that in addition to variations in the intracellular level of glutamine, there are other metabolic events of the cell by which repression and inactivation of nitrate reductase could be differently influenced.

Enzyme Activation↗

Identification of the 11,12-dihydro-11,12-dihydroxybenzo(a)pyrene as a major metabolite produced by the green alga, Selenastrum capricornutum.

Benzo(a)pyrene metabolites were isolated after incubation of [14C]-benzo(a)pyrene with the green alga, Selenastrum capricornutum. A significant amount of radioactivity chromatographed in the dihydrodiol region which did not coelute with any of the previously identified dihydrodiol metabolites isolated from this system. Following characterization by mass spectrometry, fluorescence spectroscopy, and high pressure liquid chromatography, this metabolite was identified as the cis-11,12-dihydro-11,12-dihydroxybenzo(a)pyrene. This metabolite has not been identified previously as a metabolite formed in a plant system.

Benzo(a)pyrene↗

Resonance Raman spectra of phycocyanin, allophycocyanin and phycobilisomes from blue-green alga Anacystis nidulans.

Resonance Raman spectra of native C-phycocyanin, allophycocyanin and whole, intact phycobilisomes from the blue-green alga Anacystis nidulans (Synechococcus 6301) are reported. A tentative assignment for the more prominent resonance Raman bands is suggested. The possibly sensitive regions for inter-chromophore interactions in the case of phycobilisomes are also discussed.

Cyanobacteria↗

Metabolism of 6,9,12-octadecatrienoic acid in the red alga Lithothamnion corallioides: mechanism of formation of a conjugated tetraene fatty acid.

Incubation of [1-14C]6(Z),9(Z),12(Z)-octadecatrienoic acid with an enzyme preparation from the red alga Lithothamnion corallioides Crouan led to the formation of two new compounds, i.e. the conjugated tetraene 6(Z),8(E),10(E),12(Z)-octadecatetraenoic acid and the bis-allylic hydroxy acid 11(R)-hydroxy-6(Z),9(Z),12(Z)-octadecatrienoic acid. These two compounds were formed by independent pathways and were not interconvertible by the enzyme preparation. Experiments with stereospecifically deuteriated 6,9,12-octadecatrienoic acids demonstrated that formation of 6,8,10,12-octadecatetraenoic acid was accompanied by loss of the pro-S and pro-R hydrogens from C-8 and C-11, respectively, whereas formation of 11-hydroxy-6,9,12-octadecatrienoic acid proceeded with loss of the pro-S hydrogen from C-11. Biosynthesis of 6,8,10,12-octadecatetraenoic acid was dioxygen-dependent and was accompanied by production of hydrogen peroxide. A number of artificial electron acceptors supported formation of 6,8,10,12-octadecatetraenoic acid under anaerobic conditions. The existence in Lithothamnion corallioides of a fatty acid oxidase that catalyzes the oxidation of certain poly-unsaturated fatty acids into conjugated tetraene fatty acids is postulated.

Fatty Acids, Unsaturated↗

Chemical methods for the analysis of sulphated galactans from red algae.

Methods are reported that facilitate the structural characterization of complex sulphated galactans of the red algae. Two procedures have been developed for the production of alditol acetates from carrageenans and agaroids. Both procedures generate 3,6-anhydrogalactitol acetate from the easily destroyed 3,6-anhydrogalactosyl residues in near quantitative yield. The "double hydrolysis-reduction" method involves preliminary hydrolysis under conditions sufficient to cleave all of the 3,6-anhydrogalactosidic bonds, but mild enough to avoid significant further degradation. The "reductive hydrolysis" method uses the acid-stable 4-methylmorpholine-borane to reduce the 3,6-anhydrogalactose end groups as they are released during acid hydrolysis. An alditol acetate sample can be prepared from a polysaccharide in a single tube, ready for g.l.c. analysis, in less than 2.5 h, i.e. more quickly than by any previous procedure. Problems associated with incomplete methylation of sulphated carrageenans and agaroids by the Hakomori procedure have been overcome by first converting the sulphated polysaccharide into its triethylammonium salt form. The reductive hydrolysis method is effective for the production of partially methylated alditol acetates from the methylated polysaccharides, enabling the rapid determination of the substitution pattern of these polysaccharides. These improved analytical methods have been applied successfully to kappa-, iota-, and lambda-carrageenans, as well as some agars.

Acetates↗

Studies on the post-coital contraceptive mechanisms of crude extract of Sri Lankan marine red algae, Gelidiella acerosa.

This study investigates the potential post-coital contraceptive mechanisms of the crude extract of Sri Lankan marine red algae, Gelidiella acerosa, using rats. The dose used was 1000 mg/kg/day and the route of administration was oral. The results showed that the crude extract possessed potent post-coital contraceptive activity when administered on days 7-8 of pregnancy indicating the presence of a narrow window in its action. The post-coital contraceptive activity was due to an elevated post-implantation loss (by 89%) resulting from fetal death between days 9-14 of pregnancy. The extract was neither estrogenic nor anti-estrogenic nor stressor but appeared to be anti-progestational (reduced ovarian progesterone output). It is suggested that the crude extract may reduce the ovarian progesterone release possibly via anti-platelet and PGE2-depressing activity.

Animals↗

The effects of the ionophore A23187 on pattern formation in the alga Micrasterias.

We have used the divalent cation ionophore A23187 to investigate the hypothesis that cytoplasmic localization of Ca2+ is responsible for localized growth in the alga Micrasterias. In a preliminary study we found that, of the major salts contained in the cell's medium, only CaCl2 was needed for normal development. In cells developing in the presence of A23187 and extracellular Ca2+, we postulated that the ionophore would induce a spatially uniform influx of Ca2+ that would overwhelm endogenous Ca2+ gradients. When developing cells were treated with A23187 and 2 mM CaCl2, we observed a delocalization of the cell's normal pattern of wall deposition. This effect was less pronounced when cells were exposed to A23187 and 2 mM MgCl2. These results support the hypothesis that localized regions of high Ca2+ concentration normally mediate localized expansion of tip-growing lobes in Micrasterias.

Anti-Bacterial Agents↗

The fast block against polyspermy in fucoid algae is an electrical block.

Fertilization potentials in Pelvetia fastigiata, Fucus vesiculosus, and Fucus ceranoides were studied to examine whether eggs of fucoid algae have an electrical block against polyspermy. The resting potential of eggs of all species was about -60 mV, depolarizing, respectively, to -24 +/- 5 mV (SD, n = 9) for 7.5 +/- 2.1 (n = 8) min, -26 +/- 5 (n = 9) mV for 6.4 +/- 2.3 (n = 9) min, and -24 +/- 6 (n = 5) mV for 6.7 +/- 1.9 (n = 4) min. The depolarization was slower, and the fertilization potential was about 10 mV more negative in eggs of both F. vesiculosus and Pelvetia fertilized in 45-mM Na+ ASW; many of these eggs were polyspermic. Steady current was passed through unfertilized eggs of F. vesiculosus prior to insemination to test the potential dependence of fertilization. Eggs (n = 10) bound sperm at all potentials tested (-45 to -23 mV), but fertilization was prevented if eggs were held at potentials more positive than -45 to -37 mV. Eggs underwent a second depolarization if artificially hyperpolarized to potentials more negative than -50 mV immediately after the rise of a normal fertilization potential. Thus, fucoid eggs have an electrical fast block against polyspermy. Only in F. ceranoides does the formation of the cell wall after fertilization appear to be fast enough (i.e., 3-6 min postfertilization versus at 10-15 min in F. vesiculosus and P. fastigiata) to replace the fertilization potential as a polyspermy block. Nonfertilizing fucoid sperm swim away from the egg surface by 1-3 min after rise of the fertilization potential. This suggests that there is another "intermediate block" against polyspermy.

Fertilization↗

The phototoxicity of benzo[a]pyrene in the green alga Selenastrum capricornutum.

The effects of selected polycyclic aromatic hydrocarbons (PAHs) on the growth of the green alga Selenastrum capricornutum in three light regimens were examined. In gold fluorescent light, benzo[a]pyrene (BaP) at 12 mg/liter (48 mumole/liter), benz[a]anthracene (BaA) at 40 mg/liter (175 mumole/liter), anthracene (A) at 40 mg/liter (224 mumole/liter), and 13 metabolites of BaP each at 40 micrograms/liter had no effect on algal growth. In cool-white fluorescent light, 30% inhibition of algal growth occurred with 0.1 mumole/liter BaP, 8.0 mumole/liter BaA, and 40 mumole/liter A. BaP at 0.16 mg/liter (0.64 mumole/liter) totally inhibited growth. BaP concentrations an order of magnitude lower inhibited algal growth in fluorescent blacklight. In cool-white light, 5 of 13 metabolites of BaP (each 40 micrograms/liter) inhibited algal growth: 3,6-quinone; 6-hydroxy; 9-hydroxy; 3-hydroxy; and 1,6-quinone. Based on these results, PAHs and metabolites of BaP are selectively phototoxic to S. capricornutum due to the incident light intensity below 550 nm.

Anthracenes↗

Chloroplast transit peptides from the green alga Chlamydomonas reinhardtii share features with both mitochondrial and higher plant chloroplast presequences.

Chloroplast transit peptides from the green alga Chlamydomonas reinhardtii have been analyzed and compared with chloroplast transit peptides from higher plants and mitochondrial targeting peptides from yeast, Neurospora and higher eukaryotes. In terms of length and amino acid composition, chloroplast transit peptides from C. reinhardtii are more similar to mitochondrial targetting peptides than to chloroplast transit peptides from higher plants. They also contain the potential amphiphilic alpha-helix characteristic of mitochondrial presequences. However, in similarity with chloroplast transit peptides from higher plants, they contain a C-terminal region with the potential to form an amphiphilic beta-strand. As in higher plants, transit peptides that route proteins to the thylakoid lumen consist of an N-terminal domain similar to stroma-targeting transit peptides attached to a C-terminal apolar domain that share many characteristics with secretory signal peptides.

Amino Acid Sequence↗

Light-harvesting complexes of brown algae. Biochemical characterization and immunological relationships.

The pigment composition of the light-harvesting complexes isolated from several brown algae belonging to different orders has been analysed by reverse-phase HPLC. Relative to whole chloroplasts, they were markedly enriched in Chl c, fucoxanthin and violaxanthin and conversely depleted in Chl a. The relative molar proportions of the 4 main pigments (Chl a/Chl c/fucoxanthin/violaxanthin) ranged from 100:18:76:6 to 100:30:107:17. The protein moiety of LH complexes of all the species studied were composed of one or two main polypeptide components in the range of 19-22 kDa. These polypeptide subunits were arranged in polymeric particles about 240 kDa in Laminaria saccharina. A polyclonal antibody raised against the LH polypeptide of Fucus serratus has been tested on LH apoproteins of other Chromophytes and Chlorophytes. Phylogenic implications of these results are discussed.

Amino Acids↗

Identification of 11-cis and all-trans-retinal in the photoreceptive organelle of a flagellate green alga.

Isolation of intact photoreceptive organelles (eyespot apparatuses) involved in blue-light mediated photoresponses in a flagellate green alga (Spermatozopsis similis) allowed for the first time the identification of both 11-cis- and all-trans-retinal in a plant cell. Both isomers were identified by HPLC analysis in conjunction with UV spectra. Additionally, reconstitution of a distinct absorption band, centered around 540 nm, was achieved by addition of exogenous 9-cis-retinal to bleached, isolated eyespot apparatuses.

Carrier Proteins↗