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Characterization of chlorophyll-protein complexes isolated from two marine green algae, Bryopsis maxima and Ulva pertusa, growing in the intertidal zone.

Three Chl-protein complexes were isolated from thylakoid membranes of Bryopsis maxima and Ulva pertusa, marine green algae that inhabit the intertidal zone of the Pacific Ocean off the eastern coast of Japan by dodecyl-beta-D-maltoside polyacrylamide gel electrophoresis. The slowest-moving fractions showed low Chl a/b and Chl/P-700 ratios, indicating that this fraction corresponds to complexes in PS I, which is large in both algae. The intermediate and fastest-moving fractions showed the traits of PS II complexes, with some associated Chl a/b-protein complexes and LHC II, respectively. The spectral properties of the separated Chl-proteins were also determined. The absorption spectra showed a shallow shoulder at 540 nm derived from siphonaxanthin in Bryopsis maxima, but not in Ulva pertusa. The 77 K emission spectra showed a single peak in Bryopsis maxima and two peaks in Ulva pertusa. Besides the excitation spectra indicated that the excitation energy transfer to the PS I complexes differed quite a lot higher plants. This suggested that the mechanisms of energy transfer in both of these algae differ from those of higher plants. Considering the light environment of this coastal area, the large size of the antennae of PS I complexes implies that the antennae are arranged so as to balance light absorption between the two photosystems. In addition, we discuss the relationships among the photosystem stoichiometry, the energy transfer, and the distribution between the two photosystems.

Chlorophyta↗

The Hydrobia ulvae-Maritrema subdolum association: cercarial emergence controlled by host activity.

The release of Maritrema subdolum cercariae (Digenea: Microphallidae) from the marine mud snail Hydrobia ulvae is significantly affected by temperature, salinity, light and exudates from the second intermediate amphipod host. Based on (i) previously published data on temperature-salinity dependent H. ulvae activity, (ii) new experimental data on H. ulvae activity in light and darkness as well as in the presence and absence of host exudates, and (iii) the cercarial emergence rate from free moving snails and snails prevented from crawling, the present analysis indicates that emergence of M. subdolum larvae is regulated mainly by host activity as the ultimate factor for release. The adaptive significance of such an emergence strategy is emphasized.

Adaptation, Physiological↗

Effects of solar UV-B radiation on canopy structure of Ulva communities from southern Spain.

Within the sheltered creeks of Cádiz bay, Ulva thalli form extended mat-like canopies. The effect of solar ultraviolet radiation on photosynthetic activity, the composition of photosynthetic and xanthophyll cycle pigments, and the amount of RubisCO, chaperonin 60 (CPN 60), and the induction of DNA damage in Ulva aff. rotundata Bliding from southern Spain was assessed in the field. Samples collected from the natural community were covered by screening filters, generating different radiation conditions. During daily cycles, individual thalli showed photoinhibitory effects of the natural solar radiation. This inhibition was even more pronounced in samples only exposed to photosynthetically active radiation (PAR). Strongly increased heat dissipation in these samples indicated the activity of regulatory mechanisms involved in dynamic photoinhibition. Adverse effects of UV-B radiation on photosynthesis were only observed in combination with high levels of PAR, indicating the synergistic effects of the two wavelength ranges. In samples exposed either to PAR+UV-A or to UV-B+UV-A without PAR, no inhibition of photosynthetic quantum yield was found in the course of the day. At the natural site, the top layer of the mat-like canopies is generally completely bleached. Artificially designed Ulva canopies exhibited fast bleaching of the top layer under the natural solar radiation conditions, while this was not observed in canopies either shielded from UV or from PAR. The bleached first layer of the canopies acts as a selective UV-B filter, and thus prevents subcanopy thalli from exposure to harmful radiation. This was confirmed by the differences in photosynthetic activity, pigment composition, and the concentration of RubisCO in thalli with different positions within the canopy. In addition, the induction of the stress protein CPN 60 under UV exposure and the low accumulation of DNA damage indicate the presence of physiological protection mechanisms against harmful UV-B. A mechanism of UV-B-induced inhibition of photosynthesis under field conditions is proposed.

Spain↗

The nucleotide sequences of 5S rRNAs from a multicellular green alga, Ulva pertusa, and two brown algae, Eisenia bicyclis and Sargassum fulvellum.

The nucleotide sequences of 5S rRNA from a multicellular green alga Ulva pertusa, and multicellular brown algae Eisenia bicyclis and Sargassum fulvellum, have been determined. The 5S rRNA from Ulva is composed of 120 nucleotides, and those from Eisenia and Sargassum have 118 nucleotides. The nucleotide sequence of Ulva 5S rRNA is rather similar to 5S rRNAs from unicellular green algae and higher plants, while those of Eisenia and Sargassum 5S rRNAs are unique.

Animals↗

Molecular phylogenetic evidence for a reversible morphogenetic switch controlling the gross morphology of two common genera of green seaweeds, Ulva and Enteromorpha.

Ulva and Enteromorpha are two of the most common, ubiquitous, and environmentally important genera of green seaweeds. They are widely regarded as easily distinguishable because of their dramatically different morphologies: Ulva species are flat, lettucelike blades two cell layers thick, and Enteromorpha species form hollow liquid- or gas-filled tubes one cell thick, which may also be highly branched. We present molecular phylogenetic analyses of nuclear ribosomal RNA ITS sequences from 39 samples representing 21 purported species within these two genera. The results clearly indicate that the two genera are not respectively monophyletic and that the characteristic Ulva and Enteromorpha morphologies have arisen independently several times throughout the evolutionary diversification of the group. The analyses demonstrate that this radical change in gross morphology can also happen within clades exhibiting sequence divergence typical of conspecific assemblages of this group. We suggest that this morphological flexibility is the result of some form of developmental switch that results in either blades or tubes, but that this putative switch must be activated relatively infrequently, since there is evidence that some lineages have retained their form for significant periods. This discovery suggests a possible new model system for study of the molecular mechanisms involved in the interplay between environmental stimuli and plant development.

Chlorophyta↗

Photosynthesis of Ulva sp: III. O(2) Effects, Carboxylase Activities, and the CO(2) Incorporation Pattern.

Ulva, a common green seaweed, performs at the biochemical level as a typical C(3) plant. Over 90% of label was found in glycerate 3-phosphate following a 3 second (14)C pulse in the light, and the label was subsequently transferred to sugars. Also, the level of ribulose-1,5-bisphosphate carboxylase activity in crude extracts was about 10 times higher than that of phosphoenolpyruvate carboxylase. Concerning gas exchange, photosynthetic rates of Ulva showed no O(2) sensitivity, indicating that photorespiratory CO(2) losses are repressed as in C(4) plants. This apparent anomaly could be explained by the efficient HCO(3) (-) uptake system of Ulva which might concentrate CO(2) to the chloroplasts, thus suppressing the oxygenase activity of ribulose-1,5-bisphosphate carboxylase.

Journal Article↗

Biosorption of mercury from aqueous solution by Ulva lactuca biomass.

The mercury biosorption onto non-living protonated biomass of Ulva lactuca, as an alternative method for mercury removal from aqueous solutions, was investigated. Batch equilibrium tests showed that at pH 3.5, 5.5 and 7 the maxima of mercury uptake values, according to Langmuir adsorption isotherm, were 27.24, 84.74 and 149.25 mg/g, respectively. The ability of Ulva lactuca biomass to adsorb mercury in fixed-bed column, was investigated as well. The influence of column bed height, flow rate and effluent initial concentration of metal was studied. The adsorbed metal ions were easily desorbed from the algal biomass with 0.3 N H(2)SO(4) solution. After acid desorption and regeneration with distilled water, the biomass could be reused for other biosorption assays with similar performances.

Adsorption↗

Metabolic factors influencing the sodium and potassium distribution in Ulva lactuca.

1. Methods for the use of the marine green alga, Ulva lactuca, in studies on electrolyte metabolism are described. 2. The effect of illumination and iodoacetate on the potassium and sodium content, as well as the influence of light and running sea water on the iodoacetate effect was investigated. The rate of exchange of cellular potassium ion for K(42) under conditions of light and dark at 20 and 30 degrees C. was studied. 3. Ulva maintained in the dark for long periods loses some potassium and gains sodium, both effects being reversed upon illumination. The presence of 0.001 M iodoacetate in the dark causes a marked progressive loss of potassium and gain of sodium, phenomena which do not occur when the alga is illuminated. Evidence for the penetration of the inhibitor into the cell in the presence of light is presented. The iodoacetate effect on potassium and sodium content, once established, can be "washed out" of the alga when the plant is placed in light and running sea water without the inhibitor. Illumination and increased temperature each favor a more rapid exchange of tissue for environmental potassium ion. 4. In the interpretation of these findings it is emphasized that metabolic work, perhaps in the form of ion transports, must be done by the cell to compensate for the continual flow of potassium ion and sodium ion with their respective concentration gradients and thus maintain homeostasis within the cell. Evidence is presented which indicates separate mechanisms for the distribution of sodium and potassium in this organism. It is further suggested that the degradation of phosphoglyceric acid, an important glycolytic and photosynthetic intermediate, or one of the products of its metabolism supplied the energy for these ion transports(s). The role of permeability per se is considered.

Eukaryota↗

Antihepatotoxic nature of Ulva reticulata (Chlorophyceae) on acetaminophen-induced hepatoxicity in experimental rats.

Ulva reticulata, a marine edible green alga, is a known source of proteins, vitamins, and sulfated polysaccharides. Though there are many reports in the literature regarding the composition and antiviral property of Ulva sp., studies of the antihepatotoxic property of green seaweeds in animal model are scarce. We have studied the antihepatotoxic nature of this marine green edible alga, U. reticulata, in a hot water extract (150 mg/kg of body weight for a period of 15 days) against acetaminophen- induced hepatotoxicity in experimental albino rats. The acetaminophen-induced rats showed significant elevation in levels of the serum marker enzymes aspartate transaminase and alanine transaminase and of lipid peroxides in liver tissue with decreased levels of antioxidant enzymes such as superoxide dismutase and catalase. The levels of reduced glutathione and vitamins (E and C) were also decreased in the liver tissue of acetaminophen-intoxicated rats. The oral pretreatment with a hot water extract of U. reticulata reduced the hepatotoxicity triggered by acetaminophen considerably by improving the antioxidant status in experimental animals with depleted levels of lipid peroxides. These results indicate that the oral pretreatment with a hot water extract of U. reticulata in rats is effective in reducing the hepatic oxidative stress via free radical scavenging properties, suggesting an antihepatotoxic activity.

Acetaminophen↗

Molecular characterization of a new lectin from the marine alga Ulva pertusa.

A new lectin, named UPL1, was purified from a green alga Ulva pertusa by an affinity chromatography on the bovine-thyroglobulin-Sepharose 4B column. The molecular mass of the algal lectin was about 23 kD by SDS-PAGE, and it specifically agglutinated rabbit erythrocytes. The hemagglutinating activity for rabbit erythrocytes could be inhibited by bovine thyroglobulin and N-acetyl-D-glucosamine. The lectin UPL1 required divalent cations for maintenance of its biological activity, and was heat-stable, and had higher activity within pH 6-8. The N-terminal amino acid sequence of the purified lectin was determined (P83209) and a set of degenerate primers were designed. The full-length cDNA of the lectin was cloned by rapid amplification of cDNA ends (RACE) method (AY433960). Sequence analysis of upl1 indicated it was 1084 bp long, and encoded a premature protein of 203 amino acids. The N-terminal sequence of the mature UPL1 polypeptide started at amino acid 54 of the deduced sequence from the cDNA, indicating 53 amino acids lost due to posttranslational modification. The primary structure of the Ulva pertusa lectin did not show amino acid sequence similarity with known plant and animal lectins. Hence, this protein may be the paradigm of a novel lectin family.

Acetylglucosamine↗

Disruption of quorum sensing in seawater abolishes attraction of zoospores of the green alga Ulva to bacterial biofilms.

Zoospores of the eukaryotic green seaweed Ulva respond to bacterial N-acylhomoserine lactone (AHL) quorum sensing signal molecules for the selection of surface sites for permanent attachment. In this study we have investigated the production and destruction of AHLs in biofilms of the AHL-producing marine bacterium, Vibrio anguillarum and their stability in seawater. While wild type V. anguillarum NB10 was a strong attractor of zoospores, inactivation of AHL production in this strain by either expressing the recombinant Bacillus lactonase coding gene aiiA, or by mutating the AHL biosynthetic genes, resulted in the abolition of zoospore attraction. In seawater, with a pH of 8.2, the degradation of AHL molecules was temperature-dependent, indicating that the AHLs produced by marine bacterial biofilms have short half-lives. The Ulva zoospores sensed a range of different AHL molecules and in particular more zoospores settled on surfaces releasing AHLs with longer (>six carbons) N-linked acyl chains. However, this finding is likely to be influenced by the differential diffusion rates of AHLs from the experimental surface matrix. Molecules with longer N-acyl chains, such as N-(3-oxodecanoyl)- L-homoserine lactone, diffused more slowly than those with shorter N-acyl chains such as N-(3-hydroxy-hexanoyl)- L-homoserine lactone. Image analysis using GFP-tagged V. anguillarum biofilms revealed that spores settle directly on bacterial cells and in particular on microcolonies which we show are sites of concentrated AHL production.

4-Butyrolactone↗

Functional sulfur amino acid production and seawater remediation system by sterile Ulva sp. (Chlorophyta).

Sterile Ulva, which is a macroalga, has the potential to grow stably; therefore, this seaweed is expected to be an efficient resource of functional food containing various nutrients such as sulfur amino acids, proteins, carbohydrates, and minerals. Ulva latuca was selected from the "Marine Park" in Tokyo Bay, and its growth rate (g-dry/[m2.d]) was measured using model reactors located on the land or on the surface of the sea at Yokohama. The growth rate of U. lactuca was recorded to be approx 20 g-dry/(m2.d), which is estimated to be 10 times greater than that in a natural field in the Marine Park. In addition, this growth rate was higher than that of conventional crops such as corn and rice on a farm or paddy. These data led us to newly design and propose a floating type of labor-efficient U. lactuca production system. d-Cysteinolic acid, which is included in U. lactuca as a major sulfur amino acid, inhibited the Fenton reaction, resulting in suppression of hydroxyl radical production and singlet oxygen. Addition of the sulfur amino acid (1 microM) to HepG2 cells markedly decreased the intracellular triglyceride level. Hence, this proposed facility also has the potential for industrial production of a valuable resource for the primary prevention of lifestyle-related diseases using enriched or eutrophied seawater.

Amino Acids, Sulfur↗

Solar ultraviolet radiation affects the activity of ribulose-1,5-bisphosphate carboxylase-oxygenase and the composition of photosynthetic and xanthophyll cycle pigments in the intertidal green alga Ulva lactuca L.

The effect of solar UV radiation on the physiology of the intertidal green macroalga Ulva lactuca L. was investigated. A natural Ulva community at the shore of Helgoland was covered with screening foils, excluding UV-B or UV-B + UV-A from the solar spectrum. In the sampled material, changes in the activity and concentration of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco), and the concentration of photosynthetic and xanthophyll cycle pigments were determined. Exclusion of UV radiation from the natural solar spectrum resulted in an elevated overall activity of Rubisco, related to an increase in its cellular concentration. Among the photosynthetic pigments, lutein concentration was substantially elevated under UV exclusion. In addition, marked UV effects on the xanthophyll cycle were found: exclusion of solar UV radiation (and particularly UV-B) resulted in an increased ratio of zeaxanthin concentration to the total xanthophyll content, indicating adverse effects of UV-B on the efficiency of photoprotection under high irradiances of photosynthetically active radiation. The results confirm a marked impact of present UV-B levels on macroalgal physiology under field conditions.

Chlorophyll↗

Slow 514 nm absorption phases and oxygen exchange transients in Ulva.

1. The slow 514-nm spectral changes in Ulva were studied using bright continuous 650-nm light. Transient and steady-state absorption changes were compared with changes in net rate of O2 exchange in a system designed to measure both parameters simultaneously. 2. Time courses of the 514-nm absorption change show three phases following the onset of light: one rapid increase and two slower (larger than or equal to 1 s) transient increases. Upon cessation of the light three transient absorption phases also follow: a rapid decrease and two slower (greater than 1 s) transient increases. Parallel transient phases (but opposite in sign) were found at 480 nm. 3. The kinetics of the slow 514-nm absorption transients correlate with the characteristic induction transients in net O2 exchange. 4. Similar difference spectra and the restoration kinetics of the light-on and light -off transient phases indicate that the slow 514-nm absorption changes reflect the same component(s) and process(es). 5. The experimental results are discussed in terms of the electrochromic hypothesis for the 515-nm absorption shift. We interpret the slow 514-nm absorption changes in Ulva as a reflection of relatively slow ionic readjustments across the photosynthetic membranes.

Chlorophyta↗

H blood group detection by the L-fucose binding lectin of the green marine alga Ulva lactuca.

Extracts of the green marine alga Ulva lactuca collected along the seashore of Tel-Aviv exhibit hemagglutinating activity towards papain-treated human erythrocytes. This hemagglutinating activity was shown to be inhibited by L-fucose and EDTA, and to be relatively resistant to heating at 60 degrees C, while sensitive to low pH. Like the lectin of Ulex europeus, the Ulva lectin exhibits blood group H specificity. It agglutinates most strongly erythrocytes of blood group 0(H) followed by B greater than A greater than AB. A2 and A2B erythrocytes are agglutinated by it considerably more strongly than A1 and A1B respectively. Bombay 0(hh) type erythrocytes are almost non-reactive. The lectin can be stored at -20 degrees C for years.

ABO Blood-Group System↗

Measurement of ingestion rate of Hydrobia ulvae (Pennant) on intertidal epipelic microalgae: the effect of mud snail density.

The individual mean ingestion rate of Hydrobia ulvae was measured experimentally in controlled microcosms, in the dark to avoid primary production during measurement and at constant temperature. The experimental design was based on the addition of prelabelled epipelic microalgae to microcosms in a constant proportion with unlabelled diatoms, and in such a way that algal food availability was not a limiting factor within the range of tested densities (0.3 to 4.1 snails cm(-2)). Results show that the individual mean ingestion rate decreased significantly from 26.6+/-1.1 ng Chl a snail(-1) h(-1) to 22.4+/-1.0 ng Chl a snail(-1) h(-1) between 0.7 and 3 snails cm(-2). We hypothesize that this sharp decrease (the threshold density was between 1.4 and 2.5 snails cm(-2)) may account for a density-dependent effect. We have tested this hypothesis by using a simple random walk model including basic behavioural processes such as a break in feeding activity when two individuals contact each other. The model represents quantitatively well the threshold effect, suggesting that behavioural processes have to be taken into account for estimating a global feeding activity of H. ulvae populations.

Journal Article↗

Application of trichloroacetic acid (TCA) to extraction of soft body for the determination of tissue Cd, Cu, Pb and Zn in the prosobranch Hydrobia ulvae (Pennant).

The application of trichloroacetic acid (TCA) as a shell extractant for preparation of soft body parts with reference to tissue metal concentrations (Cd, Cu, Pb, Zn) in shellfish has been evaluated on the example of the mud snail Hydrobia ulvae, a small marine prosobranch densely present in rocky and soft-bottom habitats of the eastern Atlantic. A solution of 0.1 M TCA was tested on individuals treated according to two different protocols: (1) thawed after freezing ("non-dried") and (2) thawed and air-dried to a constant weight ("dried"). Two points were investigated in detail to improve the method: individual soft tissue dry weight and tissue metal concentration following a standard digestion method. In both instances, the results were compared with those from manually dissected snails. Conditions for total shell decalcification of 60 individuals (3-4 mm long) were 5.5 h in 20 ml of 0.1 M TCA.No differences in individual soft tissue weight were observed between the treatments, indicating good efficiency of the TCA extraction with respect to weight of soft body parts. In contrast, tissue metal concentrations varied among treatments. The TCA extraction of the dried animals had a good recovery for Cd, most likely due to the lower solubility of Cd vital cellular components (proteins and mineral concretions) from the dried tissue. Satisfactory recoveries of the tissue concentrations of Cu and Pb were obtained for the non-dried individuals. This might be related to the specific distribution of metals in the organism (namely in the digestive glands and gonads) and their different chemical reactivity with TCA after the tissue was dried. Limited susceptibility of Zn-bearing protein bindings to complexing with TCA also accounts for significantly lower concentrations of Zn in the mud snail's soft tissue that was extracted. The 0.1 M TCA solution is therefore recommended for extraction of the shells of Hydrobia ulvae for tissue determination of Cd, Cu and Pb; however the treatment protocol does affect metal recovery and thus a consistent procedure should be followed. The extracted metals from the soft tissues and shells of the mud snails (on the basis of both metal concentrations and contents) were ranked in order of increasing contribution of soft body parts to the total (shell+tissue): Pb<Cd<Zn<Cu.

Animals↗

Metal accumulation in the green macroalga Ulva fasciata: effects of nitrate, ammonium and phosphate.

Coastal organisms are often exposed to both metal pollution and nutrient enrichment. The influences of major nutrients (nitrate, ammonium, and phosphate) on the accumulation of trace metals (Cd, Cr, Zn and Se) in Ulva fasciata were examined. The relative accumulation of metals was quantified by the kinetic measurements of accumulated metal concentration over a short exposure period (8 h). Our study demonstrated that macronutrients could markedly influence the rate of metal accumulation in the macroalgae. An increase in ambient nitrate concentration resulted in a significant increase in Cd accumulation rate, whereas the rate of accumulation of Cr and Zn was not greatly affected by the ambient nitrate level (between 10 and 100 microM). Zn uptake in nitrate-enriched macroalgae was, however, significantly higher than its uptake in N-starved macroalgae. The accumulation of Cd, Cr and Zn was not appreciably affected by the concentration of ammonium. Se accumulation was significantly inversely related to the ambient phosphate concentration, presumably due to the competitive inhibition by a high P concentration. Cr accumulation in the macroalgae increased significantly with increasing phosphate concentration. These data implied that the influences of major nutrients on cationic and anionic metal accumulation were highly metal-specific. The dependence of metal accumulation on major nutrients will appreciably affect our prediction of metal accumulation in macroalgae and the interpretation of biomonitoring data using the Ulva species.

Chlorophyta↗