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Growth rate of Ulva rigida in different Mediterranean eutrophicated sites.

The growth of juvenile populations of Ulva rigida C. Agardh was measured by means of immersion in in situ cages against environmental parameters (temperature, incident light, salinity, dissolved inorganic nitrogen and dissolved reactive phosphorus) in four different eutrophicated southern sites: Channel of the Thau lagoon (France), Lido, Sacca Sessola and Fusina stations (Venice lagoon, Italy). The growth curves as a function of temperature showed that, in all cases, the maximal temperature for Ulva growth was 17 degrees C (limitation in growth below 7 degrees C and above around 25 degrees C). The growth analysis of these four sites showed seasonal differences. In the least eutrophicated and calmest Lido station, grazing and dissolved reactive phosphorus (seven times lower at Lido than at Thau) played a key role. At Thau and Fusina, which are eutrophicated and turbid environments, the incident light had a strong impact on growth. Sacca Sessola, with an intermediate position between the above two mentioned situations, showed the highest growth rate. The values and relative growth rate (RGR) curves of the Mediterranean and open-sea northern sites are discussed. In particular, the temperature defines the type of growth curve (unimodal or bimodal) and the incident light is responsible for the low Mediterranean RGR values (<10% day(-1)).

Chlorophyta↗

Pseudoalteromonas ulvae sp. nov., a bacterium with antifouling activities isolated from the surface of a marine alga.

A dark-purple marine bacterium that inhibits the germination of marine algal spores and the settlement of invertebrate larvae has been characterized and assessed for taxonomic assignment. Two strains, designated UL12T and UL13, were isolated from the surface of the common marine alga Ulva lactuca. Based on 16S rDNA sequencing, UL12T and UL13 were found to show the highest similarity (97%) to members of the genus Pseudoalteromonas. DNA-DNA hybridization studies demonstrated less than 28% genomic DNA relatedness between these isolates and closely related Pseudoalteromonas species and greater than 65% homology between UL12T and UL13. The two isolates were found to display identical characteristics and are strict aerobes, motile by means of single polar flagella, exhibit non-fermentative metabolism and require sodium ions for growth. The isolates hydrolyse gelatin and can utilize citrate, maltose, mannose and glucose but not trehalose, sucrose, fructose, lactose or glycerol as sole carbon sources. The molecular evidence together with the phenotypic characteristics show that this bacterium constitutes a new species within the genus Pseudoalteromonas. The name Pseudoalteromonas ulvae sp. nov. is proposed for this bacterium and the type strain is UL12T (= UNSW 095600T = NCIMB 13762T).

Animals↗

Automatic monitoring of a circadian rhythm of change in light transmittance in ulva.

Ulva lactuca L. var. latissima (L.) DeCandolle has a circadian rhythm of visible light transmittance change which is caused by chloroplast orientation. With a continuously recording microphotometer system, clear rhythms could be monitored for up to 10 days. Measuring beam intensity effects on the free running period were seen down to 10(-7) w cm(-2). While these effects complicate the measuring process, they demonstrate that Ulva is very sensitive to light. The free running period in constant darkness at 20 C is 24 to 25 hours. The position in the cell occupied by the chloroplasts when the rhythm damps out can be influenced by light. A method is described by which the times of rhythm maxima can be calculated accurately and objectively from a relatively small number of points.

Journal Article↗

Phosphorus-31 and Nitrogen- 14 NMR Studies of the Uptake of Phosphorus and Nitrogen Compounds in the Marine Macroalgae Ulva lactuca.

Cytoplasmic phosphomonoesters and inorganic phosphate, as well as vacuolar inorganic phosphate and polyphosphates, gave rise to the major peaks in (31)P nuclear magnetic resonance (NMR) spectra of the marine macroalgae Enteromorpha sp., Ceramium sp., and Ulva lactuca which were collected from the sea. In contrast, NMR-visible polyphosphates were lacking in Pylaiella sp. and intracellular vacuolar phosphate seemed to act as the main phosphorus store in this organism. In laboratory experiments, polyphosphates decreased in growing U. lactuca which was cultivated in continuous light under phosphate-deficient conditions. In contrast, the same organism cultivated in seawater with added phosphate and ammonium, accumulated phosphate mainly in the form of polyphosphates. When nitrate was provided as the only nitrogen source, accumulation of polyphosphates in the algae decreased with increasing external nitrate concentration. From the chemical shift of the cytoplasmic Pi peak, the cytoplasmic pH of superfused preparations of Ulva was estimated at 7.2. The vacuolar pH, determined from the chemical shifts of the vacuolar Pi and the terminal polyphosphate peaks, was between 5.5 and 6.0. The intracellular nitrate and ammonium levels in U. lactuca were determined by (14)N NMR. Both nitrogen sources were taken up and stored intracellularly; however, the uptake of ammonium was much faster than that of nitrate.

Journal Article↗

Phosphorus-31 NMR Studies of Cell Wall-Associated Calcium-Phosphates in Ulva lactuca.

Phosphate concentrations in the range 0.1 to 2.0 millimolar induced the formation of extracellular amorphous calcium-phosphates in the cell wall of the marine macro algae Ulva lactuca when they were cultivated in light in seawater at 20 degrees C. A broad resonance representing these compounds as well as resonances for extracellular orthophosphate and polyphosphates could be followed by (31)P-nuclear magnetic resonance spectroscopy. The presence of the calcium-phosphate made the cells brittle and it inhibited the growth of the macro algae and caused mortality within 1 week. The formation of the calcium-phosphates was influenced by the external phosphate concentration, the extracellular pH and the nature and concentration of the external nitrogen source. Furthermore, no formation of these compounds was observed when Ulva lactuca was cultivated in the dark, at low temperatures (5 degrees C) or in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. The complex could be removed through washes with ethylenediaminetetraacetate; this treatment did not alter the intracellular pH or the orthophosphate and polyphosphate pools and it restored growth.

Journal Article↗

Flagellar roots, mating structure and gametic fusion in the green alga Ulva lactuca (Ulvales).

The slightly anisogamous gametes of Ulva lactuca exhibit a cruciate flagellar root system consisting of 4 microtubular roots (4-2-4-2 system) and an elaborate system of fibrous roots associated with the 2-stranded microtubular roots. Two fibres (32-nm striation periodicity; system I fibres) closely underlie each of the 2-stranded roots, while different fibres (150-nm striation periodicity; system II fibres) run parallel to the root microtubules, and are 150-200 nm more internally located. Female gametes have 4 system II fibres, 3 of which are combined into a compound fibre associated with one microtubular root, while the fourth fibre is associated with the opposite root. In male gametes only 2 system II fibres are present, each underlying one of the two 2-stranded roots. A special region of the plasmalemma of both gamete types about 0.5 mum away from the basal bodies and located between 2 adjacent microtubular roots is structurally specialized and acts as a mating structure in gametic fusion. The region is oval-shaped and up to I.I mum long with a maximum diameter of 0.7 mum. A continuous electron-dense boundary layer underlies the plasmalemma at the edges of the mating structure. In both gamete types the mating structure consists of a fuzzy layer of material underlying the plasmalemma and special granules (60 nm diameter) are associated with this layer on its cytoplasmic side. In addition diffuse material overlies the mating structure, especially in male gametes. The mating structure is connected to 3 different kinds of flagellar roots: the boundary layer is linked to a 2-stranded microtubular root and its associated system I fibre; the fuzzy layer of the mating structure is connected with a system II fibre; and in female gametes this is the compound system II fibre. The ultrastructural changes which occur after mixing the 2 gamete types have been followed. Mating structure activation involves contraction of system II fibres (change of striation periodicity to 100 nm), detachment of special granules from the fuzzy layer of the mating structure and their replacement by electron-transparent vesicles at the prospective cell fusion site. Furthermore, release of electron-dense contents from Golgi-derived vesicles in the anterior part of both gamete types precedes cell fusion. Cell fusion is exclusively initiated in a region delimited by the 2 mating structures. After partial dissolution the 2 plasma membranes unite within the mating structure regions. The ultrastructure of gametic fusion in Ulva lactuca is compared to that of other green algae and the significance of flagellar roots in the mating process of green algae is discussed.

Chlorophyta↗

Comparative ultrastructure of eyespot membranes in gametes and zoospores of the green alga Ulva lactuca (Ulvales).

Eyespot membranes in zoospores, and male and female gametes of the green alga Ulva lactuca, were studied comparatively by the freeze-fracture technique. The plasmalemma and the outer chloroplast envelope membrane overlying the eyespot lipid globules are specialized in all 3 types of reproductive cells. In the eyespot region the protoplasmic face (PF) of the outer chloroplast envelope membrane contains significantly more intramembraneous particles (IMP) compared to membrane areas outside the eyespot: in female gametes there are 2.5 times more IMP/micrometers 2, in zoospores 3 and in male gametes about 4. Small size-class IMP (4--6 nm diameter) are particularly abundant on both fracture faces of the outer chloroplast membrane, but size-class distribution is not significantly different between membrane areas inside and outside the eyespot region. The total number of IMP/eyespot on the PF of the outer chloroplast membrane was calculated to be 4900 in male gametes, 5500 in female gametes and 11 200 in zoospores. The results are discussed in accordance with the view that these membrane specializations participate in photoreception relating to green algal phototaxis. Evidence is presented that there is a correlation between IMP numbers per eyespot in the outer chloroplast envelope membrane and the different phototactic behaviour of gametes compared to zoospores in Ulva.

Chlorophyta↗

Role of sediment organic matter quality and feeding history in dietary absorption and accumulation of pyrene in the mud snail (Hydrobia ulvae).

Organic matter (OM) input to marine sediments varies seasonally both in quantity and quality. Because sedimentary OM (SOM) constitutes food for many benthic organisms, its properties should affect the dietary uptake of sediment-associated contaminants. We explored the effect of SOM quality/food value on short- and long-term pyrene accumulation in the mud snail (Hydrobia ulvae) and performed dual-tracer pulse-chase experiments to investigate the feeding mechanisms driving dietary pyrene uptake. The quality of the SOM was manipulated by enriching sediments either with high-quality microalgae or low-quality lignin, adding equal amounts of total organic carbon. Long- and short-term bioaccumulation increased with increasing SOM quality, as did pyrene ingestion rate (IR(pyr)), which also was affected by feeding history. By feeding selectively, snails concentrated pyrene 10-fold in ingested compared to ambient sediment, independent of SOM quality. Average pyrene absorption efficiency (AE(pyr): -65%) varied inversely with SOM quality and IR(pyr). Both AE(pyr) and gut passage time (alpha 1/IR(pyr)) agreed with theoretical models incorporating the time-dependence of absorption efficiency. Thus, SOM quality moderates dietary contaminant uptake in deposit feeders, and in H. ulvae, this occurs via OM-induced alterations of ingestion rate. Consequently, enhanced sediment-associated contaminant uptake is predicted for deposit feeders following phytoplankton blooms, principally because of OM quality-driven increases in the ingestion rate.

Animal Feed↗

[The influence of infections with trematodes Bunocotyle progenetica (Hemiuridae) and Cryptocotyle cancavum (Heterophyidae) onto mortality of littoral mollusks Hydrobia ulvae (Gastropoda: Prosobranchia) after freezing].

The mortality of prosobranch snails Hydrobia ulvae uninfected and infected with the parthenites of two species of Trematodes (Bunocotyle progenetica and Cryptocotyle cancavum) after freezing in seawater of different salinity with following thawing has been studied. Reduced salinity exceeds mortality of snails under freezing significant (p < 0.01). The mortality of mollusks increases significant (p < 0.01) with the increase of the exposure time. The mortality of females exceeds the mortality of male significant (p < 0.01) within the group of 2-years individuals. The negative effect of trematode invasion to the vitality of mollusks after freezing was not showed. Trematode infestation registered in March 2004 and in September 2003 is not significant differ (Levakin, 2004). Thus any important differential mortality of individuals H. ulvae infected with parthenites of this Trematodes is not present during a winter time. The conclusion is conformed by the results of our experiments and well accords to features of realization of life cycles of these Trematodes under conditions of the White Sea intertidial zone, where infection is possible only within the worm season.

Animals↗

A membrane-located polypeptide of Ulva sp. which may be involved in HCO3- uptake is recognized by antibodies raised against the human red-blood-cell anion-exchange protein.

Polypeptides present in a membrane fraction of the marine macroalga Ulva sp. were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and tested for cross-reactivity with antibodies raised against the human red-blood-cell anion exchanger (AE1). A polypeptide of ca. 95 kDa was identified with a monoclonal, as well as two polyclonal (one against the C-terminus and one against the whole protein) antibodies, indicating that it shares homologous domains with AE1. These findings complement an earlier study which indicated that a plasmalemma-bound, disulfonic stilbene-sensitive, protein was functionally involved in HCO3- transport into the photosynthesizing cells of Ulva (Z. Drechsler et al. 1993, Planta 191, 34-40). It is thus suggested here that a similar protein has evolved, and has been conserved, in marine photosynthetic organisms and mammalian red blood cells for the purpose of HCO3- transport.

Anion Exchange Protein 1, Erythrocyte↗

Expanding the scope of precision editing in seaweeds through the application of a novel CRISPR-associated nuclease 12a-aligned CRISPR system in Ulva prolifera.

Seaweeds, such as the fast-growing green alga Ulva prolifera, can be harnessed as valuable marine crops. The lack of scalable genome-editing tools hampers functional genomics to explore and elucidate algal molecular pathways with industrial importance. Here, we expanded precision genome modification in seaweeds by successfully demonstrating gene editing with a transgene-free AT-rich-targeting CRISPR-associated protein (Cas) system in U. prolifera. By evaluating various delivery buffers, comparing different Cas systems, and optimizing incubation temperatures, we determined suitable conditions for more widespread applicability of a novel Cas12a-aligned ST8 editor. We obtained >50 ST8-mediated knockout mutants of a toxin-based endogenous marker gene, UpAPT, at 28 &#xb0;C post-delivery incubations. Our work diversified the applicable genome-editing tools in seaweeds, advancing algal functional genomics and providing more strategies to precisely target unexplored seaweed resources.

Ulva↗

Photon- and carbon-use efficiency in Ulva rigida at different CO2 and N levels.

The seaweed Ulva rigida C. Agardh (Chlorophyta) was cultured under two CO(2) conditions supplied through the air bubbling system: non-manipulated air and 1% CO(2)-enriched aeration. These were also combined with N sufficiency and N limitation, using nitrate as the only N source. High CO(2) in U. rigida led to higher growth rates without increasing the C fixed through photosynthesis under N sufficiency. Quantum yields for charge separation at photosystem II (PSII) reaction centres (phi(PSII)) and for oxygen evolution (phi(O2)) decreased at high CO(2) even in N-sufficient thalli. Cyclic electron flow around PSII as part of a photoprotection strategy accompanied by decreased antennae size was suspected. The new re-arrangement of the photosynthetic energy at high CO(2) included reduced investment in processes other than C fixation, as well as in carbon diverted to respiration. As a result, quantum yield for new biomass-C production (phi(growth)) increased. The calculation of the individual quantum yields for the different processes involved allowed the completion of the energy flow scheme through the cell from incident light to biomass production for each of the CO(2) and N-supply conditions studied.

Carbon Dioxide↗

A novel marine algal toxicity bioassay based on sporulation inhibition in the green macroalga Ulva pertusa (Chlorophyta).

A 5-day aquatic toxicity test based on sporulation inhibition of the green macroalga Ulva pertusa Kjellman has been developed. Optimal test conditions determined for photon irradiance, salinity and temperature were 60-200 micromol photons m(-2)s(-1), 25-35 per thousand and 15-20 degrees C, respectively. Tests were conducted by exposing U. pertusa thallus disks to a reference toxicant (sodium dodecyl sulfate; SDS), metals (Cd2+, Cu2+, Zn2+, Pb2+) and elutriates of sludge collected from nine different locations. The EC(50) values for SDS was 5.35 mg L(-1). When four heavy metals were assayed, the NOECs were highest for lead (0.625 mg L(-1)) and lowest for copper (0.031 mg L(-1)). The EC50 values showed the following toxicity rankings: Cu(2+) (0.061 mg L(-1))>Cd2+ (0.326 mg L(-1))>Zn2+ (0.738 mg L(-1))>Pb2+ (0.877 mg L(-1)). The bioassay indicated also that the sporulation endpoint could be a sensitive indicator of toxicity effects of elutriates of sludge as reflected from the NOEC values equal to or lower than the lowest concentration employed (6.25%). Sporulation was significantly inhibitied in all elutriates with the greatest and least effects observed in elutriates of sludge from industrial waste (EC50 6.78%) and filtration bed (EC50 15.0%), respectively. The results of the Spearman rank correlation analysis for EC50 data versus the concentrations of toxicants in the sludge presented a significant correlation between toxicity and four heavy metals (Cd2+, Cu2+, Pb2+, Zn2+). Introduction of the concept of toxicity unit (TU) showed that these metals were the main cause of toxicity in elutriates of at least four out of nine sludge samples. Members of the order Ulvales show a wide geographic distribution and have similar reproductive characteristics, thus making it possible to apply the present test method to other algae of this taxa, elsewhere. This novel method will be a useful tool for assessing the aquatic toxicity of a wide range of toxicants, once the respective sensitivities are demonstrated.

Analysis of Variance↗

Evaluation of the marine algae Ulva fasciata and Sargassum sp. for the biosorption of Cu(II) from aqueous solutions.

In this study, the adsorption properties of two different marine algae (Ulva fasciata (green algae) and Sargassum sp. (brown algae)) were investigated. Equilibrium isotherms and kinetics were studied to evaluate the relative ability of the two algae to sequester Cu(II) from aqueous solutions. The maximum biosorption capacity obtained was 73.5 mg g(-1) for U. fasciata and 72.5 mg g(-1) for Sargassum sp. at a solution pH of 5.5 +/- 0.5. A significant fraction of the total copper(II) uptake was achieved within 30 min. The copper(II) uptake by the biosorbents was best described by pseudo-second-order rate model.

Adsorption↗

Biosorption of copper, cobalt and nickel by marine green alga Ulva reticulata in a packed column.

Biosorption of copper, cobalt and nickel by marine green alga Ulva reticulata were investigated in a packed bed up-flow column. The experiments were conducted to study the effect of important design parameters such as bed height and flow rate. At a bed height of 25 cm, the metal-uptake capacity of U. reticulata for copper, cobalt and nickel was found to be 56.3+/-0.24, 46.1+/-0.07 and 46.5+/-0.08 mgg(-1), respectively. The Bed Depth Service Time (BDST) model was used to analyze the experimental data. The computed sorption capacity per unit bed volume (N0) was 2580, 2245 and 1911 mgl(-1) for copper, cobalt and nickel, respectively. The rate constant (K(a)) was recorded as 0.063, 0.081 and 0.275 lmg(-1)h(-1) for copper, cobalt and nickel, respectively. In flow rate experiments, the results confirmed that the metal uptake capacity and the metal removal efficiency of U. reticulata decreased with increasing flow rate. The Thomas model was used to fit the column biosorption data at different flow rates and model constants were evaluated. The column regeneration studies were carried out for three sorption-desorption cycles. The elutant used for the regeneration of the biosorbent was 0.1 M CaCl2 at pH 3 adjusted using HCl. For all the metal ions, a decreased breakthrough time and an increased exhaustion time were observed as the regeneration cycles progressed, which also resulted in a broadened mass transfer zone. The pH variations during both sorption and desorption process have been reported.

Adsorption↗

[Seasonal variations of metal contents (Cd, Cu, Fe, Mn and Zn) in seaweed Ulva lactuca from the coast of El Jadida city (Morocco)].

The quality of El Jadida Atlantic coastal water was monitored from April 1998 to March 1999 by measuring hydrological parameters (dissolved oxygen, suspended particulate matter, phosphates and nitrites) and using the seaweed Ulva lactuca as a quantitative bio-indicator of cadmium, copper, iron, manganese and zinc contamination. Metal content in seaweeds, collected every month from four stations characterized by the discharge of urban and industrial waste water, showed significant variations depending on the station and sampling period. However, the seaweed of El Jadida exhibited generally lower contents compared to those of similar species from other geographical areas.

Cadmium↗

A link between lead and cadmium kinetic speciation in seawater and accumulation by the green alga Ulva lactuca.

In this work, studies on the bioaccumulation of Cd and Pb by Ulva lactuca at different sites of Gulf San Jorge (Patagonia, Argentina) are presented. Higher values of bioaccumulated Cd were found in Punta Maqueda - a site believed to serve as a control - in comparison to those in Punta Borja, a place highly exposed to urban and industrial activities. Consequently; the labile fractions of Cd and Pb in seawater were determined with a flow injection-preconcentration manifold interfaced to a graphite furnace-atomic absorption spectrometer (FI-GFAAS). The results obtained by kinetic speciation showed that the variable that correctly explains heavy metals accumulation in the alga, is the labile metal fraction in seawater. We propose to use an enhancement ratio - on the basis of the kinetically labile metal fraction - for calculation of the metal accumulated by the alga relative to its environment.

Argentina↗

Isolation and characterization of a single-copy actin gene from a sterile mutant of Ulva pertusa (Ulvales, Chlorophyta).

We constructed a cDNA library from sterile Ulva pertusa (Ulvales, Chlorophyta), and isolated and characterized a full-length cDNA clone encoding actin. The actin (ACT) cDNA consisted of 1487 nucleotides (nt) and had an open reading frame (ORF) encoding a polypeptide of 377 amino acid (AA) residues. The ACT gene had one intron in the 5'-untranslated region and three introns in the coding region. Transcription started 26 nt downstream of the putative TATA box. A potential polyadenylation signal, TGTAG, was located 100 nt downstream of the terminator codon, TAG. Amino acid alignment with actins from various algae and land plants showed that sterile U. pertusa actin was more similar to actins from Chlorophyta, Phaeophyta, Euglenophyta, and higher plants (over 76.9%) than to actins from Rhodophyta. Southern blot analysis indicated that the sterile U. pertusa genome has only a single actin-encoding gene. Thalli grown on a 12D/12L photoperiod increased in surface area some two-fold over 24 h regardless of the nutritional conditions. The growth rate of thalli during the light period was significantly higher than that during the dark period. Northern hybridization indicated that the expression of actin mRNA was induced and repressed by the light and dark treatments, respectively. These results suggest that the U. pertusa cell division cycle has a periodicity of 24 h and that the ACT gene is highly transcribed during cell growth and development in the light period.

Actins↗