Antioxidant and pro-oxidant activities of the brown algae, Laminaria digitata, Himanthalia elongata, Fucus vesiculosus, Fucus serratus and Ascophyllum nodosum

The ability of Laminaria digitata, Himanthalia elongata, Fucus vesiculosus, Fucus serratus and Ascophyllum nodosum to scavenge peroxyl radicals was investigated by kinetic studies in a model system. The thermal initiated oxidation of methyl linoleate was performed at 60°C in heptanol, with or without antioxidants. When they reached 1% of the substrate, seaweed extracts exhibited antioxidant activities by extending the induction period, but they did not suppress the rate of oxygen uptake as did vitamin E. A synergistic effect occurred when both algal extracts (1.5 g L^-1) and vitamin E (0.4 mmol L^-1) were present, and the effectiveness of the combined antioxidants during the whole induction period was vitamin E effectiveness. The synergistic effect of L. digitata, however, was subject to seasonal variations: samples collected in summer were effective synergists, whereas samples collected in winter displayed a marked negative synergism. The phospholipid fractions of F. vesiculosus, F. serratus and A. nodosum, including pigments, accounted for only 6% of the total lipid fraction, and did not exhibit a large synergistic effect. The main phospholipid was not phosphatidyl ethanolamine as usually related, but phosphatidyl inositol. Fucoxanthin had some antioxidant activity per se under our experimental conditions, but did not act as a synergist of vitamin E. The most potent synergists were recognized as chlorophyll a and related compounds by the application of liquid-liquid partition and chromatography for the identification of active components. This revised version was published online in June 2006 with corrections to the Cover Date.     

Expressed sequence tag-derived polymorphic SSR markers for Fucus serratus and amplification in other species of Fucus

The seaweed genus Fucus is a dominant component of intertidal shores throughout the North Atlantic and North Pacific and has been the focus of considerable developmental, ecological, and evolutionary research for the past century. Here, we present details of 21 expressed sequence tag-derived simple sequence repeat markers (microsatellites). All 21 were polymorphic for F. serratus, which also display considerable cross-reactivity with the sister species F. distichus (18) and the more distantly related F. vesiculosus (13), and F. spiralis (5).     

Structure of a fucoidan from the brown seaweed Fucus serratus L

A fucoidan consisting of L-fucose, sulfate and acetate in a molar proportion of 1:1:0.1 and small amounts of xylose and galactose were isolated from the brown seaweed Fucus serratus L. The fucoidan structure was investigated by 1D and 2D 1H and 13C NMR spectroscopy of its desulfated and de-O-acetylated derivatives as well as by methylation analysis of the native and desulfated polysaccharides. A branched structure was suggested for the fucoidan with a backbone of alternating 3- and 4-linked alpha-L-fucopyranose residues, -->3)-alpha-L-Fucp-(1-->4)-alpha-L-Fucp-(1-->, about half of the 3-linked residues being substituted at C-4 by trifucoside units alpha-L-Fucp-(1-->4)-alpha-L-Fucp-(1-->3)-alpha-L-Fucp-(1-->. Minor chains built up of 4-linked alpha-fucopyranose and beta-xylose residues were also detected, but their location, as well as the position of galactose residues, remained unknown. Sulfate groups were shown to occupy mainly C-2 and sometimes C-4, although 3,4-diglycosylated and some terminal fucose residues may be nonsulfated. Acetate was found to occupy C-4 of 3-linked Fuc and C-3 of 4-linked Fuc in a ratio of about 7:3.     

Assembly of the photosynthetic apparatus in embryos from Fucus serratus L

The assembly of the photosynthetic apparatus was studied during the first six days of development of Fucus serratus L. embryos. HPLC analysis revealed that oospheres and zygotes contain the same photosynthetic pigments (i.e., chlorophyll a, chlorophyll c, fucoxanthin, violaxanthin, and β-carotene) as fully developed thalli. Total pigment amount increased after fertilization, mainly due to an active synthesis of Chl a and fucoxanthin. Spectral modifications revealing the progressive integration of Chl a and Chl c in the photosynthetic units are described. In particular, a distinct emission at 705 nm, reflecting the accumulation of LHC I, was clearly detected. The emission bands at 705 nm and 725 nm were characterized by 77 K excitation fluorescence measurements. Their spectra differed by the presence of a large band at approximately 550 nm due to fucoxanthin in the excitation spectrum of F705 nm. Room temperature variable fluorescence was first observed 30 h after fertilization indicating a functional Photosystem II electron transfer at this developmental stage. This revised version was published online in August 2006 with corrections to the Cover Date.     

Isolation and Characterization of Golgi Bodies from Vegetative Tissue 1Fucus serratus

Cell fractions consisting primarily of functional and morphologicallyrecognizable Golgi bodies were prepared from the brown algaFucus serratus. Several enzyme activities were found to be associatedwith these fractions: thiamine pyrophosphatase, inosine diphosphatase,and galactosyl-transferases. The fractions catalysed the transferof galactose from UDP-galactose to endogenous acceptors, N-acetylglucosamine,ovalbumin, fucose, and fucoidan. These activities are latent,being activated by detergents (Triton X-100 or sodium deoxycholate).The chemical composition of the isolated fractions was examinedchromatographically and electrophoretically.     

Hybridization of the marine seaweeds,Fucus serratus and Fucus evanescens (Heterokontophyta: Phaeophyceae) in a 100-year-old zone of secondary contact

Historically, the intertidal seaweeds Fucus serratus (Fs) and Fucus evanescens (Fe) were sympatric only along the western coast of Norway. In the mid-1890s, Fe (monoecious) was accidentally introduced into the Oslofjord. Putative hybridization with the endemic Fs (dioecious) was observed in Oslofjord by 1977 and in the Kattegat and western Baltic Seas by 1998. At Blushøj, Denmark (Kattegat Sea) putative Fs x Fe hybrids were present only when densities of Fe and Fs exceeded 14 and 2 m(-2), respectively. All of the 58 putative hybrids that were collected in 1999 were dioecious and intermediate in morphology. Essentially all (57 out of 58) were reproductively mature, but the oogonia possessed fewer and more variably sized eggs than either parent. Examination of each parental species and putative hybrids with nuclear, mitochondrial and chloroplast molecular markers confirmed the occurrence of hybridization. Furthermore, all of the hybrids possessed Fe-type chloroplasts and mitochondria, indicating that only the Fe egg x Fs sperm pairing was successful in the field. The reciprocal cross of Fs egg x Fe sperm was absent in the field and significantly less successful in laboratory crossings. Asymmetrical hybridization has also been reported for several species of plants and animals.     

Stage-dependent sensitivity to ultraviolet radiation in zygotes of the brown alga Fucus serratus

Sensitivity to ultraviolet (UV) radiation (UV-A, lambda = 315-400 nm; plus UV-B, lambda = 280-315 nm) of zygotes of the brown alga Fucus serratus L. (Phaeophyta) has been assessed through effects on growth of developing germlings. Different stages of development were distinguished by considering 5 h periods of time after fertilisation. Both the stage of the zygote and the UV radiation condition significantly affected growth of developing germlings. The negative response of growth rate of early stages of the zygotes to UV radiation seemed to be caused by UV-B rather than UV-A radiation, as the lowest relative growth rates were always estimated for germlings developed from zygotes irradiated with UV-B radiation. As regards the stage of the zygote, those germlings that developed from zygotes irradiated at 5-10 h after fertilisation showed the strongest inhibition of growth compared with the other stages. These results point to polarisation as the most UV-sensitive process during the first 24 h of the development of the zygote. A non-linear relationship between the developmental stage of the zygote and the sensitivity to UV radiation is suggested.     

Detection and Identification of Fungi Intimately Associated with the Brown Seaweed Fucus serratus

The filamentous fungi associated with healthy and decaying Fucus serratus thalli were studied over a 1-year period using isolation methods and molecular techniques such as 28S rRNA gene PCR-denaturing gradient gel electrophoresis (DGGE) and phylogenetic and real-time PCR analyses. The predominant DGGE bands obtained from healthy algal thalli belonged to the Lindra, Lulworthia, Engyodontium, Sigmoidea/Corollospora complex, and Emericellopsis/Acremonium-like ribotypes. In the culture-based analysis the incidence of recovery was highest for Sigmoidea marina isolates. In general, the environmental sequences retrieved could be matched unambiguously to isolates recovered from the seaweed except for the Emericellopsis/Acremonium-like ribotype, which showed 99% homology with the sequences of four different isolates, including that of Acremonium fuci. To estimate the extent of colonization of A. fuci, we used a TaqMan real-time quantitative PCR assay for intron 3 of the beta-tubulin gene, the probe for which proved to be species specific even when it was used in amplifications with high background concentrations of other eukaryotic DNAs. The A. fuci sequence was detected with both healthy and decaying thalli, but the signal was stronger for the latter. Additional sequence types, representing members from the Dothideomycetes, were recovered from the decaying thallus DNA, which suggested that a change in fungal community structure had occurred. Phylogenetic analysis of these environmental sequences and the sequences of isolates and type species indicated that the environmental sequences were novel in the Dothideomycetes.     

Quantitative determination of glutamate in a Rhodophyceae (Chondrus crispus) and four Phaeophyceae (Fucus vesiculosus,Fucus serratus,Cystoseira elegans,Cystoseira barbata)

A study was made to find a better method of analyzing the glutamate pool in seaweeds than the use of HPLC, which provides unsatisfactory results with material rich in alginates and salts. A method recommended elsewhere (Inglis A, Bartone N, Finlayson J, 1988, J. Biochem. Biophys. Methods 15: 249–254) for physiological fluids has been assayed and improved for algal samples. It consisted of the addition of lithium acetate before the phenylisothiocyanate derivatization, omission of one drying step and extraction of the derivative with heptane before chromatographic analysis. Neither salt nor alginates interfered with analysis.     

Enzymatic cross-linking of a phenolic polymer extracted from the marine alga Fucus serratus

We have shown that a phenolic polymer (PP) extracted from Fucus serratus can be cross-linked using a vanadium-dependent bromoperoxidase (BPO). The methanol extracted PP was adsorbed to a quartz crystal sensor and the cross-linking was initiated by the addition of BPO, KBr, and H2O2. The decreased dissipation upon addition of the cross-linking agents, as measured with the quartz crystal microbalance with dissipation monitoring (QCM-D) method, was interpreted as intramolecular cross-links were formed between different phloroglucinol units in the PP. With surface plasmon resonance, it was shown that no desorption occurred from the sensor surface during the cross-linking. UV/vis spectroscopy verified the results achieved with QCM-D that all components, i.e., BPO, KBr, and H2O2, were necessary in order to achieve intramolecular oxidative cross-linking of the polymer.     

The physiological state of intertidal brown seaweeds Fucus serratus Linnaeus, 1753 and Fucus distichus Linnaeus, 1767 cultivated on a biofiltration system in the Barents Sea

The physiological parameters (growth rate, specific area, photosynthesis rate, metabolic activity of cells, contents of photosynthetic pigments, dry matter, and soluble phlorotannins) of the intertidal brown seaweeds Fucus serratus and F. distichus cultivated in the Barents Sea for oil biofiltration purposes were investigated. The possibility of their existence submerged to a 5 m depth is shown. The duration of the survival of the plants in the absence of a tide cycle depended on the degree of fouling and the presence of phytophages. For long-term growth of seaweeds on vertical ropes, fucoids need to be subjected to periodic desiccation.     

Geographically specific heteroplasmy of mitochondrial DNA in the seaweed, Fucus serratus (Heterokontophyta: Phaeophyceae, Fucales)

The presence of more than one type of mitochondrial DNA within the same organism (mtDNA heteroplasmy) has been reported in vertebrates, invertebrates, basidiomycetes and some angiosperms, but never in marine (macro)algae. We examined sequence differences in a 135‐base pair (bp) region of the nad11 gene in mitochondria of the intertidal rockweed, Fucus serratus, using single‐strand conformation polymorphism (SSCP). Each of 70 and 22 individuals from Blushøj (Denmark) and Oskarshamn (Sweden), respectively, displayed haplotypes 2, 3, and 4 (= mtDNA heteroplasmy), whereas only haplotype 2 was found in each of 24 individuals from locations in Spain, France, Ireland, Iceland and Norway. As Blushøj and Oskarshamn were among the last areas to emerge from ice cover during the Last Glacial Maximum (18 000–20 000 years bp ), the geographically specific heteroplasmy may represent a founder effect and therefore, a valuable marker for understanding the role of post‐Ice Age recolonization. Geographically specific heteroplasmy also has important implications in phylogeographical studies based on mtDNA sequences.     

Seasonal variation in a low shore Fucus serratus (Fucales,Phaeophyta) population and its epiphytic fauna

Seasonal variation in a Fucus serratus assemblage was studied on the Isle of Man, UK over a 13-month period. Algal cover varied greatly, related to shedding reproductive fronds and winter storms. Peak reproduction was recorded in the autumn. There was little change in density of plants, the population was dominated by a bank of germlings (<20 cm) and an adult mode between 50–70 cm. An indistinct third component of recruiting juveniles was present when gaps in the canopy formed. Seasonal variation in the algae was reflected in the population dynamics of the mobile epiphytes such as Littorina mariae and Lacuna pallidula, which exhibited annual life histories. Sessile species (such as bryozoans) utilized the algae year round, dominating the perennial stipes and lower fronds, which were not shed. Other species utilized the algae in a transient fashion: some feeding on the algae (e.g. Idotea), or other epiphytic animals (e.g. Nucella). Fucus serratus, therefore, provides a variety of spatially and temporally variable niches that explain the diversity of epiphytic animal species that exploit this alga.     

Monoclonal antibodies to sperm surface antigens of the brown alga Fucus serratus exhibit region-, gamete-, species- and genus-preferential binding

A panel of twelve monoclonal antibodies (MAbs), designated FS1 to FS12, have been raised against surface antigens of Fucus serratus sperm. The antibodies were selected on the basis that they show region-, gamete-, species- or genus-preferential binding. Indirect immunofluorescence shows that the antigens bound by the MAbs are distributed non-randomly over the cell surface. Seven MAbs (FS1, FS3, FS4, FS6, FS8, FS9, FS10) bind antigens located primarily on the cell body, while the others (FS2, FS5, FS7, FS11, FS12) bind antigens located primarily on the anterior flagellum. Of the MAbs that label the anterior flagellum, FS2, FS5, FS7 and FS12 form a halo at the perimeter of the flagellum. Electron microscopic-immunogold studies indicate that the halo results from labelling of the mastigonemes, as opposed to the flagellar plasmamembrane. Gamete-preferential binding of antibodies was detected using an enzyme-linked immunosorbent assay with egg membrane vesicles. Eight of the MAbs bind sperm antigens not common to eggs, though FS2, FS4, FS5 and FS9 bind antigens present on both sperm and eggs. In studies of species- and genus-specificity FS2, FS3, FS5, FS6, FS7, FS8, FS10, FS11 and FS12 exhibit genus-preferential binding, labelling sperm of F. serratus and F. vesiculosus more intensely than that of Ascophyllum nodosum. Only FS10 showed marked species-preferential binding, labelling sperm of F. serratus much more intensely than that of F. vesiculosus.Abbreviations Au-GAMIG gold-conjugated goat anti-mouse immunoglobulin - ELISA enzyme-linked immunosorbent assay - EM electron microscope - FITC-RAMIG fluorescein-isothiocyanate-conjugated rabbit anti-mouse immunoglobulin - IIF indirect immunofluorescence - MAb monoclonal antibody     

Isolation of protoplasts from Fucus serratus and F. vesiculosus(Fucales, Phaeophyceae): factors affecting protoplast yield

Protoplasts were isolated enzymatically from meristematic tissues of the brown algae, Fucus serratus, using a combination of 2% cellulase R-10 Onozuka, 0.5% macerozyme and 1% crude extract of gland gut of Aplysia vaccaria. The main factors affecting protoplast yield were identified. Protoplasts were produced in large quantities from apical region of thallus and from plantlets compared to mature explants. Yields were greatly improved by the addition of sodium citrate and bovine serum albumin in the enzymatic solution and could reach 5.8 × 10⁶ protoplasts per gram of fresh wt. The applicability of these optimal parameters to other species Fucus vesiculosus was shown.     

Characterization of microsatellite loci in the marine seaweeds,Fucus serratus and F. evanescens (Heterokontophyta; Fucaceae)

Fucus serratus and F. evanescens commonly occur on Northern European shores. Nine microsatellite loci were developed for F. serratus (8–22 alleles, observed heterozygosities = 0.367–0.850) and one for F. evanescens (seven alleles, observed heterozygosity = 0.804). Cross‐amplification was apparent, as five F. serratus loci were polymorphic in F. evanescens and 2–5 were polymorphic in F. vesiculosus, F. distichus, and F. spiralis.