DIFFERENCES IN LIFE-HISTORY CHARACTERISTICS ARE CONSISTENT WITH THE VERTICAL DISTRIBUTION PATTERN OF FUCUS SERRATUS AND FUCUS VESICULOSUS (FUCALES, PHAEOPHYCEAE) IN THE CENTRAL BALTIC SEA

On the rocky shores of southeast Sweden, in the central Baltic Sea, the brown seaweed species Fucus serratus L. and Fucus vesiculosus L. form dense constantly submerged stands. The relative proportion of the two species in the belt is shifting both vertically and geographically. Our hypothesis is that differences in life-history characteristics and the tolerance to physical factors such as wave force, freezing, and light may alter the competitive outcome between the two species with increasing wave exposure. This study included a combination of field studies and laboratory experiments. In sheltered locations, F. serratus and F. vesiculosus grow in mixed stands from 0.5 to 3 m deep, whereas at exposed sites F. vesiculosus grow from 0.5 to 2 m deep and F. serratus mainly from 1 to 5 m deep. The two Fucus species differed markedly in their tolerance to physical stress as well as in a number of reproductive and vegetative characteristics. Tolerance to freezing and wave action was higher in F. vesiculosus compared with F. serratus . Fucus vesiculosus also produced 10 times more eggs per receptacle, allocated significantly more tissue per frond for reproduction, and produced more reproductive fronds per individual than did F. serratus . In addition, the survival rate of F. vesiculosus germlings was slightly higher than that of F. serratus germlings during the first year. These differences in life history characteristics are suggested to partly explain the observed distribution patterns of the two Fucus species in the Baltic Sea.     

EVOLUTION OF THE FUCACEAE (PHAEOPHYCEAE) INFERRED FROM nrDNA-ITS

Sequences of the internal transcribed spacer region (ITS-1, 5.8S, and ITS-2) of nuclear ribosomal DNA were obtained from 16 species representing all six genera of Fucaceae ( Ascophyllum, Fucus, Hesperophycus, Pelvetia, Pelvetiopsis, and Xiphophora ) plus one outgroup ( Hormosira ). Parsimony analysis indicated that the family Fucaceae is monophyletic and that the northern hemisphere taxa are highly divergent from the only southern hemisphere genus, Xiphophora. The genus Pelvetia is not monophyletic because the European P. canaliculata is more closely related to Fucus, Hesperophycus, and Pelvetiopsis than to other Pelvetia species. We establish Silvetia, gen. nov. and transfer the 3 Pacific species of Pelvetia to the new genus. Fucus is monophyletic and not ancestral in the Fucaceae. The ITS sequences identified two strongly supported lineages within Fucus, one with F. serratus sister to the clade containing F. gardneri, F. distichus, and F. evanescens and a second including F. vesiculosus, F. spiralis, F. ceranoides, and F. virsoides. The ITS was not useful for resolving relationships within each of these clusters and between populations of F. vesiculosus. Within-individual variation in ITS sequences is high in Fucus, a derived genus, compared to Ascophyllum, a more ancestral genus. Mapping of the two characters that form the basis of Powell's model for speciation in the Fucaceae showed that 1) number of eggs per oogonium has not followed a gradual reduction and that 2) monoecy/dioecy has changed several times during evolution of this family.     

Arsenic binding to Fucus vesiculosus metallothionein

The seaweed Fucus vesiculosus is a member of the brown algae family. Kille and co-workers [Biochem. J. 338 (1999) 553] reported that this species contains the gene for metallothionein. Metallothionein is a metalloprotein having low molecular weight, and high cysteine content, which binds a range of metals. F. vesiculosus bioaccumulates arsenic from the aquatic environment [Mar. Chem. 18 (1986) 321]. In this paper we describe arsenic binding to F. vesiculosus metallothionein, characterized by electrospray ionization mass spectrometry. Five arsenic-MT species were detected with increasing As to protein ratios. These results provide important information about the metal-chelation behaviour of this novel algal metallothionein which is a putative model for arsenic binding to F. vesiculosus in vivo.     

Genetic entities and mating system in hermaphroditic Fucus spiralis and its close dioecious relative F. vesiculosus (Fucaceae, Phaeophyceae)

To date, molecular markers have not settled the question of the specific status of the closely related, but phylogenetically unresolved, brown seaweeds, hermaphroditic Fucus spiralis and dioecious Fucus vesiculosus, nor their propensity for natural hybridization. To test the degree of species integrity and to assess effect of the mating system on the population genetic structure, 288 individuals coming from parapatric (discontinuous) and sympatric (contiguous) spatial configurations at two sites were genotyped with five microsatellite loci. Using a Bayesian admixture analysis, our results show that F. spiralis and F. vesiculosus comprise clearly distinct genetic entities (clusters) generally characterized by cosexual and unisexual individuals, respectively. Genetic diversity within each entity suggests that F. spiralis reproduces primarily through selfing while F. vesiculosus is characterized by an endogamous breeding regime. Nevertheless, aberrant sexual phenotypes were observed in each cluster, no diagnostic alleles were revealed and 10% of study individuals were intermediate between the two genetic entities. This pattern can be explained by recent divergence of two taxa with retention of ancestral polymorphism or asymmetrical, introgressive hybridization. However, given (i) coincident monomorphism at three loci in spiralis clusters and (ii) that significantly more intermediates were observed in sympatric stations than in parapatric stations, we argue that interspecific gene flow has occurred after divergence of the two taxa. Finally, we show that whether recently separated or recently introgressive, the divergent breeding systems probably contribute to species integrity in these two taxa.     

Content of alginic acid and fucoidan in fucus algae of the Barents sea

Seasonal changes in the content of alginic acid and fucoidan have been studied in four species of fucus algae from the Barents sea: Fucus vesiculosus, F. distichus, F. serratus, and Ascophyllum nodosum. These polysaccharides are accumulated in the biomass of the algae during summer and autumn. To ensure complete processing of fucus algae it is recommended that the raw material be harvested within this period.     

Cleavage of DNA by brown algal polyphenols

Extracts of marine algae have been tested to determine their ability to cleave DNA. The species producing positive results wereAscophyllum nodosum, Fucus serratus, F. spiralis, F. vesiculosus, Halidrys siliquosa andHimanthalia elongata. Partial purification of each extract by dialysis against water revealed that the active compounds in each species were high molecular weight polyphenols.     

REPRODUCTIVE SUCCESS OF FUCUS VESICULOSUS (PHAEOPHYCEAE) IN THE BALTIC SEA

Marine organisms colonizing brackish habitats such as the Baltic Sea must cope with the negative effects of low salinities on reproductive success because these may reduce gamete viability and/or increase polyspermy. Reproductive characteristics of the marine seaweed Fucus vesiculosus L. were studied in several brackish habitats, particularly in the northern Baltic Sea, to understand its ability to reproduce where few other marine species survive. Polyspermy and fertilization success were variable at the boundary of the continuous distribution of F. vesiculosus in the Baltic Sea, and polyspermy was high (10%–30%) when fertilization was successful. A strong female bias (80%–86%, ca. 5.5:1) was found at the northernmost limit of Baltic F. vesiculosus. Electrophysiological studies showed that many eggs have a high input resistance (519 ± 150 MΩ[mean ± SE, n = 14] at Drivan, 1995), which may be helpful in preventing polyspermy in this brackish habitat. The polyspermy block remains sodium-dependent in the northern Baltic. Sperm bound quickly to northern Baltic eggs in natural water, but fertilization was delayed compared to marine F. vesiculosus. A subset of northern Baltic eggs studied during an optimal reproductive period (7–11 July 1995) had a membrane potential (E_m) of ca. −100 mV and an effective fertilization potential (FP) of ca. 2 min with a plateau of −25 mV, but repolarized too rapidly for the FP to be protective. Pronuclear migration and cell wall secretion occurred more slowly in Baltic than in marine zygotes. The reproductive success of theseboundary populations may be dependent upon windows of opportunity when there are favorable combinations of the levels of salinity, water motion, population density, and sex ratio. These factors and the short duration of the reproductive season in the northern Baltic Sea may result in reproductive failure in some years.     

Research note: Ecotype differentiation in qualitative content of water soluble organic compounds between marine and brackish Fucus vesiculosus L. (Phaeophyceae)

In the present study we compared the contents of water soluble organic compounds of the marine intertidal ecotype of Fucus vesiculosus (Phaeophyceae) from the Norwegian Sea (34 practical salinity units, psu) with the sublittoral ecotype of F. vesiculosus from the brackish Bothnian Sea (5 psu). Nuclear magnetic resonance spectra revealed that marine F. vesiculosus had additional types of water soluble organic compounds compared with brackish F. vesiculosus. The results suggested that glycine betaine in the marine ecotype could be the reason for this ecotype differentiation. Furthermore, the qualitative differences between the ecotypes were the same after one week's treatment of marine algae in brackish water and of brackish algae in marine water. These suggest that the additional types of water soluble organic compounds in marine F. vesiculosus are not caused by the salinity conditions at the growth sites. Further research concerning other environmental factors that may influence ecotype differentiation of water soluble organic compounds qualitative content and adaptation in F. vesiculosus is recommended.     

Low Divergence in rDNA ITS Sequences Among Five Species of Fucus (Phaeophyceae) Suggests a Very Recent Radiation

Sequences from the two ribosomal DNA internal transcribed spacers (ITS1 and ITS2) were compared among five species of Fucus. Based on the present taxon sampling, parsimony analysis showed that Fucus serratus is the sister-group of the remaining Fucus species when Ascophyllum nodosum was used as an outgroup. The topology of the tree was (Fucus serratus (F. lutarius (F. vesiculosus (F. spiralis+F. ceranoides)))). The extremely low variation observed suggests a very recent radiation of the genus which supports the view widely accepted that the Fucales are among the most evolutionarily advanced of the brown algae. We further note that sequence differences between Fucus and Ascophyllum were 28%: this does not rule out the utility of ITS sequences within the Fucaceae. The very low number of informative positions allows to demonstrate empirically that distance matrix methods group on the basis of symplesiomorphies. Received: 17 February 1997 / Accepted: 17 April 1997     

EVIDENCE FOR METHYL JASMONATE-INDUCED PHLOROTANNIN PRODUCTION IN FUCUS VESICULOSUS (PHAEOPHYCEAE)

The plant growth regulators jasmonic acid and methyl jasmonate (MeJA) have recently been identified in a variety of marine algae; however, their role in these organisms is currently unknown. Here we report that exposure to MeJA, during periods of tidal emergence causes the induction of polyphenolic chemical defenses (the phlorotannins) in two populations of the common rockweed Fucus vesiculosus (Linnaeus). Phlorotannin concentrations were up to 1.6 times higher in the growing apices of F. vesiculosus from both Avery Point (Connecticut, USA) and Roosevelt Inlet (Delaware, USA) within 10–14 days after a single brief exposure to airborne MeJA at concentrations ranging from 5.42 to 542 nM. The timing and magnitude of this induced increase in phlorotannin concentration are similar to that caused by real and simulated herbivory, raising the question of whether jasmonates, or their oxylipin relatives, are natural elements of antiherbivore responses in Fucus , as they are in vascular plants.     

Contents of Alginic Acid and Fuccidan in Fucus Algae of the Barents Sea

Seasonal changes in the content of alginic acid and fuccidan have been studied in four species of fucus algae from the Barents Sea: Fucus vesiculosus, F. distichus, F. serratus, and Ascophyllum nodosum. These polysaccharides are accumulated in the biomass of the algae during summer and autumn. To ensure complete processing of fucus algae it is recommended that the raw material be harvested within this period.     

Induction of phlorotannin production in a brown alga: defense or resource dynamics?

Increase of phenolic secondary metabolites, phlorotannins, in brown algae due to gastropod grazing has been interpreted as an anti-herbivore adaptation. Here we tested whether such a response could be due to changes in truly available resources for the alga, not by the grazing activity of snails as such. We allowed two species of snails, Theodoxus fluviatilis and Physa fontinalis to graze on Fucus vesiculosus . These species feed on epibiota and particulate matter on the thallus but do not eat the thallus of F. vesiculosus . We further simulated snail grazing by nutrient enhancement, removal of epibiota and by a combination of the two. Manipulations of nutrient and light availability revealed the crucial role of epibiota in mediating resource availability for F. vesiculosus . Nutrient enhancement alone increased epibiota and decreased phlorotannins. Cleaning the thallus resulted in increased growth, and together with nutrient enhancement also in a trade-off with phlorotannins. Presence of T. fluviatilis on the thallus induced phlorotannin production, a response differing from the simulations of snail grazing. However, we suggest that the increase in phlorotannins may not be an induced defense but rather a consequence of a specific way of resource manipulation by this snail species. T. fluviatilis removes hyaline hairs that facilitate nutrient uptake. P. fontinalis did not remove hyaline hairs and the response of the alga to its grazing was similar to the treatment where we mechanically removed epibiota suggesting that cleaning of the thallus is the major mechanism how this snail species affects F. vesiculosus . Genetic variation in phlorotannin concentrations highly exceeded the induced responses of simulated or real snail grazing. This casts doubt for the efficiency of induced phlorotannin production to act as a defense, but is not contradictory with the interpretation of phlorotannins responding to variation in resource availability.     

Agglutinins in marine brown algae. Dedicated to Professor Dr. H. Kn?ll on his 65th birthday

Extracts from the brown algae Fucus vesiculosus, F. serratus, and Halidrys siliquosa agglutinated human erythrocytes of groups A, B, O, and AB as well as erythrocytes from some animal species, ascites tumour cells from mice, and streptococci of many groups of the LANCEFIELD system. The extract of Chorda filum did not show haemagglutination but agglutinated ascites tumour cells and streptococci of a few groups.     

Seasonal variations in nitrate reductase activity and internal N pools in intertidal brown algae are correlated with ambient nitrate concentrations

Nitrogen metabolism was examined in the intertidal seaweeds Fucus vesiculosus, Fucus serratus, Fucus spiralis and Laminaria digitata in a temperate Irish sea lough. Internal NO(3) (-) storage, total N content and nitrate reductase activity (NRA) were most affected by ambient NO(3) (-), with highest values in winter, when ambient NO(3) (-) was maximum, and declined with NO(3) (-) during summer. In all species, NRA was six times higher in winter than in summer, and was markedly higher in Fucus species (e.g. 256 +/- 33 nmol NO(3) (-) min(-1) g(-1) in F. vesiculosus versus 55 +/- 17 nmol NO(3) (-) min(-1) g(-1) in L. digitata). Temperature and light were less important factors for N metabolism, but influenced in situ photosynthesis and respiration rates. NO(3) (-) assimilating capacity (calculated from NRA) exceeded N demand (calculated from net photosynthesis rates and C : N ratios) by a factor of 0.7-50.0, yet seaweeds stored significant NO(3) (-) (up to 40-86 micromol g(-1)). C : N ratio also increased with height in the intertidal zone (lowest in L. digitata and highest in F. spiralis), indicating that tidal emersion also significantly constrained N metabolism. These results suggest that, in contrast to the tight relationship between N and C metabolism in many microalgae, N and C metabolism could be uncoupled in marine macroalgae, which might be an important adaptation to the intertidal environment.     

Surface-associated fucoxanthin mediates settlement of bacterial epiphytes on the rockweed Fucus vesiculosus

The chemical defence against microfouling in the brown seaweed Fucus vesiculosus was investigated and an inhibitor of bacterial settlement was isolated by bioassay-guided fractionation of non-polar surface extracts. UV-vis and mass spectrometry were used to identify the compound as the carotenoid fucoxanthin. The metabolite was tested at the natural concentration (in a surface volume based assay) against the settlement of four bacterial strains isolated from F. vesiculosus and 11 strains isolated from co-occurring algae and marine sediment. Surface concentrations between 1.4 and 6 μg cm(-2) resulted in 50% inhibition of four of these isolates, which were studied in more detail using a surface area-based assay, while a fifth isolate proved to be less sensitive. The presence of fucoxanthin on the surface of F. vesiculosus was demonstrated with two different surface extraction methods. Fucoxanthin was detected at concentrations between 0.7 and 9 μg cm(-2) on the algal surface. Fucoxanthin was still present at the algal surface after removal of associated diatoms through mechanical cleaning and germanium dioxide treatment and was thus mainly produced by F. vesiculosus rather than by diatoms. Thus, the photosynthetic pigment fucoxanthin appears to be ecologically relevant as a surface-associated antimicrobial agent, acting against the settlement of bacteria on the surface of the macroalga F. vesiculosus.     

Adaptive traits are maintained on steep selective gradients despite gene flow and hybridization in the intertidal zone

Gene flow among hybridizing species with incomplete reproductive barriers blurs species boundaries, while selection under heterogeneous local ecological conditions or along strong gradients may counteract this tendency. Congeneric, externally-fertilizing fucoid brown algae occur as distinct morphotypes along intertidal exposure gradients despite gene flow. Combining analyses of genetic and phenotypic traits, we investigate the potential for physiological resilience to emersion stressors to act as an isolating mechanism in the face of gene flow. Along vertical exposure gradients in the intertidal zone of Northern Portugal and Northwest France, the mid-low shore species Fucus vesiculosus, the upper shore species Fucus spiralis, and an intermediate distinctive morphotype of F. spiralis var. platycarpus were morphologically characterized. Two diagnostic microsatellite loci recovered 3 genetic clusters consistent with prior morphological assignment. Phylogenetic analysis based on single nucleotide polymorphisms in 14 protein coding regions unambiguously resolved 3 clades; sympatric F. vesiculosus, F. spiralis, and the allopatric (in southern Iberia) population of F. spiralis var. platycarpus. In contrast, the sympatric F. spiralis var. platycarpus (from Northern Portugal) was distributed across the 3 clades, strongly suggesting hybridization/introgression with both other entities. Common garden experiments showed that physiological resilience following exposure to desiccation/heat stress differed significantly between the 3 sympatric genetic taxa; consistent with their respective vertical distribution on steep environmental clines in exposure time. Phylogenetic analyses indicate that F. spiralis var. platycarpus is a distinct entity in allopatry, but that extensive gene flow occurs with both higher and lower shore species in sympatry. Experimental results suggest that strong selection on physiological traits across steep intertidal exposure gradients acts to maintain the 3 distinct genetic and morphological taxa within their preferred vertical distribution ranges. On the strength of distributional, genetic, physiological and morphological differences, we propose elevation of F. spiralis var. platycarpus from variety to species level, as F. guiryi.     

Galactolipids rather than phlorotannins as herbivore deterrents in the brown seaweed<Emphasis Type="Italic"> Fucus vesiculosus</Emphasis>

The first investigation of seaweed chemical defense against herbivores involved the brown seaweed Fucus vesiculosus and suggested defense via phlorotannins. The first demonstration of seaweed induction of secondary metabolites in response to herbivory also involved the genus Fucus and assumed a defensive function for phlorotannins. Many other investigations correlate herbivore feeding preference with changing levels of phlorotannins in this genus and others, but few directly test the effects of phlorotannins. No studies have assessed Fucus chemical defenses using bioassay-guided separation to investigate the complete complement of compounds deterring herbivores. We investigated the deterrence of F. vesiculosus chemical extracts using herbivore bioassays to guide our chemical investigations. Although crude extracts from F. vesiculosus strongly deterred feeding by the sea urchin Arbacia punctulata, phlorotannins from this extract did not deter feeding at 2x or 4x natural concentration by dry mass. Feeding deterrence was due to: (1) a polar galactolipid in the ethyl acetate-soluble extract, and (2) a non-phenolic compound, or compounds, in the water-soluble extract. Although this is the first evidence of galactolipids deterring herbivores, such defenses could be geographically and taxonomically widespread. The galactolipid we discovered in Fucus occurs in marine dinoflagellates, and a related metabolite that deters herbivory has recently been discovered in a tropical green seaweed. We were unable to identify the second deterrent compound, but deterrence occurred in a fraction containing carbohydrates, including sulfated sugars, but no phlorotannins. Given the polarity of these chemical deterrents, they could co-occur with and confound bioassays of phlorotannins if investigators test phlorotannin-containing algal extracts without further purification.     

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.     

Density dependent grazing effects of the isopod Idotea baltica Pallas on Fucus vesiculosus L in the Baltic Sea

In the Baltic Sea, abiotic factors are often supposed to explain the distribution of the key species Fucus vesiculosus. Still, in many areas, decline of F. vesiculosus has coincided with mass occurrence of the herbivorous isopod Idotea baltica. The aim of this work was to examine whether, how and at what densities I. baltica can affect the distribution of F. vesiculosus in the central Baltic proper. Both large-scale field surveys and a two-week grazing experiment have been performed.In the field survey there was a correlation between density of I. baltica and reduction in depth penetration of F. vesiculosus. At 80 animals per 100 g F. vesiculosus wet weight, the depth penetration of the F. vesiculosus belt was reduced by 2.5 m within a year. In the grazing experiment there was a correlation between density of I. baltica and loss of F. vesiculosus biomass and meristems. In the controls biomass and number of meristems increased by 50%, while at 20 animals per 100 g of F. vesiculosus there was no net growth of F. vesiculosus. Intensity of grazing did not differ between isopod densities of 20, 40 and 60/100 g. At isopod densities of 80 and 100/100 g though, biomass and meristems decreased by 50%, indicating a threshold for the survival of F. vesiculosus in the experiment. At all densities the isopods preferred younger tissue to older.Our results indicate that grazing by Idotea baltica is an important structuring factor in the Baltic Fucus vesiculosus populations.