Variability in iodine in temperate seaweeds and iodine accumulation kinetics of Fucus vesiculosus and Laminaria digitata (Phaeophyceae,Ochrophyta)

The biogeochemistry of iodine in temperate coastal ecosystems is largely mediated by macroalgae, which act as a major biological sink and source of iodine. Their capacity to accumulate, retain and release iodine has been associated with abiotic and biotic stressors, but quantitative information is limited. We evaluated the seasonal iodine retention capacity of eleven macroalgal species belonging to different systematic groups, collected from two sites in Ireland. Iodine accumulation and retention were then further quantified in Fucus vesiculosus and Laminaria digitata in relation to I^? concentrations in seawater and temperature. In general, iodine contents were ~10¹–10² μmol · (g dw)^?1 for Laminariales, 10⁰–10¹ μmol · (g dw)^?1 for Fucales, 10^?1–10⁰ μmol · (g dw)^?1 for Rhodophyta, and 10^?1 μmol · (g dw)^?1 for Chlorophyta. Typically, algal iodine contents were above average in winter and below average in summer. Iodine accumulation in F. vesiculosus and L. digitata depended on I^? availability and followed the Michaelis‐Menten kinetic. The ratio of maximum accumulation rate to half accumulation coefficient (ρ_max: K _t) was 2.4 times higher for F. vesiculosus than for L. digitata , suggesting that F. vesiculosus was more efficient in iodine accumulation. Both species exhibited a temperature‐dependent net loss of iodine, and only an exposure to sufficient external I^? concentrations compensated for this loss. This study revealed that both environmental (e.g., I^? in seawater, temperature) and organismal (e.g., the status of the iodine storage pool) variables determine retention and variability in iodine in temperate seaweeds.     

Influence of phytohormones on morphology and chlorophyll a fluorescence parameters in embryos of Fucus vesiculosus L. (Phaeophyceae)

While a variety of plant hormones from brown algae were described, there were few studies that examined the combined effects of these hormones on morphogenesis and photosynthetic physiology in developing fucoid embryos. We evaluated the effects of phytohormones to determine the extent, to which responses were similar to those of terrestrial plants. Kinetin, IAA, ABA, GA₃, and kinetin + IAA were added to seawater at a physiological concentration (1 mg/L), and embryos of Fucus vesiculosus L. were grown for 10 days. Photosynthetic activity of single embryos or embryo cells were characterized using the following fluorescence parameters: minimum fluorescence yield (F ₀), maximum quantum yield (F _v/F _m), relative maximum rate of electron transfer to photosystem II under saturation irradiances (rETR_max), photosynthetic efficiency under non-saturating irradiances (αETR) and saturation irradiance (E _k). In addition, embryo length and diameter and apical hair length and number were determined. Morphological changes associated with hormone treatments included an increase in the embryo length in the presence of IAA, an increase in the embryo diameter in the presence of IAA, kinetin, and kinetin + IAA, an increase in the maximum hair length and number in the presence of kinetin + IAA, and a decrease in the hair length and number in the presence of ABA. With respect to fluorescence parameters, significant effects of phytohormones included an increase in the F ₀ and F _v/F _m at kinetin treatment, a synergistic effect of kinetin + IAA on F _v/F _m, rETR_max, and αETR, a promotion of F _v/F _m by GA, and a decrease of the parameters by ABA. These results are consistent with the data on responses of land plants to the same hormones and suggest that brown algae have evolved regulatory mechanisms for morphogenesis and photosynthetic regulation similar to plants.     

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.     

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.     

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.     

Photosynthesis in relation to D1, PsaA and Rubisco in marine and brackish water ecotypes of Fucus vesiculosus and Fucus radicans (Phaeophyceae)

The aim of this study was to investigate photosynthetic differences between the marine, Norwegian Sea ecotype and the brackish, Bothnian Sea ecotype of F. vesiculosus and F. radicans and to see whether photosynthetic differences could be connected with the relative amounts of D1 protein (PSII), PsaA (PSI) protein and/or Rubisco. For this purpose, we tested if a higher photosynthetic maximum (P _max) in the Atlantic Ocean ecotype of F. vesiculosus relative to the Baltic Sea ecotype, and an increase of the P _max in Baltic Sea ecotype of F. vesiculosus at higher salinity, could be due to an increase in the relative amounts of Rubisco. The proteins have been evaluated on a relative basis. Immunoblot signals showed that the amount of Rubisco was higher in both ecotypes of F. vesiculosus than in F. radicans, but no differences could be detected between the two ecotypes of F. vesiculosus. The results suggest an uneven photosystem protein stoichiometry in Fucus, with more of the PSI protein PsaA relative to the PSII protein D1. The difference in P _max between the two ecotypes of F. vesiculosus might be related to the difficulties for the algae to adapt to the environment in Bothnian Sea.     

Competitive interactions between Fucus spiralis L. and F. vesiculosus L. (Fucales,Phaeophyta)

Fucus spiralis forms a conspicuous belt in the upper intertidal on rocky shores of the NW Atlantic. The objective of this study was to determine whether competition among congeners plays a role in restricting the distribution of F. spiralis to the upper shore. A replacement series design was used to test the growth performance of F. spiralis and F. vesiculosus in monocultures and in mixtures set up in the mid shore level on an exposed rocky coast. For F. spiralis and F. vesiculosus, all individual and relative crowding coefficients were < 1 and > 1, respectively. Hence, F. vesiculosus succeeded relative to F. spiralis in all mixtures tested. This finding was confirmed by graphical analysis. Ratio diagrams showed that the outcome of long term interaction among the species is extinction of F. spiralis. The results lead to the conclusion that interaction among congeners is a major determinant of the restricted distribution of F. spiralis.     

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.     

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.     

Post-ice age recolonization and differentiation of Fucus serratus L. (Phaeophyceae; Fucaceae) populations in Northern Europe

The seaweed Fucus serratus is hypothesized to have evolved in the North Atlantic and present populations are thought to reflect recolonization from a southern refugium since the last glacial maximum 18 000-20 000 years bp. We examined genetic structure across several spatial scales by analysing seven microsatellite loci in populations collected from 21 localities throughout the species' range. Spatial auto-correlation analysis of seven microsatellite loci revealed no evidence for spatial clustering of alleles on a scale of 100 m despite limited gamete dispersal in F. serratus of approximately 2 m from parental individuals. Pairwise theta analysis suggested that the minimal panmictic unit for F. serratus was between 0.5 and 2 km. Isolation by distance was significant along some contiguous coastlines. Population differentiation was strong within the Skagerrak-Kattegat-Baltic Seas (SKB) (global theta= 0.17) despite a short history of approximately 7500 years. A neighbour-joining tree based on Reynold's distances computed from the microsatellite data revealed a central assemblage of populations on the Brittany Peninsula surrounded by four well-supported clusters consisting of the SKB, the North Sea (Ireland, Helgoland), and two populations from the northern Spanish coast. Samples from Iceland and Nova Scotia were most closely aligned with northwest Sweden and Brittany, respectively. When sample sizes were standardized (N = 41), allelic diversity was twofold higher for Brittany populations than for populations to the north and threefold higher than southern populations. The Brittany region may be a refugium or a recolonized area, whereas the Spanish populations most likely reflect present-day edge populations that have undergone repeated bottlenecks as a consequence of thermally induced cycles of recolonization and extinction.     

Inheritance patterns of ITS1, chloroplasts and mitochondria in artificial hybrids of the seaweeds Fucus serratus and F. evanescens (Phaeophyceae)

Patterns of nuclear and organelle inheritance among artificial hybrids of the seaweeds Fucus serratus and F. evanescens were detected using single-strand conformation polymorphism (SSCP). Three alleles were identified in the 231?bp rDNA-ITS1 gene (nuclear): two in F. serratus and one in F. evanescens. Alleles differed by 1–2?bp and all hybrids possessed one allele from each parent. Two haplotypes were present in the 288?bp Rubisco spacer (chloroplast), differentiated by a 33?bp indel. Two haplotypes differing by a single nucleotide were found in a 135?bp region of nad11 gene (mitochondrion). Both organelles are maternally inherited, as all hybrids contained the haplotypes of the parent contributing the egg. Although laboratory hybrids among Fucus spp. have been produced previously, this is the first time that both nuclear and cytoplasmic genetic markers have been used to document inheritance patterns. SSCPs analysed on an automated sequencer offer a rapid and powerful approach for identifying suspected hybrids from field samples, as well as a screen for intraspecific and intra-individual variation in DNA regions prior to confirmation of variations by sequencing.     

Buffering and Amplifying Interactions among OAW (Ocean Acidification & Warming) and Nutrient Enrichment on Early Life-Stage Fucus vesiculosus L. (Phaeophyceae) and Their Carry Over Effects to Hypoxia Impact

Ocean acidification and warming (OAW) are occurring globally. Additionally, at a more local scale the spreading of hypoxic conditions is promoted by eutrophication and warming. In the semi-enclosed brackish Baltic Sea, occasional upwelling in late summer and autumn may expose even shallow-water communities including the macroalga Fucus vesiculosus to particularly acidified, nutrient-rich and oxygen-poor water bodies. During summer 2014 (July–September) sibling groups of early life-stage F. vesiculosus were exposed to OAW in the presence and absence of enhanced nutrient levels and, subsequently to a single upwelling event in a near-natural scenario which included all environmental fluctuations in the Kiel Fjord, southwestern Baltic Sea, Germany (54°27 ´N, 10°11 ´W). We strove to elucidate the single and combined impacts of these potential stressors, and how stress sensitivity varies among genetically different sibling groups. Enhanced by a circumstantial natural heat wave, warming and acidification increased mortalities and reduced growth in F. vesiculosus germlings. This impact, however, was mitigated by enhanced nutrient conditions. Survival under OAW conditions strongly varied among sibling groups hinting at a substantial adaptive potential of the natural Fucus populations in the Western Baltic. A three-day experimental upwelling caused severe mortality of Fucus germlings, which was substantially more severe in those sibling groups which previously had been exposed to OAW. Our results show that global (OAW), regional (nutrient enrichment) and local pressures (upwelling), both alone and co-occurring may have synergistic and antagonistic effects on survival and/or growth of Fucus germlings. This result emphasizes the need to consider combined stress effects.     

Species richness in macroalgae and macrofauna assemblages on Fucus serratus L. (Phaeophyceae) and Zostera marina L. (Angiospermae) in Skagerrak, Norway

The macrophytes Fucus serratus and Zostera marina form similar substrates for associated flora and fauna in shallow waters in Norway. While F. serratus forms a more or less continuous belt on rocky substrate along the coast, Z. marina forms disjunct populations on sandy or muddy bottoms. This study focused on the organisms associated with these two macrophytes in two localities in the Skagerrak region. In total, 130 taxa of epiphytic organisms were identified: 22 green algae, 41 red algae, 32 brown algae, eight diatoms and 27 sessile animals. One hundred and twenty-seven taxa of mobile macrofauna were registered. The dominant group was crustaceans, with amphipods as the order containing most species. Many species of both plants and animals preferred one or the other habitat. It is concluded that coastal macrophyte systems have high species diversity.     

Role of auxin in induction of polarity in zygotes of Fucus vesiculosus L

We studied the effects of auxin (indolyl-3 acetic acid) on formation of the primary polarity axis in zygotes of the brown algae Fucus vesiculosus. Within the first 2.5 h after fertilization, the zygotes release this phytohormone in the environment. The treatment of developing zygotes with the inhibitor of indolyl-3-acetic acid transport from the cell triiodobenzoic acid at 5 mg/l arrests the auxin secretion and leads to its accumulation in the cells. This causes a significant delay in zygote polarization. The treatment of zygotes with the exogenous indolyl-3-acetic acid at 1 mg/l stimulates cell polarization and formation of a rhizoid process. When auxin was added to the medium with triiodobenzoic acid, the inhibitory effect of the latter was fully relieved. It has been proposed that the content of indolyl-3-acetic acid in the environment is a key factor in the induction of polarity of the F. vesiculosus zygotes.     

Arrested development in Fucus spiralis (Phaeophyceae) germlings exposed to copper

Exposure of Fucus spiralis germlings to precise copper concentrations (0 to 844?nM?Cu²⁺) in chemically defined medium demonstrated a relationship between ultrastructural changes and growth retardation with increasing copper concentration. Electron-translucent vesicles, present in ova, which normally disappear after fertilization, accumulated in germlings exposed to Cu²⁺ above 10.6?nM, suggesting that copper may inhibit a metabolic pathway involved in cell wall formation which is initiated by fertilization. No membrane damage was observed during the exposure period. During a post-exposure period in copper-free medium, recovery occurred (rhizoid extension, apical hair formation) in germlings previously exposed to concentrations below 106?nM?Cu²⁺ and electron-translucent vesicles became granular and disappeared. It is proposed that the electron-translucent vesicles contain a cell wall precursor and that copper inhibits its incorporation into the cell wall, preventing growth and development of the zygote.