Various extracts of the brown seaweed Cystoseira barbata with different compositions exert biostimulant effects on seedling growth of wheat.

Worldwide, where the demand for novel and greener solutions for sustainable agricultural production is increasing, the use of eco-friendly products such as seaweed-derived biostimulants as pre-sowing treatment represent a promising and important approach for the future. Cystoseira barbata, a brown seaweed species abundant in the Mediterranean Region, was collected from the Marmara Sea and subjected to water, alkali, and acidic extractions, and the biostimulant activity of these extracts was tested on wheat (Triticum durum cv. Saricanak-98) using different rates through application to the seeds or germination medium (substrate) applications. The different extracts were characterized by mineral, total phenolic, free amino acid, mannitol, polysaccharide, antioxidant concentrations and hormone-like activity. The effects of the extracts on growth parameters, root morphology, esterase activity, and mineral nutrient concentrations of wheat seedlings were investigated. Our results suggest that the substrate application was more effective in enhancing the seedling performance compared to the seed treatment. High rates of seaweed extracts applied to substrates increased the shoot length and fresh weight of wheat seedlings by up to 20 and 25%, respectively. The substrate applications enhanced the root fresh weights of wheat seedlings by up to 25% when compared to control plants. Among the biostimulant extract applications, the water extract at the highest rate yielded the most promising results in terms of the measured parameters. Cystoseira barbata extracts with different compositions can be used as effective biostimulants to boost seedling growth. The local seaweed biomass affected by mucilage problems, has great potential as a bioeconomy resource and can contribute to sustainable practices for agriculture.

Preservative Effect of Aqueous and Ethanolic Extracts of the Macroalga Bifurcaria bifurcata on the Quality of Chilled Hake (Merluccius merluccius).

This work addressed the preservative behaviour of different icing media containing extracts from the alga Bifurcaria bifurcata. A comparative study of the antimicrobial and antioxidant effects of aqueous and ethanolic extracts of this macroalga was carried out. Whole hake (Merluccius merluccius) pieces were stored in ice containing either kind of extract and analysed for quality changes throughout a 13-day storage period. A progressive loss of microbial and biochemical quality was detected in all batches as chilling time increased. A significant inhibitory effect (p < 0.05) on microbial activity could be observed as a result of including the aqueous (lowering of psychrotrophic and lipolytic counts and pH value) and ethanolic (lowering of psychrotrophic and lipolytic counts) extracts. Additionally, both kinds of extract led to a substantial inhibition (p < 0.05) in the lipid hydrolysis rate (formation of free fatty acids), greater in the case of the batch containing ethanolic extract. Concerning lipid oxidation, a similar inhibitory effect (p < 0.05) on the formation of secondary compounds (thiobarbituric acid substances) was noticed in fish specimens corresponding to both alga extracts; however, more (p < 0.05) peroxide formation was detected in fish corresponding to the ethanolic extract batch. A preservative effect can be concluded for both kinds of extract; this effect agrees with previous studies reporting the presence of hydrophilic and lipophilic bioactive compounds in B. bifurcata.

Transcriptome profiling of the meristem tissue of Saccharina japonica (Phaeophyceae, Laminariales) under severe stress of copper.

Copper (Cu) is an essential metal involved in many physiological processes of living organisms. However, beyond a certain threshold, Cu can become highly toxic. For instance, in the summer sporeling production of the economic kelp Saccharina japonica, the excess Cu accidently released from the low-quality alloys of the refrigerating machine was deadly to the seedlings and led to the failure of hatchery operations. However, the molecular basis underlying high toxicity of Cu remains unclear. In this study, juvenile sporophytes were cultured in seawater containing different concentrations of Cu2+ (10, 100, and 200 µg L-1). Bleaching was observed in the meristem of individuals in the 100 and 200 µg L-1 treatment groups on the third day, indicating that Cu has caused severe harm at these concentrations. RNA-Seq was used to profile transcriptomic changes under different Cu2+ concentrations. Compared with the control, the number of differentially expressed genes (DEGs) was 11,350 (4944 up- and 6406 down-regulated) in the 200 µg L-1 treatment group and 2868 (1075 up- and 1793 down-regulated) in the 100 µg L-1 treatment group, whereas much fewer DEGs were detected in the 10 µg L-1 treatment group. Genes coding for glutathione-S-transferase and vanadium-dependent bromoperoxidase and iodoperoxidase were found to be remarkably regulated, especially in the 200 µg L-1 treatment group. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that only down-regulated DEGs were enriched. There were 45 enriched GO terms and four enriched KEGG pathways common to the 100 and 200 µg L-1 treatment groups, which were associated with diverse essential biological processes such as photosynthesis, protein synthesis, redox activity, and metabolism and biosynthesis of functional biomolecules, among others. Suppression of these biological processes at the transcriptional level likely contributes to the observed high toxicity of Cu2+ in S. japonica.

Ecophysiological and metabolic responses to interactive exposure to nutrients and copper excess in the brown macroalga Cystoseira tamariscifolia.

Global scenarios evidence that contamination due to anthropogenic activities occur at different spatial-temporal scales, being important stressors: eutrophication, due to increased nutrient inputs; and metal pollution, mostly derived from industrial activities. In this study, we investigated ecophysiological and metabolic responses to copper and nutrient excess in the brown macroalga Cystoseira tamariscifolia. Whole plants were incubated in an indoor system under control conditions, two levels of nominal copper (0.5 and 2.0µM), and two levels of nutrient supply for two weeks. Maximal quantum yield (Fv/Fm) and maximal electron transport rate (ETRmax) increased under copper exposure. Photosynthetic pigments and phenolic compounds (PC) increased under the highest copper levels. The intra-cellular copper content increased under high copper exposure in both nutrient conditions. C. tamariscifolia from the Atlantic displayed efficient metal exclusion mechanisms, since most of the total copper accumulated by the cell was bound to the cell wall.

Effects of elevated nutrients and CO2 emission scenarios on three coral reef macroalgae.

Coral reef macroalgae are expected to thrive in the future under conditions that are deleterious to the health of reef-building corals. Here we examined how macroalgae would be affected by exposure to future CO2 emission scenarios (pCO2 and temperature), enriched nutrients and combinations of both. The species tested, Laurencia intricata (Rhodophyta), Turbinaria ornata and Chnoospora implexa (both Phaeophyceae), have active carbon-concentrating mechanisms but responded differently to the treatments. L. intricata showed high mortality under nutrient enriched RCP4.5 ("reduced" CO2 emission) and RCP8.5 ("business-as-usual" CO2 emission) and grew best under pre-industrial (PI) conditions, where it could take up carbon using external carbonic anhydrase combined, potentially, with proton extrusion. T. ornata's growth rate showed a trend for reduction under RCP8.5 but was unaffected by nutrient enrichment. In C. implexa, highest growth was observed under PI conditions, but highest net photosynthesis occurred under RCP8.5, suggesting that under RCP8.5, carbon is stored and respired at greater rates while it is directed to growth under PI conditions. None of the species showed growth enhancement under future scenarios, nutrient enrichment or combinations of both. This leads to the conclusion that under such conditions these species are unlikely to pose an increasing threat to coral reefs.

Predicting herbicide mixture effects on multiple algal species using mixture toxicity models.

The validity of the application of mixture toxicity models, concentration addition and independent action, to a species sensitivity distribution (SSD) for calculation of a multisubstance potentially affected fraction was examined in laboratory experiments. Toxicity assays of herbicide mixtures using 5 species of periphytic algae were conducted. Two mixture experiments were designed: a mixture of 5 herbicides with similar modes of action and a mixture of 5 herbicides with dissimilar modes of action, corresponding to the assumptions of the concentration addition and independent action models, respectively. Experimentally obtained mixture effects on 5 algal species were converted to the fraction of affected (>50% effect on growth rate) species. The predictive ability of the concentration addition and independent action models with direct application to SSD depended on the mode of action of chemicals. That is, prediction was better for the concentration addition model than the independent action model for the mixture of herbicides with similar modes of action. In contrast, prediction was better for the independent action model than the concentration addition model for the mixture of herbicides with dissimilar modes of action. Thus, the concentration addition and independent action models could be applied to SSD in the same manner as for a single-species effect. The present study to validate the application of the concentration addition and independent action models to SSD supports the usefulness of the multisubstance potentially affected fraction as the index of ecological risk. Environ Toxicol Chem 2017;36:2624-2630. © 2017 SETAC.

High-density genetic map and identification of QTLs for responses to temperature and salinity stresses in the model brown alga Ectocarpus.

Deciphering the genetic architecture of adaptation of brown algae to environmental stresses such as temperature and salinity is of evolutionary as well as of practical interest. The filamentous brown alga Ectocarpus sp. is a model for the brown algae and its genome has been sequenced. As sessile organisms, brown algae need to be capable of resisting the various abiotic stressors that act in the intertidal zone (e.g. osmotic pressure, temperature, salinity, UV radiation) and previous studies have shown that an important proportion of the expressed genes is regulated in response to hyposaline, hypersaline or oxidative stress conditions. Using the double digest RAD sequencing method, we constructed a dense genetic map with 3,588 SNP markers and identified 39 QTLs for growth-related traits and their plasticity under different temperature and salinity conditions (tolerance to high temperature and low salinity). GO enrichment tests within QTL intervals highlighted membrane transport processes such as ion transporters. Our study represents a significant step towards deciphering the genetic basis of adaptation of Ectocarpus sp. to stress conditions and provides a substantial resource to the increasing list of tools generated for the species.

Increased pCO2 and temperature reveal ecotypic differences in growth and photosynthetic performance of temperate and Arctic populations of Saccharina latissima.

MAIN CONCLUSION: The Arctic population of the kelp Saccharina latissima differs from the Helgoland population in its sensitivity to changing temperature and CO 2 levels. The Arctic population does more likely benefit from the upcoming environmental scenario than its Atlantic counterpart. The previous research demonstrated that warming and ocean acidification (OA) affect the biochemical composition of Arctic (Spitsbergen; SP) and cold-temperate (Helgoland; HL) Saccharina latissima differently, suggesting ecotypic differentiation. This study analyses the responses to different partial pressures of CO2 (380, 800, and 1500 µatm pCO2) and temperature levels (SP population: 4, 10 °C; HL population: 10, 17 °C) on the photophysiology (O2 production, pigment composition, D1-protein content) and carbon assimilation [Rubisco content, carbon concentrating mechanisms (CCMs), growth rate] of both ecotypes. Elevated temperatures stimulated O2 production in both populations, and also led to an increase in pigment content and a deactivation of CCMs, as indicated by 13C isotopic discrimination of algal biomass (ε p) in the HL population, which was not observed in SP thalli. In general, pCO2 effects were less pronounced than temperature effects. High pCO2 deactivated CCMs in both populations and produced a decrease in the Rubisco content of HL thalli, while it was unaltered in SP population. As a result, the growth rate of the Arctic ecotype increased at elevated pCO2 and higher temperatures and it remained unchanged in the HL population. Ecotypic differentiation was revealed by a significantly higher O2 production rate and an increase in Chl a, Rubisco, and D1 protein content in SP thalli, but a lower growth rate, in comparison to the HL population. We conclude that both populations differ in their sensitivity to changing temperatures and OA and that the Arctic population is more likely to benefit from the upcoming environmental scenario than its Atlantic counterpart.

Modulatory effect of the exudates released by the brown kelp Lessonia spicata on the toxicity of copper in early developmental stages of ecologically related organisms.

Lessonia spicata is a key dominant species along the Pacific coast of South America, providing a habitat for many organisms. However, this role can be affected by abiotic stress, such as metals. To counteract the toxic effect, L. spicata, among other seaweeds, releases exudates that bind metals. In this study, tolerances to copper of organisms related to the kelp forest (spores of Ulva lactuca (Chlorophyceae) and L. spicata (Phaeophyceae) and Zoea I of Taliepus dentatus (Milne-Edwards, Crustacea)) were studied; then, exudates are assessed by their protective effect. Exudates increase the 48-h 50% effective concentration (EC50) of the germination of spores from 8 to 23 µg Cu L-1 for U. lactuca and from 119 to 213 µg Cu L-1 for L. spicata and the survival of the larvae Zoea I 48-h 50% of lethal concentration (LC50) from 144 to 249 µg Cu L-1. Results indicated that exudates had a protective effect. Each species is specifically sensitive to copper. Crab larvae Zoea I were able to support higher doses, and exposure before hatching increased their tolerance.

Co-contamination of Cu and Cd in paddy fields: Using periphyton to entrap heavy metals.

The ubiquitous native periphyton was used to entrap Cu and Cd from paddy fields. Results showed that Cu- and Cd-hydrate species such as CuOH(+), Cu2(OH)2(2+), CdOH(+), and Cu3(OH)4(2+) decreased with time in the presence of periphyton. When the initial concentrations of Cu and Cd were 10mg/L, the heavy metal content in the periphyton fluctuated from 145.20mg/kg to 342.42 mg/kg for Cu and from 101.75 mg/kg to 236.29 mg/kg for Cd after 2h exposure. The concentration of Cd in periphytic cells varied from 42.93 mg/kg to 174 mg/kg after 2h. The dominant periphyton microorganism species shifted from photoautotrophs to heterotrophs during the exposure of periphyton to Cu and Cd co-contamination. Although Cu and Cd could inhibit periphyton photosynthesis and carbon utilization, the periphyton was able to adapt to the test conditions. Cu and Cd accumulation in rice markedly decreased in the presence of periphyton while the number of rice seeds germinating was higher in the periphyton treatments. These results suggest that the inclusion of native periphyton in paddy fields provides a promising buffer to minimize the effects of Cu and Cd pollution on rice growth and food safety.

A novel field transplantation technique reveals intra-specific metal-induced oxidative responses in strains of Ectocarpus siliculosus with different pollution histories.

A novel field transplantation technique, in which seaweed material is incorporated into dialysis tubing, was used to investigate intra-specific responses to metals in the model brown alga Ectocarpus siliculosus. Metal accumulation in the two strains was similar, with higher concentrations in material deployed to the metal-contaminated site (Ventanas, Chile) than the pristine site (Quintay, Chile). However, the oxidative responses differed. At Ventanas, strain Es147 (from low-polluted site) underwent oxidative damage whereas Es524 (from highly polluted site) was not affected. Concentrations of reduced ascorbate (ASC) and reduced glutathione (GSH) were significantly higher in Es524. Activities of the antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), and glutathione reductase (GR) all increased in Es524, whereas only SOD increased in Es147. For the first time, employing a field transplantation technique, we provide unambiguous evidence of inter-population variation of metal-tolerance in brown algae and establish that antioxidant defences are, in part, responsible.

Optimization of volatile fatty acids and hydrogen production from Saccharina japonica: acidogenesis and molecular analysis of the resulting microbial communities.

Response surface methodology (RSM) was used to optimize the production of volatile fatty acids (VFAs) and hydrogen from mixed anaerobic cultures of Saccharina japonica with respect to two independent variables: methanogenic inhibitor concentration and temperature. The effects of four methanogenic inhibitors on acidogenic processes were tested, and qualitative microbial analyses were carried out. Escherichia, Acinetobacter, and Clostridium were the most predominant genera in samples treated with chloroform (CHCl3), iodoform (CHI3), 2-bromoethanesulfonate (BES), or ß-cyclodextrin (ß-CD), respectively. RSM showed that the production of VFAs reached a peak of 12.5 g/L at 38.6 °C in the presence of 7.4 g/L ß-CD; these were the conditions under which hydrogen production was also nearly maximal. The quantitative polymerase chain reaction (qPCR) showed that shifts in the bacterial community population correlated with the concentrations of ß-CD indicating that this compound effectively inhibited methanogens.

Response differences between Ectocarpus siliculosus populations to copper stress involve cellular exclusion and induction of the phytochelatin biosynthetic pathway.

Some populations of brown seaweed species inhabit metal-polluted environments and can develop tolerance to metal stress, but the mechanisms by which this is accomplished are still to be elucidated. To address this, the responses of two strains of the model brown alga Ectocarpus siliculosus isolated from sites with different histories of metal contamination exposed to total copper (CuT) concentrations ranging between 0 and 2.4 µM for 10 days were investigated. The synthesis of the metal-chelator phytochelatin (PCs) and relative levels of transcripts encoding the enzymes γ-glutamylcysteine synthetase (γ-GCS), glutathione synthase (GS) and phytochelatin synthase (PCS) that participate in the PC biosynthetic pathway were measured, along with the effects on growth, and adsorption and uptake of Cu. Growth of strain LIA, from a pristine site in Scotland, was inhibited to a greater extent, and at lower concentrations, than that of Es524, isolated from a Cu-contaminated site in Chile. Concentrations of intra-cellular Cu were higher and the exchangeable fraction was lower in LIA than Es524, especially at the highest exposure levels. Total glutathione concentrations increased in both strains with Cu exposure, whereas total PCs levels were higher in Es524 than LIA; PC2 and PC3 were detected in Es524 but PC2 only was found in LIA. The greater production and levels of polymerisation of PCs in Es524 can be explained by the up-regulation of genes encoding for key enzymes involved in the synthesis of PCs. In Es524 there was an increase in the transcripts of γ-GCS, GS and PCS, particularly under high Cu exposure, whereas in LIA4 transcripts of γ-GCS1 increased only slightly, γ-GCS2 and GS decreased and PCS did not change. The consequences of higher intra-cellular concentrations of Cu, lower production of PCs, and lower expression of enzymes involved in GSH-PCs synthesis may be contributing to an induced oxidative stress condition in LIA, which explains, at least in part, the observed sensitivity of LIA to Cu. Therefore, responses to Cu exposure in E. siliculosus relate to the contamination histories of the locations from where the strains were isolated and differences in Cu exclusion and PCs production are in part responsible for the development of intra-specific resistance.

Copper-induced intra-specific oxidative damage and antioxidant responses in strains of the brown alga Ectocarpus siliculosus with different pollution histories.

Inter- and intra-specific variation in metal resistance has been observed in the ecologically and economically important marine brown macroalgae (Phaeophyceae), but the mechanisms of cellular tolerance are not well elucidated. To investigate inter-population responses of brown seaweeds to copper (Cu) pollution, the extent of oxidative damage and antioxidant responses were compared in three strains of the filamentous brown seaweed Ectocarpus siliculosus, the model organism for the algal class Phaeophyceae that diverged from other major eukaryotic groups over a billion year ago. Strains isolated from locations with different pollution histories (i.e. LIA, from a pristine site in Scotland; REP and Es524 from Cu-contaminated sites in England and Chile, respectively) were exposed to total dissolved Cu concentrations (CuT) of up to 2.4 µM (equivalent to 128 nM Cu(2+)) for 10 d. LIA exhibited oxidative stress, with increases in hydrogen peroxide (H2O2) and lipid peroxidation (measured as TBARS levels), and decreased concentrations of photosynthetic pigments. Es524 presented no apparent oxidative damage whereas in REP, TBARS increased, revealing some level of oxidative damage. Adjustments to activities of enzymes and antioxidant compounds concentrations in Es524 and REP were strain and treatment dependent. Mitigation of oxidative stress in Es524 was by increased activities of superoxide dismutases (SOD) at low CuT, and catalase (CAT) and ascorbate peroxidase (APX) at all CuT, accompanied by higher levels of antioxidants (ascorbate, glutathione, phenolics) at higher CuT. In REP, only APX activity increased, as did the antioxidants. For the first time evidence is presented for distinctive oxidative stress defences under excess Cu in two populations of a species of brown seaweed from environments contaminated by Cu.

Effects of ocean acidification on the brown alga Padina pavonica: decalcification due to acute and chronic events.

Since the industrial revolution, anthropogenic CO2 emissions have caused ocean acidification, which particularly affects calcified organisms. Given the fan-like calcified fronds of the brown alga Padina pavonica, we evaluated the acute (short-term) effects of a sudden pH drop due to a submarine volcanic eruption (October 2011-early March 2012) affecting offshore waters around El Hierro Island (Canary Islands, Spain). We further studied the chronic (long-term) effects of the continuous decrease in pH in the last decades around the Canarian waters. In both the observational and retrospective studies (using herbarium collections of P. pavonica thalli from the overall Canarian Archipelago), the percent of surface calcium carbonate coverage of P. pavonica thalli were contrasted with oceanographic data collected either in situ (volcanic eruption event) or from the ESTOC marine observatory data series (herbarium study). Results showed that this calcified alga is sensitive to acute and chronic environmental pH changes. In both cases, pH changes predicted surface thallus calcification, including a progressive decalcification over the last three decades. This result concurs with previous studies where calcareous organisms decalcify under more acidic conditions. Hence, Padina pavonica can be implemented as a bio-indicator of ocean acidification (at short and long time scales) for monitoring purposes over wide geographic ranges, as this macroalga is affected and thrives (unlike strict calcifiers) under more acidic conditions.

Biochemical composition of temperate and Arctic populations of Saccharina latissima after exposure to increased pCO2 and temperature reveals ecotypic variation.

Previous research suggested that the polar and temperate populations of the kelp Saccharina latissima represent different ecotypes. The ecotypic differentiation might also be reflected in their biochemical composition (BC) under changing temperatures and pCO2. Accordingly, it was tested if the BC of Arctic (Spitsbergen) and temperate S. latissima (Helgoland) is different and if they are differently affected by changes in temperature and pCO2. Thalli from Helgoland grown at 17 °C and 10 °C and from Spitsbergen at 10 °C and 4 °C were all tested at either 380, 800, or 1,500 µatm pCO2, and total C-, total N-, protein, soluble carbohydrate, and lipid content, as well as C/N-ratio were measured. At 10 °C, the Arctic population had a higher content of total C, soluble carbohydrates, and lipids, whereas the N- and protein content was lower. At the lower tested temperature, the Arctic ecotype had particularly higher contents of lipids, while content of soluble carbohydrates increased in the Helgoland population only. In Helgoland-thalli, elevated pCO2 caused a higher content of soluble carbohydrates at 17 °C but lowered the content of N and lipids and increased the C/N-ratio at 10 °C. Elevated pCO2 alone did not affect the BC of the Spitsbergen population. Conclusively, the Arctic ecotype was more resilient to increased pCO2 than the temperate one, and both ecotypes differed in their response pattern to temperature. This differential pattern is discussed in the context of the adaptation of the Arctic ecotype to low temperature and the polar night.

Transcriptomic and metabolomic analysis of copper stress acclimation in Ectocarpus siliculosus highlights signaling and tolerance mechanisms in brown algae.

BACKGROUND: Brown algae are sessile macro-organisms of great ecological relevance in coastal ecosystems. They evolved independently from land plants and other multicellular lineages, and therefore hold several original ontogenic and metabolic features. Most brown algae grow along the coastal zone where they face frequent environmental changes, including exposure to toxic levels of heavy metals such as copper (Cu). RESULTS: We carried out large-scale transcriptomic and metabolomic analyses to decipher the short-term acclimation of the brown algal model E. siliculosus to Cu stress, and compared these data to results known for other abiotic stressors. This comparison demonstrates that Cu induces oxidative stress in E. siliculosus as illustrated by the transcriptomic overlap between Cu and H2O2 treatments. The common response to Cu and H2O2 consisted in the activation of the oxylipin and the repression of inositol signaling pathways, together with the regulation of genes coding for several transcription-associated proteins. Concomitantly, Cu stress specifically activated a set of genes coding for orthologs of ABC transporters, a P1B-type ATPase, ROS detoxification systems such as a vanadium-dependent bromoperoxidase, and induced an increase of free fatty acid contents. Finally we observed, as a common abiotic stress mechanism, the activation of autophagic processes on one hand and the repression of genes involved in nitrogen assimilation on the other hand. CONCLUSIONS: Comparisons with data from green plants indicate that some processes involved in Cu and oxidative stress response are conserved across these two distant lineages. At the same time the high number of yet uncharacterized brown alga-specific genes induced in response to copper stress underlines the potential to discover new components and molecular interactions unique to these organisms. Of particular interest for future research is the potential cross-talk between reactive oxygen species (ROS)-, myo-inositol-, and oxylipin signaling.

Physiological plasticity of Dictyota kunthii (Phaeophyceae) to copper excess.

The brown alga Dictyota kunthii is one of the dominant species in the coastal areas of northern Chile affected by copper enrichment due to accumulated mining wastes. To assess its physiological plasticity in handling copper-mediated oxidative stress, 4-days copper exposure (ca. 100 µg/L) experiments were conducted with individuals from a copper impacted area and compared with the responses of plants from a non-impacted site. Several biochemical parameters were then evaluated and compared between populations. Results showed that individuals from the copper-impacted population normally displayed higher levels of copper content and antioxidant enzymes activity (catalase (CAT), ascorbate peroxidase (AP), dehydroascorbate reductase (DHAR), glutathione peroxidase (GP) and peroxiredoxins (PRX)). After copper exposure, antioxidant enzyme activity increased significantly in plants from the two selected sites. In addition, we found that copper-mediated oxidative stress was associated with a reduction of glutathione reductase (GR) activity. Moreover, metabolic profiling of extracellular metabolites from both populations showed a significant change after plants were exposed to copper excess in comparison with controls, strongly suggesting a copper-induced release of metabolites. The copper-binding capacity of those exudates was determined by anodic stripping voltammetry (ASV) and revealed an increased ligand capacity of the medium with plants exposed to copper excess. Results indicated that D. kunthii, regardless their origin, counteracts copper excess by various mechanisms, including metal accumulation, activation of CAT, AP, DHAR, GP and PRX, and an induced release of Cu binding compounds. Thus, plasticity in copper tolerance in D. kunthii seems constitutive, and the occurrence of a copper-tolerant ecotype seems unlikely.

Surface-bound iron: a metal ion buffer in the marine brown alga Ectocarpus siliculosus?

Although the iron uptake and storage mechanisms of terrestrial/higher plants have been well studied, the corresponding systems in marine algae have received far less attention. Studies have shown that while some species of unicellular algae utilize unique mechanisms of iron uptake, many acquire iron through the same general mechanisms as higher plants. In contrast, the iron acquisition strategies of the multicellular macroalgae remain largely unknown. This is especially surprising since many of these organisms represent important ecological and evolutionary niches in the coastal marine environment. It has been well established in both laboratory and environmentally derived samples, that a large amount of iron can be 'non-specifically' adsorbed to the surface of marine algae. While this phenomenon is widely recognized and has prompted the development of experimental protocols to eliminate its contribution to iron uptake studies, its potential biological significance as a concentrated iron source for marine algae is only now being recognized. This study used an interdisciplinary array of techniques to explore the nature of the extensive and powerful iron binding on the surface of both laboratory and environmental samples of the marine brown alga Ectocarpus siliculosus and shows that some of this surface-bound iron is eventually internalized. It is proposed that the surface-binding properties of E. siliculosus allow it to function as a quasibiological metal ion 'buffer', allowing iron uptake under the widely varying external iron concentrations found in coastal marine environments.

Non-cell autonomous regulation of life cycle transitions in the model brown alga Ectocarpus.

The model brown alga Ectocarpus has a haploid-diploid life cycle, involving alternation between two independent multicellular generations, the gametophyte and the sporophyte. Recent work has shown that alternation of generations is not determined by ploidy but is rather under genetic control, involving at least one master regulatory locus, OUROBOROS (ORO). Using cell biology approaches combined with measurements of generation-specific transcript abundance we provide evidence that alternation of generations can also be regulated by non-cell autonomous mechanisms. The Ectocarpus sporophyte produces a diffusible factor that causes major developmental reprogramming in gametophyte cells. Cells become resistant to reprogramming when the cell wall is synthetized, suggesting that the cell wall may play a role in locking an individual into the developmental program that has been engaged. A functional ORO gene is necessary for the induction of the developmental switch. Our results highlight the role of the cell wall in maintaining the differentiated generation stage once the appropriate developmental program has been engaged and also indicate that ORO is a key member of the developmental pathway triggered by the sporophyte factor. Alternation between gametophyte and sporophyte generations in Ectocarpus is surprisingly labile, perhaps reflecting an adaptation to the variable seashore environment inhabited by this alga.