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.

Highly restricted dispersal in habitat-forming seaweed may impede natural recovery of disturbed populations.

Cystoseira sensu lato (Class Phaeophyceae, Order Fucales, Family Sargassaceae) forests play a central role in marine Mediterranean ecosystems. Over the last decades, Cystoseira s.l. suffered from a severe loss as a result of multiple anthropogenic stressors. In particular, Gongolaria barbata has faced multiple human-induced threats, and, despite its ecological importance in structuring rocky communities and hosting a large number of species, the natural recovery of G. barbata depleted populations is uncertain. Here, we used nine microsatellite loci specifically developed for G. barbata to assess the genetic diversity of this species and its genetic connectivity among fifteen sites located in the Ionian, the Adriatic and the Black Seas. In line with strong and significant heterozygosity deficiencies across loci, likely explained by Wahlund effect, high genetic structure was observed among the three seas (ENA corrected FST = 0.355, IC = [0.283, 0.440]), with an estimated dispersal distance per generation smaller than 600 m, both in the Adriatic and Black Sea. This strong genetic structure likely results from restricted gene flow driven by geographic distances and limited dispersal abilities, along with genetic drift within isolated populations. The presence of genetically disconnected populations at small spatial scales (< 10 km) has important implications for the identification of relevant conservation and management measures for G. barbata: each population should be considered as separated evolutionary units with dedicated conservation efforts.

Production and monitoring of biomass and fucoxanthin with brown microalgae under outdoor conditions.

The effect of light on biomass and fucoxanthin (Fx) productivities was studied in two microalgae, Tisochrysis lutea and Phaeodactylum tricornutum. High and low biomass concentrations (1.1 and 0.4 g L-1 ) were tested in outdoor pilot-scale flat-panel photobioreactors at semi-continuous cultivation mode. Fluorescence spectroscopy coupled with chemometric modeling was used to develop prediction models for Fx content and for biomass concentration to be applied for both microalgae species. Prediction models showed high R2 for cell concentration (.93) and Fx content (.77). Biomass productivity was lower for high biomass concentration than low biomass concentration, for both microalgae (1.1 g L-1 : 75.66 and 98.14 mg L-1 d-1 , for T. lutea and P. tricornutum, respectively; 0.4 g L-1 : 129.9 and 158.47 mg L-1 d-1 , T. lutea and P. tricornutum). The same trend was observed in Fx productivity (1.1 g L-1 : 1.14 and 1.41 mg L-1 d-1 , T. lutea and P. tricornutum; 0.4 g L-1 : 2.09 and 1.73 mg L-1 d-1 , T. lutea and P. tricornutum). These results show that biomass and Fx productivities can be set by controlling biomass concentration under outdoor conditions and can be predicted using fluorescence spectroscopy. This monitoring tool opens new possibilities for online process control and optimization.

Impairment of microbial and meiofaunal ecosystem functions linked to algal forest loss.

Habitat loss is jeopardizing marine biodiversity. In the Mediterranean Sea, the algal forests of Cystoseira spp. form one of the most complex, productive and vulnerable shallow-water habitats. These forests are rapidly regressing with negative impact on the associated biodiversity, and potential consequences in terms of ecosystem functioning. Here, by comparing healthy Cystoseira forests and barren grounds (i.e., habitats where the macroalgal forests disappeared), we assessed the effects of habitat loss on meiofaunal and nematode biodiversity, and on some ecosystem functions (here measured in terms of prokaryotic and meiofaunal biomass). Overall, our results suggest that the loss of Cystoseira forests and the consequent barren formation is associated with the loss of meiofaunal higher taxa and a decrease of nematode biodiversity, leading to the collapse of the microbial and meiofaunal variables of ecosystem functions. We conclude that, given the very limited resilience of these ecosystems, active restoration of these vulnerable habitats is needed, in order to recover their biodiversity, ecosystem functions and associated services.

Genome-wide analysis of the Saccharina japonica sulfotransferase genes and their transcriptional profiles during whole developmental periods and under abiotic stresses.

BACKGROUND: As a unique sulfated polysaccharide, fucoidan is an important component of cell wall in brown seaweeds. Its biochemical properties are determined by the positions and quantity of sulfate groups. Sulfotransferases (STs) catalyze the sulfation process, which transfer the sulfuryl groups to carbohydrate backbones and are crucial for fucoidan biosynthesis. Nevertheless, the structures and functions of STs in brown seaweeds are rarely investigated. RESULTS: There are a total of 44 ST genes identified from our genome and transcriptome analysis of Saccharina japonica, which were located in the 17 scaffolds and 11 contigs. The S. japonica ST genes have abundant introns and alternative splicing sites, and five tandem duplicated gene clusters were identified. Generally, the ST genes could be classified into five groups (Group I ~ V) based on phylogenetic analysis. Accordingly, the ST proteins, which were encoded by genes within the same group, contained similar conserved motifs. Members of the S. japonica ST gene family show various expression patterns in different tissues and developmental stages. Transcriptional profiles indicate that the transcriptional levels of more than half of the ST genes are higher in kelp basal blades than in distal blades. Except for ST5 and ST28, most ST genes are down-regulated with the kelp development stages. The expression levels of nine ST genes were detected by real-time quantitative PCR, which demonstrates that they responded to low salinity and drought stresses. CONCLUSIONS: Various characteristics of the STs allow the feasibilities of S. japonica to synthesize fucoidans with different sulfate groups. This enables the kelp the potential to adapt to the costal environments and meet the needs of S. japonica growth.

Three-dimensional growth: a developmental innovation that facilitated plant terrestrialization.

One of the most transformative events in the history of life on earth was the transition of plants from water to land approximately 470 million years ago. Within the Charophyte green algae, the closest living relatives of land plants, body plans have evolved from those that comprise simple unicells to those that are morphologically complex, large and multicellular. The Charophytes developed these broad ranging body plans by exploiting a range of one-dimensional and two-dimensional growth strategies to produce filaments, mats and branches. When plants were confronted with harsh conditions on land, they were required to make significant changes to the way they shaped their body plans. One of the fundamental developmental transitions that occurred was the evolution of three-dimensional growth and the acquisition of apical cells with three or more cutting faces. Plants subsequently developed a range of morphological adaptations (e.g. vasculature, roots, flowers, seeds) that enabled them to colonise progressively drier environments. 3D apical growth also evolved convergently in the brown algae, completely independently of the green lineage. This review summarises the evolving developmental complexities observed in the early divergent Charophytes all the way through to the earliest conquerors of land, and investigates 3D apical growth in the brown algae.

Design Principles of Branching Morphogenesis in Filamentous Organisms.

The radiation of life on Earth was accompanied by the diversification of multicellular body plans in the eukaryotic kingdoms Animalia, Plantae, Fungi and Chromista. Branching forms are ubiquitous in nature and evolved repeatedly in the above lineages. The developmental and genetic basis of branch formation is well studied in the three-dimensional shoot and root systems of land plants, and in animal organs such as the lung, kidney, mammary gland, vasculature, etc. Notably, recent thought-provoking studies combining experimental analysis and computational modeling of branching patterns in whole animal organs have identified global patterning rules and proposed unifying principles of branching morphogenesis. Filamentous branching forms represent one of the simplest expressions of the multicellular body plan and constitute a key step in the evolution of morphological complexity. Similarities between simple and complex branching forms distantly related in evolution are compelling, raising the question whether shared mechanisms underlie their development. Here, we focus on filamentous branching organisms that represent major study models from three distinct eukaryotic kingdoms, including the moss Physcomitrella patens (Plantae), the brown alga Ectocarpus sp. (Chromista), and the ascomycetes Neurospora crassa and Aspergillus nidulans (Fungi), and bring to light developmental regulatory mechanisms and design principles common to these lineages. Throughout the review we explore how the regulatory mechanisms of branching morphogenesis identified in other models, and in particular animal organs, may inform our thinking on filamentous systems and thereby advance our understanding of the diverse strategies deployed across the eukaryotic tree of life to evolve similar forms.

A key role for sex chromosomes in the regulation of parthenogenesis in the brown alga Ectocarpus.

Although evolutionary transitions from sexual to asexual reproduction are frequent in eukaryotes, the genetic bases of these shifts remain largely elusive. Here, we used classic quantitative trait analysis, combined with genomic and transcriptomic information to dissect the genetic basis of asexual, parthenogenetic reproduction in the brown alga Ectocarpus. We found that parthenogenesis is controlled by the sex locus, together with two additional autosomal loci, highlighting the key role of the sex chromosome as a major regulator of asexual reproduction. We identify several negative effects of parthenogenesis on male fitness, and different fitness effects of parthenogenetic capacity depending on the life cycle generation. Although allele frequencies in natural populations are currently unknown, we discuss the possibility that parthenogenesis may be under both sex-specific selection and generation/ploidally-antagonistic selection, and/or that the action of fluctuating selection on this trait may contribute to the maintenance of polymorphisms in populations. Importantly, our data provide the first empirical illustration, to our knowledge, of a trade-off between the haploid and diploid stages of the life cycle, where distinct parthenogenesis alleles have opposing effects on sexual and asexual reproduction and may help maintain genetic variation. These types of fitness trade-offs have profound evolutionary implications in natural populations and may structure life history evolution in organisms with haploid-diploid life cycles.

Algal Sex Determination and the Evolution of Anisogamy.

Algae are photosynthetic eukaryotes whose taxonomic breadth covers a range of life histories, degrees of cellular and developmental complexity, and diverse patterns of sexual reproduction. These patterns include haploid- and diploid-phase sex determination, isogamous mating systems, and dimorphic sexes. Despite the ubiquity of sexual reproduction in algae, their mating-type-determination and sex-determination mechanisms have been investigated in only a limited number of representatives. These include volvocine green algae, where sexual cycles and sex-determining mechanisms have shed light on the transition from mating types to sexes, and brown algae, which are a model for UV sex chromosome evolution in the context of a complex haplodiplontic life cycle. Recent advances in genomics have aided progress in understanding sexual cycles in less-studied taxa including ulvophyte, charophyte, and prasinophyte green algae, as well as in diatoms.

Diverse responses of sporophytic photochemical efficiency and gametophytic growth for two edible kelps, Saccharina japonica and Undaria pinnatifida, to ocean acidification and warming.

Ocean acidification and warming represent major environmental threats to kelp mariculture. In this study, sporophytic photochemical efficiency and gametophytic growth of Saccharina japonica and Undaria pinnatifida were evaluated under different pCO2 levels (360, 720, and 980 ppmv) and temperatures (5, 10, 15, and 20 °C for sporophytes; 15 and 20 °C for gametophytes). Sporophytic photochemical efficiencies of both kelps were significantly greater at 720 ppmv than at 360 and 980 ppmv. Female gametophytes of both kelps grew significantly better at 360 ppmv than at higher pCO2 levels. The growth of U. pinnatifida gametophytes was significantly greater at 20 °C than at 15 °C, while no significant difference was observed for the growth of S. japonica. These results indicate that increased pCO2 stimulated sporophytic photochemical efficiency while inhibited gametophytic growth of these kelps, which might negatively affect their seedling cultivation. U. pinnatifida exhibited higher productivity in warmer ocean than S. japonica.

Climate change induced range shifts in seaweeds distributions in Europe.

There are evidences of how climate change is affecting seaweeds distribution and the ecosystems services they provide. Therefore, it is necessary to consider these impacts when managing marine areas. One of the most applied tools in recent years to deal with this are species distribution models, however there are still some challenges to solve, such as the inclusion of hydrodynamic predictors and the application of effective, transferable and user-oriented methodologies. Five species (Saccorhiza polyschides, Gelidium spinosum, Sargassum muticum, Pelvetia canaliculata and Cystoseira baccata) in Europe and 15 variables were considered. Nine of them were projected to the RCPs 4.5 and 8.5 for the mid-term (2040-2069) and the long term (2070-2099). Algorithms for each species were applied to generate models that were assessed by comparison of probabilities and observations (area under the curve, true skill statistics, Boyce index, sensitivity, correct classification rate), niches overlap (Schoener's D, Hellinger's I), geographical similarity (interquartile range) and ecological realism. Models built demonstrated very good predictive accuracy and sensitivity, without overfitting risk. A medium overlap in the historical and RCPs environmental conditions were obtained, therefore the models can be considered transferable and results accurate because only some isolated points were detected as outliers, corresponding to low probabilities. The areas of S. polyschides and G. spinosum have been identified to be dramatically reduced, meanwhile S. muticum and C. baccata were predicted to expand their range. P. canaliculata was expected to keep its sites of presence but with a decrease in its probability of occurrence. For all species it was remarkable the importance of hydrodynamic variables and parameters representing extreme conditions. Spatially predictions of the potential species and areas at risk are decisive for defining management strategies and resource allocation. The performance and usefulness of the approach applied in this study have been demonstrated for algae with different ecological requirements (from upper littoral to subtidal) and distributional patterns (native and invasive), therefore results can be used by marine planners with different goals: marine protected areas designation, monitoring efforts guiding, invasions risk assessment or aquaculture facilities zonation.

Pretreatment Processes of Biomass for Biorefineries: Current Status and Prospects.

This article seeks to be a handy document for the academy and the industry to get quickly up to speed on the current status and prospects of biomass pretreatment for biorefineries. It is divided into two biomass sources: vegetal and animal. Vegetal biomass is the material produced by plants on land or in water (algae), consuming sunlight, CO2, water, and soil nutrients. This includes residues or main products from, for example, intensive grass crops, forestry, and industrial and agricultural activities. Animal biomass is the residual biomass generated from the production of food from animals (e.g., manure and whey). This review does not mean to include every technology in the area, but it does evaluate physical pretreatments, microwave-assisted extraction, and water treatments for vegetal biomass. A general review is given for animal biomass based in physical, chemical, and biological pretreatments.

Rapid turnover of life-cycle-related genes in the brown algae.

BACKGROUND: Sexual life cycles in eukaryotes involve a cyclic alternation between haploid and diploid phases. While most animals possess a diploid life cycle, many plants and algae alternate between multicellular haploid (gametophyte) and diploid (sporophyte) generations. In many algae, gametophytes and sporophytes are independent and free-living and may present dramatic phenotypic differences. The same shared genome can therefore be subject to different, even conflicting, selection pressures during each of the life cycle generations. Here, we analyze the nature and extent of genome-wide, generation-biased gene expression in four species of brown algae with contrasting levels of dimorphism between life cycle generations. RESULTS: We show that the proportion of the transcriptome that is generation-specific is broadly associated with the level of phenotypic dimorphism between the life cycle stages. Importantly, our data reveals a remarkably high turnover rate for life-cycle-related gene sets across the brown algae and highlights the importance not only of co-option of regulatory programs from one generation to the other but also of a role for newly emerged, lineage-specific gene expression patterns in the evolution of the gametophyte and sporophyte developmental programs in this major eukaryotic group. Moreover, we show that generation-biased genes display distinct evolutionary modes, with gametophyte-biased genes evolving rapidly at the coding sequence level whereas sporophyte-biased genes tend to exhibit changes in their patterns of expression. CONCLUSION: Our analysis uncovers the characteristics, expression patterns, and evolution of generation-biased genes and underlines the selective forces that shape this previously underappreciated source of phenotypic diversity.

Convergent recruitment of TALE homeodomain life cycle regulators to direct sporophyte development in land plants and brown algae.

Three amino acid loop extension homeodomain transcription factors (TALE HD TFs) act as life cycle regulators in green algae and land plants. In mosses these regulators are required for the deployment of the sporophyte developmental program. We demonstrate that mutations in either of two TALE HD TF genes, OUROBOROS or SAMSARA, in the brown alga Ectocarpus result in conversion of the sporophyte generation into a gametophyte. The OUROBOROS and SAMSARA proteins heterodimerise in a similar manner to TALE HD TF life cycle regulators in the green lineage. These observations demonstrate that TALE-HD-TF-based life cycle regulation systems have an extremely ancient origin, and that these systems have been independently recruited to regulate sporophyte developmental programs in at least two different complex multicellular eukaryotic supergroups, Archaeplastida and Chromalveolata.

The brown algal mode of tip growth: Keeping stress under control.

Tip growth has been studied in pollen tubes, root hairs, and fungal and oomycete hyphae and is the most widely distributed unidirectional growth process on the planet. It ensures spatial colonization, nutrient predation, fertilization, and symbiosis with growth speeds of up to 800 µm h-1. Although turgor-driven growth is intuitively conceivable, a closer examination of the physical processes at work in tip growth raises a paradox: growth occurs where biophysical forces are low, because of the increase in curvature in the tip. All tip-growing cells studied so far rely on the modulation of cell wall extensibility via the polarized excretion of cell wall-loosening compounds at the tip. Here, we used a series of quantitative measurements at the cellular level and a biophysical simulation approach to show that the brown alga Ectocarpus has an original tip-growth mechanism. In this alga, the establishment of a steep gradient in cell wall thickness can compensate for the variation in tip curvature, thereby modulating wall stress within the tip cell. Bootstrap analyses support the robustness of the process, and experiments with fluorescence recovery after photobleaching (FRAP) confirmed the active vesicle trafficking in the shanks of the apical cell, as inferred from the model. In response to auxin, biophysical measurements change in agreement with the model. Although we cannot strictly exclude the involvement of a gradient in mechanical properties in Ectocarpus morphogenesis, the viscoplastic model of cell wall mechanics strongly suggests that brown algae have evolved an alternative strategy of tip growth. This strategy is largely based on the control of cell wall thickness rather than fluctuations in cell wall mechanical properties.

Effects of a co-culture of marine algae and shrimp (Litopenaeus vannamei) on the growth, survival and immune response of shrimp infected with Vibrio parahaemolyticus and white spot virus (WSSV).

In aquaculture, fighting infectious diseases is a necessity. This study measured the immuno-stimulating effect of live macroalgae consumption on Litopenaeus vannamei against Vibrio parahaemolyticus and WSSV infection in two independent bioassays. Shrimps and macroalgae were cultivated in a co-culture with two species of macroalgae separately (Gracilaria vermiculophylla and Dictyota dichotoma), and later, shrimp were infected with V. parahaemolyticus. In another bioassay, shrimp and macroalgae (G. vermiculophylla, D. dichotoma and Ulva lactuca) were grown and subsequently infected with WSSV. For both bioassays, survival after 120 h was determined, the total hemocyte count (TCH) was measured and the activity of superoxide dismutase (SOD) and catalase (CAT) in tissue were measured. The results indicate that the use of macroalgae in co-culture with L. vannamei provides a nutritional benefit that achieves higher growth than the control organisms, as well as improvements of the ammonium concentration and immune response after infection with V. parahaemolyticus and WSSV. A better immune response was obtained in organisms cultured with macroalgae in both bioassays at a ratio of 1.6-1.9 for organisms infected with bacteria and 1.4 to 1.6 times for organisms infected with the virus. In turn, the enzymatic activity of SOD and CAT were higher in the treated organisms relative to the controls in both experiments.

Auxin Function in the Brown Alga Dictyota dichotoma.

Auxin controls body plan patterning in land plants and has been proposed to play a similar role in the development of brown algae (Phaeophyta) despite their distant evolutionary relationship with land plants. The mechanism of auxin action in brown algae remains controversial because of contradicting conclusions derived from pharmacological studies on Fucus In this study, we used Dictyota dichotoma as a model system to show that auxin plays a role during the apical-basal patterning of the embryo of brown algae. Indole-3-acetic acid was detectable in D. dichotoma germlings and mature tissue. Although two-celled D. dichotoma zygotes normally develop a rhizoid from one pole and a thallus meristem from the other, addition of exogenous auxins to one-celled embryos affected polarization, and both poles of the spheroidal embryo developed into rhizoids instead. The effect was strongest at lower pH and when variable extrinsic informational cues were applied. 2-[4-(diethylamino)-2-hydroxybenzoyl]benzoic acid, an inhibitor of the ABC-B/multidrug resistance/P-glycoprotein subfamily of transporters in land plants, affected rhizoid formation by increasing rhizoid branching and inducing ectopic rhizoids. An in silico survey of auxin genes suggested that a diverse range of biosynthesis genes and transport genes, such as PIN-LIKES, and the ATP-binding cassette subfamily (ABC-B/multidrug resistance/P-glycoprotein) transporters from land plants have homologs in D. dichotoma and Ectocarpus siliculosus Together with reports on auxin function in basal lineages of green algae, these results suggest that auxin function predates the divergence between the green and brown lineage and the transition toward land plants.

Functional biodiversity loss along natural CO2 gradients.

The effects of environmental change on biodiversity are still poorly understood. In particular, the consequences of shifts in species composition for marine ecosystem function are largely unknown. Here we assess the loss of functional diversity, i.e. the range of species biological traits, in benthic marine communities exposed to ocean acidification (OA) by using natural CO2 vent systems. We found that functional richness is greatly reduced with acidification, and that functional loss is more pronounced than the corresponding decrease in taxonomic diversity. In acidified conditions, most organisms accounted for a few functional entities (i.e. unique combination of functional traits), resulting in low functional redundancy. These results suggest that functional richness is not buffered by functional redundancy under OA, even in highly diverse assemblages, such as rocky benthic communities.

Increased temperature but not pCO2 levels affect early developmental and reproductive traits of the economically important habitat-forming kelp Lessonia trabeculata.

The effects of ocean warming and ocean acidification on developmental and reproductive traits of Lessonia trabeculata were evaluated. Meiospores were cultured for 35 days in an experimental mesocosm where temperature (~15 and 19 °C) and partial CO2 pressure (pCO2, ~400 and 1300 µatm) were controlled. The results indicate that germination was reduced at 19 °C, whereas the increase of pCO2 only had effects at 15 °C. Likewise, the increase in temperature significantly affected the vegetative growth of female gametophytes. Sex ratio was not affected significantly by any of the variables studied. Fertility and reproductive success decreased by about 50% at 19 °C. The pCO2 levels had no significant effects on most early developmental traits. The results suggest that ocean warming or periodic warming events (e.g. an El Niño event) might affect the recruiting capacity of this or other similar species by affecting their early developmental stages.

Ex situ cultivation protocol for Cystoseira amentacea var. stricta (Fucales, Phaeophyceae) from a restoration perspective.

Due to multiple impacts, Cystoseira forests are experiencing a significant decline, which is affecting the ecosystem services they provide. Despite conservation efforts, there is an urgent need to develop best practices and large-scale restoration strategies. To implement restoration actions, we developed an ex situ protocol for the cultivation of Cystoseira. amentacea var. stricta, aimed at reducing the time needed for laboratory culture, thus avoiding prolonged maintenance and minimizing costs. Specifically, we tested the effects of temperature, light and substratum on settlement and growth of early life stages using a factorial experiment. Temperature (20 and 24°C) and photoperiod (15L:9D) were selected to reflect the conditions experienced in the field during the reproductive period. Two light intensities (125 and 250 µmol photons m-2s-1) were selected to mimic the condition experienced in the absence of canopy (i.e. barren-higher light intensity) or in the understory (lower light intensity) during gamete release. The tested substrata were flat polished pebbles and rough clay tiles. The release of gametes and the successive survival and development of embryo and germlings were followed for two weeks. Regardless of the culture conditions, rougher tiles showed higher zygote settlement, but the substrata did not affect the successive development. Zygote mortality after one week averaged 50% and at the end of the second week, embryonic survival was higher under lower light and temperature conditions, which also determined the growth of larger embryos.