Characterization and comparison of bacterial communities of an invasive and two native Caribbean seagrass species sheds light on the possible influence of the microbiome on invasive mechanisms

Invasive plants, including marine macrophytes, are one of the most important threats to biodiversity by displacing native species and organisms depending on them. Invasion success is dependent on interactions among living organisms, but their study has been mostly limited to negative interactions while positive interactions are mostly underlooked. Recent studies suggested that microorganisms associated with eukaryotic hosts may play a determinant role in the invasion process. Along with the knowledge of their structure, taxonomic composition, and potential functional profile, understanding how bacterial communities are associated with the invasive species and the threatened natives (species-specific/environmentally shaped/tissue-specific) can give us a holistic insight into the invasion mechanisms. Here, we aimed to compare the bacterial communities associated with leaves and roots of two native Caribbean seagrasses (Halodule wrightii and Thalassia testudinum) with those of the successful invader Halophila stipulacea, in the Caribbean island Curaçao, using 16S rRNA gene amplicon sequencing and functional prediction. Invasive seagrass microbiomes were more diverse and included three times more species-specific core OTUs than the natives. Associated bacterial communities were seagrass-specific, with higher similarities between natives than between invasive and native seagrasses for both communities associated with leaves and roots, despite their strong tissue differentiation. However, with a higher number of OTUs in common, the core community (i.e., OTUs occurring in at least 80% of the samples) of the native H. wrightii was more similar to that of the invader H. stipulacea than T. testudinum, which could reflect more similar essential needs (e.g., nutritional, adaptive, and physiological) between native and invasive, in contrast to the two natives that might share more environment-related OTUs. Relative to native seagrass species, the invasive H. stipulacea was enriched in halotolerant bacterial genera with plant growth-promoting properties (like Halomonas sp. and Lysinibacillus sp.) and other potential beneficial effects for hosts (e.g., heavy metal detoxifiers and quorum sensing inhibitors). Predicted functional profiles also revealed some advantageous traits on the invasive species such as detoxification pathways, protection against pathogens, and stress tolerance. Despite the predictive nature of our findings concerning the functional potential of the bacteria, this investigation provides novel and important insights into native vs. invasive seagrasses microbiome. We demonstrated that the bacterial community associated with the invasive seagrass H. stipulacea is different from native seagrasses, including some potentially beneficial bacteria, suggesting the importance of considering the microbiome dynamics as a possible and important influencing factor in the colonization of non-indigenous species. We suggest further comparison of H. stipulacea microbiome from its native range with that from both the Mediterranean and Caribbean habitats where this species has a contrasting invasion success. Also, our new findings open doors to a more in-depth investigation combining meta-omics with bacterial manipulation experiments in order to confirm any functional advantage in the microbiome of this invasive seagrass.CEECINST/00114/2018info:eu-repo/semantics/publishedVersio

Some don't like it hot: microhabitat-dependent thermal and water stresses in a trailing edge population

The distributional limits of species in response to environmental change are usually studied at large temporal and/or geographical scales. However, organismal scale habitat variation can be overlooked when investigating large-scale averages of key factors such as temperature. We examine how microhabitat thermal conditions relate to physiological limits, which may contribute to recent range shifts in an intertidal alga. We defined the onset and maximum temperatures of the heat-shock response (HSR) for a southern edge population of Fucus vesiculosus, which has subsequently become extinct. The physiological threshold for resilience (assayed using chlorophyll fluorescence) coincided with declining HSR, determined from the temperature-dependent induction of seven heat-shock protein transcripts. In intertidal habitats, temperature affects physiology directly by controlling body temperature and indirectly through evaporative water loss. We investigated the relationship between the thermal environment and in situ molecular HSR at microhabitat scales. Over cm to m scales, four distinct microhabitats were defined in algal patches (canopy surface, patch edge, subcanopy, submerged channels), revealing distinct thermal and water stress environments during low-tide emersion. The in situ HSR agreed with estimated tissue temperatures in all but one microhabitat. Remarkably, in the most thermally extreme microhabitat (canopy surface), the HSR was essentially absent in desiccated tissue, providing a potential escape from the cellular metabolic costs of thermal stress. Meteorological records, microenvironmental thermal profiles and HSR data indicate that the maximum HSR is approached or exceeded in hydrated tissue during daytime low tides for much of the year. Furthermore, present-day summer seawater temperatures are sufficient to induce HSR during high-tide immersion, preventing recovery and resulting in continuous HSR during daytime low-tide cycles over the entire summer. HSR in the field matched microhabitat temperatures more closely than local seawater or atmospheric data, suggesting that the impacts of climatic change are best understood at the microhabitat scale, particularly in intertidal areas.FCT - Portuguese Science Foundation [POCTI/MAR/61105/2004, EXCL/AAG-GLO/0661/2012, SFRH/BPD/63/03/2009, SFRH/BD/74436/2010]info:eu-repo/semantics/publishedVersio

Seaweed loads cause stronger bacterial community shifts in coastal lagoon sediments than nutrient loads

The input of nutrients from anthropogenic sources is the leading cause of coastal eutrophication and is usually coupled with algal/seaweed blooms. Effects may be magnified in semi-enclosed systems, such as highly productive coastal lagoon ecosystems. Eutrophication and seaweed blooms can lead to ecosystem disruption. Previous studies have considered only one of these factors, disregarding possible interactive effects and the effect of the blooming species' identity on sediment bacterial communities. We tested the effect of experimental nutrient loading and two common blooming seaweeds (Ulva rigida and Gracilaria vermiculophylla) in coastal lagoon sediments, on the structure of bacterial communities (using 16S rRNA amplicon sequencing) and corresponding putative functional potential (using PiCRUSt). At the Operational Taxonomic Unit (OTU) level, the addition of nutrients reduced bacterial community α-diversity and decreased the abundance of sulfate reducers (Desulfobacterales) compared to sulfur oxidizers/denitrifiers (Chromatiales and Campylobacterales), whereas this was not the case at the order level. Seaweed addition did not change bacterial α-diversity and the effect on community structure depended on the taxonomic level considered. The addition of Gracilaria increased the abundance of orders and OTUs involved in sulfate reduction and organic matter decomposition (Desulfobacterales, Bacteroidales, and Clostridiales, respectively), an effect which was also detected when only Ulva was added. Nutrients and the seaweeds combined only interacted for Ulva and nutrients, which increased known sulfide oxidizers and denitrifiers (order Campylobacterales). Seaweed enrichment affected putative functional profiles; a stronger increase of sulfur cycling KEGG pathways was assigned to nutrient-disturbed sediments, particularly with the seaweeds and especially Ulva. In contrast, nitrogen and sulfur cycle pathways showed a higher abundance of genes related to dissimilatory nitrate reduction to ammonium (DNRA) in Ulva+nutrients treatments. However, the other seaweed treatments increased the nitrogen fixation genes. Thiosulfate reduction, performed by sulfate-reducing bacteria, increased in seaweed treatments except when Ulva was combined with nutrients. In conclusion, the in situ addition of nutrients and the seaweeds to intertidal sediments affected the bacterial communities differently and independently. The predicted functional profile suggests a shift in relative abundances of putative pathways for nitrogen and sulfur cycles, in line with the taxonomic changes of the bacterial communities.FCT - Foundation for Science and Technology UID/Multi/04326/2019 UID/Multi/04326/2016 SEAS-ERA/0001/2012 SFRH/BPD/107878/2015 SFRH/BPD/116774/2016 research priority area Systems Biology of the University of Amsterdaminfo:eu-repo/semantics/publishedVersio

Metagenomic analysis shows the presence of bacteria related to free-living forms of sulfur-Oxidizing Chemolithoautotrophic Symbionts in the rhizosphere of the seagrass Zostera marina

Seagrasses play an important role as ecosystem engineers; they provide shelter to many animals and improve water quality by filtering out nutrients and by controlling pathogens. Moreover, their rhizosphere promotes a myriad of microbial interactions and processes, which are dominated by microorganisms involved in the sulfur cycle. This study provides a detailed insight into the metabolic sulfur pathways in the rhizobiome of the seagrass Zostera marina, a dominant seagrass species across the temperate northern hemisphere. Shotgun metagenomic sequencing revealed the relative dominance of Gamma- and Deltaproteobacteria, and comparative analysis of sulfur genes identified a higher abundance of genes related to sulfur oxidation than sulfate reduction. We retrieved four high-quality draft genomes that are closely related to the gill symbiont of the clam Solemya velum, which suggests the presence of putative free-living forms of symbiotic bacteria. These are potentially highly versatile chemolithoautotrophic bacteria, able to alternate their metabolism between parallel pathways of sulfide oxidation (via sqr and fcc), nitrate reduction (denitrification or DNRA) and carbon fixation (via CBB or TCA cycle), depending on the environmental availability of sulfide. Our results support the hypothesis that seagrass meadows might function as a source of symbionts for invertebrates that inhabit within or around seagrass meadows. While providing ideal conditions for the proliferation of these free-living forms of symbionts, seagrasses would benefit from their genetic versatility, which contributes to sulfide detoxification and ammonium production, the seagrasses' preferred nitrogen source.European Union ERC 322551 European Science Foundation ConGenOmics program 6349 Fundacao para a Ciencia e Tecnologia (FCT) SFRH/BPD/63/03/2009 SFRH/BPD/107878/2015info:eu-repo/semantics/publishedVersio

Interactions between seagrasses and seaweeds during surge nitrogen acquisition determine interspecific competition

Seagrasses dominate shallow coastal environments where nitrogen (N) availability in the water column is often sporadic and mainly in the form of pulses. We investigated the N uptake competition between seagrasses and seaweeds through a series of N-15 surge uptake experiments combining single-species and mixed incubations across ammonium concentrations. N surge uptake rates of seagrasses were 2 to 14-fold higher than those of seaweeds in the majority of combinations, showing that seagrasses are generally in a competitive advantage over seaweeds in N-poor environments with N-pulses. No threshold concentration of ammonium was found beyond which seaweeds performed better than seagrasses. Mixed incubations revealed interspecific interactions that affected rates positively and negatively. Uptake rates obtained in single-species incubations, therefore, cannot always be used to predict the outcome of uptake competition. Only two (Zostera marina vs. Ulva rotundata and Zostera marina vs. Codium decorticatum) of the nine combinations tested (Z. marina, Z. noltei and Cymodocea nodosa vs. U. rotundata, C. decorticatum and Dictyota dichotoma) were found to enhance macroalgal uptake. Our results showed that the surge uptake capacity of seagrasses represents an important mechanism in their N acquisition strategy that justifies their dominance in shallow oligotrophic environments.Portuguese Foundation for Science and Technology (FCT) [PTDC/MAR/098069/2008, SFRH/BPD/63703/2009]; FCT - Foundation for Science and Technology [CCMAR/Multi/04326/2013

Summer shifts of bacterial communities associated with the invasive brown seaweed Sargassum muticum are location and tissue dependent

Seaweed-associated microbiota experience spatial and temporal shifts in response to changing environmental conditions and seaweed physiology. These shifts may result in structural, functional and behavioral changes in the host with potential consequences for its fitness. They, thus, may help the host to adapt to changing environmental conditions. The current knowledge of seasonal variation of seaweed-associated microbiota is however still limited. In this study, we explored temporal and spatial variation of microbial communities associated with the invasive brown seaweed S. muticum. We sampled in northern and southern Portugal, in September, March and July-August (summer). In addition, as (pseudo-)perennial seaweeds display seasonal reproductive phenology, we sampled various parts of the individuals to disentangle the effect of temporal changes from those due to structural development variations. The diversity and structure of associated microbial communities were determined using next generation sequencing of the variable regions V5-7 of the 16S rDNA. We expected to find differentiation in associated microbial communities between regions and sampling months, but with differences depending on the seaweed structure examined. As expected, the study revealed substantial temporal shifts in S. muticum microbiome, for instance with large abundance of Rhodobacteraceae and Loktanella in September-March but prevalence of Pirellulales during the summer months. Variations between regions and tissues were also observed: in northern Portugal and on basal structures, bacterial diversity was higher as compared to the South and apical parts. All examined seaweed structures showed temporal differences in associated microbial community structure over time, except for holdfasts between September and March. Bacteria contributing to these changes varied spatially. Conversely to all other structures, the holdfast also did not show differences in associated community structure between southern and northern regions. Our study highlights the importance of structural microscale differentiations within seaweeds hosts with regard to their associated microbial communities and their importance across temporal and spatial dimensions.Erasmus Mundus Doctoral Programme MARES on Marine Ecosystem Health Conservation - MARES_13_08 FCT (Portugal) - SFRH/BPD/107878/2015, SFRH/BPD/116774/2016 project EU SEAS-ERA INVASIVES - SEAS-ERA/0001/2012 project MARFOR - FCT-BIODIVERSA/004/2015 CCMAR/Multi/04326/2013info:eu-repo/semantics/publishedVersio

Differential frond growth in the isomorphic haploid–diploid red seaweed Agarophyton chilense by long‐term in situ monitoring (1)

Conditional differentiation between haploids and diploids has been proposed to drive the evolutionary stability of isomorphic biphasic life cycles. The cost of producing and maintaining genetic information has been posed as a possible driver of this conditional differentiation. Under this hypothesis, haploids benefit over diploids in resource-limited environments by halving the costs of producing and maintaining DNA. Spared resources can be allocated to enhance survival, growth or fertility. Here we test in the field whether indeed haploids have higher growth rates than diploids. Individuals of the red seaweed Agarophyton chilense, were mapped and followed during 2 years with 4-month census intervals across different stands within the Valdivia River estuary, Chile. As hypothesized, haploids grew larger and faster than diploids, but this was sex-dependent. Haploid (gametophyte) females grew twice as large and 15% faster than diploids (tetrasporophytes), whereas haploid males only grew as large and as fast as the maximum obtained by diploids in summer. However, haploid males maintained their maximum sizes and growth rates constant year-round, while diploids were smaller and had lower growth rates during the winter. In conclusion, our results confirm the conditional differentiation in size and growth between haploids and diploids but also identified important differences between males and females. Besides understanding life cycle evolution, the dynamics of A. chilense frond growth reported informs algal farmers regarding production optimization and should help in determining best planting and harvesting strategies.CONICYT FONDECYT: 1090360/ 1170541.FCT: UIDB/EEA/50009/2020/ SFRH/BPD/107878/2015/ UID/Multi/04326/2020info:eu-repo/semantics/publishedVersio

Response of kelps from different latitudes to consecutive heat shock

Although extensive work has focused on kelp responses to constant temperature, little is known about their response to the consecutive temperature shocks they are often exposed to in the shallow subtidal and intertidal pools. Here we characterized the responses of the two southernmost forest-forming kelp species in the Northeast Atlantic, Laminaria ochroleuca De La Pylaie and Saccorhiza polyschides (Lightf.) Batt. to multiple cycles of thermal stress. Individuals from the upper vertical limit of the geographical distribution edges where the two species co-occur forming forests, France and Portugal, were exposed to 4 consecutive cycles of thermal shock simulating a spring tide. A 24 h cycle consisted of culture at 15 degrees C, plus 1 h heat shock at one of five levels (20, 22.5, 25, 27.5 or 30 degrees C). The maximum quantum yield (Fv/Fm) of chlorophyll fluorescence of photosystem 2 (PS2) was used to detect impaired reaction center function, as a proxy for individual fitness costs, during recovery from heat shock. Both species showed resilience to temperatures from 20 to 25 degrees C. While exposure to 27.5 degrees C caused no inhibition to Fv/Fm of S. polyschides, a threshold was met above this temperature and exposure to 30 degrees C caused the death of all individuals. In contrast, L ochroleuca from France was damaged but able to survive 30 degrees C shocks and individuals from Portugal showed complete resilience to this treatment. In both species, blade elongation decreased with increasing temperature, with necrosis surpassing growth at higher temperatures. Resilience to high temperature exposure may confer an advantage to L ochroleuca to colonize intertidal pools on the Portuguese coast, in agreement with the observation that both species recruit in tide pools but only L ochroleuca reach adulthood. Our results indicate that as summer temperatures increase with climate change, the disappearance of S. polyschides from intertidal pools and a decrease in the density of L ochroleuca can be expected. (C) 2014 Elsevier B.V. All rights reserved.ASSEMBLE (ASSociation of European Marine Biological Laboratories) [227799]; Portuguese Science and Technology Foundation (FCT); FCT [PTDC/AAC-CLI/109108/2008, EXCL/AAG-GLO/0661/2012]info:eu-repo/semantics/publishedVersio

Differentiation in fitness-related traits in response to elevated temperatures between leading and trailing edge populations of marine macrophytes

The nature of species distribution boundaries is a key subject in ecology and evolution. Edge populations are potentially more exposed to climate-related environmental pressures. Despite research efforts, little is known about variability in fitness-related traits in leading (i.e., colder, high latitude) versus trailing (i.e., warmer, low latitude) edge populations. We tested whether the resilience, i.e. the resistance and recovery, of key traits differs between a distributional cold (Greenland) and warm (Portugal) range edge population of two foundation marine macrophytes, the intertidal macroalga Fucus vesiculosus and the subtidal seagrass Zostera marina. The resistance and recovery of edge populations to elevated seawater temperatures was compared under common experimental conditions using photosynthetic efficiency and expression of heat shock proteins (HSP). Cold and warm edge populations differed in their response, but this was species specific. The warm edge population of F. vesiculosus showed higher thermal resistance and recovery whereas the cold leading edge was less tolerant. The opposite was observed in Z. marina, with reduced recovery at the warm edge, while the cold edge was not markedly affected by warming. Our results confirm that differentiation of thermal stress responses can occur between leading and trailing edges, but such responses depend on local population traits and are thus not predictable just based on thermal pressures.FCT (Portuguese Science Foundation) [BIODIVERSA/0004/2015, PTDC/MAR-EST/6053/2014, UID/Multi/04326/2013, SFRH/BPD/63/03/2009, SFRH/BPD/107878/2015, SFRH/BD/74436/2010]; European Commission (ATP) [226248

An individual-based Model of the Red Alga Agarophyton chilense unravels the complex demography of Its intertidal stands

Algal demographic models have been developed mainly to study their life cycle evolution or optimize their commercial exploitation. Most commonly, structured-aggregated population models simulate the main life cycle stages considering their fertility, growth and survival. Their coarse resolution results in weak predictive abilities since neglected details may still impact the whole. In our case, we need a model of Agarophyton chilense natural intertidal populations that unravels the complex demography of isomorphic biphasic life cycles and be further used for: (i) introduction of genetics, aimed at studying the evolutionary stability of life cycles, (ii) optimizing commercial exploitation, and (iii) adaptation for other species. Long-term monitoring yield 6,066 individual observations and 40 population observations. For a holistic perspective, we developed an Individual-Based Model (IBM) considering ploidy stage, sex stage, holdfast age and survival, frond size, growth and breakage, fecundity, spore survival, stand biomass, location and season. The IBM was calibrated and validated comparing observed and estimated sizes and abundances of gametophyte males, gametophyte females and tetrasporophytes, stand biomass, haploid:dipoid ratio (known as H:D or G:T), fecundity and recruitment. The IBM replicated well the respective individual and population properties, and processes such as winter competition for light, self-thinning, summer stress from desiccation, frond breakage and re-growth, and different niche occupation by haploids and diploids. Its success depended on simulating with precision details such as the holdfasts' dynamics. Because "details" often occur for a reduced number of individuals, inferring about them required going beyond statistically significant evidences and integrating these with parameter calibration aimed at maximized model fit. On average, the population was haploid-dominated (H:D > 1). In locations stressed by desiccation, the population was slightly biased toward the diploids and younger individuals due to the superior germination and survival of the diploid sporelings. In permanently submerged rock pools the population was biased toward the haploids and older individuals due to the superior growth and survival of the haploid adults. The IBM application demonstrated that conditional differentiation among ploidy stages was responsible for their differential niche occupation, which, in its turn, has been argued as the driver of the evolutionary stability of isomorphic biphasic life cycles.info:eu-repo/semantics/publishedVersio