Dissolved Nitrogen Acquisition in the Symbioses of Soft and Hard Corals With Symbiodiniaceae: A Key to Understanding Their Different Nutritional Strategies?

Nitrogen is one of the limiting nutrients for coral growth and primary productivity. Therefore, the capacity of different associations between corals and their algal symbionts (Symbiodiniaceae) to efficiently exploit the available nitrogen sources will influence their distribution and abundance. Recent studies have advanced our understanding of nitrogen assimilation in reef-building scleractinian (hard) coral-Symbiodiniaceae symbioses. However, the nutrient metabolism of other coral taxa, such as Alcyoniina (soft corals), remains underexplored. Using stable isotope labeling, we investigated the assimilation of dissolved nitrogen (i.e., ammonium, nitrate, and free amino acids) by multiple species of soft and hard corals sampled in the Gulf of Aqaba in shallow (8-10 m) and mesophotic (40-50 m) reefs. Our results show that dissolved nitrogen assimilation rates per tissue biomass were up to 10-fold higher in hard than in soft coral symbioses for all sources of nitrogen. Although such differences in assimilation rates could be linked to the Symbiodiniaceae density, Symbiodiniaceae species, or the C:N ratio of the host and algal symbiont fractions, none of these parameters were different between the two coral taxa. Instead, the lower assimilation rates in soft coral symbioses might be explained by their different nutritional strategy: whereas soft corals may obtain most of their nitrogen via the capture of planktonic prey by the coral host (heterotrophic feeding), hard corals may rely more on dissolved nitrogen assimilation by their algal symbionts to fulfill their needs. This study highlights different nutritional strategies in soft and hard coral symbioses. A higher reliance on heterotrophy may help soft corals to grow in reefs with higher turbidity, which have a high concentration of particles in suspension in seawater. Further, soft corals may benefit from lower dissolved nitrogen assimilation rates in areas with low water quality.

Low levels of ultra-violet radiation mitigate the deleterious effects of nitrate and thermal stress on coral photosynthesis.

Reef ecosystems are under increasing pressure from global and local stressors. Rising seawater temperature and high ultraviolet radiation (UVR) levels are the main drivers of the disruption of the coral-dinoflagellate symbiosis (bleaching). Bleaching can also be exacerbated by nitrate contamination in coastal reefs. However, the underlying physiological mechanisms are still poorly understood. Here, we assessed the physiological and oxidative state of the scleractinian coral Pocillopora damicornis, maintained eight weeks in a crossed-factorial design including two temperatures (26 °C or 30 °C), and two nitrate (0.5 and 3 µM-enriched), and UVR (no UVR and 25/1.5 Wm-2 UVA/B) levels. Nitrate enrichment, and high temperature, significantly impaired coral photosynthesis. However, UVR alleviated the nitrate and temperature-induced decrease in photosynthesis, by increasing the coral's antioxidant capacity. The present study contributes to our understanding of the combined effects of abiotic stressors on coral bleaching susceptibility. Such information is urgently needed to refine reef management strategies.

Symbiont regulation in Stylophora pistillata during cold stress: an acclimation mechanism against oxidative stress and severe bleaching.

Widespread coral bleaching and mortality, leading to coral reef decline, have been mainly associated with climate-change-driven increases in sea surface temperature. However, bleaching and mortality events have also been related to decreases in sea surface temperature, with cold stress events (e.g. La Niña events) being expected to increase in frequency or intensity as a result of a changing climate. Cold stress creates physiological symptoms in symbiotic reef-building corals similar to those observed when they are heat stressed, and the biochemical mechanisms underpinning cold stress in corals have been suggested to be related to an oxidative stress condition. However, up to now, this hypothesis had not been tested. This study assessed how short and long cold excursions in seawater temperature affect the physiology and biochemical processes related to oxidative stress in the reef-building coral Stylophora pistillata We provide, for the first time, direct evidence that the mechanisms underpinning cold stress and bleaching are related to the production of reactive oxygen species, and that rapid expulsion of a significant proportion of the symbiont population by the host during cooling conditions is an acclimation mechanism to avoid oxidative stress and, ultimately, severe bleaching. Furthermore, this study is one of the first to show that upwelling conditions (short-term cold stress+nutrient enrichment) can provoke a more severe oxidative stress condition in corals than cold stress alone.

Artificial light at night (ALAN) alters the physiology and biochemistry of symbiotic reef building corals.

Artificial Light at Night (ALAN), which is the alteration of natural light levels as the result of anthropogenic light sources, has been acknowledged as an important factor that alters the functioning of marine ecosystems. Using LEDs light to mimic ALAN, we studied the effect on the physiology (symbiont and chlorophyll contents, photosynthesis, respiration, pigment profile, skeletal growth, and oxidative stress responses) of two scleractinian coral species originating from the Red Sea. ALAN induced the photoinhibition of symbiont photosynthesis, as well as an overproduction of reactive oxygen species (ROS) and an increase in oxidative damage to lipids in both coral species. The extent of the deleterious effects of ALAN on the symbiotic association and coral physiology was aligned with the severity of the oxidative stress condition experienced by the corals. The coral species Sylophora pistillata, which experienced a more severe oxidative stress condition than the other species tested, Turbinaria reniformis, also showed a more pronounced bleaching (loss of symbionts and chlorophyll content), enhanced photoinhibition and decreased photosynthetic rates. Findings of the present study further our knowledge on the biochemical mechanisms underpinning the deleterious impacts of ALAN on scleractinian corals, ultimately shedding light on the emerging threat of ALAN on coral reef ecology. Further, considering that global warming and light pollution will increase in the next few decades, future studies should be taken to elucidate the potential synergetic effects of ALAN and global climate change stressors.

Unravelling the different causes of nitrate and ammonium effects on coral bleaching.

Mass coral bleaching represents one of the greatest threats to coral reefs and has mainly been attributed to seawater warming. However, reduced water quality can also interact with warming to increase coral bleaching, but this interaction depends on nutrient ratios and forms. In particular, nitrate (NO3-) enrichment reduces thermal tolerance while ammonium (NH4+) enrichment tends to benefit coral health. The biochemical mechanisms underpinning the different bleaching responses of corals exposed to DIN enrichment still need to be investigated. Here, we demonstrated that the coral Stylophora pistillata underwent a severe oxidative stress condition and reduced aerobic scope when exposed to NO3- enrichment combined with thermal stress. Such condition resulted in increased bleaching intensity compared to a low-nitrogen condition. On the contrary, NH4+ enrichment was able to amend the deleterious effects of thermal stress by favoring the oxidative status and energy metabolism of the coral holobiont. Overall, our results demonstrate that the opposite effects of nitrate and ammonium enrichment on coral bleaching are related to the effects on corals' energy/redox status. As nitrate loading in coastal waters is predicted to significantly increase in the future due to agriculture and land-based pollution, there is the need for urgent management actions to prevent increases in nitrate levels in seawater. In addition, the maintenance of important fish stocks, which provide corals with recycled nitrogen such as ammonium, should be favoured.

Productivity and carbon fluxes depend on species and symbiont density in soft coral symbioses.

Soft corals often constitute one of the major benthic groups of coral reefs. Although they have been documented to outcompete reef-building corals following environmental disturbances, their physiological performance and thus their functional importance in reefs are still poorly understood. In particular, the acclimatization to depth of soft corals harboring dinoflagellate symbionts and the metabolic interactions between these two partners have received little attention. We performed stable isotope tracer experiments on two soft coral species (Litophyton sp. and Rhytisma fulvum fulvum) from shallow and upper mesophotic Red Sea coral reefs to quantify the acquisition and allocation of autotrophic carbon within the symbiotic association. Carbon acquisition and respiration measurements distinguish Litophyton sp. as mainly autotrophic and Rhytisma fulvum fulvum as rather heterotrophic species. In both species, carbon acquisition was constant at the two investigated depths. This is a major difference from scleractinian corals, whose carbon acquisition decreases with depth. In addition, carbon acquisition and photosynthate translocation to the host decreased with an increase in symbiont density, suggesting that nutrient provision to octocoral symbionts can quickly become a limiting factor of their productivity. These findings improve our understanding of the biology of soft corals at the organism-scale and further highlight the need to investigate how their nutrition will be affected under changing environmental conditions.

Response of the temperate scleractinian coral Cladocora caespitosa to high temperature and long-term nutrient enrichment.

Anthropogenic nutrient enrichment and increased seawater temperatures are responsible for coral reef decline. In particular, they disrupt the relationship between corals and their dinoflagellate symbionts (bleaching). However, some coral species can afford either high temperatures or nutrient enrichment and their study can bring new insights into how corals acclimate or adapt to stressors. Here, we focused on the role of the nutrient history in influencing the response of the Mediterranean scleractinian coral Cladocora caespitosa to thermal stress. Colonies living naturally in nutrient-poor (<0.5 µM nitrogen, <0.2 µM phosphorus, LN) and nutrient-rich (ca. 10-20 µM nitrogen, 0.4 µM phosphorus, HN) locations were sampled, maintained under the right nutrient conditions, and exposed to a temperature increase from 17 °C to 24 °C and 29 °C. While both HN and LN colonies decreased their concentrations of symbionts and/or photosynthetic pigments, HN colonies were able to maintain significant higher rates of net and gross photosynthesis at 24 °C compared to LN colonies. In addition, while there was no change in protein concentration in HN corals during the experiment, proteins continuously decreased in LN corals with increased temperature. These results are important in that they show that nutrient history can influence the response of scleractinian corals to thermal stress. Further investigations of under-studied coral groups are thus required in the future to understand the processes leading to coral resistance to environmental perturbations.

Seasonal Stability in the Microbiomes of Temperate Gorgonians and the Red Coral Corallium rubrum Across the Mediterranean Sea.

Populations of key benthic habitat-forming octocoral species have declined significantly in the Mediterranean Sea due to mass mortality events caused by microbial disease outbreaks linked to high summer seawater temperatures. Recently, we showed that the microbial communities of these octocorals are relatively structured; however, our knowledge on the seasonal dynamics of these microbiomes is still limited. To investigate their seasonal stability, we collected four soft gorgonian species (Eunicella singularis, Eunicella cavolini, Eunicella verrucosa and Leptogorgia sarmentosa) and the precious red coral (Corallium rubrum) from two coastal locations with different terrestrial impact levels in the Mediterranean Sea, and used next-generation amplicon sequencing of the 16S rRNA gene. The microbiomes of all soft gorgonian species were dominated by the same 'core microbiome' bacteria belonging to the Endozoicomonas and the Cellvibrionales clade BD1-7, whereas the red coral microbiome was primarily composed of 'core' Spirochaetes, Oceanospirillales ME2 and Parcubacteria. The associations with these bacterial taxa were relatively consistent over time at each location for each octocoral species. However, differences in microbiome composition and seasonal dynamics were observed between locations and could primarily be attributed to locally variant bacteria. Overall, our data provide further evidence of the intricate symbiotic relationships that exist between Mediterranean octocorals and their associated microbes, which are ancient and highly conserved over both space and time, and suggest regulation of the microbiome composition by the host, depending on local conditions.

Effects of temperature and UVR on organic matter fluxes and the metabolic activity of Acropora muricata.

Coral bleaching events are predicted to occur more frequently in the coming decades with global warming. The susceptibility of corals to bleaching during thermal stress episodes depends on many factors, including the magnitude of thermal stress and irradiance. The interactions among these two factors, and in particular with ultra-violet radiation (UVR), the most harmful component of light, are more complex than assumed, and are not yet well understood. This paper explores the individual and combined effects of temperature and UVR on the metabolism of Acropora muricata, one of the most abundant coral species worldwide. Particulate and dissolved organic matter (POM/DOM) fluxes and organic matter (OM) degradation by the mucus-associated bacteria were also monitored in all conditions. The results show that UVR exposure exacerbated the temperature-induced bleaching, but did not affect OM fluxes, which were only altered by seawater warming. Temperature increase induced a shift from POM release and DOM uptake in healthy corals to POM uptake and DOM release in stressed ones. POM uptake was linked to a significant grazing of pico- and nanoplankton particles during the incubation, to fulfil the energetic requirements of A. muricata in the absence of autotrophy. Finally, OM degradation by mucus-associated bacterial activity was unaffected by UVR exposure, but significantly increased under high temperature. Altogether, our results demonstrate that seawater warming and UVR not only affect coral physiology, but also the way corals interact with the surrounding seawater, with potential consequences for coral reef biogeochemical cycles and food webs.

The c-Jun N-terminal kinase prevents oxidative stress induced by UV and thermal stresses in corals and human cells.

Coral reefs are of major ecological and socio-economic interest. They are threatened by global warming and natural pressures such as solar ultraviolet radiation. While great efforts have been made to understand the physiological response of corals to these stresses, the signalling pathways involved in the immediate cellular response exhibited by corals remain largely unknown. Here, we demonstrate that c-Jun N-terminal kinase (JNK) activation is involved in the early response of corals to thermal and UV stress. Furthermore, we found that JNK activity is required to repress stress-induced reactive oxygen species (ROS) accumulation in both the coral Stylophora pistillata and human skin cells. We also show that inhibiting JNK activation under stress conditions leads to ROS accumulation, subsequent coral bleaching and cell death. Taken together, our results suggest that an ancestral response, involving the JNK pathway, is remarkably conserved from corals to human, protecting cells from the adverse environmental effects.

Physiological response of the cold-water coral Desmophyllum dianthus to thermal stress and ocean acidification.

Rising temperatures and ocean acidification driven by anthropogenic carbon emissions threaten both tropical and temperate corals. However, the synergistic effect of these stressors on coral physiology is still poorly understood, in particular for cold-water corals. This study assessed changes in key physiological parameters (calcification, respiration and ammonium excretion) of the widespread cold-water coral Desmophyllum dianthus maintained for ∼8 months at two temperatures (ambient 12 °C and elevated 15 °C) and two pCO2 conditions (ambient 390 ppm and elevated 750 ppm). At ambient temperatures no change in instantaneous calcification, respiration or ammonium excretion rates was observed at either pCO2 levels. Conversely, elevated temperature (15 °C) significantly reduced calcification rates, and combined elevated temperature and pCO2 significantly reduced respiration rates. Changes in the ratio of respired oxygen to excreted nitrogen (O:N), which provides information on the main sources of energy being metabolized, indicated a shift from mixed use of protein and carbohydrate/lipid as metabolic substrates under control conditions, to less efficient protein-dominated catabolism under both stressors. Overall, this study shows that the physiology of D. dianthus is more sensitive to thermal than pCO2 stress, and that the predicted combination of rising temperatures and ocean acidification in the coming decades may severely impact this cold-water coral species.

Among-species variation in the energy budgets of reef-building corals: scaling from coral polyps to communities.

The symbiosis between corals and dinoflagellates promotes the rapid growth of corals in shallow tropical oceans, and the high overall productivity of coral reefs. The aim of this study was to quantify and understand variation in carbon acquisition and allocation among coral species. We measured multiple physiological traits (including symbiont density, calcification, photosynthesis and tissue composition) for the same coral fragments to facilitate direct comparisons between species (Stylophora pistillata, Pocillopora damicornis, Galaxea fascicularis, Turbinaria reniformis and Acropora sp.). Tissue protein content was highly sensitive to the availability of particulate food, increasing in fed colonies of all species. Despite among-species variation in physiology, and consistent effects of feeding on some traits, overall energy allocation to tissue compared with skeleton growth did not depend on food availability. Extrapolating from our results, estimated whole-assemblage carbon uptake varied >20-fold across different coral assemblages, but this variation was largely driven by differences in the tissue surface area of different colony morphologies, rather than by differences in surface-area-specific physiological rates. Our results caution against drawing conclusions about reef productivity based solely on physiological rates measured per unit tissue surface area. Understanding the causes and consequences of among-species variation in physiological energetics provides insight into the mechanisms that underlie the fluxes of organic matter within reefs, and between reefs and the open ocean.

Thermally tolerant corals have limited capacity to acclimatize to future warming.

Thermal stress affects organism performance differently depending on the ambient temperature to which they are acclimatized, which varies along latitudinal gradients. This study investigated whether differences in physiological responses to temperature are consistent with regional differences in temperature regimes for the stony coral Oculina patagonica. To resolve this question, we experimentally assessed how colonies originating from four different locations characterized by >3 °C variation in mean maximum annual temperature responded to warming from 20 to 32 °C. We assessed plasticity in symbiont identity, density, and photosynthetic properties, together with changes in host tissue biomass. Results show that, without changes in the type of symbiont hosted by coral colonies, O. patagonica has limited capacity to acclimatize to future warming. We found little evidence of variation in overall thermal tolerance, or in thermal optima, in response to spatial variation in ambient temperature. Given that the invader O. patagonica is a relatively new member of the Mediterranean coral fauna, our results also suggest that coral populations may need to remain isolated for a long period of time for thermal adaptation to potentially take place. Our study indicates that for O. patagonica, mortality associated with thermal stress manifests primarily through tissue breakdown under moderate but prolonged warming (which does not impair symbiont photosynthesis and, therefore, does not lead to bleaching). Consequently, projected global warming is likely to cause repeat incidents of partial and whole colony mortality and might drive a gradual range contraction of Mediterranean corals.

The response of the scleractinian coral Turbinaria reniformis to thermal stress depends on the nitrogen status of the coral holobiont.

The physiological response of the scleractinian coral Turbinaria reniformis to ammonium enrichment (3 µmol l(-1)) was examined at 26°C as well as during a 7 day increase in temperature to 31°C (thermal stress). At 26°C, ammonium supplementation had little effect on the coral physiology. It induced a decrease in symbiont density, compensated by an increase in chlorophyll content per symbiont cell. Organic carbon release was reduced, likely because of a better utilization of the photosynthesized carbon (i.e. incorporation into proteins, kept in the coral tissue). The δ(15)N signatures of the ammonium-enriched symbionts and host tissue were also significantly decreased, by 4 and 2‰, respectively, compared with the non-enriched conditions, suggesting a significant uptake of inorganic nitrogen by the holobiont. Under thermal stress, coral colonies that were not nitrogen enriched experienced a drastic decrease in photosynthetic and photoprotective pigments (chlorophyll a, ß-carotene, diadinoxanthin, diatoxanthin and peridinin), followed by a decrease in the rates of photosynthesis and calcification. Organic carbon release was not affected by this thermal stress. Conversely, nitrogen-enriched corals showed an increase in their pigment concentrations, and maintained rates of photosynthesis and calcification at ca. 60% and 100% of those measured under control conditions, respectively. However, these corals lost more organic carbon into the environment. Overall, these results indicate that inorganic nitrogen availability can be important to determining the resilience of some scleractinian coral species to thermal stress, and can have a function equivalent to that of heterotrophic feeding concerning the maintenance of coral metabolism under stress conditions.

Controlling effects of irradiance and heterotrophy on carbon translocation in the temperate coral Cladocora caespitosa.

Temperate symbiotic corals, such as the Mediterranean species Cladocora caespitosa, live in seasonally changing environments, where irradiance can be ten times higher in summer than winter. These corals shift from autotrophy in summer to heterotrophy in winter in response to light limitation of the symbiont's photosynthesis. In this study, we determined the autotrophic carbon budget under different conditions of irradiance (20 and 120 µmol photons m(-2) s(-1)) and feeding (fed three times a week with Artemia salina nauplii, and unfed). Corals were incubated in H(13)CO(3) (-)-enriched seawater, and the fate of (13)C was followed in the symbionts and the host tissue. The total amount of carbon fixed by photosynthesis and translocated was significantly higher at high than low irradiance (ca. 13 versus 2.5-4.5 µg cm(-2) h(-1)), because the rates of photosynthesis and carbon fixation were also higher. However, the percent of carbon translocation was similar under the two irradiances, and reached more than 70% of the total fixed carbon. Host feeding induced a decrease in the percentage of carbon translocated under low irradiance (from 70 to 53%), and also a decrease in the rates of carbon translocation per symbiont cell under both irradiances. The fate of autotrophic and heterotrophic carbon differed according to irradiance. At low irradiance, autotrophic carbon was mostly respired by the host and the symbionts, and heterotrophic feeding led to an increase in host biomass. Under high irradiance, autotrophic carbon was both respired and released as particulate and dissolved organic carbon, and heterotrophic feeding led to an increase in host biomass and symbiont concentration. Overall, the maintenance of high symbiont concentration and high percentage of carbon translocation under low irradiance allow this coral species to optimize its autotrophic carbon acquisition, when irradiance conditions are not favourable to photosynthesis.