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Coral restoration efforts have rapidly increased worldwide, including the development of several programmes on the Great Barrier Reef (GBR) in recent years. While many restoration programmes utilise in-water nurseries to accelerate coral biomass yields, the impact of nursery environments on propagule quality has not been examined despite the importance of coral fitness for ensuring resistant populations. Here, we investigated two fitness indicators (lipid diversity and tissue protein abundance) of Acropora millepora adults and eggs grown on coral nurseries versus native reef on the GBR, with adults assessed at two sites (Blue Lagoon and Rayban) and eggs assessed at one site (Blue Lagoon). Lipid profiles of adult colonies varied by site and origin (nursery versus wild reef), with adult nursery corals exhibiting an elevated relative abundance of storage lipids (diacylglycerols and triacylglycerols) and lipid classes responsible for regulating membrane structure (phosphatidylcholines and sterol esters), while wild corals were characterised by a greater relative abundance of fatty acids and classes involved in immunoregulation. Comparing eggs from different origins, nursery offspring were richer in energy-storing triacylglycerols, as well as ceramides and phosphatidylcholines essential for membrane structure, while wild eggs had a greater relative abundance of wax ester species also important for energy storage. No differences were found in total protein abundance (adult or eggs) or egg physical characteristics (count and size) between nursery and wild origins. Variations in lipid profiles are consistent with differences in environmental conditions between reef sites and origin (nursery versus wild), highlighting the need to consider site selection and propagation conditions when planning restoration projects. Importantly, these findings demonstrate that the lipid classes with the highest relative abundance in A. millepora nursery and wild eggs differed from those in adults from the same origin, suggesting that propagation origin is more important for driving lipid profiles in coral eggs compared to parental effects. [ABSTRACT FROM AUTHOR]

Coral reefs are highly diverse ecosystems of immense ecological, economic, and aesthetic importance built on the calcium-carbonate-based skeletons of stony corals. The formation of these skeletons is threatened by increasing ocean temperatures and acidification, and a deeper understanding of the molecular mechanisms involved may assist efforts to mitigate the effects of such anthropogenic stressors. In this study, we focused on the role of the predicted bicarbonate transporter SLC4γ, which was suggested in previous studies to be a product of gene duplication and to have a role in coral-skeleton formation. Our comparative-genomics study using 30 coral species and 15 outgroups indicates that SLC4γ is present throughout the stony corals, but not in their non-skeleton-forming relatives, and apparently arose by gene duplication at the onset of stony-coral evolution. Our expression studies show that SLC4γ, but not the closely related and apparently ancestral SLC4ß, is highly upregulated during coral development coincident with the onset of skeleton deposition. Moreover, we show that juvenile coral polyps carrying CRISPR/Cas9-induced mutations in SLC4γ are defective in skeleton formation, with the severity of the defect in individual animals correlated with their frequencies of SLC4γ mutations. Taken together, the results suggest that the evolution of the stony corals involved the neofunctionalization of the newly arisen SLC4γ for a unique role in the provision of concentrated bicarbonate for calcium-carbonate deposition. The results also demonstrate the feasibility of reverse-genetic studies of ecologically important traits in adult corals. [ABSTRACT FROM AUTHOR]

Reproductive traits such as fecundity (i.e., the number of gametes produced) and the size and age of coral colonies at reproductive onset can vary in predictable ways among life history strategies. However, most studies on the onset of reproductive maturity in corals only report the presence or absence of oocytes with little known about variation in fecundity across size and age classes. This study aimed to determine the colony size and fecundity at the onset of reproductive maturity across size classes of two scleractinian corals with contrasting life history strategies, Acropora millepora (competitive) and Favites colemani (stress-tolerant). Colonies at a site in northwestern Philippines were sampled to determine the smallest colony size class with mature oocytes and to estimate fecundity across size classes. Histological slides were also prepared to verify the presence of mature gametes. Colonies were able to produce mature oocytes when they had attained colony diameters of 4.7 cm for A. millepora and 1.5 cm for F. colemani. A. millepora had lower fecundity, but larger oocytes compared to F. colemani. Although small colonies can contribute to the larval pool, the proportion of mature colonies increased for larger size classes, suggesting that larger colonies make a disproportionately greater contribution to population reproductive output. These findings contribute to our understanding of coral population dynamics, particularly in parameterizing population and demographic models for different coral life histories. [ABSTRACT FROM AUTHOR]

Significant genetic differentiation between Symbiodiniaceae populations in coral hosts can be induced by a range of factors including geography, latitude, depth, temperature and light utilisation. The conventional method of measuring Symbiodiniaceae diversity involving the ITS2 region of rDNA has several limitations, stemming from insufficient genetic resolution and the multi-copy nature of the marker. This could be improved by using higher throughput whole genome sequencing to identify fine-scale population genetic differences and provide new insight into factors influencing coral-Symbiodiniaceae associations. The aim of this study was to investigate the genetic diversity of Symbiodiniaceae populations using low-coverage whole genome sequencing in sympatric populations of Acropora cf. secale and allopatric populations of Acropora millepora that reproduce in different seasons in Western Australia. Genetic diversity of Symbiodiniaceae populations in these two species was examined using principal coordinates analysis and permutational analysis of variance. This analysis revealed that while all colonies were dominated by Cladocopium , there was a significant genetic difference between Symbiodiniaceae populations in both species. In A. millepora , this variation could be due to the latitudinal variation between populations or differences in reproductive seasonality, but in sympatric populations of A. cf. secale , genetic differences between Symbiodiniaceae populations were clearly aligned with the reproductive seasonality of the coral host. The use of whole genome sequencing improved the sensitivity to detect Symbiodiniaceae genetic population structure between coral populations, which increases our ability to identify genetic and potentially functional differences associated with variation in Symbiodiniaceae populations., Competing Interests: The authors declare no conflicts of interest., (© 2025 The Author(s). Ecology and Evolution published by John Wiley & Sons Ltd.)

Reef-building corals are among the largest producers of dimethylsulfoniopropionate (DMSP), an essential compound in marine biogeochemical cycles. DMSP can be catabolised in coral mucus by a wide diversity of coral-associated bacteria, where it can either be demethylated, leading to the incorporation of sulfur and carbon into bacterial biomass – or cleaved by lyases, releasing the climatically-active gas dimethyl sulfide (DMS). It has been demonstrated that thermal stress increases DMSP concentrations in many coral species, however the effect of increased DMSP availability on coral-associated bacteria has not been explored. Here we performed thermal stress experiments to examine how changes in DMSP availability impact bacterial degradation pathways in the mucus of Acropora millepora. DMSP concentrations increased with temperature, reaching a maximum of 177.3 μM after 10 days of heat stress, which represents the highest concentration of DMSP recorded in any environment to date. Bacterial communities in coral mucus were significantly different from the surrounding seawater, yet they did not vary significantly between temperature or time. However, during thermal stress, when DMSP concentrations increased, a significant increase in the abundance of both the demethylation gene dmdA and the cleavage gene dddP were recorded. Importantly, our results show that for the highest DMSP concentrations recorded (above 30 μM), the cleavage pathway became more abundant than the demethylation pathway. This suggests that under high DMSP concentrations characteristic of heat stress, a larger fraction of the DMSP pool in the coral mucus is likely catabolised through the DMS-producing cleavage pathway.

Studies of broadcast spawning in corals are fundamental to our understanding of early life history characteristics, reproductive biology, restoration etc. Spawning of corals for research is routinely conducted, but this is mostly restricted to sites adjacent to reefs and from broodstock collected from the wild just prior to gamete release. Only recently has it been possible to induce predictable broadcast spawning in an ex situ environment, and nobody has successfully closed the life cycle (i.e., production of an F2 generation) of these corals. Here, for the first time, we closed the life cycle of the broadcast spawning coral Acropora millepora in a fully closed ex situ mesocosm. This breakthrough has numerous implications for our understanding of reproductive biology, specifically it offers potential to deepen our understanding of the genetic influence on adaptive traits such as heat tolerance, growth and disease resistance over multiple generations. [ABSTRACT FROM AUTHOR]

Coral reefs are declining globally as climate change and local water quality press environmental conditions beyond the physiological tolerances of holobionts—the collective of the host and its microbial symbionts. To assess the relationship between symbiont composition and holobiont stress tolerance, community diversity metrics were quantified for dinoflagellate endosymbionts (Family: Symbiodiniaceae) from eight Acropora millepora genets that thrived under or responded poorly to various stressors. These eight selected genets represent the upper and lower tails of the response distribution of 40 coral genets that were exposed to four stress treatments (and control conditions) in a 10‐day experiment. Specifically, four 'best performer' coral genets were analyzed at the end of the experiment because they survived high temperature, high pCO2, bacterial exposure, or combined stressors, whereas four 'worst performer' genets were characterized because they experienced substantial mortality under these stressors. At the end of the experiment, seven of eight coral genets mainly hosted Cladocopium symbionts, whereas the eighth genet was dominated by both Cladocopium and Durusdinium symbionts. Symbiodiniaceae alpha and beta diversity were higher in worst performing genets than in best performing genets. Symbiont communities in worst performers also differed more after stress exposure relative to their controls (based on normalized proportional differences in beta diversity), than did best performers. A generalized joint attribute model estimated the influence of host genet and treatment on Symbiodiniaceae community composition and identified strong associations among particular symbionts and host genet performance, as well as weaker associations with treatment. Although dominant symbiont physiology and function contribute to host performance, these findings emphasize the importance of symbiont community diversity and stochasticity as components of host performance. Our findings also suggest that symbiont community diversity metrics may function as indicators of resilience and have potential applications in diverse disciplines from climate change adaptation to agriculture and medicine. [ABSTRACT FROM AUTHOR]

Abstract Background Climate change causes the breakdown of the symbiotic relationships between reef-building corals and their photosynthetic symbionts (genus Symbiodinium), with thermal anomalies in 2015–2016 triggering the most widespread mass coral bleaching on record and unprecedented mortality on the Great Barrier Reef. Targeted studies using specific coral stress indicators have highlighted the complexity of the physiological processes occurring during thermal stress, but have been unable to provide a clear mechanistic understanding of coral bleaching. Results Here, we present an extensive multi-trait-based study in which we compare the thermal stress responses of two phylogenetically distinct and widely distributed coral species, Acropora millepora and Stylophora pistillata, integrating 14 individual stress indicators over time across a simulated thermal anomaly. We found that key stress responses were conserved across both taxa, with the loss of symbionts and the activation of antioxidant mechanisms occurring well before collapse of the physiological parameters, including gross oxygen production and chlorophyll a. Our study also revealed species-specific traits, including differences in the timing of antioxidant regulation, as well as drastic differences in the production of the sulfur compound dimethylsulfoniopropionate during bleaching. Indeed, the concentration of this antioxidant increased two-fold in A. millepora after the corals started to bleach, while it decreased 70% in S. pistillata. Conclusions We identify a well-defined cascading response to thermal stress, demarking clear pathophysiological reactions conserved across the two species, which might be central to fully understanding the mechanisms triggering thermally induced coral bleaching. These results highlight that bleaching is a conserved mechanism, but specific adaptations linked to the coral’s antioxidant capacity drive differences in the sensitivity and thus tolerance of each coral species to thermal stress.

Abstract Background Dimethylsulfoniopropionate (DMSP) is a small sulphur compound which is produced in prodigious amounts in the oceans and plays a pivotal role in the marine sulfur cycle. Until recently, DMSP was believed to be synthesized exclusively by photosynthetic organisms; however we now know that corals and specific bacteria can also produce this compound. Corals are major sources of DMSP, but the molecular basis for its biosynthesis is unknown in these organisms. Results Here we used salinity stress, which is known to trigger DMSP production in other organisms, in conjunction with transcriptomics to identify coral genes likely to be involved in DMSP biosynthesis. We focused specifically on both adults and juveniles of the coral Acropora millepora: after 24 h of exposure to hyposaline conditions, DMSP concentrations increased significantly by 2.6 fold in adult corals and 1.2 fold in juveniles. Concomitantly, candidate genes enabling each of the necessary steps leading to DMSP production were up-regulated. Conclusions The data presented strongly suggest that corals use an algal-like pathway to generate DMSP from methionine, and are able to rapidly change expression of the corresponding genes in response to environmental stress. However, our data also indicate that DMSP is unlikely to function primarily as an osmolyte in corals, instead potentially serving as a scavenger of ROS and as a molecular sink for excess methionine produced as a consequence of proteolysis and osmolyte catabolism in corals under hypo-osmotic conditions.

Climate change threatens organisms in a variety of interactive ways that requires simultaneous adaptation of multiple traits. Predicting evolutionary responses requires an understanding of the potential for interactions among stressors and the genetic variance and covariance among fitness‐related traits that may reinforce or constrain an adaptive response. Here we investigate the capacity of Acropora millepora, a reef‐building coral, to adapt to multiple environmental stressors: rising sea surface temperature, ocean acidification, and increased prevalence of infectious diseases. We measured growth rates (weight gain), coral color (a proxy for Symbiodiniaceae density), and survival, in addition to nine physiological indicators of coral and algal health in 40 coral genets exposed to each of these three stressors singly and combined. Individual stressors resulted in predicted responses (e.g., corals developed lesions after bacterial challenge and bleached under thermal stress). However, corals did not suffer substantially more when all three stressors were combined. Nor were trade‐offs observed between tolerances to different stressors; instead, individuals performing well under one stressor also tended to perform well under every other stressor. An analysis of genetic correlations between traits revealed positive covariances, suggesting that selection to multiple stressors will reinforce rather than constrain the simultaneous evolution of traits related to holobiont health (e.g., weight gain and algal density). These findings support the potential for rapid coral adaptation under climate change and emphasize the importance of accounting for corals' adaptive capacity when predicting the future of coral reefs. [ABSTRACT FROM AUTHOR]

Highlights from the article: Key words: Acropora millepora, Acropora digitifera, genome, WGS. (f, g) Circos plots showing relationships between the A. millepora and A. digitifera genomes. 420-Mb genome size estimate for A. digitifera based on The genome assembly and gene model data set, the CEGMA Acropora millepora, Acropora digitifera, genome, WGS.

Coral-dinoflagellate symbiosis is the key biological interaction enabling existence of modern-type coral reefs, but the mechanisms regulating initial host–symbiont attraction, recognition and symbiont proliferation thus far remain largely unclear. A common reef-building coral, Acropora millepora, displays conspicuous fluorescent polymorphism during all phases of its life cycle, due to the differential expression of fluorescent proteins (FPs) of the green fluorescent protein family. In this study, we examine whether fluorescent variation in young coral juveniles exposed to natural sediments is associated with the uptake of disparate Symbiodinium assemblages determined using ITS-2 deep sequencing. We found that Symbiodinium assemblages varied significantly when redness values varied, specifically in regards to abundances of clades A and C. Whether fluorescence was quantified as a categorical or continuous trait, clade A was found at higher abundances in redder juveniles. These preliminary results suggest juvenile fluorescence may be associated with Symbiodinium uptake, potentially acting as either an attractant to ecologically specific types or as a mechanism to modulate the internal light environment to control Symbiodinium physiology within the host.

The success of corals in nutrient poor environments is largely attributed to the symbiosis between the cnidarian host and its intracellular alga. Warm water anomalies have been shown to destabilize this symbiosis, yet detailed analysis of the effect of temperature and expulsion on cell-specific carbon and nutrient allocation in the symbiont is limited. Here, we exposed colonies of the hard coral Acropora millepora to heat stress and using synchrotron-based infrared microspectroscopy measured the biomolecular profiles of individual in hospite and expelled symbiont cells at an acute state of bleaching. Our results showed symbiont metabolic profiles to be remarkably distinct with heat stress and expulsion, where the two effectors elicited opposing metabolic adjustments independent of treatment or cell type. Elevated temperature resulted in biomolecular changes reflecting cellular stress, with relative increases in free amino acids and phosphorylation of molecules and a concomitant decline in protein content, suggesting protein modification and degradation. This contrasted with the metabolic profiles of expelled symbionts, which showed relative decreases in free amino acids and phosphorylated molecules, but increases in proteins and lipids, suggesting expulsion lessens the overall effect of heat stress on the metabolic signature of the algal symbionts. Interestingly, the combined effects of expulsion and thermal stress were additive, reducing the overall shifts in all biomolecules, with the notable exception of the significant accumulation of lipids and saturated fatty acids. This first use of a single-cell metabolomics approach on the coral symbiosis provides novel insight into coral bleaching and emphasizes the importance of a single-cell approach to demark the cell-to-cell variability in the physiology of coral cellular populations.

Coral populations continue to decline in many regions because of natural and anthropogenic disturbances; and the persistence of coral populations mainly relies on natural recruitment through coral reproduction. However, studies on the reproduction of many important reef-building coral species in one of the most biodiverse region, the Coral Triangle, are very limited. Here, we investigated the reproductive biology of the coral Acropora millepora in Bolinao-Anda Reef Complex, northwestern Philippines from February 2014 to March 2015, through in situ assessment of gamete maturity, histological examination of gamete development and ex situ spawning observations. Results showed that A. millepora exhibited an annual gametogenic cycle. Formation of oocytes took place within 8 months, whereas spermatocytes developed in 3 months before the gamete release. Abundance and size of eggs in A. millepora were greatest during the months nearest to the spawning period, which coincides with the rapid increase in sea surface temperature. Ex situ broadcast spawning occurred around the months of February-March, which suggests an extended spawning pattern. Results provide additional information on coral reproduction in the region and reveal the potential of A. millepora in providing coral propagules for the replenishment and recovery of degraded coral populations. [ABSTRACT FROM AUTHOR]

Fire corals of the hydrocoral genus Millepora provide an important ecological role as framework builders of coral reefs in the Indo-Pacific and the Atlantic. Recent works have demonstrated the incongruence between molecular data and the traditional taxonomy of Millepora spp. based on overall skeleton growth form and pores. In an attempt to establish a reliable and standardized approach for defining species boundaries in Millepora, we focused on those from the Red Sea. In this region, three species are currently recognized: the fan-shaped branching M. dichotoma, the blade-like M. platyphylla, and the massive/encrusting M. exaesa. A total of 412 colonies were collected from six localities. Two mitochondrial marker genes (COI and 16S rDNA) were sequenced to obtain phylogeny reconstructions and haplotype networks. Eight morphological traits of pores and the nematocysts of both polyp and eumedusoid stages were measured to determine whether significant morphological differences occur among the three species. Both markers clearly resolved M. dichotoma, M. platyphylla, and M. exaesa as distinct, monophyletic lineages in the Red Sea. Nevertheless, they also revealed deep genetic breaks with Southwestern Indian Ocean populations of the three species. In the Red Sea, the three species were further distinguished based on their pore and nematocyst features. A discriminant analysis revealed dactylopore density, number of dactylopores per gastropore, dactylopore distance, and gastropore diameter as the most informative discriminative characters. The heteronemes, the large and small stenoteles of polyps, and the distribution of mastigophores of eumedusoids also showed significant interspecific differences. An integrated morpho-molecular approach proved to be decisive in defining species boundaries of Millepora supported by a combination of pore and nematocyst characters, which may be phylogenetically informative. [ABSTRACT FROM AUTHOR]

Abstract Elevated suspended sediment concentrations (SSCs) often impact coral fertilisation success, but sediment composition can influence effect thresholds, which is problematic for accurately predicting risk. Here, we derived concentration–response thresholds and cause-effect pathways for SSCs comprising a range of realistic mineral and organic compositions on coral fertilisation success. Effect concentration thresholds (EC 10 : 10% fertilisation inhibition) varied markedly, with fertilisation highly sensitive to inshore organic-clay rich sediments and bentonite clay at <5 mg L−1. Mineral clays and organic matter within these sediments likely promoted flocculation of the coral sperm, which in turn reduced fertilisation. In contrast, sediments lacking these properties bound less sperm, leading to higher SSC thresholds for coral fertilisation (EC 10 > 40 mg L−1). The effect thresholds for relevant sediment types were combined with in situ turbidity data from locations near dredging operations to assess the risks posed by dredging to coral fertilisation at these locations. Highlights • Sediment type strongly influences the effect concentration thresholds for coral fertilisation success. • Mineral clays and organic matter promote the sinking of sperm via sediment-sperm flocculation. • "Cohesive" sediments pose the greatest risk to coral fertilisation during dredging or natural high turbidity events. • The risk posed by less 'cohesive' sediments during dredging could be alleviated with a spatial separation distance of >1 km. • Sediment type characterisation at proposed dredging sites would improve risk assessments for coral fertilisation impacts. [ABSTRACT FROM AUTHOR]

Scleratinian corals (hard corals) and their symbiotic algae are the ecological engineers of biodiverse and geological important coral reef habitats. The complex, linked physiological processes that enable the holobiont (coral + algae) to calcify and generate reef structures are consequently of great interest. However, the mechanism of calcification is difficult to study for several reasons including the small spatial scales of the processes and the close coupling between the symbiont and host. In this study, we explore the utility of pH and Ca 2+ microelectrodes for constraining the rates and spatial distribution of photosynthesis, respiration, and calcification. The work focuses on vertical profiles of pH and Ca 2+ through the coelenteron cavity, a semi-isolated compartment of modified seawater amenable to quantitative interpretation. In two studied species, Turbinaria reniformis and Acropora millepora , Ca 2+ concentrations decreased in a roughly linear manner from the mouth to the base of the coelenteron, indicating the primary physiological process affecting Ca 2+ concentration is removal for calcification below the coelenteron. In contrast, the H + concentration remained relatively constant over much of the coelenteron cavity before it increased sharply toward the base of the coelenteron, indicative of proton-pumping from the calcification fluid below. The estimated H + gradient between the coelenteron cavity and the calcification site was >10 times higher than previously predicted. Consequently, the energy required to export protons from the calcifying fluid was estimated to be ~3 times higher than previously calculated. A one-dimensional reaction-diffusion model was used to interpret the pH profile considering the effects of photosynthesis, respiration, and calcification. This model provided a good fit to the observed pH profile and helped to constrain the rates and spatial distribution of these processes. Our modeling results also suggested that adult corals with deeper polyps may be more sensitive to ocean acidification (OA) because of enhanced difficulty to transport H + out of the coelenteron and into the surrounding seawater. [ABSTRACT FROM AUTHOR]

Abstract: Reef‐building corals can increase their resistance to heat‐induced bleaching through adaptation and acclimatization and/or by associating with a more thermo‐tolerant strain of algal symbiont (Symbiodinium sp.). Here, we show that these two adaptive pathways interact. We collected Acropora millepora corals from two contrasting thermal environments on the Great Barrier Reef: cooler, mid‐latitude Orpheus Island, where all corals hosted a heat‐sensitive clade C Symbiodinium, and warmer, low‐latitude Wilkie Island, where corals hosted either a clade C or a more thermo‐tolerant clade D. Corals were kept in a benign common garden to reveal differences in baseline gene expression, reflecting prior adaptation/long‐term acclimatization. Model‐based analysis identified gene expression differences between Wilkie and Orpheus corals that were negatively correlated with previously described transcriptome‐wide signatures of heat stress, signifying generally elevated thermotolerance of Wilkie corals. Yet, model‐free analyses of gene expression revealed that Wilkie corals hosting clade C were distinct from Wilkie corals hosting clade D, whereas Orpheus corals were more variable. Wilkie corals hosting clade C symbionts exhibited unique functional signatures, including downregulation of histone proteins and ion channels and upregulation of chaperones and RNA processing genes, putatively representing constitutive “frontloading” of stress response genes. Furthermore, clade C Symbiodinium exhibited constitutive expression differences between Wilkie and Orpheus, indicative of contrasting life history strategies. Our results demonstrate that hosting alternative Symbiodinium types is associated with different pathways of local adaptation for the coral host. These interactions could play a significant role in setting the direction of genetic adaptation to global warming in the two symbiotic partners. [ABSTRACT FROM AUTHOR]

Coral-dinoflagellate symbiosis is the key biological interaction enabling existence of modern-type coral reefs, but the mechanisms regulating initial host-symbiont attraction, recognition and symbiont proliferation thus far remain largely unclear. A common reef-building coral, Acropora millepora, displays conspicuous fluorescent polymorphism during all phases of its life cycle, due to the differential expression of fluorescent proteins (FPs) of the green fluorescent protein family. In this study, we examine whether fluorescent variation in young coral juveniles exposed to natural sediments is associated with the uptake of disparate Symbiodinium assemblages determined using ITS-2 deep sequencing. We found that Symbiodinium assemblages varied significantly when redness values varied, specifically in regards to abundances of cladesAand C. Whether fluorescence was quantified as a categorical or continuous trait, clade A was found at higher abundances in redder juveniles. These preliminary results suggest juvenile fluorescence may be associated with Symbiodinium uptake, potentially acting as either an attractant to ecologically specific types or as a mechanism to modulate the internal light environment to control Symbiodinium physiology within the host. [ABSTRACT FROM AUTHOR]

Reef-building corals are critically important species that are threatened by anthropogenic stresses including climate change. In attempts to understand corals' responses to stress and other aspects of their biology, numerous genomic and transcriptomic studies have been performed, generating a variety of hypotheses about the roles of particular genes and molecular pathways. However, it has not generally been possible to test these hypotheses rigorously because of the lack of genetic tools for corals. Here, we demonstrate efficient genome editing using the CRISPR/Cas9 system in the coral Acropora millepora. We targeted the genes encoding fibroblast growth factor 1a (FGF1a), green fluorescent protein (GFP), and red fluorescent protein (RFP). After microinjecting CRISPR/Cas9 ribonucleoprotein complexes into fertilized eggs, we detected induced mutations in the targeted genes using changes in restriction-fragment length, Sanger sequencing, and high-throughput Illumina sequencing. We observed mutations in ~50% of individuals screened, and the proportions of wild-type and various mutant gene copies in these individuals indicated that mutation induction continued for at least several cell cycles after injection. Although multiple paralogous genes encoding green fluorescent proteins are present in A. millepora, appropriate design of the guide RNA allowed us to induce mutations simultaneously in more than one paralog. Because A. millepora larvae can be induced to settle and begin colony formation in the laboratory, CRISPR/Cas9- based gene editing should allow rigorous tests of gene function in both larval and adult corals. [ABSTRACT FROM AUTHOR]

Coral reefs are essential to many nations, and are currently in global decline. Although climate models predict decreases in seawater pH (∼0.3 units) and oxygen saturation (∼5 percentage points), these are exceeded by the current daily pH and oxygen fluctuations on many reefs (pH 7.8–8.7 and 27–241% O2 saturation). We investigated the effect of oxygen and pH fluctuations on coral calcification in the laboratory using the model species Acropora millepora. Light calcification rates were greatly enhanced (+178%) by increased seawater pH, but only at normoxia; hyperoxia completely negated this positive effect. Dark calcification rates were significantly inhibited (51–75%) at hypoxia, whereas pH had no effect. Our preliminary results suggest that within the current oxygen and pH range, oxygen has substantial control over coral growth, whereas the role of pH is limited. This has implications for reef formation in this era of rapid climate change, which is accompanied by a decrease in seawater oxygen saturation owing to higher water temperatures and coastal eutrophication.

Heterotrophy plays an important role in improving coral growth rate in aquaculture. While a lack of sufficient food will reduce growth and delay production timelines, excessive food deteriorates water quality and represents an additional production cost without benefit. Currently, there is limited information about the maximum feeding rates of corals. Using the curvilinear Michaelis-Menten model with six different concentrations of Artemia during day and night feeding, we estimated the maximum feeding rates for Acropora millepora (4.6 ind cm − 2 h − 1 ), Hydnophora rigida (20.4 ind cm − 2 h − 1 ) and Duncanopsammia axifuga (22.8 ind polyp − 1 h − 1 ) and the necessary concentrations of food to reach the maximum levels of feeding. Overall, the three species increased their feeding rates with increasing Artemia densities until the concentration reached 120 ind ml − 1 , after which there was no change to feeding rate. No significant differences were detected between day and night feeding rates for the three corals species. We demonstrate that the concentration to reach the maximum Artemia feeding rates for these commercially important coral species was above 50 ind ml − 1 . [ABSTRACT FROM AUTHOR]

Rising seawater temperature and ocean acidification threaten the survival of coral reefs. The relationship between coral physiology and its microbiome may reveal why some corals are more resilient to these global change conditions. Here, we conducted the first experiment to simultaneously investigate changes in the coral microbiome and coral physiology in response to the dual stress of elevated seawater temperature and ocean acidification expected by the end of this century. Two species of corals, Acropora millepora containing the thermally sensitive endosymbiont C21a and Turbinaria reniformis containing the thermally tolerant endosymbiont Symbiodinium trenchi, were exposed to control (26.5°C and pCO2 of 364 μatm) and treatment (29.0°C and pCO2 of 750 μatm) conditions for 24 days, after which we measured the microbial community composition. These microbial findings were interpreted within the context of previously published physiological measurements from the exact same corals in this study (calcification, organic carbon flux, ratio of photosynthesis to respiration, photosystem II maximal efficiency, total lipids, soluble animal protein, soluble animal carbohydrates, soluble algal protein, soluble algal carbohydrate, biomass, endosymbiotic algal density, and chlorophyll a). Overall, dually stressed A. millepora had reduced microbial diversity, experienced large changes in microbial community composition, and experienced dramatic physiological declines in calcification, photosystem II maximal efficiency, and algal carbohydrates. In contrast, the dually stressed coral T. reniformis experienced a stable and more diverse microbiome community with minimal physiological decline, coupled with very high total energy reserves and particulate organic carbon release rates. Thus, the microbiome changed and microbial diversity decreased in the physiologically sensitive coral with the thermally sensitive endosymbiotic algae but not in the physiologically tolerant coral with the thermally tolerant endosymbiont. Our results confirm recent findings that temperature-stress tolerant corals have a more stable microbiome, and demonstrate for the first time that this is also the case under the dual stresses of ocean warming and acidification. We propose that coral with a stable microbiome are also more physiologically resilient and thus more likely to persist in the future, and shape the coral species diversity of future reef ecosystems. [ABSTRACT FROM AUTHOR]

Little is known about the potential for acclimatization or adaptation of corals to ocean acidification and even less about the molecular mechanisms underpinning these processes. Here, we examine global gene expression patterns in corals and their intracellular algal symbionts from two replicate population pairs in Papua New Guinea that have undergone long-term acclimatization to natural variation in pCO2. In the coral host, only 61 genes were differentially expressed in response to pCO2 environment, but the pattern of change was highly consistent between replicate populations, likely reflecting the core expression homeostasis response to ocean acidification. Functional annotations highlight lipid metabolism and a change in the stress response capacity of corals as key parts of this process. Specifically, constitutive downregulation of molecular chaperones was observed, which may impact response to combined climate change-related stressors. Elevated CO2 has been hypothesized to benefit photosynthetic organisms but expression changes of in hospite Symbiodinium in response to acidification were greater and less consistent among reef populations. This population-specific response suggests hosts may need to adapt not only to an acidified environment, but also to changes in their Symbiodinium populations that may not be consistent among environments, adding another challenging dimension to the physiological process of coping with climate change. [ABSTRACT FROM AUTHOR]

Scleractinian corals are colonial organisms comprising multiple physiologically integrated polyps and branches. Colonialism in corals is highly beneficial, and allows a single colony to undergo several life processes at once through physiological integra- tion and compartmentalised functioning. Elucidating differences in the biochemical composition of intra-colonial branch positions will provide valuable insight into the nutritional reserves underlying different regions in individual coral colonies. This will also ascertain prudent harvesting strategies of wild donor-colonies to generate coral stock with high survival and vigour prospects for reef-rehabilitation efforts and captive husbandry. This study examined the effects of colony branch position on the nutritional profile of two different colony sizes of the common scleractinian, Acropora millepora. For smaller colonies, branches were sampled at three locations: the colony centre (S-centre), 50% of the longitudinal radius length (LRL) (S-50), and the colony edge (S-edge). For larger colonies, four locations were sampled: the colony centre (L-centre), 33.3% of the LRL (L-33), 66.6% of the LRL (L-66), and the edge (L-edge). Results demonstrate significant branch position effects, with the edge regions containing higher protein, likely due to increased tissue synthesis and calcification. Meanwhile, storage lipid and total fatty acid concentrations were lower at the edges, possibly reflecting catabolism of high-energy nutrients to support proliferating cells. Results also showed a significant effect of colony size in the two classes examined. While the major protein and structural lipid sink was exhibited at the edge for both sizes, the major sink for high-energy lipids and fatty acids appeared to be the L-66 position of the larger colonies and the S-centre and S-50 positions for the smaller colonies. These results confirm that the scleractinian coral colony is not nutritionally homogeneous, and while different regions of the coral colony are functionally specialised, so too are their nutritional profiles geared toward meeting specific energetic demands. [ABSTRACT FROM AUTHOR]

Successful recruitment in corals is important for the sustenance of coral reefs, and is considered a demographic bottleneck in the recovery of reef populations following disturbance events. Yet several factors influence larval settlement behaviour, and here we quantified thresholds associated with light attenuation and accumulated sediments on settlement substrates. Sediments deposited on calcareous red algae (CRA) directly and indirectly impacted coral settlement patterns. Although not avoiding direct contact, Acropora millepora larvae were very reluctant to settle on surfaces layered with sediments, progressively shifting their settlement preference from upward to downward facing (sediment-free) surfaces under increasing levels of deposited sediment. When only upward-facing surfaces were presented, 10% of settlement was inhibited at thresholds from 0.9 to 16 mg cm − 2 (EC 10 ), regardless of sediment type (carbonate and siliciclastic) or particle size (fine and coarse silt). These levels equate to a very thin (< 150 μm) veneer of sediment that occurs within background levels on reefs. Grooves within settlement surfaces slightly improved options for settlement on sediment-coated surfaces (EC 10 : 29 mg cm − 2 ), but were quickly infilled at higher deposited sediment levels. CRA that was temporarily smothered by sediment for 6 d became bleached (53% surface area), and inhibited settlement at ~ 7 mg cm − 2 (EC 10 ). A minor decrease in settlement was observed at high and very low light intensities when using suboptimal concentrations of a settlement inducer (CRA extract); however, no inhibition was observed when natural CRA surfaces along with more realistic diel-light patterns were applied. The low deposited sediment thresholds indicate that even a thin veneer of sediment can have consequences for larval settlement due to a reduction of optimal substrate. And while grooves and overhangs provide more settlement options in high deposition areas, recruits settling at these locations may be subject to ongoing stress from shading, competition, and sediment infilling. [ABSTRACT FROM AUTHOR]

Background: Climate change causes the breakdown of the symbiotic relationships between reef-building corals and their photosynthetic symbionts (genus Symbiodinium), with thermal anomalies in 2015-2016 triggering the most widespread mass coral bleaching on record and unprecedented mortality on the Great Barrier Reef. Targeted studies using specific coral stress indicators have highlighted the complexity of the physiological processes occurring during thermal stress, but have been unable to provide a clear mechanistic understanding of coral bleaching. Results: Here, we present an extensive multi-trait-based study in which we compare the thermal stress responses of two phylogenetically distinct and widely distributed coral species, Acropora millepora and Stylophora pistillata, integrating 14 individual stress indicators over time across a simulated thermal anomaly. We found that key stress responses were conserved across both taxa, with the loss of symbionts and the activation of antioxidant mechanisms occurring well before collapse of the physiological parameters, including gross oxygen production and chlorophyll a. Our study also revealed species-specific traits, including differences in the timing of antioxidant regulation, as well as drastic differences in the production of the sulfur compound dimethylsulfoniopropionate during bleaching. Indeed, the concentration of this antioxidant increased two-fold in A. millepora after the corals started to bleach, while it decreased 70% in S. pistillata. Conclusions: We identify a well-defined cascading response to thermal stress, demarking clear pathophysiological reactions conserved across the two species, which might be central to fully understanding the mechanisms triggering thermally induced coral bleaching. These results highlight that bleaching is a conserved mechanism, but specific adaptations linked to the coral's antioxidant capacity drive differences in the sensitivity and thus tolerance of each coral species to thermal stress. [ABSTRACT FROM AUTHOR]

This study was conducted to investigate the effects of temperature, pH, ammonia nitrogen content ad salinity on the bleaching of Acropora millepora and Palythoa sp. This study included two experiments: Experiment 1 was designed to test the effects of different gradients of temperature, pH, ammonia nitrogen content and salinity at a time point (6h) on Palythoa sp. breeding; and Experiment 2 tested the effects of different levels of temperature, pH, ammonia nitrogen content and salinity at three time points (2, 48 and 72h) on A. millepora bleaching. Before the experiments, A. millepora and Palythoa sp. were washed with filtered seawater (0.45 µm microporous membrane filtration) and then reared in interior seawater aquarium cylinder. The experiments were carried out when the coral tentacles extended and the growth condition were good. The results showed that under high temperature (32-40 °C ), both A. millepora and Palythoa sp. showed drastic reactions in the course of time, reflecting considerably significant stress responses. The number of releasing zooxathellae increased with increase or decrease of the pH value ( pH 0, 4, 6, 10, 12) over time. Meanwhile, a relatively thick layer of mucus was found on the upper zone of the seawater and the corals started to bleach. As the concentration of ammonia nitrogen ( NH4+, 0 ammo/L) increased in the course of time, the number of zooxathellae floating on A. millepora and Palythoa sp. had the tendency to rise. When the salinity level were at 15%c ad 40%c for A. mlllepora and below 34%c for Palythoa sp., a considerable number of zooxathellae floated out and had rising trend in their number over time at 2 - 72 h and 6 h, respectively. In this occasion, the corals stopped growing and started to bleach. In conclusion, it revealed that temperature, pH, ammonia nitrogen content and salinity are the potential impact factors on coral breeding. It is suggested that, as temperature, pH value and salinity decrease or increase, as well as ammonia nitrogen content rises, the quantity of phycobiont would be raised, and the bleaching would be more severe. [ABSTRACT FROM AUTHOR]

Background: Dimethylsulfoniopropionate (DMSP) is a small sulphur compound which is produced in prodigious amounts in the oceans and plays a pivotal role in the marine sulfur cycle. Until recently, DMSP was believed to be synthesized exclusively by photosynthetic organisms; however we now know that corals and specific bacteria can also produce this compound. Corals are major sources of DMSP, but the molecular basis for its biosynthesis is unknown in these organisms. Results: Here we used salinity stress, which is known to trigger DMSP production in other organisms, in conjunction with transcriptomics to identify coral genes likely to be involved in DMSP biosynthesis. We focused specifically on both adults and juveniles of the coral Acropora millepora: after 24 h of exposure to hyposaline conditions, DMSP concentrations increased significantly by 2.6 fold in adult corals and 1.2 fold in juveniles. Concomitantly, candidate genes enabling each of the necessary steps leading to DMSP production were up-regulated. Conclusions: The data presented strongly suggest that corals use an algal-like pathway to generate DMSP from methionine, and are able to rapidly change expression of the corresponding genes in response to environmental stress. However, our data also indicate that DMSP is unlikely to function primarily as an osmolyte in corals, instead potentially serving as a scavenger of ROS and as a molecular sink for excess methionine produced as a consequence of proteolysis and osmolyte catabolism in corals under hypo-osmotic conditions. [ABSTRACT FROM AUTHOR]

Coral reefs surrounding the islands lying close to the coast are unique to the Great Barrier Reef (GBR) in that they are frequently exposed to disturbance events including floods caused by cyclonic rainfall, strong winds and occasional periods of prolonged above-average temperatures during summer. In one such group of islands in the southern GBR, the Keppel Island archipelago, climate-driven disturbances frequently result in major coral mortality. Whilst these island reefs have clearly survived such dramatic disturbances in the past, the consequences of extreme mortality events may include the loss of genetic diversity, and hence adaptive potential, and a reduction in fitness due to inbreeding, especially if new recruitment from external sources is limited. Here we examined the level of isolation of the Keppel Island group as well as patterns of gene flow within the Keppel Islands using 10 microsatellite markers in nine populations of the coral, Acropora millepora. Bayesian cluster analysis and assignment tests indicated gene flow is restricted, but not absent, between the outer and inner Keppel Island groups, and that extensive gene flow exists within each of these island groups. Comparison of the Keppel Island data with results from a previous GBR-wide study that included a single Keppel Island population, confirmed that A. millepora in the Keppel Islands is genetically distinct from populations elsewhere on the GBR, with exception of the nearby inshore High Peak Reef just north of the Keppel Islands. We compared patterns of genetic diversity in the Keppel Island populations with those from other GBR populations and found them to be slightly, but significantly lower, consistent with the archipelago being geographically isolated, but there was no evidence for recent bottlenecks or deviation from mutation-drift equilibrium. A high incidence of private alleles in the Keppel Islands, particularly in the outer islands, supports their relative isolation and contributes to the conservation value of the archipelago. The lack of evidence for genetic erosion, in combination with our observation that the North Keppel Island population samples collected in 2002 and 2008, respectively, exhibited a pairwise genetic distance of zero, supports previous published work indicating that, following bleaching, Acropora corals in the Keppel Islands predominantly recover from regrowth of small amounts of remaining live tissue in apparently dead coral colonies. This is likely supplemented by recruitment of larvae from genetically similar, less disturbed populations at nearby reefs, particularly following extreme flood events.

Connectivity underpins the persistence and recovery of marine ecosystems. The Great Barrier Reef (GBR) is the world's largest coral reef ecosystem and managed by an extensive network of no-take zones; however, information about connectivity was not available to optimize the network's configuration. We use multivariate analyses, Bayesian clustering algorithms and assignment tests of the largest population genetic data set for any organism on the GBR to date (Acropora tenuis, >2500 colonies; >50 reefs, genotyped for ten microsatellite loci) to demonstrate highly congruent patterns of connectivity between this common broadcast spawning reef-building coral and its congener Acropora millepora (~950 colonies; 20 reefs, genotyped for 12 microsatellite loci). For both species, there is a genetic divide at around 19°S latitude, most probably reflecting allopatric differentiation during the Pleistocene. GBR reefs north of 19°S are essentially panmictic whereas southern reefs are genetically distinct with higher levels of genetic diversity and population structure, most notably genetic subdivision between inshore and offshore reefs south of 19°S. These broadly congruent patterns of higher genetic diversities found on southern GBR reefs most likely represent the accumulation of alleles via the southward flowing East Australia Current. In addition, signatures of genetic admixture between the Coral Sea and outer-shelf reefs in the northern, central and southern GBR provide evidence of recent gene flow. Our connectivity results are consistent with predictions from recently published larval dispersal models for broadcast spawning corals on the GBR, thereby providing robust connectivity information about the dominant reef-building genus Acropora for coral reef managers. [ABSTRACT FROM AUTHOR]

Coral reefs are facing major global and local threats due to climate change-induced increases in dissolved inorganic carbon (DIC) and because of land-derived increases in organic and inorganic nutrients. Recent research revealed that high availability of labile dissolved organic carbon (DOC) negatively affects scleractinian corals. Studies on the interplay of these factors, however, are lacking, but urgently needed to understand coral reef functioning under present and near future conditions. This experimental study investigated the individual and combined effects of ambient and high DIC (pCO2 403 μatm/ pHTotal 8.2 and 996 μatm/pHTotal 7.8) and DOC (added as Glucose 0 and 294 μmol L-1, background DOC concentration of 83 μmol L-1) availability on the physiology (net and gross photosynthesis, respiration, dark and light calcification, and growth) of the scleractinian coral Acropora millepora (Ehrenberg, 1834) from the Great Barrier Reef over a 16 day interval. High DIC availability did not affect photosynthesis, respiration and light calcification, but significantly reduced dark calcification and growth by 50 and 23%, respectively. High DOC availability reduced net and gross photosynthesis by 51% and 39%, respectively, but did not affect respiration. DOC addition did not influence calcification, but significantly increased growth by 42%. Combination of high DIC and high DOC availability did not affect photosynthesis, light calcification, respiration or growth, but significantly decreased dark calcification when compared to both controls and DIC treatments. On the ecosystem level, high DIC concentrations may lead to reduced accretion and growth of reefs dominated by Acropora that under elevated DOC concentrations will likely exhibit reduced primary production rates, ultimately leading to loss of hard substrate and reef erosion. It is therefore important to consider the potential impacts of elevated DOC and DIC simultaneously to assess real world scenarios, as multiple rather than single factors influence key physiological processes in coral reefs. [ABSTRACT FROM AUTHOR]

The Spratly Island archipelago is a remote network of coral reefs and islands in the South China Sea that is a likely source of coral larvae to the greater region, but about which little is known. Using a particle-tracking model driven by oceanographic data from the Coral Triangle region, we simulated both spring and fall spawning events of Acropora millepora, a common coral species, over a 46-yr period (1960-2005). Simulated population biology of A. millepora included the acquisition and loss of competency, settlement over appropriate benthic habitat, and mortality based on experimental data. The simulations aimed to provide insights into the connectivity of reefs within the Spratly Islands, the settlement of larvae on reefs of the greater South China Sea, and the potential dispersal range of reef organisms from the Spratly Islands. Results suggest that (1) the Spratly Islands may be a significant source of A. millepora larvae for the Palawan reefs (Philippines) and some of the most isolated reefs of the South China Sea; and (2) the relatively isolated western Spratly Islands have limited source reefs supplying them with larvae and fewer of their larvae successfully settling on other reefs. Examination of particle dispersal without biology (settlement and mortality) suggests that larval connectivity is possible throughout the South China Sea and into the Coral Triangle region. Strong differences in the spring versus fall larval connectivity and dispersal highlight the need for a greater understanding of spawning dynamics of the region. This study confirms that the Spratly Islands are likely an important source of larvae for the South China Sea and Coral Triangle region. [ABSTRACT FROM AUTHOR]

Background: Research into various aspects of coral biology has greatly increased in recent years due to anthropogenic threats to coral health including pollution, ocean warming and acidification. However, knowledge of coral early development has lagged. The present paper describes the embryonic development of two previously uncharacterized robust corals, Favia lizardensis (a massive brain coral) and Ctenactis echinata (a solitary coral) and compares it to that of the previously characterized complex coral, Acropora millepora, both morphologically and in terms of the expression of a set of key developmental genes. Results: Illumina sequencing of mixed age embryos was carried out, resulting in embryonic transcriptomes consisting of 40605 contigs for C.echinata (N50 = 1080 bp) and 48536 contigs for F.lizardensis (N50 = 1496 bp). The transcriptomes have been annotated against Swiss-Prot and were sufficiently complete to enable the identification of orthologs of many key genes controlling development in bilaterians. Developmental series of images of whole mounts and sections reveal that the early stages of both species contain a blastocoel, consistent with their membership of the robust clade. In situ hybridization was used to examine the expression of the developmentally important genes brachyury, chordin and forkhead. The expression of brachyury and forkhead was consistent with that previously reported for Acropora and allowed us to confirm that the pseudo-blastopore sometimes seen in robust corals such as Favia spp. is not directly associated with gastrulation. C.echinata chordin expression, however, differed from that seen in the other two corals. Conclusions: Embryonic transcriptomes were assembled for the brain coral Favia lizardensis and the solitary coral Ctenactis echinata. Both species have a blastocoel in their early developmental stages, consistent with their phylogenetic position as members of the robust clade. Expression of the key developmental genes brachyury, chordin and forkhead was investigated, allowing comparison to that of their orthologs in Acropora, Nematostella and bilaterians and demonstrating that even within the Anthozoa there are significant differences in expression patterns. [ABSTRACT FROM AUTHOR]