Machine-Learning-Based Proteomic Predictive Modeling with Thermally-Challenged Caribbean Reef Corals

Coral health is currently diagnosed retroactively; colonies are deemed “stressed” upon succumbing to bleaching or disease. Ideally, health inferences would instead be made on a pre-death timescale that would enable, for instance, environmental mitigation that could promote coral resilience. To this end, diverse Caribbean coral (Orbicella faveolata) genotypes of varying resilience to high temperatures along the Florida Reef Tract were exposed herein to elevated temperatures in the laboratory, and a proteomic analysis was taken with a subset of 20 samples via iTRAQ labeling followed by nano-liquid chromatography + mass spectrometry; 46 host coral and 40 Symbiodiniaceae dinoflagellate proteins passed all stringent quality control criteria, and the partial proteomes of biopsies of (1) healthy controls, (2) sub-lethally stressed samples, and (3) actively bleaching corals differed significantly from one another. The proteomic data were then used to train predictive models of coral colony bleaching susceptibility, and both generalized regression and machine-learning-based neural networks were capable of accurately forecasting the bleaching susceptibility of coral samples based on their protein signatures. Successful future testing of the predictive power of these models in situ could establish the capacity to proactively monitor coral health.

Transcriptomics analysis reveals intracellular mutual regulation of coral-zooxanthella holobionts

Corals should make excellent models for cross-kingdom regulation research because of their natural animal-photobiont holobiont composition, yet a lack of studies and experimental data restricts their use. Here we integrate new full-length transcriptomes and small RNAs of four common reef-building corals with the published Symbiodinium C1 genome to gain deeper insight into mutual gene regulation in coral-zooxanthella holobionts. We show that zooxanthellae secrete miRNA to downregulate rejection from host coral cells, and that a potential correlation exists between miRNA diversity and physiological activity. Convergence of these holobionts' biological functions in different species is also revealed, which implies the low gene impact on bottom ecological niche organisms. This work provides evidence for the early origin of cross-kingdom regulation as a mechanism of self-defense autotrophs can use against heterotrophs, sheds more light on coral-zooxanthella holobionts, and contributes valuable data for further coral research.

Diversification and distribution of gall crabs (Brachyura: Cryptochiridae: Opecarcinus) associated with Agariciidae corals

Coral reefs are home to the greatest diversity of marine life, and many species on reefs live in symbiotic associations. Studying the historical biogeography of symbiotic species is key to unravelling (potential) coevolutionary processes and explaining species richness patterns. Coral-dwelling gall crabs (Cryptochiridae) live in obligate symbiosis with a scleractinian host, and are ideally suited to study the evolutionary history between heterogeneous taxa involved in a symbiotic relationship. The genus Opecarcinus Kropp and Manning, 1987, like its host coral family Agariciidae, occurs in both Indo-Pacific and Caribbean seas, and is the only cryptochirid genus with a circumtropical distribution. Here, we use mitochondrial and nuclear DNA gene fragments of Opecarcinus specimens sampled from 21 Indo-Pacific localities and one Atlantic (Caribbean) locality. We applied several species delimitation tests to characterise species diversity, inferred a Bayesian molecular-clock time-calibrated phylogeny to estimate divergence times and performed an ancestral area reconstruction. Time to the most recent common ancestor (tMRCA) of Opecarcinus is estimated at 15−6 Mya (middle Miocene—late Miocene). The genus harbours ~ 15 undescribed species as well as several potential species complexes. There are indications of strict host-specificity patterns in certain Opecarcinus species in the Indo-Pacific and Atlantic, however, a robust phylogeny reconstruction of Agariciidae corals—needed to test this further—is currently lacking. The Indo-West Pacific was inferred to be the most probable ancestral area, from where the Opecarcinus lineage colonised the Western Atlantic and subsequently speciated into O. hypostegus. Opecarcinus likely invaded from the Indo-West Pacific across the East Pacific Barrier to the Atlantic, before the full closure of the Isthmus of Panama. The subsequent speciation of O. hypostegus, is possibly associated with newly available niches in the Caribbean, in combination with genetic isolation following the closure of the Panama Isthmus.

Whole-genome transcriptome analyses of native symbionts reveal host coral genomic novelties for establishing coral-algae symbioses

Reef-building corals and photosynthetic, endosymbiotic algae of the family Symbiodiniaceae establish mutualistic relationships that are fundamental to coral biology, enabling coral reefs to support a vast diversity of marine species. Although numerous types of Symbiodiniaceae occur in coral reef environments, Acropora corals select specific types in early life stages. In order to study molecular mechanisms of coral-algal symbioses occurring in nature, we performed whole-genome transcriptomic analyses of Acropora tenuis larvae inoculated with Symbiodinium microadriaticum strains isolated from Acropora. In order to identify genes specifically involved in symbioses with native symbionts in early life stages, we also investigated transcriptomic responses of Acropora larvae exposed to closely related, non-symbiotic, and occasionally symbiotic Symbiodinium strains. We found that the number of differentially expressed genes was largest when larvae acquired native symbionts. Repertoires of differentially expressed genes indicated that corals reduced amino acid, sugar, and lipid metabolism, such that metabolic enzymes performing these functions were derived primarily from S. microadriaticum rather than from A. tenuis. Up-regulated gene expression of transporters for those metabolites occurred only when coral larvae acquired their natural symbionts, suggesting active utilization of native symbionts by host corals. We also discovered that in Acropora, genes for sugar and amino acid transporters, prosaposin-like, and Notch ligand-like, were up-regulated only in response to native symbionts, and included tandemly duplicated genes. Gene duplications in coral genomes may have been essential to establish genomic novelties for coral-algae symbiosis.

Size-specific recolonization success by coral-dwelling damselfishes moderates resilience to habitat loss

Increasing degradation of coral reef ecosystems and specifically, loss of corals is causing significant and widespread declines in the abundance of coral reef fishes, but the proximate cause(s) of these declines are largely unknown. Here, we examine specific responses to host coral mortality for three species of coral-dwelling damselfishes (Dascyllus aruanus, D. reticulatus, and Pomacentrus moluccensis), explicitly testing whether these fishes can successfully move and recolonize nearby coral hosts. Responses of fishes to localized coral loss was studied during population irruptions of coral feeding crown-of-thorns starfish, where starfish consumed 29 (34%) out of 85 coral colonies, of which 25 (86%) were occupied by coral-dwelling damselfishes. Damselfishes were not tagged or individually recognizable, but changes in the colonization of different coral hosts was assessed by carefully assessing the number and size of fishes on every available coral colony. Most damselfishes (> 90%) vacated dead coral hosts within 5 days, and either disappeared entirely (presumed dead) or relocated to nearby coral hosts. Displaced fishes only ever colonized corals already occupied by other coral-dwelling damselfishes (mostly conspecifics) and colonization success was strongly size-dependent. Despite movement of damselfishes to surviving corals, the local abundance of coral-dependent damselfishes declined in approximate accordance with the proportional loss of coral habitat. These results suggest that even if alternative coral hosts are locally abundant, there are significant biological constraints on movement of coral-dwelling damselfishes and recolonization of alternative coral habitats, such that localized persistence of habitat patches during moderate or patchy disturbances do not necessarily provide resilience against overall habitat loss.

Examining the Effect of Heat Stress on Montastraea cavernosa (Linnaeus 1767) from a Mesophotic Coral Ecosystem (MCE)

Coral reefs are under increasing pressure from global warming. Little knowledge, however, exists regarding heat induced stress on deeper mesophotic coral ecosystems (MCEs). Here, we examined the effect of acute (72 h) and chronic (480 h) heat stress on the host coral Montastraea cavernosa (Linnaeus 1767) collected from an upper MCE (~30 m) in Florida, USA. We examined six immune/stress-related genes: ribosomal protein L9 (RpL9), ribosomal protein S7 (RpS7), B-cell lymphoma 2 apoptosis regulator (BCL-2), heat shock protein 90 (HSP90), catalase, and cathepsin L1, as a proxy for coral response to heat stress. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to evaluate the gene expression. Overall, both acute and chronic heat stress treatments elicited a response in gene expression relative to control samples. Acute heat exposure resulted in up-regulation of catalase, BCL-2, and HSP90 at all time points from hour 24 to 48, suggesting the activation of an oxidative protective enzyme, molecular chaperone, and anti-apoptotic protein. Fewer genes were up-regulated in the chronic experiment until hour 288 (30 °C) where catalase, RpL9, and RpS7 were significantly up-regulated. Chronic heat exposure elicited a physiological response at 30 °C, which we propose as a heat-stress threshold for Montastraea cavernosa (M. cavernosa) collected from an MCE.

Deep-sea corals near cold seeps associate with chemoautotrophic bacteria that are related to the symbionts of cold seep and hydrothermal vent mussels

Cnidarians are known for their symbiotic relationships, yet no known association exists between corals and chemoautotrophic microbes. Deep-sea corals, which support diverse animal communities in the Gulf of Mexico, are often found on authigenic carbonate in association with cold seeps. Sulfur-oxidizing chemoautotrophic bacteria of the SUP05 cluster are dominant symbionts of bathymodiolin mussels at cold seeps and hydrothermal vents around the world and have also been found in association with sponges. Therefore, we investigated whether other basal metazoans, corals, also associate with bacteria of the SUP05 cluster and report here that such associations are widespread. This was unexpected because it has been proposed that cnidarians would not form symbioses with chemoautotrophic bacteria due to their high oxygen demand and their lack of specialized respiratory structures. We screened corals, water, and sediment for SUP05 using 16S metabarcoding and found SUP05 phylotypes associated with corals at high relative abundance (10 – 91%). These coral-associated SUP05 phylotypes were coral host specific, absent in water samples, and rare or not detected in sediment samples. The genome of one SUP05 phylotype associated with Paramuricea sp. type B3, contained the genetic potential to oxidize reduced sulfur compounds and fix carbon and these pathways were transcriptionally active. Finally, the relative abundance of this SUP05 phylotype was positively correlated with chemoautotrophically-derived carbon and nitrogen input into the coral holobiont based on stable carbon and nitrogen isotopic compositions. We propose that SUP05 may supplement the diet of its host coral through chemoautotrophy or may provide nitrogen, essential amino acids, or vitamins. This is the first documented association between a chemoautotrophic symbiont and a cnidarian, broadening the known symbioses of corals and may represent a novel interaction between coral communities and cold seeps.

Diverse symbiont bleaching responses are evident from 2-degree heating week bleaching conditions as thermal stress intensifies in coral

Coral bleaching is the dysfunction of the coral–algal endosymbiosis and is characterised as a loss of Symbiodiniaceae cells from host tissues or the loss of photosynthetic pigments. This breakdown of symbiosis occurs as a result of elevated temperature beyond the organism’s thermal threshold. The thermal tipping points within the symbiosis have not yet been well resolved, and the mechanisms underlying the various cellular processes of the corals bleaching response remain unknown. This study characterised the cellular responses of the symbiont Cladocopium sp. (syn. clade C3) within the host coral Acropora aspera during exposure to thermal stress. Exposure to temperatures between 2 and 3°C below the bleaching threshold, equating to 2-degree heating weeks (DHWs), results in changes to the symbiont cell morphology and cell division rates. Once corals were exposed to 4 DHWs, over 90% of the symbiont cells showed signs of degradation. Although sub-bleaching thermal stress is not sufficient to trigger bleaching alerts at an ecological scale, this stressor substantially affects the coral symbiosis. It is therefore vital that we begin to quantify how sub-bleaching thermal stress affects the fitness of Symbiodiniacea populations, their coral hosts and subsequently reefs worldwide.