Cecilia D'Angelo

Contributions of host and symbiont pigments to the coloration of reef corals

For a variety of coral species, we have studied the molecular origin of their coloration to assess the contributions of host and symbiont pigments. For the corals Catalaphyllia jardinei and an orange‐emitting color morph of Lobophyllia hemprichii, the pigments belong to a particular class of green fluorescent protein‐like proteins that change their color from green to red upon irradiation with ∼400 nm light. The optical absorption and emission properties of these proteins were characterized in detail. Their spectra were found to be similar to those of phycoerythrin from cyanobacterial symbionts. To unambiguously determine the molecular origin of the coloration, we performed immunochemical studies using double diffusion in gel analysis on tissue extracts, including also a third coral species, Montastrea cavernosa, which allowed us to attribute the red fluorescent coloration to green‐to‐red photoconvertible fluorescent proteins. The red fluorescent proteins are localized mainly in the ectodermal tissue and contribute up to 7.0% of the total soluble cellular proteins in these species. Distinct spatial distributions of green and cyan fluorescent proteins were observed for the tissues of M. cavernosa. This observation may suggest that differently colored green fluorescent protein‐like proteins have different, specific functions. In addition to green fluorescent protein‐like proteins, the pigments of zooxanthellae have a strong effect on the visual appearance of the latter species.

Symbiodinium thermophilum sp. nov., a thermotolerant symbiotic alga prevalent in corals of the world's hottest sea, the Persian/Arabian Gulf

Coral reefs are in rapid decline on a global scale due to human activities and a changing climate. Shallow water reefs depend on the obligatory symbiosis between the habitat forming coral host and its algal symbiont from the genus Symbiodinium (zooxanthellae). This association is highly sensitive to thermal perturbations and temperatures as little as 1°C above the average summer maxima can cause the breakdown of this symbiosis, termed coral bleaching. Predicting the capacity of corals to survive the expected increase in seawater temperatures depends strongly on our understanding of the thermal tolerance of the symbiotic algae. Here we use molecular phylogenetic analysis of four genetic markers to describe Symbiodinium thermophilum, sp. nov. from the Persian/Arabian Gulf, a thermally tolerant coral symbiont. Phylogenetic inference using the non-coding region of the chloroplast psbA gene resolves S. thermophilum as a monophyletic lineage with large genetic distances from any other ITS2 C3 type found outside the Gulf. Through the characterisation of Symbiodinium associations of 6 species (5 genera) of Gulf corals, we demonstrate that S. thermophilum is the prevalent symbiont all year round in the worlds hottest sea, the southern Persian/Arabian Gulf.

It's cheap to be colorful. Anthozoans show a slow turnover of GFP-like proteins.

Pigments homologous to the green fluorescent protein (GFP) contribute up to ∼ 14% of the soluble protein content of many anthozoans. Maintenance of such high tissue levels poses a severe energetic penalty to the animals if protein turnover is fast. To address this as yet unexplored issue, we established that the irreversible green‐to‐red conversion of the GFP‐like pigments from the reef corals Montastrea cavernosa (mcavRFP) and Lobophyllia hemprichii (EosFP) is driven by violet–blue radiation in vivo and in situ. In the absence of photoconverting light, we subsequently tracked degradation of the red‐converted forms of the two proteins in coral tissue using in vivo spectroscopy and immunochemical detection of the post‐translational peptide backbone modification. The pigments displayed surprisingly slow decay rates, characterized by half‐lives of ∼ 20 days. The slow turnover of GFP‐like proteins implies that the associated energetic costs for being colorful are comparatively low. Moreover, high in vivo stability makes GFP‐like proteins suitable for functions requiring high pigment concentrations, such as photoprotection.

Local adaptation constrains the distribution potential of heat-tolerant Symbiodinium from the Persian/ Arabian Gulf

The symbiotic association of corals and unicellular algae of the genus Symbiodinium in the southern Persian/Arabian Gulf (PAG) display an exceptional heat tolerance, enduring summer peak temperatures of up to 36 °C. As yet, it is not clear whether this resilience is related to the presence of specific symbiont types that are exclusively found in this region. Therefore, we used molecular markers to identify the symbiotic algae of three Porites species along >1000 km of coastline in the PAG and the Gulf of Oman and found that a recently described species, Symbiodinium thermophilum, is integral to coral survival in the southern PAG, the world’s hottest sea. Despite the geographic isolation of the PAG, we discovered that representatives of the S. thermophilum group can also be found in the adjacent Gulf of Oman providing a potential source of thermotolerant symbionts that might facilitate the adaptation of Indian Ocean populations to the higher water temperatures expected for the future. However, corals from the PAG associated with S. thermophilum show strong local adaptation not only to high temperatures but also to the exceptionally high salinity of their habitat. We show that their superior heat tolerance can be lost when these corals are exposed to reduced salinity levels common for oceanic environments elsewhere. Consequently, the salinity prevailing in most reefs outside the PAG might represent a distribution barrier for extreme temperature-tolerant coral/Symbiodinium associations from the PAG.

A Green Fluorescent Protein with Photoswitchable Emission from the Deep Sea

A colorful variety of fluorescent proteins (FPs) from marine invertebrates are utilized as genetically encoded markers for live cell imaging. The increased demand for advanced imaging techniques drives a continuous search for FPs with new and improved properties. Many useful FPs have been isolated from species adapted to sun-flooded habitats such as tropical coral reefs. It has yet remained unknown if species expressing green fluorescent protein (GFP)-like proteins also exist in the darkness of the deep sea. Using a submarine-based and -operated fluorescence detection system in the Gulf of Mexico, we discovered ceriantharians emitting bright green fluorescence in depths between 500 and 600 m and identified a GFP, named cerFP505, with bright fluorescence emission peaking at 505 nm. Spectroscopic studies showed that ∼15% of the protein bulk feature reversible ON/OFF photoswitching that can be induced by alternating irradiation with blue und near-UV light. Despite being derived from an animal adapted to essentially complete darkness and low temperatures, cerFP505 maturation in living mammalian cells at 37°C, its brightness and photostability are comparable to those of EGFP and cmFP512 from shallow water species. Therefore, our findings disclose the deep sea as a potential source of GFP-like molecular marker proteins.

Fluorescent protein‐mediated colour polymorphism in reef corals: multicopy genes extend the adaptation/acclimatization potential to variable light environments

The genomic framework that enables corals to adjust to unfavourable conditions is crucial for coral reef survival in a rapidly changing climate. We have explored the striking intraspecific variability in the expression of coral pigments from the green fluorescent protein (GFP) family to elucidate the genomic basis for the plasticity of stress responses among reef corals. We show that multicopy genes can greatly increase the dynamic range over which corals can modulate transcript levels in response to the light environment. Using the red fluorescent protein amilFP597 in the coral Acropora millepora as a model, we demonstrate that its expression increases with light intensity, but both the minimal and maximal gene transcript levels vary markedly among colour morphs. The pigment concentration in the tissue of different morphs is strongly correlated with the number of gene copies with a particular promoter type. These findings indicate that colour polymorphism in reef corals can be caused by the environmentally regulated expression of multicopy genes. High‐level expression of amilFP597 is correlated with reduced photodamage of zooxanthellae under acute light stress, supporting a photoprotective function of this pigment. The cluster of light‐regulated pigment genes can enable corals to invest either in expensive high‐level pigmentation, offering benefits under light stress, or to rely on low tissue pigment concentrations and use the conserved resources for other purposes, which is preferable in less light‐exposed environments. The genomic framework described here allows corals to pursue different strategies to succeed in habitats with highly variable light stress levels. In summary, our results suggest that the intraspecific plasticity of reef corals’ stress responses is larger than previously thought.

Water column productivity and temperature predict coral reef regeneration across the Indo-Pacific

Predicted increases in seawater temperatures accelerate coral reef decline due to mortality by heat-driven coral bleaching. Alteration of the natural nutrient environment of reef corals reduces tolerance of corals to heat and light stress and thus will exacerbate impacts of global warming on reefs. Still, many reefs demonstrate remarkable regeneration from past stress events. This paper investigates the effects of sea surface temperature (SST) and water column productivity on recovery of coral reefs. In 71 Indo-Pacific sites, coral cover changes over the past 1-3 decades correlated negative-exponentially with mean SST, chlorophyll a, and SST rise. At six monitoring sites (Persian/Arabian Gulf, Red Sea, northern and southern Galápagos, Easter Island, Panama), over half of all corals were <31 years, implying that measured environmental variables indeed shaped populations and community. An Indo-Pacific-wide model suggests reefs in the northwest and central Indian Ocean, as well as the central west Pacific, are at highest risk of degradation, and those at high latitudes the least. The model pinpoints regions where coral reefs presently have the best chances for survival. However, reefs best buffered against temperature and nutrient effects are those that current studies suggest to be most at peril from future ocean acidification.

An experimental mesocosm for long-term studies of reef corals

Molecular biological methods including genomic and proteomic approaches hold a specific promise to provide new insights into the stress physiology of corals. However, to fully exploit the power of these techniques, aquarium setups are required that allow biological assays under tightly controlled laboratory conditions. Here, details are provided about the successful development of a closed coral mesocosm at the National Oceanography Centre, Southampton. The system can be operated without access to natural seawater and allows long-term observations and experimental studies of reef corals. The individual experimental tanks allow the corals to be exposed, for example, to different light and/or temperature conditions without the need to disconnect them from the system.

Diversity of zooxanthellae from corals and sea anemones after long-term aquarium culture

Aquarium systems allow technically sophisticated experiments that promise new opportunities to answer urgent questions about reef coral biology, for instance assessing the responses to decreasing environmental pH and/or increased temperatures. Over recent years, long-term culture and (predominantly asexual) propagation of corals has become possible in such systems. At present however, only limited data are available that clarify whether or not responses of the coral holobiont are dominated by the acclimatization to life in captivity or continue to reflect, for example, taxonomic differences seen in nature. We studied the diversity of the symbiotic algae in corals and sea anemones after long-term aquaculture by analysis of their small subunit (SSU) ribosomal DNA gene. A field sample of Acropora clathrata from the Arabian Gulf which was used as a control contained clade C zooxanthellae. The aquarium corals also harboured clade C symbionts, but sequencing of the SSU DNA suggested that the analysed animals host different subclades. A prevalence of clade C is also found among corals from the Indo-Pacific region, the origin of most of the aquarium samples. An individual of the temperate sea anemone Anemonia sulcata (viridis) contained clade A symbionts, similar to those found in nature, even after nearly 10 years of co-culture with sea anemones (Entacmaea quadricolor) and corals hosting clade C symbionts. The results indicate that the specific host–symbiont association occurring in nature appears to persist over >2 years timescales in captivity, with no mixing of symbionts between hosts maintained in the same aquarium or apparent selection of stress-tolerant symbiont strains such as clade D.

Phosphate deficiency promotes coral bleaching and is reflected by the ultrastructure of symbiotic dinoflagellates

Enrichment of reef environments with dissolved inorganic nutrients is considered a major threat to the survival of corals living in symbiosis with dinoflagellates (Symbiodinium sp.). We argue, however, that the direct negative effects on the symbiosis are not necessarily caused by the nutrient enrichment itself but by the phosphorus starvation of the algal symbionts that can be caused by skewed nitrogen (N) to phosphorus (P) ratios. We exposed corals to imbalanced N:P ratios in long-term experiments and found that the undersupply of phosphate severely disturbed the symbiosis, indicated by the loss of coral biomass, malfunctioning of algal photosynthesis and bleaching of the corals. In contrast, the corals tolerated an undersupply with nitrogen at high phosphate concentrations without negative effects on symbiont photosynthesis, suggesting a better adaptation to nitrogen limitation. Transmission electron microscopy analysis revealed that the signatures of ultrastructural biomarkers represent versatile tools for the classification of nutrient stress in symbiotic algae. Notably, high N:P ratios in the water were clearly identified by the accumulation of uric acid crystals.