Coral photosymbiosis: Linking phylogenetic identity to single cell metabolic activity in mixed symbiont populations

Tropical reef-building corals live in symbiosis with a wide range of micro-organisms, including unicellular Symbiodinium dinoflagellates also called zooxanthellae. These photosynthetic endosymbionts live inside the coral gastroderm cells. Although corals can feed on planktonic preys, the dinoflagellates significantly contribute to the nutrition of their host by transferring a large fraction (up to 90%) of photosynthates that are produced through the fixation of dissolved inorganic carbon (DIC) and nitrogen (nitrate or ammonium). Nine major clades (A-I) of Symbiodinium have been identified by molecular genetic analyses. Each clade presents distinct physiological features and the specific association between coral species and Symbiodinium clade determines the phenotype of the holobiont. Differences in the photosynthetic response to irradiance, rates of carbon fixation, and thermal tolerance can be attributed to symbiont clade. Several Symbiodinium clades can simultaneously exist within a single coral and the host can dynamically modify the proportion between dominant and background clades to adapt to changing environmental conditions. Investigating at the cellular level the metabolic exchanges between coral and Symbiodinium is of great interest to understand e.g. the bleaching process (loss of zooxanthellae) which often leads to coral death. We are aiming at quantitatively image the differential metabolic activity between symbiont clades in the same host, in the intact symbiosis. Previous studies have used mass bulk techniques to investigate the metabolism of symbionts at the colony scale. However, such studies cannot determine the specific contribution of the individual cells, as a function of their distribution in the coral host tissue. We have developed a SIMS-ISH method combining nanoscale secondary ion mass spectrometry (NanoSIMS) and in situ hybridization (ISH) for the simultaneous in situ identification of Symbiodinium genotype, and visualization of symbiont-host metabolic exchange at the level of individual cell. We focus on two reef-building coral species Pocillopora damicornis and Stylophora pistillata for which a large amount of complementary metabolic data exists. We designed specific fluorescent DNA probes to identify clade C Symbiodinium in P. damicornis and clade A in S. pistillata, and in mixed cultures. We combined the probes with pulse-chase experiments using isotopically labeled seawater (13C-bicarbonate and 15N-nitrate) to attribute a particular metabolism to a specific clade. This combined method enables us to phylogenetically identify metabolically active cells from a NanoSIMS isotopic/elemental image. The precise correlation between TEM and the NanoSIMS isotopic maps allows us to follow the turnover and translocation of metabolites with sub-cellular precision in both the symbionts and the host. Due to the complex nature of the coral symbiosis, the ability to discriminate the phylogenetic identity and metabolic role of specific Symbiodinium populations in situ is crucial to understand the effects of environmental stress on the coral holobiont plasticity. Moreover, analyzing symbiotic associations in situ provides a unique insight into the spatio-temporal patterns of metabolic interactions in holobionts. This analytical breakthrough promises to open entirely new areas of research focused on understanding the dynamics of interactions between animals and the microbial world

Cell proliferation and migration during early development of a symbiotic scleractinian coral

In scleractinian reef-building corals, patterns of cell self-renewal, migration and death remain virtually unknown, limiting our understanding of cellular mechanisms underlying initiation of calcification, and ontogenesis of the endosymbiotic dinoflagellate relationship. In this study, we pulse-labelled the coral Stylophora pistillata for 24 h with BrdU at four life stages (planula, early metamorphosis, primary polyp and adult colony) to investigate coral and endosymbiont cell proliferation during development, while simultaneously recording TUNEL-positive (i.e. apoptotic) nuclei. In the primary polyp, the fate of BrdU-labelled cells was tracked during a 3-day chase. The pharynx and gastrodermis were identified as the most proliferative tissues in the developing polyp, and BrdU-labelled cells accumulated in the surface pseudostratified epithelium and the skeletogenic calicodermis during the chase, revealing cell migration to these epithelia. Surprisingly, the lowest cell turnover was recorded in the calicodermis at all stages, despite active, ongoing skeletal deposition. In dinoflagellate symbionts, DNA synthesis was systematically higher than coral host gastrodermis, especially in planula and early metamorphosis. The symbiont to host cell ratio remained constant, however, indicating successive post-mitotic control mechanisms by the host of its dinoflagellate density in early life stages, increasingly shifting to apoptosis in the growing primary polyp

Evidence for Rhythmicity Pacemaker in the Calcification Process of Scleractinian Coral

Reef-building scleractinian (stony) corals are among the most efficient bio-mineralizing organisms in nature. The calcification rate of scleractinian corals oscillates under ambient light conditions, with a cyclic, diurnal pattern. A fundamental question is whether this cyclic pattern is controlled by exogenous signals or by an endogenous 'biological-clock' mechanism, or both. To address this problem, we have studied calcification patterns of the Red Sea scleractinian coral Acropora eurystoma with frequent measurements of total alkalinity (AT) under different light conditions. Additionally, skeletal extension and ultra-structure of newly deposited calcium carbonate were elucidated with Sr-86 isotope labeling analysis, combined with NanoSIMS ion microprobe and scanning electron microscope imaging. Our results show that the calcification process persists with its cyclic pattern under constant light conditions while dissolution takes place within one day of constant dark conditions, indicating that an intrinsic, light-entrained mechanism may be involved in controlling the calcification process in photosymbiotic corals

Highly dynamic host regulation of Symbiodinium population during the metamorphosis of the scleractinian coral larva

Highly dynamic host regulation of Symbiodinium population during the metamorphosis of the scleractinian coral larv

Data from: Cell proliferation and migration during early development of a symbiotic scleractinian coral

In scleractinian reef-building corals, patterns of cell self-renewal, migration and death remain virtually unknown, limiting our understanding of cellular mechanisms underlying initiation of calcification, and ontogenesis of the endosymbiotic dinoflagellate relationship. In this study we pulse-labeled the coral Stylophora pistillata for 24 h with BrdU at four life stages (planula, early metamorphosis, primary polyp, and adult colony) to investigate coral and endosymbiont cell proliferation during development, while simultaneously recording TUNEL-positive, i.e. apoptotic, nuclei. In the primary polyp, the fate of BrdU-labeled cells was tracked during a 3 days chase. The pharynx and gastrodermis were identified as the most proliferative tissues in the developing polyp, and BrdU-labeled cells accumulated in the surface pseudostratified epithelium and the skeletogenic calicodermis during the chase, revealing cell migration to these epithelia. Surprisingly, the lowest cell turnover was recorded in the calicodermis at all stages, despite active, ongoing skeletal deposition. In dinoflagellate symbionts, DNA synthesis was systematically higher than in coral host gastrodermis, especially in planula and early metamorphosis. The symbiont to host cell ratio remained however constant, indicating successive post-mitotic control mechanisms by the host of its dinoflagellate density in early life stages, increasingly shifting to apoptosis in the growing primary polyp

Skeletal growth dynamics linked to trace-element composition in the scleractinian coral Pocillopora damicornis

The micro-and ultra-structural skeletal growth dynamics of the scleractinian coral Pocillopora damicornis (Linnaeus 1758) was studied with pulsed Sr-86-labeling and high spatial resolution NanoSIMS isotopic imaging. Average extension rates for the two basic ultra-structural components of the skeleton, Rapid Accretion Deposits (RAD) and Thickening Deposits (TD), were compared between corallite wall, spines and dissepiments. The RAD, forming the basal part of the dissepiment, were found to form extremely fast compared with RAD and TD in other parts of the skeleton. Trace element compositions (i.e., Mg/Ca and Sr/Ca ratios) obtained for each ultra-structural component reveal the full range of chemical variation on the scale of the individual corallite. The Mg/Ca ratio was found to vary about a factor of 6, from similar to 2.2 mmol/mol in slow growing ornamental spines to similar to 13 mmol/mol in the fast forming dissepiment RAD. A positive relationship between Mg/Ca and inferred average extension rate was observed. Sr/Ca, on the other hand, although it varies substantially in the range between similar to 6.5 and similar to 9.5 mmol/mol, does not show any relationship with inferred average extension rate, nor does is correlate with the Mg/Ca ratio. Rayleigh fractionation models are not capable of explaining the trace element variations. The results presented in this study imply that the coral is capable of controlling its biomineralization activity with great temporal and spatial precision. Spatial heterogeneity in coral tissue activity should be carefully investigated in the development of biomineralization models for scleractinian corals. (C) 2012 Elsevier Ltd. All rights reserved

Assessing calcareous sponges and their associated bacteria for the discovery of new bioactive natural products.

International audienceAn overview of the chemistry and microbiology of calcareous sponges (Calcispongiae) is provided, highlighting the potential of these sessile filter-feeding marine invertebrates and their associated bacteria for the discovery of new bioactive natural products. 103 compounds are presented and 116 references cited