Project description:Despite their early evolutionary divergence, reef-building corals exhibit complex circadian responses to diurnal, lunar and annual changes in the conditions around them. Understanding circadian regulation in reef-building corals is, however, complicated by the presence of photosynthetic endosymbionts that have a profound physiochemical influence on the intracellular environment. How corals tune their animal-based clock machinery to respond to external cues while at the same time responding to internal physiological changes imposed by the symbiont is not clear. We explore this issue using microarray analysis to dissect genes governed directly by the circadian machinery from those responding indirectly as a consequence of changing internal oxygen tensions.
Project description:A mutualistic relationship between reef-building corals and endosymbiotic algae (Symbiodinium spp.) forms the basis for the existence of coral reefs. Genotyping tools for Symbiodinium spp. have added a new level of complexity to studies concerning cnidarian growth, nutrient acquisition, and stress. For example, the response of the coral holobiont to thermal stress is connected to the host-Symbiodinium genotypic combination, as different partnerships can have different bleaching susceptibilities. If, and to what extent, differences in algal symbiont clade contents can exert effects on the coral host transcriptome is currently unknown. In this study, we monitored algal physiological parameters and profiled the coral host transcriptional responses in acclimated, thermally stressed, and recovered coral fragments using a custom cDNA gene expression microarray. Combining these analyses with results from algal and host genotyping revealed a striking symbiont effect on both the acclimated coral host transcriptome and the magnitude of the thermal stress response. This is the first study that links coral host transcriptomic patterns to the clade content of their algal symbiont community. Our data provide a critical step to elucidating the molecular basis of the apparent variability seen among different coral-algal partnerships.
Project description:Despite their early evolutionary divergence, reef-building corals exhibit complex circadian responses to diurnal, lunar and annual changes in the conditions around them. Understanding circadian regulation in reef-building corals is, however, complicated by the presence of photosynthetic endosymbionts that have a profound physiochemical influence on the intracellular environment. How corals tune their animal-based clock machinery to respond to external cues while at the same time responding to internal physiological changes imposed by the symbiont is not clear. We explore this issue using microarray analysis to dissect genes governed directly by the circadian machinery from those responding indirectly as a consequence of changing internal oxygen tensions. Three coral colonies were sampled at 4 hr intervals during two consecutive days under an ambient light/dark (LD) cycle and under constant darkness (DD). In total 72 arrays were hybridized, as each array represented a sample from a treatment and a time point (n=3).
Project description:Abstract The coral–dinoflagellate symbiosis is increasingly disrupted by global and local anthropogenic stressors. Coral bleaching is primarily a result of high sea surface temperatures, while eutrophication is associated with reef ecosystem degradation. Excess inorganic nitrogen relative to phosphate has been proposed to directly sensitise corals to thermal bleaching and accelerate reef decline. We assessed the proteomic response of the dinoflagellate coral symbiont Symbiodinium microadriaticum to elevated temperatures under multiple nutrient conditions by mass spectrometry. Elevated temperatures resulted in reductions of many chloroplast proteins, particularly light-harvesting complexes, with simultaneous increases in chaperone proteins. N:P imbalance had a larger effect on the proteome than temperature, but the biological processes and proteins responding to each stressor largely overlapped. The proteomes were highly similar at low N:P ratios but were strongly affected by phosphate starvation. High N:P ratios inhibited cell division, reflected by changes in proteins involved in protein translation. Imbalanced N:P did not increase sensitivity to high temperatures as measured by physiological means; however, imbalanced N:P strongly upregulated cell redox homeostasis proteins at high temperatures. As redox balance is critical during thermal bleaching, these data provide insight into the mechanisms of cellular responses to thermal and multiple stresses in the coral–dinoflagellate symbiosis.
Project description:Animal and plant genomes produce numerous small RNAs (smRNAs) regulating gene expression affecting metabolism, development, and epigenetic inheritance. In order to characterize the repertoire of endogenous microRNAs and potential gene targets, we conducted smRNA and mRNA expression profiling over nine experimental treatments of cultures from the dinoflagellate Symbiodinium sp. A1, a photosynthetic symbiont of scleractinian corals. We identified a total of 75 novel smRNAs in Symbiodinum sp. A1 that share stringent key features with functional microRNAs from other model organisms. A subset of 38 smRNAs was predicted independently over all nine treatments and their putative gene targets were identified. We found 3,187 animal-like target sites in the 3'UTRs of 12,858 mRNAs and 53 plant-like target sites in 51,917 genes. We assembled a transcriptome of 58,649 genes and determined differentially expressed genes (DEGs) between treatments. Heat stress was found to produce a much larger number of DEGs than other treatments. Analysis of DEGs also revealed that minicircle-encoded photosynthesis proteins seem to be common targets of transcriptional regulation. Furthermore, we identified the core RNAi protein machinery in Symbiodinium. Integration of smRNA and mRNA expression profiling identified a variety of processes that could be under microRNA control, e.g. regulation of translation, DNA modification, and chromatin silencing. Given that Symbiodinium seems to have a paucity of transcription factors and differentially expressed genes, identification and characterization of its smRNA repertoire establishes the possibility of a range of gene regulatory mechanisms in dinoflagellates acting post-transcriptionally. Symbiodinium cultures were treated with nine different conditions: cold shock, cold stress, heat stress, heat shock, hyposalinity, hypersalinity, dark stress, dark cycle and control.
Project description:Animal and plant genomes produce numerous small RNAs (smRNAs) regulating gene expression affecting metabolism, development, and epigenetic inheritance. In order to characterize the repertoire of endogenous microRNAs and potential gene targets, we conducted smRNA and mRNA expression profiling over nine experimental treatments of cultures from the dinoflagellate Symbiodinium sp. A1, a photosynthetic symbiont of scleractinian corals. We identified a total of 75 novel smRNAs in Symbiodinum sp. A1 that share stringent key features with functional microRNAs from other model organisms. A subset of 38 smRNAs was predicted independently over all nine treatments and their putative gene targets were identified. We found 3,187 animal-like target sites in the 3'UTRs of 12,858 mRNAs and 53 plant-like target sites in 51,917 genes. We assembled a transcriptome of 58,649 genes and determined differentially expressed genes (DEGs) between treatments. Heat stress was found to produce a much larger number of DEGs than other treatments. Analysis of DEGs also revealed that minicircle-encoded photosynthesis proteins seem to be common targets of transcriptional regulation. Furthermore, we identified the core RNAi protein machinery in Symbiodinium. Integration of smRNA and mRNA expression profiling identified a variety of processes that could be under microRNA control, e.g. regulation of translation, DNA modification, and chromatin silencing. Given that Symbiodinium seems to have a paucity of transcription factors and differentially expressed genes, identification and characterization of its smRNA repertoire establishes the possibility of a range of gene regulatory mechanisms in dinoflagellates acting post-transcriptionally. Symbiodinium cultures were treated with nine different conditions: cold shock, cold stress, heat stress, heat shock, hyposalinity, hypersalinity, dark stress, dark cycle and control.
Project description:Using transcriptomics, we show that Symbiodinium acclimation to elevated temperature involves up-regulated expression of meiosis genes followed by up-regulated expression of numerous reactive oxygen species scavenging genes and molecular chaperone genes. Our study connects Symbiodinium transcriptional regulation with physiological heat stress responses as well as known bleaching responses of corals harboring these same Symbiodinium. By uncovering these critical links, we greatly advance understanding of the bleaching susceptibility of corals, which is a key process responsible for global coral reef health.