Project description:Corals especially the reef-building species are very important to marine ecosystems. Proteomics has been used for researches on coral diseases, bleaching and responses to the environment change. Corals especially the reef-building species are very important to marine ecosystems. Proteomics has been used for researches on coral diseases, bleaching and responses to the environment change. In the present study, five protocols were compared for protein extraction from stony corals.
Project description:Despite the ecological and economic significance of stony corals (Scleractinia), a robust understanding of their phylogeny remains elusive due to patchy taxonomic and genetic sampling, as well as the limited availability of informative markers. To increase the number of genetic loci available for phylogenomic analyses in Scleractinia, we designed 15,919 DNA enrichment baits targeting 605 orthogroups (mean 565 ± SD 366 bp) over 1,139 exon regions. A further 236 and 62 barcoding baits were designed for COI and histone H3 genes respectively for quality and contamination checks. Hybrid capture using these baits was performed on 18 coral species spanning the presently understood scleractinian phylogeny, with two corallimorpharians as outgroup. On average, 74% of all loci targeted were successfully captured for each species. Barcoding baits were matched unambiguously to their respective samples and revealed low levels of cross-contamination in accordance with expectation. We put the data through a series of stringent filtering steps to ensure only scleractinian and phylogenetically informative loci were retained, and the final probe set comprised 13,479 baits, targeting 452 loci (mean 531 ± SD 307 bp) across 865 exon regions. Maximum likelihood, Bayesian and species tree analyses recovered maximally supported, topologically congruent trees consistent with previous phylogenomic reconstructions. The phylogenomic method presented here allows for consistent capture of orthologous loci among divergent coral taxa, facilitating the pooling of data from different studies and increasing the phylogenetic sampling of scleractinians in the future.
Project description:Scleractinian corals acquire autotrophic nutrients via the photosynthetic activity of their symbionts and the subsequent transfer of photosynthates. Zooplankton predation by the animal (heterotrophy) is an additional food source. Under stress events, corals loose their symbionts, a phenomena known as bleaching, which eventually leads to starvation, unless corals increase their heterotrophic capacities. Molecular mechanisms by which heterotrophy sustains metabolism in stressed corals remain elusive. Here for the first time, we identify specific genes expressed in heterotrophically fed and unfed corals maintained under normal and light-stress conditions inducing bleaching. Physiological parameters and gene expression profiling showed ominously that fed corals better resisted the stress than unfed corals, by presenting less oxidative damage and protein/DNA degradation. Light stressed and unfed/starved corals (HLS) up-regulated by 140 and 13 times two genes (CP2U1 and CP1A2), which belong to the Cytochrome P450 superfamily, while these genes remained almost unchanged in fed corals (HLF). Other genes of redox regulation, DNA damage response, molecular chaperones, and protein degradation were also up-regulated in HLS corals, presenting higher bleaching, and strong decrease of the photosynthesis performance compared to HLF corals. Several pivotal genes associated with the calcification apparatus such as carbonic anhydrases, calcium-transporting ATPase, calcium channel subunit, and bone morphogenetic proteins (BMPs), were significantly down-regulated only in HLS corals. A parallel decrease in the calcification rates of these later corals was also observed. All together, these results show clearly that heterotrophy helps preventing oxidative stress in corals, and thus avoid the cascade of metabolic problems downstream this stress.
Project description:Oxylipins are well-established lipid mediators in plants and animals. In mammals, arachidonic acid (AA)-derived eicosanoids control inflammation, fever, blood coagulation, pain perception and labor, and, accordingly, are used as drugs, while lipoxygenases (LOX), as well as cyclooxygenases (COX) serve as therapeutic targets for drug development. In soft corals, eicosanoids are synthesized on demand from AA by LOX, COX, and catalase-related allene oxide synthase-lipoxygenase (cAOS-LOX) and hydroperoxide lyase-lipoxygenase (cHPL-LOX) fusion proteins. Reef-building stony corals are used as model organisms for the stress-related genomic studies of corals. Yet, the eicosanoid synthesis capability and AA-derived lipid mediator profiles of stony corals have not been determined. In the current study, the genomic and transcriptomic data about stony coral LOXs, AOS-LOXs, and COXs were analyzed and the eicosanoid profiles and AA metabolites of three stony corals, Acropora millepora, A. cervicornis, and Galaxea fascicularis, were determined by reverse-phase high-performance liquid chromatography (RP-HPLC) coupled with MS-MS and a radiometric detector. Our results confirm that the active LOX and AOS-LOX pathways are present in Acropora sp., which correspond to the genomic/sequence data reported earlier. In addition, LOX, AOS-LOX, and COX products were detected in the closely related species G. fascicularis. In conclusion, the functional 8R-LOX and/or AOS-LOX pathways are abundant among corals, while COXs are restricted to certain soft and stony coral lineages.
Project description:Syringe samples of field corals within Broward County were collected. Included are apparently healthy, apparently recovered, and corals visually identified as infected with Stony Coral Tissue Loss Disease. The extracts of these samples were analyzed with LC-MS/MS to identify compounds detected within the aforementioned samples. Included are extracts of cultured Symbiodiniaceae including Breviolum sp., Durusdinium sp., and Symbiodinium sp.