Project description:We report the full transcriptome (RNA-Seq) of Vibrio fischeri ES114 in rich medium, seawater, and after venting from the Hawaiian bobtail squid Euprymna scolopes. We also report the effects of ribodepletion on low-biomass samples, down to input amount of 1ng total RNA.
Project description:The marine bacterium Vibrio fischeri requires flagellar motility to undergo symbiotic initiation with its host, the Hawaiian bobtail squid Euprymna scolopes. We sought to identify the genes activated by the sigma54-dependent flagellar master regulator, FlrA, in V. fischeri, thereby determining the flagellar regulon in this model symbiont.
Project description:Raw data and Metabolomics of hemocytes from the Hawaiian bobtail squid E. scolopes exposed to Vibrio fischeri and magnetic nanoparticles
Project description:The marine bacterium Vibrio fischeri requires flagellar motility to undergo symbiotic initiation with its host, the Hawaiian bobtail squid Euprymna scolopes. We sought to identify the genes activated by the sigma54-dependent flagellar master regulator, FlrA, in V. fischeri, thereby determining the flagellar regulon in this model symbiont. We performed microarray analysis on wild-type Vibrio fischeri ES114 and a flrA deletion mutant, DM159, grown to mid-log phase in seawater tryptone, a condition in which cells are highly motile (two biological replicates per condition).
Project description:The association with photosymbiotic algae is crucial for the proliferation of many coral reef organisms, but increases their sensitivity to environmental changes. Large benthic foraminifera (LBF) are a diverse group of carbonate producers harboring algal photosymbionts. They act as key ecological engineers and are widely used as bioindicators. As in corals, elevated temperatures and light intensities are known to induce bleaching in LBF, but the combined effects of ocean acidification and warming remain unclear. To shed light into the adaptive physiology of LBF, we linked the assessment of the holobiont and photosymbiont physiological condition (mortality, growth, coloration, and chlorophyll a) to a bottom-up proteomics approach that allows the examination of cellular responses of host and symbionts simultaneously. In a two-months experiment, we exposed Amphistegina lobifera to the combined effects of ocean acidification (400, 1000 and 2800 ppm pCO2) and warming (28-control and 31°C). More than 1,000 proteins were identified by label-free mass spectrometry-based whole proteome analysis and assigned to the host or photosymbionts. Photopigment concentrations declined in response to elevated pCO2, visible by discoloration. These indicate the reduction of photosymbiont densities under ocean acidification, despite the fertilizing effects suggested for high inorganic carbon availability, and imply metabolic adjustments. Increases of proteolytic proteins suggest active host regulation of photosymbiont density in order to maintain homeostasis with its algal photosymbionts. Growth rates, however, were unaffected by elevated pCO2 levels at control temperatures, but high pCO2 levels (2800 ppm, pH 7.52) combined with thermal stress (31°C) impaired growth, though mortality and shell dissolution was negligible. While growth was unaffected by intermediate pCO2 levels (1000 ppm, pH 7.98) combined with ocean warming, this treatment induced the most distinct proteome responses. These include the regulation of ion transporters and host cytoplasmic proteins that likely abet calcification under ocean acidification. This study reveals a highly complex cellular response in both the host and the photosymbiont, which appears to facilitate a high resilience potential of A. lobifera to end of the century ocean conditions. Nevertheless, our results imply that when pCO2 levels rise above 1000 ppm during persistent ocean warming or extreme heating events these adaptive mechanisms become disrupted.
Project description:Increasing atmospheric CO2 raises sea surface temperatures and results in ocean acidification, which will impact upon calcifying marine organisms, such as the commercially and ecologically important Pacific oyster (Crassostrea gigas). Larval stages of development are particularly sensitive to such stressors and may represent population bottlenecks. A two-dimensional electrophoresis (2-DE) proteomic approach was used to investigate the response of 40 hour C. gigas larvae to ocean acidification and warming, and to relate protein expression to phenotypic variation in size and calcification. Larvae were reared at two pHs (8.1 and 7.9) and two temperatures (20°C and 22°C), and comparisons carried out between the four possible treatment combinations. In total 22 differentially expressed spots, corresponding to 18 proteins, were identified by nano-liquid chromatography tandem mass spectrometry. These proteins had roles in metabolism, biomineralisation, intra- and extra-cellular matrix formation and as molecular chaperones. Thirteen of these spots responded to acidification, of which 11 showed reduced expression during acidification. Declines in ATP synthase, arginine kinase and other metabolic proteins suggest metabolic depression occurred during acidification and reduced protein synthesis. In contrast, 6 of 7 proteins that were differentially expressed during warming showed increased expression. Among these were molecular chaperones including protein disulphide isomerase (PDI) and Grp78. Concurrent acidification and warming appeared to mitigate some proteomic changes and negative phenotypic effects observed in acidification at 20°C; however, differential expression of nine proteins and other temperature-independent effects on calcification phenotypes suggest that larval responses to multiple stressors will be complex.
2022-03-01 | PXD002316 | Pride
Project description:Reproductive organogenesis in the Hawaiian bobtail squid linked to symbiotic bacteria
Project description:Construction of a comprehensive spectral library for the coral reef fish, Acanthochromis polyacanthus, from both DIA and DDA MS runs. The spectral library was then used to quantify proteomes of individual fish exposed to different environmental conditions including ocean acidification and ocean warming. Proteomes were measured for both liver and brain tissue and differential expression between environmental conditions was analyzed.