ABSTRACT: Microcosms made of filtered seawater innoculated with Enterococcus faecalis v583 were exposed to artificial sunlight to investigate photoinactivation mechanisms. Microcosms exposed to artificial sunlight were compared to dark controls. Three experiments were done on three separate days. During every experiment, the light and dark microcosms were samples at the begining (time = 0 hours) and then at 2, 6, 12 and 24 hours.
Project description:This experiment was designed to uncover the transcriptomic changes that would occur in rice after 24 h of supplemental FR treatment, in an accession dependant manner. To do so, 6 different varieties of rice (IR64, Nipponbare, Luk Takhar, M Blatec, Mudgo, Sabharaj and Zhenshan) were grown in the greenhouse facilities of the Botanical Gardens, Utrecht University, in The Netherlands, in summer and autumn of 2021. After five days in WL (400 µmol m-2 s-1 of combined sunlight and artificial light), the treatment group was exposed to supplemental FR (500 µmol m-2 s-1 FR, R:FR of 0.2) light for 24 hours. The whole shoot was sampled, with four plants pooled in one sample. The experiment was repeated four times, resulting in 48 samples (6 varieties x 2 treatments x 4 replicates). Same samples were run on two individual lanes.
Project description:We investigated the transcriptional response of cand. Pelagibacter ubique cultures to light and dark when grown under carbon replete and deplete conditions. Results: During exponential phase, few genes were differentially transcribed between light:dark growth conditions. In stationary phase, a high proportion (9.7%) of coding sequences were found differentially expressed between treatments; 4.7% being up-regulated in the light (n=64) and 4.9% being up-regulated in the dark (n=67). Several components of the oxidative phosphorylation pathway were up-regulated in the dark, supporting physiological data showing higher respiration rates in darkness than in the light under starvation conditions. We also observed up-regulation of a proton translocating pyrophosphate synthase (SAR11_1040), which may be an additional energy production mechanism utilized by SAR11 cells in the dark. Finally, we noted the up-regulation of pili formation genes in the array data, consistent with the observation of pili in dark grown cells via electron microscopy. Triplicate biological replicates were collected from five growth conditions (n=15 total): 1) exponential phase, 24 hour light (n=3) 2) exponential phase, 24 hour dark (n=3) 3) stationary phase, 12:12 hour light:dark cycles (n=3) 4) stationary phase, 24 hour dark (n=3) 5) stationary phase, 6 hours after transfer to darkness (n=3)
Project description:Recent studies have shown that Pelagibacter oxidize a wide range of one carbon (C1) and methylated compounds that are ubiquitous in the oceans. However, the metabolic pathways used to oxidize and assimilate these compounds are complex and have been only partly described. To understand the metabolism of these compounds in Pelagibacter and to identify candidate genes involved in these pathways, we used microarray to study changes in gene expression in response to five different compounds (trimethylamine N-oxide (TMAO), methylamine, dimethylsulfoniopropionate (DMSP), methanol, and glycine betaine (GBT)) in Pelagibacter strain HTCC1062. This project will examine the transcriptional response of the marine microorganism Pelagibacter HTCC1062 to five methylated compounds. To do this, 18 flasks were innoculated with cells - 3 flasks without any methylated compounds and rest of 15 treated with different methylated compounds respectively (TMAO (trtmA), methylamine (trtmB), DMSP (trtmC), methanol (trtmD) and glycine betaine (trtmE)). Cells were all harvested at the same timepoint in the exponential phase.
Project description:This study presents statistical analyses of gene expression in 5, 10 and 15 day post-fertilization (dpf) embryos of the teleost Fundulus heteroclitus treated with control vehicle (DMSO) or a potent non-ortho-PCB (PCB-126; 3,3’,4,4’,5-pentachlorobiphenyl). The embryos were from two populations: a clean, reference population (SC, Scorton Creek, MA USA) and a polluted Superfund population (N, New Bedford Harbor, MA USA). For each site, eggs from 8 females (~1100 total) were fertilized using minced testes from 5 males. After non-fertile eggs were culled, embryos were exposed to vehicle (DMSO; 0.1%) or PCB-126 (50 nM) in filtered seawater (salinity 25 part per thousand, 65 embros per 20 ml in glass petri dish) for 4 hr at 20º C. After exposure, the embryos were washed in filtered seawater and incubated at 20º C under a 14-h light, 10-h dark cycle. At 5-, 10-, and 15-dpf, embryos were collected as three pools of 20 embryos from each treatment group and flash frozen in liquid nitrogen and stored at -80º C until used for RNA isolation. A loop design was used for the microarray hybridizations where each sample is hybridized to 2 arrays using both Cy3 and Cy5 labeled fluorophores. We used three loops in which each loop consisted of Cy3- and Cy5-labeled embryo aRNAs from 12 samples: one sample from each population-treatment-time combination. Within a population-treatment-time, embryos were randomly assigned to one of the three loops. In total, 36 embryos were hybridized to 36 microarrays. The loops formed were SC5C→ SC5P → NBH5C→ NBH5P →SC10C→ SC10P→ NBH10C→ NBH10P → SC15C→ SC15P → NBH15C → NBH15P → SC5C, where each arrow represents a separate hybridization (array) with the biological sample at the base of the arrow labeled with Cy3 and the biological sample at the head of the arrow labeled with Cy5. SC represents the Scorton Creek, Sandwich, MA population (reference), NBH represents the New Bedford Harbor, MA population, C represents control dose (DMSO), P represents the PCB-126 dose, 5 represents 5 dpf, 10 represents 10 dpf and 15 represents 15 dpf.
Project description:Mouse lung tissue transcriptome at postnatal day 10. Samples include 9 germ-free, 9 monocultured Bifidobacterium innoculated, and 9 full caecium microbiota innoculated. Plus 5 adult lung tissue as reference.
Project description:The interaction of animals with microbes relies on the specific recognition of microbial-derived molecules by receptors of the immune system. Sponges (phylum Porifera), as sister group of the Eumetazoa, provide insights into conserved mechanisms for animal-microbe crosstalk, but empirical data is limited. Here we aimed to characterize the immune response of sponges upon microbial stimuli by RNA-Seq. Two sponges species from the Mediterranean Sea, Aplysina aerophoba and Dysidea avara, were challenged with microbial-associated molecular patterns (lipopolysaccharide and peptidoglycan) or sterile artificial seawater (control) in aquarium experiments. Sponge tissue samples were collected 1h, 3h, and 5h after treatment. The response of the sponges to the treatments was assessed by differential gene expression analysis of RNA-Seq data. For each species, we compared the transcriptomic profiles of the samples in MAMP treatment to control within each time point.
Project description:The anadromous Atlantic salmon undergo preparatory physiological transformations before seawater entry, referred to as smoltification. Little is known about the photoperiod-influence and genome regulatory processes driving smoltification such as the large scale changes in lipid metabolism and energy homeostasis in the developing smolt liver. To shed light on this, we performed a smoltification trial using contrasting photoperiod regimes and generate ATAC-seq data from livers throughout smoltification and after seawater transfer to assess the differences in chromatin accessibility. In this experiment Atlantic salmon were reared for a total of 46 weeks from the parr stage, through smoltification, and seawater transfer. After 21 week from first feeding, the group was given artificial winter photoperiod (8 hours light, 16 hours dark) for 8 weeks to induce smoltification before returning to constant light. Liver tissue was sampled from individuals first at week 1, 21 weeks after first feeding, then again at week 10, after the winter period, at week 19, after the expected smoltification time, and lastly at week 25, after transfer to seawater.
Project description:An adult P. rus colony was imported from Indonesia following the CITES protocols (Permit number 14846/IV/SATS-LN/2007) and kept in the Animal Facilities of the Justus Liebig University, Giessen. The colony was maintained in a circulating artificial seawater system at approximately 26 °C with 20-40 µmol photons m-2s-1 (T5 light) of photosynthetically active radiation on a 10:14 h light-dark cycle. A fragment of approx. 9 cm2 was separated from the colony and used as a source of tissue for hologenomic DNA/RNA isolation. Tissue was removed by scraping the fragment´s surface with a sterilized razor blade.
Project description:The anadromous Atlantic salmon undergo preparatory physiological transformations before seawater entry, referred to as smoltification. Little is known about the photoperiod-influence and genome regulatory processes driving smoltification such as the large scale changes in lipid metabolism and energy homeostasis in the developing smolt liver. To shed light on this, we performed a smoltification trial using contrasting photoperiod regimes and generate a transcriptome data from livers throughout smoltification and after seawater transfer. In this experiment two groups of Atlantic salmon were reared for a total of 46 weeks from the parr stage, through smoltification, and seawater transfer. After 21 week from first feeding, the experiment group was given artificial winter photoperiod (8 hours light, 16 hours dark) for 8 weeks to induce smoltification before returning to constant light. The second control group received constant light throughout the experiment. Liver tissue was sampled from individuals first at week 1, 21 weeks after first feeding, then again at week 10, after the winter period, at week 19, after the expected smoltification time, and lastly at week 25, after transfer to seawater.
Project description:Transcriptional profiling of E. faecalis V583 during intracellular survival compared to V583 grown to mid-log phase (OD 600 = 0.05) in THB + 1% glucose. In this study we report the complete intracellular E. faecalis V583 transcriptome following infection of RAW264.7 macrophages. During intracellular survival, approximately 45% of the V583 genome was differentially regulated including numerous genes involved in the oxidative stress, heat shock and SOS responses. We observed that the E. faecalis-containing phagosome was limited for glycolytic substrates, nucleotides, amino acids and numerous ions necessary for growth and protein function. Approximately 35% of the genes differentially regulated during survival within macrophages were of hypothetical/unknown function, suggesting that the V583 response to phagocytosis involves many previously unstudied loci. Here, we provide the first comprehensive study elucidating the transcriptional response of E. faecalis to phagocytosis, which may provide new targets for future studies.