Project description:Investigation of mRNA expression (using HiSeq 2500) in response to treatment of Daphnia magna to pyriproxyfen, wetland water, or stormwater samples.
Project description:Methanotrophs, which help regulate atmospheric levels of methane, are active in diverse natural and man-made environments. This range of habitats and the feast-famine cycles seen by many environmental methanotrophs suggest that methanotrophs dynamically mediate rates of methane oxidation. Global methane budgets require ways to account for this variability in time and space. Functional gene biomarker transcripts are increasingly being studied to inform the dynamics of diverse biogeochemical cycles. Previously, per-cell transcript levels of the methane oxidation biomarker, pmoA, were found to vary quantitatively with respect to methane oxidation rates in model aerobic methanotroph, Methylosinus trichosporium OB3b. In the present study, these trends were explored for two additional aerobic methanotroph pure cultures, Methylocystis parvus OBBP and Methylomicrobium album BG8. At steady-state conditions, per cell pmoA mRNA transcript levels strongly correlated with per cell methane oxidation across the three methanotrophs across many orders of magnitude of activity (R2 = 0.91). Additionally, genome-wide expression data (RNA-seq) were used to explore transcriptomic responses of steady state M. album BG8 cultures to short-term CH4 and O2 limitation. These limitations induced regulation of genes involved in central carbon metabolism (including carbon storage), cell motility, and stress response.
Project description:The synthetic microbial community used in this study was composed of the major functional guilds (cellulolytic fermenter, sulfate reducer, hydrogenotrophic methanogen and acetoclastic methanogen) that mediate the anaerobic conversion of cellulosic biomass to CH4 and CO2 in wetland soils. The choice of a facultative sulfate-reducing bacterium (Desulfovibrio vulgaris Hildenborough) introduced metabolic versatility and enabled investigations into the community response to sulfate intrusion. The growth status of these multi-species cultures was measured over a week by daily analysis of substrate consumption and product accumulation. The quad-cultures were analyzed with metaproteomics at the end of experiment to characterize the community structure and metabolic activities.
