Project description:Sequencing the metatranscriptome can provide information about the response of organisms to varying environmental conditions. We present a methodology for obtaining random whole-community mRNA from a complex microbial assemblage using Pyrosequencing. The metatranscriptome had, with minimum contamination by ribosomal RNA, significant coverage of abundant transcripts, and included significantly more potentially novel proteins than in the metagenome. Keywords: metatranscriptome, mesocosm, ocean acidification This experiment is part of a much larger experiment. We have produced 4 454 metatranscriptomic datasets and 6 454 metagenomic datasets. These were derived from 4 samples. The experiment is an ocean acidification mesocosm set up in a Norwegian Fjord in 2006. We suspended 6 bags containing 11,000 L of sea water in a Coastal Fjord and then we bubbled CO2 through three of these bags to simulate ocean acidification conditions in the year 2100. The other three bags were bubbled with air. We then induced a phytoplankton bloom in all six bags and took measurements and performed analyses of phytoplankton, bacterioplankton and physiochemical characteristics over a 22 day period. We took water samples from the peak of the phytoplankton bloom and following the decline of the phytoplankton bloom to analyses using 454 metagenomics and 454 metatranscriptomics. Day 1, High CO2 Bag and Day 1, Present Day Bag, refer to the metatranscriptomes from the peak of the bloom. Day 2, High CO2 Bag and Day 2, Present Day Bag, refer to the metatranscriptomes following the decline of the bloom. Obviously High CO2 refers to the ocean acidification mesocosm and Present Day refers to the control mesocosm. Raw data for both the metagenomic and metatranscriptomic components are available at NCBI's Short Read Archive at ftp://ftp.ncbi.nlm.nih.gov/sra/Studies/SRP000/SRP000101
Project description:Prymnesium parvum is regarded as one of the most notorious harmful algal bloom (HAB) species worldwide. In recent years, it has frequently formed toxic blooms in coastal and brackish waters of America, Europe, Australia, Africa and Asia, causing large-scale mortalities of wild and cultured fish and other gill-breathing animals. In the last decade, blooms of P. parvum have expanded to inland fresh waters in the USA, presumably due to changes in environmental conditions. The aim of the experiment was to establish the gill transcriptomic responses to P. parvum in rainbow trout. We used 2 different concentrations of P. parvum and identified fish with low and moderate responses to the algae. Based on the dose of and the fish response, fish were classified into 4 groups with high exposure/moderate response (HM), high exposure/low response (HL), low exposure/low response (LL) and control group (C) with no exposure/no response. Gene expression profiling of the gill tissue was performed using a microarray platform developed and validated for rainbow trout.
Project description:In this project, the metaproteome of the marine bacterioplankton was analyzed to assess its respone towards an algal bloom in the southern North Sea in spring 2010. Proteins were extracted applying two different methods: (i) applying chemical cell lysis using trifluoroethanol in combination with in-solution digest and (ii) mechanical cell lysis applying bead beating, SDS-PAGE prefractionation and in-gel digest. Both samples were analyzed by nanoLC and ESI-iontrap MS. In case of the TFE lysis samples, also nanoLC-MALDI-TOF MS was applied.