The central questions this project aims to answer by functional investigation of the microbial communities are those regarding the activity and identity of the microbes present, or, to put it simply, “who is doing what, where and when?” Also of importance are measurements of methylotrophic activity to develop a better understanding of the impact of these processes in marine ecosystems. Also, the interactions of methylotrophs with other organisms and the influence of environmental parameters on their activity are being investigated.
The project makes extensive use of both metagenomics combined with DNA Stable Isotope Probing (DNA-SIP) and single cell ecophysiological techniques (Raman-FISH) for functional investigations of marine microbial populations. The incorporation of the heavy isotopes into microbial DNA is being analysed to follow carbon and nitrogen fluxes through microbial communities, from methylotrophs to heterotrophs. 13C labeled DNA of methylotrophs is then used for high-throughput sequencing to reconstruct genomes of these marine organisms. Key functional genes are being analysed by PCR and qPCR. We are therefore able to identify active methylotrophs in complex marine microbial communities, and quantify their contribution to marine nutrient fluxes. Raman microspectroscopy in combination with optical tweezers based on specific FISH probes is also being used for isolation of potential novel methylotrophs and single cell genomics.
This project is revealing novel insights into marine microbial communities driven by methylotrophic metabolism. The identification of novel methylotrophs and new variants of enzymes involved in methylotrophic processes allows a better understanding of these metabolic processes. Knowledge of key organisms and their pathways is yielding valuable insights into the impact of methylotrophs on carbon and nitrogen cycling in marine ecosystems.
Gordon and Betty Moore Foundation (GBMF)
Prof JC Murrell, UEA (PI)
Prof Y Chen, Warwick (Co-PI)
Dr. Carolina Grob
Dr. Martin Taubert
Dr Jo Dixon (Plymouth Marine Laboratory)
Prof Wei Huang (University of Oxford)