Project proposal form Project title: From seagrass debris to natural bioreactors Project code: Host institution: University of Warwick Theme: theme 1, environmental sustainability; and theme 4, microbial ecology and biogeochemistry Key words: Marine ecology, microbial community, biopolymer degradation, meta-genomics/proteomics Supervisory team (including institution & email address): Joseph A. Christie-Oleza, University of Warwick, J.Christie-Oleza@warwick.ac.uk Elizabeth Wellington, University of Warwick, E.M.H.Wellington@warwick.ac.uk Project Highlights: Tonnes of seagrass debris are annually generated and washed to the shore where they accumulate and decompose in large stacks. Who can degrade it and how? What can be learned from these natural bioreactors? Can they have a biotechnological use? Overview: Humankind has historically copied nature and will continue to exploit its biotechnological capabilities for many years to come. This PhD project aims to learn the biological processes that take place during seagrass degradation on marine coasts, from the ecological understanding of microbial communities to the molecular characterisation of hydrolytic enzymes and generation of biogas. Debris generated from the vascular angiosperm Posidonia oceanica will be used as a case study. This seagrass is found in the Mediterranean Sea and in South Australia when coastal ecosystems reach ecological climax. These coastal “sea-forests” produce large amounts of debris that is washed to shore where it accumulates. Despite its inherent protective action avoiding sand beaches to be washed away during winter storms, local authorities remove it from touristic beaches as it is considered an unpleasant solid waste with consequent economic cost. In unmanaged beaches, this debris can accumulate year after year generating large stratified piles or be covered by sand causing anaerobic digestion. The natural bioreactors that are developed in these ecosystems are environments with an enormous enzymatic potential for hydrolysing biopolymers (e.g. lignin and cellulose) and ideal locations for the isolation of extremophile microbes with biotechnological uses (e.g. high salinity). Figure 1: Meadow of Posidonia oceanica and large debris deposits on coastal environments. Methodology: This PhD Project aims to answer key ecological questions on the biodegradation of seagrass debris in marine systems. The successful student will analyse the microorganisms that carry out such vital process in these natural bioreactors, investigating the enzymes involved and the processes that take place. Side products such as depolymerised compounds or generation of biogas will be monitored. The microbial community of natural samples will be compared to those developed in mini-bioreactors setup in the lab under different conditions. The student will analyse microbial community evolution, metagenomic analysis (using high-throughput sequencing), metaproteomic analysis of the exoproteome (looking for relevant secreted enzymes; using high-throughput proteomics), biodegradation of the polymers (light scattering and chromatography systems to determine the kinetics of degradation) and, ultimately, isolation and characterisation of microbial degraders and fermenters. Finally, the student will setup a controlled synthetic community that will mimic the natural reactor and that could be re-wired for biotechnological purposes. Training and skills: CENTA students are required to complete 45 days training throughout their PhD including a 10 day placement. In the first year, students will be trained as a single cohort on environmental science, research methods and core skills. Throughout the PhD, training will progress from core skills sets to master classes specific to the student's projects and themes. This multidisciplinary PhD project will offer a unique opportunity for the student to learn state-of-the-art techniques in environmental –OMICS (such as highthroughput proteomics and genomics), microbial ecology and biochemistry, and analytical chemistry of biopolymer degradation. The laboratories of supervisors Dr Christie-Oleza and Prof Wellington (School of Life Sciences) are excellently equipped to carry out this cutting-edge project. Applicants: We are looking for enthusiastic applicants with BSc or MSci in Biology, Biochemistry or related fields. Applicants with 1st class degrees and laboratory experience will be valued. Partners and collaboration (including CASE): This project has a high potential in becoming CASE, building collaborations with the biopolymer and biotechnology industry. Possible timeline: Year 1: Field sampling with our collaborators in the Balearic Islands (Spain) and microbial community analysis of both natural and lab-based seagrass biodegradation reactors. Isolation of key players with interesting biodegradative capabilities. Year 2: Analysis of community evolution under lab conditions, metagenomics and exoproteomics analysis. Search for hydrolytic exoenzymes both in natural communities and microbial isolates. Evaluation of biopolymer degradation under different incubation conditions. Year 3: Setup of synthetic communities that will carry out the key processes in the biodegradation of seagrass and generate genetically modified organisms to re-direct the use of nutrients. Further reading: Christie-Oleza et al (2015) Proteomics, doi: 10.1002/pmic.201400562. Christie-Oleza et al (2012) Mol Cel Proteomics, 11: M111.013110. Johnson-Rollings et al (2014) ISME J, 8: 2148-2150. Panno et al (2013) N Biotechnol, 30: 685–694. Cuomo et al (1987) Biochem System Ecol, 15: 635637. Pilavtepe et al (2013) Indust Crops Prod, 51: 348– 354. Further details: Enthusiastic applicants with 1st class BSc or MSci degrees in Biology, Biochemistry or related fields and laboratory experience are welcome to apply. Contact: j.christie-oleza@warwick.ac.uk