This is a Small Grant for Exploratory Research (SGER) to develop novel methodology for high-throughput preparation of single cell whole genome amplification products from natural aquatic microbial assemblages. The project has an element of risk in that the experimental approaches will push cell sorting and molecular biological tools to new extremes. The investigators will combine fluorescence-activated cell sorting, with whole-genome amplification and sequencing methods. If successful, these results will have a significant impact in the following areas: 1) rapid availability of partial genomes of yet-uncultured microorganisms, including rare species; 2) high-throughput culture-independent function assignment; and 3) genetic variation studies at organismal rather than population level.
While not without risk, this project could lead to high payoffs in basic and applied microbial science. Environmental genomic studies have revealed enormous, previously unknown phylogenetic and metabolic diversity of prokaryotes, providing new insights into microbial evolution, global biogeochemical cycles, and sources of products for biotechnology and pharmacology. However, even very large 'metagenome' sequencing projects have not resulted in complete genome assembly of even the most abundant microorganisms. This poses a critical obstacle in assigning metabolic functions to yet-uncultured microorganisms and in deconstructing entire metabolic pathways that may find practical applications in biotechnology and pharmacology, particularly from rare, uncultured species. The approach to be taken in the current SGER project is designed to solve these issues. Methodology developed through this project may have a major impact on the fields of microbial ecology, environmental microbiology, and bioprospecting and will enhance the transfer of knowledge between academia and industry. The dissemination of education-relevant findings will be conducted through collaboration with the Center for Ocean Sciences Education Excellence (COSEE) Ocean Systems outreach programs.
