A Mutant Resource to Transform Reverse Genetics in Chlamydomonas

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Intellectual Merit: This project will generate the first genome-wide collection of mutants in the single-celled green alga Chlamydomonas reinhardtii. At least one mutant will be isolated for nearly every nuclear gene. Individual clones, as well as the complete library, will be distributed to the scientific community through the Chlamydomonas Resource Center. The mutant library will allow for the isolation and study of mutants in nearly all genes required for photosynthesis in green algae and plants. Furthermore, the library will also yield mutants in many highly conserved plant genes with putative roles in protein maturation and degradation, signaling, small molecule transport, redox regulation, lipid metabolism, and trace metal homeostasis. More broadly, half of the most widely conserved green plant genes remain completely uncharacterized. This resource will yield mutants in nearly all of these genes, and the study of these mutants will likely reveal entirely new pathways whose critical functions in plant biology are still not appreciated. This resource will also enable breakthroughs in other fundamental processes, including the green algal carbon concentrating mechanism, phototaxis, and the assembly and function of eukaryotic flagella.Broader Impacts: Photosynthetic organisms provide energy, oxygen and organic building blocks for nearly all of the life on Earth. Plants are the origin of most of our food and many of our basic materials, and have important potential as a source of renewable energy and industrial chemicals. This mutant resource will lead to advances in our understanding of the basic mechanisms of plant biology, which will enable future increases in crop yields, the development of next-generation biofuels, and more informed policy decisions as human activities increasingly impact our environment. This resource will dramatically expand our knowledge of green algae, which will facilitate efforts to engineer these organisms for the large-scale production of recombinant proteins and renewable fuels, and for bioremediation. Finally, use of this resource will provide countless opportunities for training early-stage biologists in genetics and the application of high-throughput, genome-wide approaches to basic plant biology questions.