A Mutant Resource to Transform Reverse Genetics in Chlamydomonas

智力优势：该项目将产生单细胞绿藻莱茵衣藻突变体的第一个全基因组收集。几乎每个核基因至少会分离出一个突变体。单个克隆以及完整的文库将通过衣藻资源中心分发给科学界。突变文库将允许对绿藻和植物光合作用所需的几乎所有基因的突变进行分离和研究。此外，该文库还将获得许多高度保守的植物基因的突变体，这些基因可能在蛋白质成熟和降解、信号传导、小分子运输、氧化还原调节、脂质代谢和微量金属稳态中发挥作用。更广泛地说，一半最广泛保守的绿色植物基因仍然完全没有特征。这种资源将产生几乎所有这些基因的突变体，对这些突变体的研究可能会揭示出在植物生物学中仍未被认识到的关键功能的全新途径。这一资源还将推动其他基础过程的突破，包括绿藻碳浓缩机制、趋光性以及真核鞭毛的组装和功能。更广泛的影响：光合生物为地球上几乎所有的生命提供能量、氧气和有机组成部分。植物是我们大多数食物和许多基本材料的来源，并且作为可再生能源和工业化学品的来源具有重要潜力。这种突变资源将导致我们对植物生物学基本机制的理解的进步，这将使未来作物产量的增加，下一代生物燃料的开发，以及随着人类活动对环境的影响越来越大，更明智的政策决策成为可能。这一资源将极大地扩展我们对绿藻的认识，这将促进我们为大规模生产重组蛋白质和可再生燃料以及生物修复而设计这些生物的努力。最后，利用这一资源将提供无数的机会，培训早期生物学家在遗传学和应用高通量，全基因组的方法来解决基本的植物生物学问题。

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.