A Mutant Resource to Transform Reverse Genetics in Chlamydomonas

学术成就：该项目将在单细胞绿色衣藻中产生第一个全基因组突变体集合。几乎每一个核基因至少会分离出一个突变体。单个克隆，以及完整的图书馆，将通过衣原体资源中心分发给科学界。 突变体库将允许分离和研究绿色藻类和植物中几乎所有光合作用所需基因的突变体。此外，该文库还将产生许多高度保守的植物基因的突变体，这些基因在蛋白质成熟和降解、信号传导、小分子转运、氧化还原调节、脂质代谢和微量金属稳态中具有推定的作用。更广泛地说，最广泛保守的绿色植物基因中有一半仍然完全没有特征。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将可能揭示全新的途径，其在植物生物学中的关键功能尚未得到重视。这一资源还将使其他基本过程取得突破，包括绿色藻类碳浓缩机制、趋光性以及真核生物鞭毛的组装和功能。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、氧气和有机构件。植物是我们大部分食物和许多基本材料的来源，并具有作为可再生能源和工业化学品来源的重要潜力。这种突变资源将导致我们对植物生物学基本机制的理解取得进展，这将使未来的作物产量增加，下一代生物燃料的开发以及更明智的政策决策，因为人类活动越来越多地影响我们的环境。这一资源将极大地扩展我们对绿色藻类的了解，这将有助于对这些生物进行工程改造，以大规模生产重组蛋白质和可再生燃料，以及进行生物修复。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，以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.