A wide range of plants are grown for food, feed or fuel, and biotechnology has the potential to improve the quality, yield, and sustainability of these plants. Biotechnology applications include improving nutrition, enhancing resilience to stress, carbon sequestration, production of bioplastics/biomaterials/bioproducts/biofuels, and space exploration. Yet, technology barriers currently prevent construction of plants with complex new features. This project will generate a novel, first-of-its-kind platform – a chassis – that will enable the design and fabrication of plants with novel properties. These properties include 1) the production of high value chemicals such as pesticides, bio-materials, bio-plastics, and pharmaceuticals, 2) improved carbon sequestration to address excess global carbon dioxide levels, 3) enhanced sustainability such as the reduced need for fertilizer or insecticides and 4) improved resiliency and adaptation to drought and temperature stress. To encourage participation in science and to convey to a younger generation that science can address global challenges, this project will directly engage undergraduates in the characterization of the chassis. To broaden the diversity of participants in science, this project will recruit and train members of under-represented groups in emerging biotechnology and computational methods, thereby aiding development of a 21st century workforce in scientific disciplines. In the 21st century, the nexus of genomics and biotechnology will be the development of plants with improved yield, nutrition, and market quality, enhanced resilience to biotic and abiotic stressors, increased sustainability, and/or novel phenotypes such as the ability to sequester carbon or synthesize bio-products, biomaterials, and new-to-nature molecules. To achieve this “Third Green Revolution” in agriculture, we need the ability to engineer complex pathways and processes to harness the sustainable production platform of plants. Development of minimal bacterial and yeast genomes have permitted the rewriting of genomes and enhanced the utility of engineered microbes for biotechnology. Advancing plant genome engineering methods and resources are essential for plants to be utilized as sustainable platforms in the bioeconomy. One barrier to either singly or combinatorial engineering complex metabolic pathways and developmental processes is the sheer redundancy of gene function due to whole/segmental genome and gene duplication in all angiosperms. This project will develop a gene editing platform at scale that will be used to generate novel plant genotypes with reduced gene content and redundancy, which can serve as chassis (i.e., a platform) to rapidly engineer plants with new phenotypes and test heterologous genes relevant to biological processes of interest. This project will be the first phase in creating a true minimal plant genome, such that we can begin to rewrite the genome for new-to-nature molecules and biological processes. Due to its small stature, rapid life cycle, and amenability to transformation, the logical choice for creating a minimal plant genome is Arabidopsis thaliana, the mouse-ear cress. Our specific aims include a design phase and two cycles of build-test-learn in which we target 324 protein-coding genes for deletion across 18 5 Mb modules that span the Arabidopsis genome using state-of-the-art multiplexed gene editing approaches. The presence of synthetic lethal pairs will be tested through pairwise and higher order crosses of the modules. Machine learning approaches will be used with empirical and bioinformatic data to develop and improve models that predict gene dispensability. This project will include a Vertically Integrated Project in which all personnel are engaged across the project and institutions and in which a larger number of undergraduates can participate via a custom course entitled Building a Minimal Plant Genome to Enable a Sustainable Bioeconomy. This project will engage under-represented groups to advance participation in STEM disciplines relevant for the 21st century.
Funder: National Science Foundation
PI: Carol Buell, College of Agricultural and Environmental Sciences