B5: Bigger Better Brassicaceae Biofuels and Bioproducts—An Overview
Doug Allen1,2, Malia Gehan1, Philip D. Bates3, Timothy P. Durrett4, Ruth Welti4, Jerome M. Fox5, Chaofu Lu6, Michael J. Smanski7, Trupti Joshi8, Jay J. Thelen8, Dong Xu8, and Edgar B. Cahoon9* (firstname.lastname@example.org)
1Donald Danforth Plant Science Center; 2Agricultural Research Center, U.S. Department of Agriculture; 3Washington State University; 4Kansas State University; 5University of Colorado–Boulder; 6Montana State University; 7University of Minnesota; 8University of Missouri; and 9University of Nebraska–Lincoln
The project addresses three goals:
- Systems-guided interrogation of the plastid bio-factory for enhanced production of fatty acids and predictable production of fatty acids with tailored chain-lengths
- Synthetic biology tool development for predictable and high-throughput oilseed crop engineering
- Integration of the redesigned plastid bio-factory with extra-plastidial metabolism for enhanced oils and biocontainment.
B5 will address the imperative need for sustainable liquid fuels and oils of defined structures desired by the U.S. bioenergy and oleochemical sectors. The project will integrate fundamental knowledge generation and synthetic biology tool development to predictably and more rapidly develop non-food Brassicaceae oilseeds that produce high quantities of oils and oils with tailored fatty acid compositions (Figure 1). B5’s multidisciplinary team will interrogate plastid metabolic circuitry for carbon flux through fatty acid biosynthesis in seeds of the Brassicaceae, pennycress, and camelina. Focus on both species will generate “rules” for next-generation metabolic engineering of Brassicaceae oilseeds and provide higher-value and broader cover/rotation crop options for U.S. farmers. B5 efforts will be guided by mathematical models as well as biochemical data acquired from seeds of metabolically “extreme” species that produce exceptionally high levels of medium-chain fatty acids. In concert, B5 will develop synthetic biology tools to deliver transgene combinations into defined genome regions and advanced gene-editing methods for tunable up- or down-regulation or replacement of endogenous genes. Aided by a comprehensive analytical learning platform and computational models, B5 will integrate data and toolsets to develop enhanced pennycress and camelina germplasm through design, build, test, learn (DBTL) cycles. Given the central metabolic role of fatty acids in the cell, robust and integrated DBTL cycles will be key to discovering how plants “fight back” against lipid metabolic remodeling. The high-quality genome sequences, plethora of genomic resources, existing metabolic engineering toolbox, and simple Agrobacterium-based floral infiltration transformation systems make both pennycress and camelina ideal crops for modified oil production. These attributes will also accelerate synthetic biology chassis optimization and introduction of genetic biocontainment technology for safe, sustainable production on marginal and underutilized land across wide portions of the United States. The availability of U.S. Department of Agriculture–Animal and Plant Health Inspection Service regulated field sites and a high-throughput camera-based phenotyping system will facilitate agronomic evaluation of engineered germplasm under diverse environments. In addition to significant fundamental and translational outputs, B5 will further develop extant databases (e.g., FatPlants, ARALIP) for the scientific community and train nine undergraduates, six graduate students, and 11 postdoctoral scientists as the next generation of investigators to tackle U.S. and global energy security, natural resources, and environmental challenges.
Figure 1. B5: Bigger and better oil through integrated approaches across aims. Courtesy B5 Team.
This research was supported by the DOE Office of Science, Biological and Environmental Research (BER) Program, grant no. DE-SC0023142.