Performing Department
(N/A)
Non Technical Summary
This project aims to improve agronomic traits of pennycress to enhance its potential as an emerging cash cover crop through a unique accelerated breeding approach that leverages computational simulations and gene-editing technologies. Cover crops can reduce soil erosion and improve soil health, but they adoption of cover cropping as management practice is hindered by implementation cost and potential cash crop yield loss. Pennycress has the potential to provide soil health benefits and generate 879 liters/ha of seed oil that can be converted into biodiesel and bio-jet fuels. However, pennycress is only partially domesticated, and efforts to improve agronomic traits such as flowering time, seed quality, and stress tolerance are ongoing.To accelerate the development of commercially viable pennycress, this project will leverage computational protein simulation methods to identify beneficial variants to the target gene, the E1 component of the ODGH enzyme complex, that result in earlier flowering times, larger seed sizes, and improved root system architecture. Potential gene variants will be introduced into pennycress plants using CRISPR-CAS gene editing technologies to generate plant varieties with the precise gene variant of interest. These plant varieties will then be evaluated at seedling stage and at maturity to evaluate the impact of the gene edit on plant aboveground and belowground growth as well as oil quality. Agronomic traits such as flowering time and seed size will also be measured and evaluated. Improved pennycress varieties produced from this work has the potential to bring pennycress closer to market as a cash cover crop and enable a circular bioeconomy for sustainable energy production.
Animal Health Component
50%
Research Effort Categories
Basic
25%
Applied
50%
Developmental
25%
Goals / Objectives
Overall Goal: The goal of this project is to identify beneficial mutations to the E1 component of oxoglutarate dehydrogenase that improves flowering time, seed size, and root system architecture in the emerging cash cover crop pennycress. The project will accomplish this goal by establishing a combined computation and experimental pipeline that enables protein mutations to be computationally screened prior to gene editing in plants to enable an accelerated and more predictive breeding framework.Objectives:Establish a computational-experimental pipeline for screening single amino-acid substitutions at the E1-OGDH enzymatically active site.Identify and generate mutant lines of pennycress with the top 5 and bottom 5 candidate mutations from simulation using CRISPR-CAS gene editing technologies.Compare early shoot and root development by X-ray CT in pennycress seedlings of mutant lines to wild type, gene knockouts, and overexpression lines.Comparing root and shoot development of mature pennycress plants grown in mesocosms.Assess biochemical properties of pennycress tissues and seeds in mutant lines compared to wild type, gene knockouts, and overexpression lines.
Project Methods
This project will utilize protein simulations, namely NAMD Free Energy Perturbation simulations, to identify single amino-acid substitutions to the active site of the E1 component of OGDH that increase and lower enzymatic rate. Candidate mutations will be introduced into pennycress plants utilizing CRISPR-CAS gene editing technologies, prime and base editing. Homozygous mutant lines will be evaluated for changes in root system architecture using a gel-based imaging system. Mutant lines with beneficial mutations that induce morphological changes in planta will be further screened at seedling and reproductive stages using x-ray CT imaging and photogrammetry to better understand changes in root growth behavior. Root traits will be extracted from 3D root models using custom code. Plant tissues and seeds will be evaluated by LC-MS for changes in plant metabolism.