De novo domestication of wild Brassica species using CRISPR/Cas9 gene editing for resilient crop development
A genome editing platform that transforms wild and feral Brassica crop relatives adapted to harsh growing conditions into new crops requiring less water, pesticides, and fertilizers. The approach uses CRISPR/Cas9 to target MADS-box genes, beginning with the CLF curly leaf gene, building on proven de novo domestication methods demonstrated in tomato and eggplant relatives.
Overview
This research program applies CRISPR/Cas9 genome editing to enable de novo domestication of wild and feral Brassica species adapted to harsh growing conditions. The goal is to develop new crop varieties that thrive in regions where traditional crops are maladapted, reducing requirements for water, pesticides, and fertilizers. The approach builds on recently demonstrated de novo domestication methods in wild relatives of tomato and eggplant, extending these techniques to Brassica hilarionis, a wild species that naturally grows in challenging environments. By targeting MADS-box genes such as CLF (curly leaf), the program aims to produce domesticated Brassica crops with improved agronomic traits.
Technical specifications
Key research elements:
- CRISPR/Cas9 gene editing applied to MADS-box genes, with initial focus on the CLF gene, which produces easily identifiable curled-leaf and reduced-size phenotypes
- Two-phase transformation pipeline: protocol optimization first in Brassica oleracea DH1012 (a line proven transformable via Agrobacterium co-culture of cotyledons), then application to wild Brassica hilarionis
- Tissue culture and regeneration: cotyledon-based Agrobacterium co-culture with herbicide selection to identify successful transformants
- Selfing de novo domestication platform: Brassica hilarionis optimized as a self-pollinating platform to accelerate trait fixation
- Genome assembly: developing a reference genome for Brassica hilarionis to accelerate the domestication pipeline
- Validation methods: sequencing-based verification of transformants, herbicide and GFP screening, and tracking of efficiency and heritability in offspring
Current proof-of-concept results:
- First batch: 100 cotyledons screened, 11 surviving herbicide selection, with 6 regenerated controls from 20
- Brassica hilarionis selfing platform established
Technology readiness level
The technology is currently at an early research and proof-of-concept stage. Protocol optimization is underway in the model line Brassica oleracea DH1012, with initial tissue culture regeneration demonstrating feasibility. Transformant verification by sequencing is the next immediate step. Future validation will involve screening hundreds of cotyledons for herbicide resistance and GFP expression, sequencing verification, and tracking heritability across generations in DH1012 before repeating the process in wild B. hilarionis. Parallel work will optimize gRNA design for MADS-box genes. Funding would support a semester of salary, genome assembly, and reagents needed to advance this research toward a functional de novo domestication platform for resilient Brassica crops.
About University of Missouri, Columbia
Founded as Missouri’s flagship land‑grant, the University of Missouri–Columbia is a comprehensive public research university serving a large student body and partners statewide. Industry engages on campus through co‑located research cores, a research park and incubator in Columbia, and access to a high‑power university research reactor supporting isotope production and advanced testing. An integrated academic health system enables clinical studies and translation, while a statewide extension and agricultural research network links companies to field sites, producers, and community testbeds across Missouri. Research is supported by competitive federal funding from agencies such as NIH, NSF, USDA, and DOE. A dedicated tech transfer office supports IP, licensing, and sponsored‑research agreements.
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