Production of High Oil, Transgene Free Camelina sativa Plants through Genome Editing

Camelina sativa (‘false flax,’ Brassicaceae) is a traditional European oilseed that is now an engineering target as a dedicated bioenergy crop. Camelina oil’s high unsaturated fatty acid content makes it uniquely suited for biodiesel production; however, low overall seed yield limits agricultural profitability. The hexaploid Camelina genome has been sequenced and efficient transformation is achieved through Agrobacterium-mediated gene transfer via floral dip. We have used the CRISPR/Cas9 gene editing system to target genes involved in the regulation of carbon assimilation, transport, and fatty acid biosynthesis pathways in order to increase seed yield and oil content. The use of CRISPR/Cas9 genome editing has allowed the generation of custom mutants for gene function studies, which are used in applied work on yield improvement for increased profitability of this crop. We have also established an amplicon deep-sequencing genotyping method to identify edited sites, which allows us to overcome both background SNPs in the allohexaploid plant and variation in CRISPR edits across plants. To date using this method as well as conventional methods we have genotyped up to the third transgenic generation of multiple lines, and are currently assessing yield and oil content.

Acknowledgement: This research is funded by BETO award from the Department of Energy in collaboration between North Carolina State University and Yield 10 (Formerly MetaboliX)