Enhancing the bio-industrial application of Brassica carinata using metabolic engineering and genome editing

Abstract Description: Brassica carinata commonly called Abyssinian/Ethiopian mustard is considered as a new crop platform dedicated to bio-industrial applications such as for biofuel and oleo-chemical industries. The presence of long chain fatty acid (VLCFA) especially erucic acid and other features of the crop make B.carinata as new potential feedstock for sustainable aviation fuel (SAF). The use of SAF in turn contribute to the reduction of greenhouse gas (GHG) emission or CO2 emission caused by the fastest growing aviation industries in the globe. Moreover, other beneficial coproduct molecules can be generated for oleo-chemical industries during conversion of the B. carinata oil into biofuel.  The aim of our study is to enhance the role B.carinata for low carbon bio-economy through developing genotypes with super-oil quality for bio-industrial application using biotechnological tools. So far, we found six B.carinata lines engineered for high erucic acid content (up to 52.7%) and five lines that can produce wax esters ranges from 12 to 25.6% using metabolic engineering. Studies are underway for further enhancement of the oil oxidative stability of B.carinata oil using DNA-free genome editing. We demonstrated PEG-mediated transfection of protoplast cells using ribonucleoprotein (RNP)-mediated CRISPR/Cas9. Our target gene was fatty acid desaturase (BcFAD2) whereby four potential sgRNAs were designed using the target sites of the gene. Among the four guides, we found two guides sgRNA2 and sgRNA4 that showed higher editing efficiency of 54.75% and 75%, respectively.  The output of the aforementioned studies will contribute to the efforts of improving B.carinata for bio-industrial applications especially for the current demands to shift from fossil fuel towards to a sustainable aviation fuel. The relevance of the project can also be looked from SDG agendas that encourages the access of affordable, reliable and modern renewable energy sources in the globe.   

Equity and Inclusion: As we explained the overall aim of the project, this project proposal focuses on addressing the common agenda of the globe to generate high-quality products that address the global issue of a carbon economy. The research will engage research groups of diverse research background in Brassica carinata and will include other expertise from chemical industries and related fields. The final output of the study will benefit all individuals and will also touch one of the common goals of SDG, getting affordable, reliable and modern renewable energy .