Tissue Culture Optimization and Construction of a Suitable Vector for Engineering Very Long-Chain Fatty Acids in Camelina sativa L.

Oil quality from oilseeds is determined by the fatty acid (FA) content. Fatty Acid Elongase ۱ (FAE۱) is an essential FA gene that has been genetically modified to change the constitution of fatty acids in oilseed plants. In this study, recombinant vector construction for genome editing of the CsFAE۱B, CsFAE۱A, and CsFAE۱C genes using CRISPR/Cas۹ was used to improve the quality of Camelina Sativa oil (which contains about ۵۰% oil and ۴% erucic acid). Additionally, camelina tissue culture optimization for recombinant vector transfer was performed concurrently. Finally, gRNA was designed to target three copies of the CsFAE۱ genes, and after transfer to the pFGC-pcoCas۹ vector, the recombinant vector was confirmed by PCR and enzymatic digestion. The tissue culture optimization results indicated that hormone ratios of ۰.۵ mg L-۱ NAA and ۳ mg L-۱ BAP for cotyledon and hypocotyl in the Gamborg medium induced embryogenic calluses three weeks after cultivation. Additionally, hormone ratios of ۰.۵ NAA, ۲ BAP, and ۱ (mg L-۱) Kin led to direct regeneration in cotyledon explants. In future studies, tissue culture optimization and recombinant vector construction for genome editing of FAEs genes could improve oil quality with the genetic transformation of camelina.

کلیدواژه ها:

CRISPR-Cas۹ ، CsFAE۱ ، gRNA ، Recombinant Vector