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Abstract
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Oil quality from oilseeds is determined by the fatty acid (FA) content. Fatty Acid Elongase 1 (FAE1) 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 CsFAE1B, CsFAE1A, and
CsFAE1C genes using CRISPR/Cas9 was used to improve the quality of C. sativa oil (which contains
about 50% oil and 4% erucic acid). Additionally, camelina tissue culture optimization for recombinant
vector transfer was performed concurrently. Finally, gRNA was designed to target three copies of the
CsFAE1 genes, and after transfer to the pFGC-pcoCas9 vector, the recombinant vector was confirmed by
PCR and enzymatic digestion. The tissue culture optimization results indicated that hormone ratios of 0.5
mg L-1 NAA and 3 mg L-1 BAP for cotyledon and hypocotyl in the Gamborg medium induced
embryogenic calluses three weeks after cultivation. Additionally, hormone ratios of 0.5 NAA, 2 BAP, and
1 (mg L-1) 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.
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