METHOD OF INCREASING ERUCIC ACID PRODUCTION BY HAIRPIN RNA FAD2 GENE SILENCING AND FAE GENE OVEREXPRESSION

摘要

The 3'-UTR of the FAD2 gene from Brassica carinata was cloned by PCR and used to prepare an intron-spliced hairpin RNA (ihpRNA) construct. This construct, when expressed in B. carinata, resulted in a high degree of FAD2 gene silencing accompanied with a strong increase in oleic and erucic acid content by up to 267% and 27% respectively, compared to that of the wild type background (WT). A second construct containing ihpRNA targeted to the endogenous FAD2 gene in addition to the heterologous Crambe abyssinica FAE gene under the control of seed-specific napin promoter, was used to transform B. carinata. This approach resulted in a more dramatic increase in erucic acid content, by up to 41 % in T1 segregating seeds as compared to that of WT control.