Decreasing erucic acid level by RNAi-mediated silencing of fatty acid elongase 1 (BnFAE1.1) in rapeseeds (Brassica napus L.)
The β-ketoacyl CoA synthase encoded by fatty acid elongase 1 gene (BnFAE1.1) is a rate-limiting enzyme regulating biosynthesis of erucic acid in rapeseeds (Brassica napus). To develop low level of erucic acid in rapeseeds by intron-spliced hairpin RNA, an inverted repeat unit of a partial BnFAE1.1 gene interrupted by a spliceable intron was cloned into pCAMBIA3301, and a seed-specific (Napin) promoter was used to control the transcription of the transgene. Four transgenic plants harboring a single copy of transgene were generated. Expression of endogenous BnFAE1.1 gene in developing T3 seeds was significantly reduced. In mature T3 seeds, erucic acid was decreased by 60.8 to 99.1% compared with wild type seeds, and accounted for 0.36 to 15.56% of total fatty acids. The level of eicosenoic acid was also greatly decreased. Furthermore, it resulted in a significant increase in the level of oleic acid, but total fatty acid content in T3 seeds was the same with that in wild type seeds. In conclusion, the expression of endogenous BnFAE1.1 was efficiently silenced by the designed RNAi silencer, causing a significant down-regulation in the level of erucic acid. Therefore, the RNAi-mediated post-transcriptional silencing of FAE1 gene to reduce oleic acid in rapeseeds was an efficient method to breed some new B. napus lines.
Key words: Brassica napus L., fatty acid elongase, intron-spliced hairpin RNA, down-regulation, erucic acid.