Expression Of BnFAD2-C5 Gene In Brassica Napus L. And Functional Analysis Of Its Regulatory Sequences

High oleic rapeseed breeding and the formation of oleic acid has become a central issue after finding the important economic value of rapeseed oil with high oleic acid.The fatty acid dehydrogenase gene (FAD2 gene) is a key enzyme gene to control oleic acid content, but the regulation of FAD2 gene is not well understood. We chose the BnFAD2-C5 gene to make an in-depth analysis. The expression of BnFAD2-C5 gene has been obtained by qRT-PCR, showing that some internal factors will introduce the gene expressing in 20days after flowering. Promoter and intron analysis of BnFAD2-C5 gene using ?-glucuronidase (GUS) reporter and green fluorescent protein (GFP)reporter system demonstrated that BnFAD2-C5 promoter is a seed-specific inducible promoter, and the inducible elements are existed in-1257?-1020bp and -319?-bp regions and play an important role in the early developing seeds. The+526?+631bp region of intron also has some strong inducible cis-elements and make functions to induce the BnFAD2-C5 gene express in early time of developing seed namely the fatty acid synthesis time. Furthermore, the intron confers the enhancement of GFP expression in transgenic Arabidopsis seeds as compared with intron-less controls. Prerequisite cis-elements for the BnFAD2-C5intron-mediated enhancement of gene expression was located in +631?+1033bp region, and it can enhance not only the BnFAD2-C5 promoter but also the CAMV35S promoter. All of the positive cis-elements in promoter and intron plays function in early time of developing seeds and negative cis-elements late time.