DOI: 10.3724/SP.J.1006.2016.01471

Acta Agronomica Sinica (作物学报) 2016/42:10 PP.1471-1478

Functional Analysis of BnFAD2-C5 Promoter and Intron at Expression Level in Brassica napus

High oleic rapeseed breeding and the formation mechanism of oleic acid have become a central issue after finding the important economic value of rapeseed oil with high oleic acid. The fatty acid dehydrogenase gene (FAD2) is a key enzyme gene to control oleic acid content, but the regulation of FAD2 gene is not well understood. According to the homology between rapeseed and oleracea, the BnFAD2-C5 promoter sequence of 1257 bp was cloned. Promoter and intron of BnFAD2-C5 gene were analyzed using β-glucuronidase (GUS) reporter and green fluorescent protein (GFP) reporter system to construct deleted vectors and trans-form Arabidopsis thaliana. Deletion analysis of BnFAD2-C5 promoter through GUS stainning revealed that -319 to -1 bp was the minimum promoter region. And deletion analysis of BnFAD2-C5 promoter and intron through GFP reporter system using western technique showed that -1257 to -1020 bp and -319 to -1 bp regions of BnFAD2-C5 promoter could induce expression of reporter genes effectively in transgenic Arabidopsis seed in the mid stage of seed development, while BnFAD2-C5 intron could confer the enhancement of promoter's function and the intron-mediated enhancement region was mainly located in +631 to +1033 bp.

Key words:Brassica napus,BnFAD2-C5 gene,Intron-mediated enhancement,Arabidopsis thalina

ReleaseDate:2017-01-12 13:30:45

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