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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (10): 1471-1478.doi: 10.3724/SP.J.1006.2016.01471

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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

LIU Rui-Yang,LIU Fang,ZHANG Zhen-Qian,GUAN Chun-Yun   

  1. College of Agronomy, Hunan Agricultural University / National Oilseed Crops Improvement Center in Hunan, Changsha 410128, China
  • Received:2016-02-16 Revised:2016-05-09 Online:2016-10-12 Published:2016-06-06
  • Contact: 官春云, E-mail: guancy2011@yahoo.com.cn E-mail:ruiyang_liu2007@126.com
  • Supported by:

    This study was supported by Graduate Innovation Foundation of Hunan (CX2013A012) and the Major State Basic Research Development Program of China (2015CB150200).

Abstract:

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 transform 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 regionsof 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

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