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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (11): 1629-1637.

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

Cloning and Functional Analysis of Peanut Δ9-Stearoyl-Acyl Carrier Protein Desaturase Promoter

SHI Lei, MIAO Li-Juan, QI Fei-Yan, ZHANG Zhong-Xin, GAO Wei, SUN Zi-Qi, HUANG Bing-Yan, DONG Wen-Zhao, TANG Feng-Shou, and ZHANG Xin-You*   

  1. Industrial Crops Research Institute, Henan Academy of Agricultural Sciences / Key Laboratory of Oil Crops in Huanghuaihai Plains, Ministry of Agriculture / Henan Provincial Key Laboratory for Oil Crops Improvement, Zhengzhou 450002, China
  • Received:2016-02-29 Revised:2016-07-11 Online:2016-11-12 Published:2016-08-11
  • Contact: Zhang Xinyou,E-mail:haasz@126.com,Tel:0371-65729560 E-mail:leis100@163.com
  • Supported by:
    This study was supported by National High-Tech R&D Program of China (863 Program) (2013AA102602-6), the Key Project of Science and Technology of Henan Province (141100110600), the China Agriculture Research System (CARS-14), the Henan Provincial Agriculture Research System (S2012-5), and the Scientific Foundation for Excellent Young Scholars of Henan Academy of Agricultural Sciences (2013YQ11).

Abstract:

The Δ9-stearoyl-acyl carrier protein desaturase (SAD) plays key roles in determining the ratio of saturated to unsaturated fatty acids in higher plants. The 720 bp 5′ flanking sequence of peanut SAD (AhSAD) gene was isolated from the genomic DNA of peanut cultivar Yuhua 9326 by nested PCR using genomic walking method. The fragment of 5′ UTR was obtained using 5′ RACE (Rapid Amplification of cDNA End), confirming that the 720 bp 5′ flanking sequence is promoter fragment based on sequence alignments. Bioinformatics analysis indicated that AhSAD promoter contained several light, hormone responsive elements and enhancer-like elements as well as CAAT box and TATA box. To study the function of this promoter, we constructed a binary expression vector pBI-PAhSAD by replacing CaMV35S promoter of pBI121 with the AhSAD promoter, which was introduced into Arabidopsis and transiently expressed in peanut, respectively, by Agrobacterium-mediated transformation. Histochemical staining analysis showed that the GUS gene mainly expressed in roots, stem, leaves, cotyledon and peanut pegs. The histochemical staining was observed in peanut whole leaves, while only in Arabidopsis veins of cauline leaves.

Key words: Peanut(Arachis hypogaea L.), Δ9-Stearoyl-ACPdesaturase(SAD), Promoter, Genomicwalking, GUSreportergene

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