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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (12): 2139-2146.doi: 10.3724/SP.J.1006.2012.02139

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

Molecular Cloning of Sucrose Synthase Gene and Expression Analysis under Drought Stress in Peanut (Arachis hypogaea L.)

HE Mei-Jing,LIU Li-Feng*,MU Guo-Jun,HOU Ming-Yu,CHEN Huan-Ying,CUI Shun-Li   

  1. Key Laboratory of Crop Germplasm Resources of Hebei Province / College of Agronomy, Agricultural University of Hebei, Baoding 071001, China
  • Received:2012-07-15 Revised:2012-07-15 Online:2012-12-12 Published:2012-10-08
  • Contact: 刘立峰, E-mail: liulifeng@hebau.edu.cn

Abstract:

Sucrose synthase (SuSy), a key enzyme in sucrose metabolism, plays an important role in both plant growth and development and osmotic adjustment. Here, a cDNA sequence of sucrose synthase gene, designated AhSuSy (GenBank accession number: JF346233), was isolated from peanut using homologous cloning, RACE and TAIL-PCR. The full-length of AhSuSy cDNA is 2 790 bp, contained an ORF of 2 421 bp, the 5′UTR and 3′UTR of 57 bp and 312 bp, respectively. A peptide of 806 amino acid residues was deduced according to the ORF, which showed more than 75% homologous to the corresponding proteins in soybean, Arabidopsis and maize. The AhSuSy was ligated into pET-32a(+) and transformed into E. coli competent cell. After induced by IPTG, the fusion protein was obtained and its relative molecular weight was 92 kD, which was consistent with the theoretical value. The tissue specific expression analysis by using semi-quantitative RT-PCR assays indicated that AhSuSy was expressed in root, stem and leaf. After treated with 10%PEG, seedlings of peanut were used to determine transcript level for AhSuSy, sucrose synthase activity and sucrose content. The result showed that all of the three parameters were gradually increased during 4h to 12 h after treatment and the correlation coefficient between sucrose synthase activity and sucrose content was 0.993 (P=0.007<0.01) during this period, then declined from 12h to 24 h after treatment, which indicated that AhSuSy may play an important role in adaptation of plants to drought stress.

 

Key words: Peanut, Sucrose synthase gene, Cloning, Semi-quantitative RT-PCR, Expression analysis

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