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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (04): 636-643.doi: 10.3724/SP.J.1006.2014.00636


Cloning and Expression Analysis of Copper and Zinc Superoxide Dismutase (Cu/Zn-SOD) Gene from Brassica campestris L.

ZENG Xiu-Cun1,2,LIU Zi-Gang2,SHI Peng-Hui2,XU Yao-Zhao1,SUN Jia3,FANG Yan4,YANG Gang2,WU Jun-Yan2,KONG De-Jing2,SUN Wan-Cang2,*   

  1. 1 College of Agronomy and Biotechnology, Hexi University, Zhangye 734000, China; 2 The College of Agronomy Gansu Agricultural University / Gansu Engineering Research Center of Rapeseed, Lanzhou 730070, China; 3 Department of Plant Sciences, University of Saskatchewan, Saskatoon, Saskatchewan S7N5A8, Canada; 4 Research and Testing Center of Gansu Agricultural University, Lanzhou 730070, China
  • Received:2013-07-07 Revised:2013-11-24 Online:2014-04-12 Published:2014-01-16
  • Contact: 孙万仓, E-mail: 18293121851@163.com


Superoxide Dismutase (SOD) is a key enzyme eliminating reactive oxygen species (ROS), and the copper and zinc superoxide dismutase (Cu/Zn-SOD) is the most important active oxygen scavenger in SOD family, which can improve plant tolerance to environmental stresses. The cDNA of Brassica campestris L. cultivar Longyou 7 was cloned by RT-PCR, using the primers designed according to the published crucifer Cu/Zn-SOD cDNA sequences. The sequence of Cu/Zn-SOD from B. campestris L. was 459 bp, encoding a predicted protein of 152 amino acid residues. Bioinformatics analysis showed that amino sequence similarity with Brassica napus was 99%, and the predicted Cu/Zn-SOD protein was a hydrophilic protein without signal-peptide as well as transmembrane region. It contained specific sequence characteristics and conserved domain of copper and zinc superoxide dismutase (Cu/Zn-SOD) superfamily. The expression analysis of Cu/Zn-SOD gene using semi-quantitative RT-PCR and results of SOD activity in response to lower temperature showed that the Cu/Zn-SOD was a differential expressed gene induced by lower temperature. Results of separating low-temperature-induced proteins from Brassica campestris L. cultivar Longyou 6 by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry further showed that Cu/Zn-SOD was a stress-responsed gene, whose expression was induced by lower temperature.

Key words: Winter rape (Brassica compestris L.), Cu/Zn-SOD gene cloning, Lower temperature, Expression analysis, Activity of SOD, Low-temperature-induced protein

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