Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (21): 4603-4611.doi: 10.3864/j.issn.0578-1752.2013.21.024

• RESEARCH NOTES • Previous Articles     Next Articles

Cloning and Expression Analysis of Fe Superoxide Dismutase (Fe-SOD) Gene from Winter Turnip Rape (Brassica rapa L.)

 ZENG  Xiu-Cun-1, 2 , SUN  Wan-Cang-1, SUN  Jia-3, XU  Yao-Zhao-2, FANG  Yan-1, SHI  Peng-Hui-1, YANG  Gang-1, KONG  De-Jing-1, WU  Jun-Yan-1, LIU  Zi-Gang-1   

  1. 1.Agricultural College, Gansu Agricultural University, Lanzhou 730070, China
    2.College of Agronomy and Biotechnology, Hexi University, Zhangye 734000, Gansu, China
    3.Department of Plant Sciences, University of Saskatchewan, Saskatoon, Saskatchewan,  Canada S7N5A8
  • Received:2013-06-18 Online:2013-11-01 Published:2013-08-14

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Abstract: 【Objective】The objectives of the present study were to clone the Fe-SOD gene of SOD family from an extremely low temperature (-32℃) resistant winter turnip rape (B. rapa L.) cultivar Longyou 7 and analyze its expression under low temperature conditions .【Method】The cDNA sequence of Fe-SOD was isolated by RT-PCR, and the obtained cDNA sequence and the deduced amino acid sequence was analyzed. Semi-quantitative and real time RT-PCR were used to assess the expression of Fe-SOD in response to low temperature stress. The superoxide dismutase enzyme activity was measured by NBT deoxidization method in leaves and roots. 【Result】The Fe-SOD gene was isolated from winter turnip rape (GenBank accession number. KF178713). The cDNA sequence of this gene was 645 bp in length, containing a 639 bp opening reading frame (ORF) , encoded a polypeptide of 212 amino acid，and with 99 % of amino sequence similarity to a vegetable cultivar of B. rapa (Chiifu). The protein encoded by this gene was a hydrophilic protein without signal-peptide and transmembrane region. The prediction of the second structures indicated that the Fe-SOD was a steady protein with more α-helices. Semi-quantitative and real time RT-PCR result showed that Fe-SOD was expressed upregulatedly in response to early low temperature stress (4℃). However, the expression of this gene was inhibited at the lower temperature stress (-4℃ and -8℃). The result of measured enzyme activity showed that superoxide dismutase enzyme activity in roots was higher than that of leaves，which could make winter turnip rape successfully overwinter. 【Conclusion】The Fe-SOD gene cloned from winter turnip rape had genetic characteristics similar with other known species and it might play a role in cold tolerance of the B. rapa cultivar Longyou 7.

Key words: winter rape (Brassica rapa L.) , lower temperature , Fe-SOD , expression analysis , activity of SOD

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