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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (08): 1400-1408.doi: 10.3724/SP.J.1006.2013.01400

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

Cloning, Expression, and Activity Analysis of Ascorbate Peroxidase (APX) Gene from Winter Turnip Rape (Brassica campestris L.)

ZENG Xiu-Cun1,2,SUN Wan-Cang1,*,FANG Yan1,LIU Zi-Gang1,DONG Yun3,SUN Jia4,WU Jun-Yan1,ZHANG Peng-Fei1,SHI Peng-Hui1,KONG De-Jing1,ZHANG Teng-Guo5,HE Li1,ZHAO Cai-Xia1   

  1. 1 Agricultural College, Gansu Agricultural University, Lanzhou 730070, China; 2 College of Agronomy and Biotechnology, Hexi University, Zhangye 734000, China; 3 Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070 China; 4 Department of Plant Sciences, University of Saskatchewan, Saskatoon, Saskatchewan, Canada. S7N5A8; 5 College of Life Science, Northwest Normal University, Lanzhou 730070, China
  • Received:2012-12-23 Revised:2013-04-22 Online:2013-08-12 Published:2013-05-22
  • Contact: 孙万仓, E-mail: wangcangsun@yahoo.com.cn

Abstract:

Ascorbate Peroxidase (APX) is a key enzyme which eliminates oxygen free radicals and increases plant resistance in adverse circumstances. The complete open reading frame of APX gene was cloned from an extremely low temperature (–32℃) resistant winter turnip rape (Brassica campestris L.) cultivar Longyou 7 using the method of reverse transcription PCR (RT-PCR). The primers were designed according to the published APX cDNA sequences. The sequence of APX from B. campestris L. was 753 bp, encoding a protein of 250 amino acid residues with a predicted molecular weight of 27.7 kD and a theoretical pI of 5.58. Bioinformatics analysis showed that the predicted APX protein contained a conserved amino acid sequence corresponding to the plant peroxidase (POD) superfamily. The APX had no signal peptide and was a hydrophilic protein locating in cytoplasm. The prediction of the second structures indicated theAPX was a steady protein with more random coils and α-helices. The expression analysis of APX gene using real time RT-PCR and the activity of APX in response to lower temperature showed that the APX could be induced and the activity of APX could be changed by lower temperature. In conclusion, APX gene might play a role in cold tolerance of the B. campestris L. cultivar Longyou 7.

Key words: Winter rape (Brassica compestris L.), Lower temperature, APX gene cloning, Expression analysis, Activity of APX

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