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Acta Agron Sin ›› 2009, Vol. 35 ›› Issue (1): 71-78.doi: 10.3724/SP.J.1006.2009.00071

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

Cloning of Cu/Zn-Superoxide Dismutase of Brassica napus and Its Induced Expression by Sclerotinia slerotiorum

YANY Yang-Yang,LI Yun,DING Yong,XI Chun-Lei,ZHANG Cheng-Gui,LIU Ying,GAN Li   

  1. College of Plant Science and Technology,Huazhong Agricultural University, Wuhan 430070,China
  • Received:2008-04-20 Revised:2008-06-15 Online:2009-01-12 Published:2008-11-17
  • Contact: YANY Yang-Yang E-mail:yangyang401@126.com

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Abstract:

Bythe primers designed from conserved homologous sequences of the genes for superoxide dismutase (SOD) among Arabidopsis thaliana, Brassica juncea and B. rapa ssp. pekinensis, Cu/ZnSOD and FeSOD genes in B. napus were cloned using the methods of homology-based candidate gene and RACE-PCR. After cloning andsequencing, the full lengths of cDNA of Cu/ZnSOD and FeSOD genes (accession no. AY970822 and EF634058 in GenBank) were 756 and 1 037 bp, respectively. Their corresponding genomic sequences were 1 322 bp for Cu/ZnSOD gene and 1 659 bp for FeSOD gene with accession nos. DQ431853 and EF634057 in GenBank, respectively. Bioinformatic analyses showed that the ORF of Cu/ZnSOD gene with 459 bp contained six introns with seven extrons and a polyA tail and could express a protein of 152 amino acids; and the one of FeSOD gene with 792 bp contained seven introns with eight extrons and could determine a protein of 263 amino acids. With cDNA of Cu/ZnSOD gene as probe, Northern blotting analysis showed that its mRNA expression in B. napus cultivars increased after the infection by Sclerotinia sclerotiorum, the expression amount in resistant or tolerant cultivars was higher than that in sensitive ones. The same trend was observed for the SOD activity in leaves. These results suggested that SOD genes were related with the resistance to the disease.

Key words: Cu/ZnSOD, FeSOD, Brassica napus, Sclerotinia sclerotionrum, Gene expression analysis

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