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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (04): 565-573.doi: 10.3724/SP.J.1006.2015.00565

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

Cloning of BnADH3 Gene from Brassica napus L. and Submergence Tolerance of BnADH3 Transgenic Arabidopsis

LÜ Yan-Yan,FU San-Xiong,CHEN Song,ZHANG Wei,QI Cun-Kou*   

  1. Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences / Jiangsu Collaborative Innovation Center for Modern Crop Production, Nanjing 210014, China
  • Received:2014-07-11 Revised:2015-02-05 Online:2015-04-12 Published:2015-03-02
  • Contact: 戚存扣, E-mail: qck9898@sina.com

Abstract:

BnADH3 gene was highly homologous to BoADH3 from Brassica oleracea and AtADH3 from Arabidopsis, with the BnADH3  expression was induced by submergence and the up-regulation occurred since 6-hour post treatment. The BnADH3 transgenic Arabidopsis was obtained and the transgenic seedBnADH3 gene was cloned from submergence-tolerant line WR-4 of Brassica napus L. using RT-PCR technique. The full-length open reading frame is 1137 bp, encoding 379 amino acids. Homology analysis showed that BnADH3 gene was highly homologous to BoADH3 from Brassica oleracea and AtADH3 from Arabidopsis, with the 96% and 91% similarity, respectively. Quantitative RT-PCR assay was carried out to compare BnADH3 expression between the submergence-tolerant line WR-4 and the submergence-susceptible line WR-24. The result showed that the BnADH3  expression was induced by submergence and the up-regulation occurred since 6-hour post treatment. The BnADH3 transgenic Arabidopsis was obtained and the transgenic seedlings were exposed to three-day submergence stress. Overexpression of BnADH3 resulted in higher ADH activity in leaf and root of transgenic Arabidopsis compared to that of the wild type. The four- and six-week seedlings of T2 generation showed higher tolerance to submergence stress after three-day submergence treatment and most T2 seedlings were recovered with normal growth when the stress was relieved for three days. However, the wild-type seedlings withered until death. After five-day submergence, the survival ratios were 26.7% for the wild type, 80.0% for transgenic line ADH33, and 66.7% for transgenic line ADH44.

Key words: Brassica napus L., Gene BnADH3, Transgenic Arabidopsis, Flood tress, Flood tolerance

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