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This article in CS

  1. Vol. 46 No. 2, p. 706-711
     
    Received: Apr 14, 2005
    Published: Mar, 2006


    * Corresponding author(s): xupeilin@hotmail.com
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doi:10.2135/cropsci2005.0290

Use of aroA-M1 as a Selectable Marker for Brassica napus Transformation

  1. J. X. Wang,
  2. F. Y. Zhao and
  3. P. Xu *
  1. Biotechnology Research Center, The Key Laboratory of Gene Engineering of Education Ministry, Zhongshan University, Guangzhou, 510275, The People's Republic of China

Abstract

The aroA-M1 mutant consisting of an N-terminal portion of Escherichia coli 5-enolpyruvylshkimate-3-phosphate synthase (EPSPS) and a C-terminal portion of Salmonella typhimurium EPSPS confers high resistance to glyphosate because of a decrease in the K m (PEP) and an increase in the K i (glyphosate). In this paper, we constructed a binary vector carrying the aroA-M1 mutant gene under the control of the CaMV 35S promoter for Agrobacterium-mediated gene transfer to Brassica napus L. Transgenic Brassica plants with increased resistance to glyphosate were obtained by cocultivation of hypocotyl explants with Agrobacterium tumefaciens and direct selection on media containing glyphosate. Southern blot and western blot analyses confirmed gene integration and expression in T0 transgenic Brassica plants. Transgenic plants exhibited increased resistance to glyphosate compared with untransformed plants. Our results also indicated the aroA-M1 mutant gene may be a useful selectable marker for the transformation of plants and for the production of herbicide-resistant plants.

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