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Published on September 11, 2003; 10.1104/pp.103.026245

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Received May 15, 2003
Returned for revision July 1, 2003
Accepted July 19, 2003

Development of Protoporphyrinogen Oxidase as an Efficient Selection Marker for Agrobacterium tumefaciens-Mediated Transformation of Maize

Xianggan Li *, Sandy L. Volrath , David B.G. Nicholl , Charles E. Chilcott , Marie A. Johnson , Eric R. Ward , and Marcus D. Law

Syngenta Biotechnology, Inc., P.O. Box 12257, 3054 Cornwallis Road, Research Triangle Park, North Carolina 27709-2257

* Corresponding author; email: Xianggan.Li{at}syngenta.com.

In this article, we report the isolation of plant protoporphyrinogen oxidase (PPO) genes and the isolation of herbicide-tolerant mutants. Subsequently, an Arabidopsis double mutant (Y426M + S305L) was used to develop a selectable marker system for Agrobacterium tumefaciens-mediated transformation of maize (Zea mays) and to obtain multiple events tolerant to the PPO family of herbicides. Maize transformants were produced via butafenacil selection using a flexible light regime to increase selection pressure. Butafenacil selection per se did not change transgene copy number distribution relative to other selectable marker systems, but the most tolerant events identified in the greenhouse were more likely to contain multiple copies of the introduced mutant PPO gene. To date, more than 2,500 independent transgenic maize events have been produced using butafenacil selection. The high frequency of A. tumefaciens-mediated transformation via PPO selection enabled us to obtain single-copy transgenic maize lines tolerant to field levels of butafenacil.




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