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Development of phosphinothricin resistant fertile transgenic tobacco plants

Development of phosphinothricin resistant fertile transgenic tobacco plants

Acta Agronomica Hungarica 46(2): 97-104

This study was performed to develop transgenic tobacco plants (Nicotiana tabacum, Samsun) resistant to broad spectrum herbicides based on phosphinothricin (PPT). The bar gene that is responsible for the synthesis of the phosphinothricin N-acetyl-transferase (BAR) enzyme, which detoxifies PPT through acetylation, was transferred to tobacco plants. The LBA4404 strain of Agrobacterium tumefaciens harbouring the binary vector pDHB321.1 was used in the transformation experiments. Gene transfer was performed using the leaf disc transformation system. At the end of the transformation experiments, callus stem and root formation were observed at 10 mg/L PPT, which was the lethal dose. Following transfer to soil, the regenerated primary transgenic plants were not affected by the application of 1% (v/v) BASTA (30 L/ha) preparations equivalent to 2.0 g/L gluphosinate ammonium. The control plants, grown under the same conditions and treated with the same dose of herbicide, died within 15-20 days of the herbicide application. The expression of the bar gene was verified by electrophoretic studies where the 23 kD BAR enzyme was shown to be absent in the controls. Progeny obtained from selfed transgenic plants were able to germinate and develop in the presence of 10 mg/L PPT, indicating the transfer and expression of the gene in F1 transgenic plants.

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