Abstract
In a study to optimize the use of the bar gene as a selectable marker gene in plant transformation, the selection mechanism of phosphinothricin in vitro was studied. It was found that tissues with a high anabolic activity were more sensitive to the toxicity of ammonium than tissues with a low anabolic activity. However, tissues with a low anabolic and high catabolic activity were more sensitive to glutamine deprivation. The metabolic (anabolic and catabolic) activity of callusing hypocotyl explants of Brassica napus L. was influenced by the carbon source of the medium and the light condition. The expression of the bar gene regulated by the light-inducible promoter derived from the gene coding for the small subunit of ribulose-1,5-bisphosphate carboxylase-oxygenase was also influenced by both light condition and carbon source. Finally, a bioassay to screen for, and to quantify the resistance of expiants to, phosphinothricin is presented.
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Abbreviations
- PAT:
-
phosphinothricin acetyl transferase
- PPT:
-
phosphinothricin
- p ssu :
-
promoter derived from the Arabidopsis gene coding for the small subunit of Rubisco
- Rubisco:
-
ribulose-1,5-biphosphate carboxylase-oxygenase
- TTC:
-
oxidized form of 2,3,5-triphenyltetrazoliumchloride
- TTC-H:
-
reduced form of 2,3,5-triphenyltetrazoliumchloride
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We thank Dr. M. Williams (Plant Genetic Systems, Gent, Belgium) for the critical reading of the manuscript.
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De Block, M., De Sonville, A. & Debrouwer, D. The selection mechanism of phosphinothricin is influenced by the metabolic status of the tissue. Planta 197, 619–626 (1995). https://doi.org/10.1007/BF00191569
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DOI: https://doi.org/10.1007/BF00191569