Abstract
Phosphinothricin (PPT) inhibits glutamine synthetase in plant cells, resulting in an accumulation of ammonia that can be toxic. Bar, from Streptomyces hygroscopicus is a gene that codes for phosphinothricin acetyl-transferase, an enzyme that detoxifies PPT by acetylation. We used polyethylene-glycol-mediated transformation to introduce bar into protoplasts of maize (Zea mays L.). With a novel assay utilizing the pH indicator chlorophenol red we identified transformants after only two to four weeks of selection on PPT.
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Abbreviations
- BT:
-
Bacillus thuringiensis
- CR assay:
-
chloro-phenol-red assay
- EDTA:
-
(ethylenedinitro)tetraacetic acid disodium salt
- PAT:
-
phosphinothricin acetyltransferase
- PCR:
-
polymerase chain reaction
- PEG:
-
polyethylene glycol
- PPT:
-
phosphinothricin
- TRIS:
-
(hydroxymethyl)aminomethane
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The authors thank G. Pace for critical reading of the manuscript and Jeanette Tyler for technical assistance.
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Kramer, C., DiMaio, J., Carswell, G.K. et al. Selection of transformed protoplast-derived Zea mays colonies with phosphinothricin and a novel assay using the pH indicator chlorophenol red. Planta 190, 454–458 (1993). https://doi.org/10.1007/BF00224783
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DOI: https://doi.org/10.1007/BF00224783