Articles
Transformation of Nicotiana tabacum using the herbicide resistance bar gene as a selectable marker
Article number
1297_60
Pages
455 – 462
Language
English
Abstract
Transgenic plants of Nicotiana tabacum were regenerated by leaf disc transformation using a bar gene as a selectable marker.
The bar encodes for phosphinothricin acetyl transferase that detoxifies Phosphinothricin (PPT), the active ingredient of herbicides such as BASTA. A transformation procedure with the bar gene as a selectable marker established via Agrobacterium-mediated transformation was reported in this study.
Transgenic plants of N. tabacum were regenerated by leaf disc transformation using a bar gene as a selectable marker.
A primary experiment with different concentrations of PPT as the active ingredient of BASTA was done and 0.1 mM PPT was found as the best concentration of selective medium.
Transgenic plantlets were selected in medium supplemented with PPT (up to 0.1 mM). Transgenic N. tabacum plants resistant to PPT were positive upon PCR by bar gene-specific primers.
All regenerated plants were fertile and without any somaclonal variations.
The transgenic plants transferred to a greenhouse proved to be resistant to 12 mM PPT by spraying BASTA solution.
The leaves of To plants and that of the progeny having the bar gene were resistant to the application of BASTA. Thus, the bar gene has proven to be a useful selectable and screenable marker for the transformation of tobacco plants in this study.
The lack of somaclonal variation, ease of PPT application and low cost of PPT makes this protocol an attractive alternative for regeneration of transgenic N. tabacum. The production of PPT herbicide-resistant N. tabacum plants may have significant commercial applications in crop production.
The bar encodes for phosphinothricin acetyl transferase that detoxifies Phosphinothricin (PPT), the active ingredient of herbicides such as BASTA. A transformation procedure with the bar gene as a selectable marker established via Agrobacterium-mediated transformation was reported in this study.
Transgenic plants of N. tabacum were regenerated by leaf disc transformation using a bar gene as a selectable marker.
A primary experiment with different concentrations of PPT as the active ingredient of BASTA was done and 0.1 mM PPT was found as the best concentration of selective medium.
Transgenic plantlets were selected in medium supplemented with PPT (up to 0.1 mM). Transgenic N. tabacum plants resistant to PPT were positive upon PCR by bar gene-specific primers.
All regenerated plants were fertile and without any somaclonal variations.
The transgenic plants transferred to a greenhouse proved to be resistant to 12 mM PPT by spraying BASTA solution.
The leaves of To plants and that of the progeny having the bar gene were resistant to the application of BASTA. Thus, the bar gene has proven to be a useful selectable and screenable marker for the transformation of tobacco plants in this study.
The lack of somaclonal variation, ease of PPT application and low cost of PPT makes this protocol an attractive alternative for regeneration of transgenic N. tabacum. The production of PPT herbicide-resistant N. tabacum plants may have significant commercial applications in crop production.
Authors
M. Jamshidnia, S.K. Kazemitabar, C. Lindermayr, H. Najafi Zarini
Keywords
Nicotiana tabacum, transformation, bar gene, somaclonal variation, phosphinothricin acetyl transferase, selectable marker
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