Targeted biolistics for improved transformation of impatiens balsamina
A transgenesis programme has been developed for Impatiens balsamina that will allow elucidation of the roles played by individual genes in the flower reversion phenomenon shown by this model species. The lack of explants exhibiting adventitious shooting in I. balsamina hinders Agrobacterium-based tr...
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my.utm.360382017-02-16T00:29:17Z http://eprints.utm.my/id/eprint/36038/ Targeted biolistics for improved transformation of impatiens balsamina Wetten, Andy C. Thomas, Jean Luc Wagiran, Alina Chiurugwi, Tinashe R Medicine (General) A transgenesis programme has been developed for Impatiens balsamina that will allow elucidation of the roles played by individual genes in the flower reversion phenomenon shown by this model species. The lack of explants exhibiting adventitious shooting in I. balsamina hinders Agrobacterium-based transformation, but the multiple shoots that arise from cotyledonary nodes present a suitable target for biolistics. These tissues can be disrupted by the helium blast effect associated with conventional biolistic devices, so we have utilised modifications to the PDS 1000/He equipment originally developed for transformation of fragile insect tissues. By loading microcarriers on to a rigid, rather than flexible, macrocarrier, the blast effect is largely eliminated, and the use of a focussing nozzle allows the bombardment to be concentrated on the target tissues. This approach reduces waste of plasmid DNA and gold microcarriers and achieves transfection at lower, less disruptive helium pressures than would otherwise be necessary to efficiently penetrate below the shoot epidermis and generate heritable transgenic lines. Springer 2012 Article PeerReviewed Wetten, Andy C. and Thomas, Jean Luc and Wagiran, Alina and Chiurugwi, Tinashe (2012) Targeted biolistics for improved transformation of impatiens balsamina. Methods in Molecular Biology, 847 . pp. 255-265. ISSN 1064-3745 http://dx.doi.org/10.1007/978-1-61779-558-9_22 DOI:10.1007/978-1-61779-558-9_22 |
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R Medicine (General) Wetten, Andy C. Thomas, Jean Luc Wagiran, Alina Chiurugwi, Tinashe Targeted biolistics for improved transformation of impatiens balsamina |
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A transgenesis programme has been developed for Impatiens balsamina that will allow elucidation of the roles played by individual genes in the flower reversion phenomenon shown by this model species. The lack of explants exhibiting adventitious shooting in I. balsamina hinders Agrobacterium-based transformation, but the multiple shoots that arise from cotyledonary nodes present a suitable target for biolistics. These tissues can be disrupted by the helium blast effect associated with conventional biolistic devices, so we have utilised modifications to the PDS 1000/He equipment originally developed for transformation of fragile insect tissues. By loading microcarriers on to a rigid, rather than flexible, macrocarrier, the blast effect is largely eliminated, and the use of a focussing nozzle allows the bombardment to be concentrated on the target tissues. This approach reduces waste of plasmid DNA and gold microcarriers and achieves transfection at lower, less disruptive helium pressures than would otherwise be necessary to efficiently penetrate below the shoot epidermis and generate heritable transgenic lines. |
| format |
Article |
| author |
Wetten, Andy C. Thomas, Jean Luc Wagiran, Alina Chiurugwi, Tinashe |
| author_facet |
Wetten, Andy C. Thomas, Jean Luc Wagiran, Alina Chiurugwi, Tinashe |
| author_sort |
Wetten, Andy C. |
| title |
Targeted biolistics for improved transformation of impatiens balsamina |
| title_short |
Targeted biolistics for improved transformation of impatiens balsamina |
| title_full |
Targeted biolistics for improved transformation of impatiens balsamina |
| title_fullStr |
Targeted biolistics for improved transformation of impatiens balsamina |
| title_full_unstemmed |
Targeted biolistics for improved transformation of impatiens balsamina |
| title_sort |
targeted biolistics for improved transformation of impatiens balsamina |
| publisher |
Springer |
| publishDate |
2012 |
| url |
http://eprints.utm.my/id/eprint/36038/ http://dx.doi.org/10.1007/978-1-61779-558-9_22 |
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1643649885798400000 |
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