Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants
The green fluorescent protein (GFP) from Aequorea victoria has been introduced into three different citrus genotypes [Citrus aurantium L., C. aurantifolia (Christm.) Swing. and C. sinensis L. Osbeck x Poncirus trifoliata (L.) Raf.] which are considered recalcitrant to transformation, mainly due to l...
| Autores principales: | , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
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2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/5274 |
| _version_ | 1855032252280340480 |
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| author | Ghorbel, R. Juárez, José Navarro, Luis Pena, Leandro |
| author_browse | Ghorbel, R. Juárez, José Navarro, Luis Pena, Leandro |
| author_facet | Ghorbel, R. Juárez, José Navarro, Luis Pena, Leandro |
| author_sort | Ghorbel, R. |
| collection | ReDivia |
| description | The green fluorescent protein (GFP) from Aequorea victoria has been introduced into three different citrus genotypes [Citrus aurantium L., C. aurantifolia (Christm.) Swing. and C. sinensis L. Osbeck x Poncirus trifoliata (L.) Raf.] which are considered recalcitrant to transformation, mainly due to low transformation frequencies and to the regeneration of escape shoots at high frequencies from the Agrobacterium-inoculated explants. High-level GFP expression was detected in transgenic cells, tissues and plants. Using GFP as a vital marker has allowed us to localize the sites of transgene expression in specific cells, always occurring in callus tissue formed from the cambium of the cut ends of explants. Whereas green fluorescent shoots regenerated in all cases from this callus, most escapes regenerated directly from explants with almost no callus formation. Thus, development of callus from cambium is a prerequisite for citrus transformation. Furthermore, in vivo monitoring of GFP expression permitted a rapid and easy discrimination of transgenic and escape shoots. The selection of transgenic shoots could be easily favored by eliminating the escapes and/or by performing shoot-tip grafting of the transgenic buds soon after their origin. GFP-expressing shoots have also been observed in citrus explants cocultivated with Agrobacterium but cultured in a medium without the selective agent kanamycin. This opens the possibility to rescue the transgenic sectors and to regenerate transgenic plants without using selectable marker genes conferring antibiotic or herbicide resistance, which is currently a topic of much discussion for the commercialization of transgenic plants. |
| format | article |
| id | ReDivia5274 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia52742025-04-25T14:45:49Z Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants Ghorbel, R. Juárez, José Navarro, Luis Pena, Leandro The green fluorescent protein (GFP) from Aequorea victoria has been introduced into three different citrus genotypes [Citrus aurantium L., C. aurantifolia (Christm.) Swing. and C. sinensis L. Osbeck x Poncirus trifoliata (L.) Raf.] which are considered recalcitrant to transformation, mainly due to low transformation frequencies and to the regeneration of escape shoots at high frequencies from the Agrobacterium-inoculated explants. High-level GFP expression was detected in transgenic cells, tissues and plants. Using GFP as a vital marker has allowed us to localize the sites of transgene expression in specific cells, always occurring in callus tissue formed from the cambium of the cut ends of explants. Whereas green fluorescent shoots regenerated in all cases from this callus, most escapes regenerated directly from explants with almost no callus formation. Thus, development of callus from cambium is a prerequisite for citrus transformation. Furthermore, in vivo monitoring of GFP expression permitted a rapid and easy discrimination of transgenic and escape shoots. The selection of transgenic shoots could be easily favored by eliminating the escapes and/or by performing shoot-tip grafting of the transgenic buds soon after their origin. GFP-expressing shoots have also been observed in citrus explants cocultivated with Agrobacterium but cultured in a medium without the selective agent kanamycin. This opens the possibility to rescue the transgenic sectors and to regenerate transgenic plants without using selectable marker genes conferring antibiotic or herbicide resistance, which is currently a topic of much discussion for the commercialization of transgenic plants. 2017-06-01T10:12:02Z 2017-06-01T10:12:02Z 1999 JUL article Ghorbel, R., Juarez, J., Navarro, L., Pena, L. (1999). Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants. Theoretical and Applied Genetics, 99(1-2), 350-358. 0040-5752 http://hdl.handle.net/20.500.11939/5274 10.1007/s001220051244 en openAccess Impreso |
| spellingShingle | Ghorbel, R. Juárez, José Navarro, Luis Pena, Leandro Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants |
| title | Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants |
| title_full | Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants |
| title_fullStr | Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants |
| title_full_unstemmed | Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants |
| title_short | Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants |
| title_sort | green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants |
| url | http://hdl.handle.net/20.500.11939/5274 |
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