Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration
Genetic transformation of mature trees is difficult because adult tissues are recalcitrant to Agrobacterium tumefaciens infection and transformation and because transgenic mature events are less competent for regeneration. We have shown that reinvigoration allows manipulation of the vegetative phase...
| Main Authors: | , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
2017
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| Online Access: | http://hdl.handle.net/20.500.11939/5017 |
| _version_ | 1855491852619218944 |
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| author | Cervera, Magdalena Navarro, Antonio Navarro, Luis Pena, Leandro |
| author_browse | Cervera, Magdalena Navarro, Antonio Navarro, Luis Pena, Leandro |
| author_facet | Cervera, Magdalena Navarro, Antonio Navarro, Luis Pena, Leandro |
| author_sort | Cervera, Magdalena |
| collection | ReDivia |
| description | Genetic transformation of mature trees is difficult because adult tissues are recalcitrant to Agrobacterium tumefaciens infection and transformation and because transgenic mature events are less competent for regeneration. We have shown that reinvigoration allows manipulation of the vegetative phase to increase the potential for transformation and regeneration without loss of competence for flowering and fruiting. To produce transgenic plants from clementine mandarin (Citrus clementina hort. ex Tanaka), we optimized the conditions of the source material both ex vitro and in vitro. Grafting of mature buds on juvenile rootstocks in the spring and preventing multiple bud sprouting by removing all but one bud permitted selection of vigorous first flushes for in vitro culture. Use of additional virulence genes from A. tumefaciens to increase transformation frequency and optimization of culture media and conditions to enhance explant cell competence for T-DNA integration and organogenesis resulted in efficient and reliable transgenic plant production. Transformed regenerants from explants, cultured in media without antibiotics, were identified by a screenable marker (either beta-glucuronidase or green fluorescent protein (GFP)), creating the possibility of generating transgenic clementine plants without antibiotic resistance marker genes. Stable integration of foreign genes was demonstrated by Southern blot analysis, and expression of these foreign genes was confirmed by detection of GFP fluorescence in leaves, floral organs and fruits of the transgenic plants. |
| format | Artículo |
| id | ReDivia5017 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia50172025-04-25T14:45:10Z Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration Cervera, Magdalena Navarro, Antonio Navarro, Luis Pena, Leandro Genetic transformation of mature trees is difficult because adult tissues are recalcitrant to Agrobacterium tumefaciens infection and transformation and because transgenic mature events are less competent for regeneration. We have shown that reinvigoration allows manipulation of the vegetative phase to increase the potential for transformation and regeneration without loss of competence for flowering and fruiting. To produce transgenic plants from clementine mandarin (Citrus clementina hort. ex Tanaka), we optimized the conditions of the source material both ex vitro and in vitro. Grafting of mature buds on juvenile rootstocks in the spring and preventing multiple bud sprouting by removing all but one bud permitted selection of vigorous first flushes for in vitro culture. Use of additional virulence genes from A. tumefaciens to increase transformation frequency and optimization of culture media and conditions to enhance explant cell competence for T-DNA integration and organogenesis resulted in efficient and reliable transgenic plant production. Transformed regenerants from explants, cultured in media without antibiotics, were identified by a screenable marker (either beta-glucuronidase or green fluorescent protein (GFP)), creating the possibility of generating transgenic clementine plants without antibiotic resistance marker genes. Stable integration of foreign genes was demonstrated by Southern blot analysis, and expression of these foreign genes was confirmed by detection of GFP fluorescence in leaves, floral organs and fruits of the transgenic plants. 2017-06-01T10:11:33Z 2017-06-01T10:11:33Z 2008 JAN article Cervera, M., Navarro, A., Navarro, L., Pena, L. (2008). Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration. Tree physiology, 28(1), 55-66. 0829-318X http://hdl.handle.net/20.500.11939/5017 10.1093/treephys/28.1.55 en openAccess Impreso |
| spellingShingle | Cervera, Magdalena Navarro, Antonio Navarro, Luis Pena, Leandro Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration |
| title | Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration |
| title_full | Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration |
| title_fullStr | Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration |
| title_full_unstemmed | Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration |
| title_short | Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration |
| title_sort | production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration |
| url | http://hdl.handle.net/20.500.11939/5017 |
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