Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration
We have previously developed procedures for the efficient production of sweet orange (Citrus sinensis L. Osbeck) and Carrizo citrange (C. sinensis L. OsbeckxPoncirus trifoliata L. Raf.) transgenic plants using an Agrobacterium tumefaciens-mediated transformation and shoot tip grafting in vitro regen...
| Main Authors: | , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
2017
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| Online Access: | http://hdl.handle.net/20.500.11939/4334 |
| _version_ | 1855491734717333504 |
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| author | Pena, Leandro Cervera, Magdalena Juárez, José Navarro, Antonio Pina, José A. Navarro, Luis |
| author_browse | Cervera, Magdalena Juárez, José Navarro, Antonio Navarro, Luis Pena, Leandro Pina, José A. |
| author_facet | Pena, Leandro Cervera, Magdalena Juárez, José Navarro, Antonio Pina, José A. Navarro, Luis |
| author_sort | Pena, Leandro |
| collection | ReDivia |
| description | We have previously developed procedures for the efficient production of sweet orange (Citrus sinensis L. Osbeck) and Carrizo citrange (C. sinensis L. OsbeckxPoncirus trifoliata L. Raf.) transgenic plants using an Agrobacterium tumefaciens-mediated transformation and shoot tip grafting in vitro regeneration system. We now report on the optimization of the cocultivation, regeneration and selection conditions for efficient and reliable production of transgenic lime (C. aurantifolia Swing.) plants. Improved transformation frequencies were obtained by co-cultivating the explants with Agrobacterium on feeder plates. Optimum regeneration of transgenic shoots was obtained by exposing the explants to darkness for 2 weeks and by using kanamycin at 100 mg/l as selective agent. Attempts to use geneticin as selection antibiotic were not successful. Shoot tip grafting of regenerated shoots on Troyer citrange seedlings resulted in 100% successful production of transgenic plants. The presence and expression of the transferred genes in the regenerated plants was verified by beta-glucuronidase histochemical and fluorimetric assays, neomycin phosphotransferase ELISA assays, PCR and Southern analyses. |
| format | Artículo |
| id | ReDivia4334 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia43342025-04-25T14:42:52Z Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration Pena, Leandro Cervera, Magdalena Juárez, José Navarro, Antonio Pina, José A. Navarro, Luis We have previously developed procedures for the efficient production of sweet orange (Citrus sinensis L. Osbeck) and Carrizo citrange (C. sinensis L. OsbeckxPoncirus trifoliata L. Raf.) transgenic plants using an Agrobacterium tumefaciens-mediated transformation and shoot tip grafting in vitro regeneration system. We now report on the optimization of the cocultivation, regeneration and selection conditions for efficient and reliable production of transgenic lime (C. aurantifolia Swing.) plants. Improved transformation frequencies were obtained by co-cultivating the explants with Agrobacterium on feeder plates. Optimum regeneration of transgenic shoots was obtained by exposing the explants to darkness for 2 weeks and by using kanamycin at 100 mg/l as selective agent. Attempts to use geneticin as selection antibiotic were not successful. Shoot tip grafting of regenerated shoots on Troyer citrange seedlings resulted in 100% successful production of transgenic plants. The presence and expression of the transferred genes in the regenerated plants was verified by beta-glucuronidase histochemical and fluorimetric assays, neomycin phosphotransferase ELISA assays, PCR and Southern analyses. 2017-06-01T10:09:52Z 2017-06-01T10:09:52Z 1997 SEP article Pena, L., Cervera, M., Juarez, J., Navarro, A., Pina, JA, Navarro, L. (1997). Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration. Plant Cell Reports, 16(11), 731-737. 0721-7714 http://hdl.handle.net/20.500.11939/4334 10.1007/s002990050311 en openAccess Impreso |
| spellingShingle | Pena, Leandro Cervera, Magdalena Juárez, José Navarro, Antonio Pina, José A. Navarro, Luis Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration |
| title | Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration |
| title_full | Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration |
| title_fullStr | Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration |
| title_full_unstemmed | Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration |
| title_short | Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration |
| title_sort | genetic transformation of lime citrus aurantifolia swing factors affecting transformation and regeneration |
| url | http://hdl.handle.net/20.500.11939/4334 |
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