Early events in Agrobacterium-mediated genetic transformation of citrus explants
Background and Aims Genetic transformation of plants relies on two independent but concurrent processes: integration of foreign DNA into plant cells and regeneration of whole plants from these transformed cells. Cell competence for regeneration and for transformation does not always fall into the sa...
| 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/20.500.11939/4338 |
| _version_ | 1855032103769473024 |
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| author | Pena, Leandro Perez, Rosa Cervera, Magdalena Juárez, José Navarro, Luis |
| author_browse | Cervera, Magdalena Juárez, José Navarro, Luis Pena, Leandro Perez, Rosa |
| author_facet | Pena, Leandro Perez, Rosa Cervera, Magdalena Juárez, José Navarro, Luis |
| author_sort | Pena, Leandro |
| collection | ReDivia |
| description | Background and Aims Genetic transformation of plants relies on two independent but concurrent processes: integration of foreign DNA into plant cells and regeneration of whole plants from these transformed cells. Cell competence for regeneration and for transformation does not always fall into the same cell type/developmental stage, and this is one of the main causes of the so-called recalcitrance for transformation of certain plant species. In this study, a detailed examination of the first steps of morphogenesis from citrus explants after co-cultivation with Agrobacterium tumefaciens was performed, and an investigation into which cells and tissues are competent for regeneration and transformation was carried out. Moreover, the role of phytohormones in the co-cultivation medium as possible enhancers of gene transfer was also studied. Methods A highly responsive citrus genotype and well-established culture conditions were used to perform a histological analysis of morphogenesis and cell competence for transformation after co-cultivation of citrus epicotyl segments with A. tumefaciens. In addition, the role of phytohormones as transformation enhancers was investigated by flow cytometry. Key Results It is demonstrated that cells competent for transformation are located in the newly formed callus growing from the cambial ring. Conditions conducive to further development of this callus, such as treatment of explants in a medium rich in auxins, resulted in a more pronounced formation of cambial callus and a slower shoot regeneration process, both in Agrobacteritum-inoculated and non-inoculated explants. Furthermore, co-cultivation in a medium rich in auxins caused a significant increase in the rate of actively dividing cells in S-phase, the stage in which cells are more prone to integrate foreign DNA. Conclusions Use of proper co-cultivation medium and conditions led to a higher number of stably transformed cells and to an increase in the final number of regenerated transgenic plants. |
| format | article |
| id | ReDivia4338 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia43382025-04-25T14:42:53Z Early events in Agrobacterium-mediated genetic transformation of citrus explants Pena, Leandro Perez, Rosa Cervera, Magdalena Juárez, José Navarro, Luis Background and Aims Genetic transformation of plants relies on two independent but concurrent processes: integration of foreign DNA into plant cells and regeneration of whole plants from these transformed cells. Cell competence for regeneration and for transformation does not always fall into the same cell type/developmental stage, and this is one of the main causes of the so-called recalcitrance for transformation of certain plant species. In this study, a detailed examination of the first steps of morphogenesis from citrus explants after co-cultivation with Agrobacterium tumefaciens was performed, and an investigation into which cells and tissues are competent for regeneration and transformation was carried out. Moreover, the role of phytohormones in the co-cultivation medium as possible enhancers of gene transfer was also studied. Methods A highly responsive citrus genotype and well-established culture conditions were used to perform a histological analysis of morphogenesis and cell competence for transformation after co-cultivation of citrus epicotyl segments with A. tumefaciens. In addition, the role of phytohormones as transformation enhancers was investigated by flow cytometry. Key Results It is demonstrated that cells competent for transformation are located in the newly formed callus growing from the cambial ring. Conditions conducive to further development of this callus, such as treatment of explants in a medium rich in auxins, resulted in a more pronounced formation of cambial callus and a slower shoot regeneration process, both in Agrobacteritum-inoculated and non-inoculated explants. Furthermore, co-cultivation in a medium rich in auxins caused a significant increase in the rate of actively dividing cells in S-phase, the stage in which cells are more prone to integrate foreign DNA. Conclusions Use of proper co-cultivation medium and conditions led to a higher number of stably transformed cells and to an increase in the final number of regenerated transgenic plants. 2017-06-01T10:09:52Z 2017-06-01T10:09:52Z 2004 JUL article Pena, L., Perez, R., Cervera, M., Juarez, J. & Navarro, L. (2004). Early events in Agrobacterium-mediated genetic transformation of citrus explants. Annals of Botany, 94(1), 67-74. 0305-7364 http://hdl.handle.net/20.500.11939/4338 10.1093/aob/mch117 en openAccess Impreso |
| spellingShingle | Pena, Leandro Perez, Rosa Cervera, Magdalena Juárez, José Navarro, Luis Early events in Agrobacterium-mediated genetic transformation of citrus explants |
| title | Early events in Agrobacterium-mediated genetic transformation of citrus explants |
| title_full | Early events in Agrobacterium-mediated genetic transformation of citrus explants |
| title_fullStr | Early events in Agrobacterium-mediated genetic transformation of citrus explants |
| title_full_unstemmed | Early events in Agrobacterium-mediated genetic transformation of citrus explants |
| title_short | Early events in Agrobacterium-mediated genetic transformation of citrus explants |
| title_sort | early events in agrobacterium mediated genetic transformation of citrus explants |
| url | http://hdl.handle.net/20.500.11939/4338 |
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