Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus
The coat protein gene of citrus tristeza virus (CTV) has been introduced into Mexican lime (Citrus aurantifolia Swing.) plants by using an improved Agrobacterium-mediated genetic transformation system. Internodal stem segments from greenhouse-grown seedlings were co-cultivated with A. tumefaciens st...
| Autores principales: | , , , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/5148 |
| _version_ | 1855032233371369472 |
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| author | Domínguez, Alfons Guerri, José Cambra, Mariano Navarro, Luis Moreno, Pedro Pena, Leandro |
| author_browse | Cambra, Mariano Domínguez, Alfons Guerri, José Moreno, Pedro Navarro, Luis Pena, Leandro |
| author_facet | Domínguez, Alfons Guerri, José Cambra, Mariano Navarro, Luis Moreno, Pedro Pena, Leandro |
| author_sort | Domínguez, Alfons |
| collection | ReDivia |
| description | The coat protein gene of citrus tristeza virus (CTV) has been introduced into Mexican lime (Citrus aurantifolia Swing.) plants by using an improved Agrobacterium-mediated genetic transformation system. Internodal stem segments from greenhouse-grown seedlings were co-cultivated with A. tumefaciens strain EHA 105 carrying the binary plasmid pBI 121/CTV-CP in a medium rich in auxins that provided the explant cells with the proper treatment to shift them to a competent state for transformation. The transformation frequency was enhanced, and this allowed us to recover 42 transgenic plants from 1200 explants. Regenerated shoots were identified as transformants by performing beta-glucuronidase (GUS) assays and subsequently by PCR amplifications of the CTV-CP transgene. Southern analyses revealed that at least one copy of the CTV-CP gene was integrated in all PCR positive plants. Interestingly, 70% of them had linked T-DNAs arranged at one locus. Copy number of the CTV-CP gene varied from one to six among the transgenic lines. Half of them showed truncated T-DNAs in which the left border was lost. Expression of the CTV-CP transgene was demonstrated in 38 out of 42 plants by western analysis and DASI-ELISA. No correlation was found between coat protein expression and transgene copy number or integration pattern. |
| format | article |
| id | ReDivia5148 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia51482025-04-25T14:45:30Z Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus Domínguez, Alfons Guerri, José Cambra, Mariano Navarro, Luis Moreno, Pedro Pena, Leandro The coat protein gene of citrus tristeza virus (CTV) has been introduced into Mexican lime (Citrus aurantifolia Swing.) plants by using an improved Agrobacterium-mediated genetic transformation system. Internodal stem segments from greenhouse-grown seedlings were co-cultivated with A. tumefaciens strain EHA 105 carrying the binary plasmid pBI 121/CTV-CP in a medium rich in auxins that provided the explant cells with the proper treatment to shift them to a competent state for transformation. The transformation frequency was enhanced, and this allowed us to recover 42 transgenic plants from 1200 explants. Regenerated shoots were identified as transformants by performing beta-glucuronidase (GUS) assays and subsequently by PCR amplifications of the CTV-CP transgene. Southern analyses revealed that at least one copy of the CTV-CP gene was integrated in all PCR positive plants. Interestingly, 70% of them had linked T-DNAs arranged at one locus. Copy number of the CTV-CP gene varied from one to six among the transgenic lines. Half of them showed truncated T-DNAs in which the left border was lost. Expression of the CTV-CP transgene was demonstrated in 38 out of 42 plants by western analysis and DASI-ELISA. No correlation was found between coat protein expression and transgene copy number or integration pattern. 2017-06-01T10:11:47Z 2017-06-01T10:11:47Z 2000 MAR article Dominguez, A., Guerri, J., Cambra, M., Navarro, L., Moreno, P., Pena, L. (2000). Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus. Plant Cell Reports, 19(4), 427-433. 0721-7714 http://hdl.handle.net/20.500.11939/5148 10.1007/s002990050751 en openAccess Impreso |
| spellingShingle | Domínguez, Alfons Guerri, José Cambra, Mariano Navarro, Luis Moreno, Pedro Pena, Leandro Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus |
| title | Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus |
| title_full | Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus |
| title_fullStr | Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus |
| title_full_unstemmed | Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus |
| title_short | Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus |
| title_sort | efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus |
| url | http://hdl.handle.net/20.500.11939/5148 |
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