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Genetic transformation of Vitis vinifera via organogenesis

BACKGROUND: Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. The current methods for the production of transgenic grape plants are based on Agrobacterium-mediated transformation followed...

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Autores principales: Mezzetti, Bruno, Pandolfini, Tiziana, Navacchi, Oriano, Landi, Lucia
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2002
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC130035/
https://www.ncbi.nlm.nih.gov/pubmed/12354328
http://dx.doi.org/10.1186/1472-6750-2-18
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author Mezzetti, Bruno
Pandolfini, Tiziana
Navacchi, Oriano
Landi, Lucia
author_facet Mezzetti, Bruno
Pandolfini, Tiziana
Navacchi, Oriano
Landi, Lucia
author_sort Mezzetti, Bruno
collection PubMed
description BACKGROUND: Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. The current methods for the production of transgenic grape plants are based on Agrobacterium-mediated transformation followed by regeneration from embryogenic callus. However, grape embryogenic calli are laborious to establish and the phenotype of the regenerated plants can be altered. RESULTS: Transgenic grape plants (V. vinifera, table-grape cultivars Silcora and Thompson Seedless) were produced using a method based on regeneration via organogenesis. In vitro proliferating shoots were cultured in the presence of increasing concentrations of N(6)-benzyl adenine. The apical dome of the shoot was removed at each transplantation which, after three months, produced meristematic bulk tissue characterized by a strong capacity to differentiate adventitious shoots. Slices prepared from the meristematic bulk were used for Agrobacterium-mediated transformation of grape plants with the gene DefH9-iaaM. After rooting on kanamycin containing media and greenhouse acclimatization, transgenic plants were transferred to the field. At the end of the first year of field cultivation, DefH9-iaaM grape plants were phenotypically homogeneous and did not show any morphological alterations in vegetative growth. The expression of DefH9-iaaM gene was detected in transgenic flower buds of both cultivars. CONCLUSIONS: The phenotypic homogeneity of the regenerated plants highlights the validity of this method for both propagation and genetic transformation of table grape cultivars. Expression of the DefH9-iaaM gene takes place in young flower buds of transgenic plants from both grape cultivars.
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spelling pubmed-1300352002-10-25 Genetic transformation of Vitis vinifera via organogenesis Mezzetti, Bruno Pandolfini, Tiziana Navacchi, Oriano Landi, Lucia BMC Biotechnol Research Article BACKGROUND: Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. The current methods for the production of transgenic grape plants are based on Agrobacterium-mediated transformation followed by regeneration from embryogenic callus. However, grape embryogenic calli are laborious to establish and the phenotype of the regenerated plants can be altered. RESULTS: Transgenic grape plants (V. vinifera, table-grape cultivars Silcora and Thompson Seedless) were produced using a method based on regeneration via organogenesis. In vitro proliferating shoots were cultured in the presence of increasing concentrations of N(6)-benzyl adenine. The apical dome of the shoot was removed at each transplantation which, after three months, produced meristematic bulk tissue characterized by a strong capacity to differentiate adventitious shoots. Slices prepared from the meristematic bulk were used for Agrobacterium-mediated transformation of grape plants with the gene DefH9-iaaM. After rooting on kanamycin containing media and greenhouse acclimatization, transgenic plants were transferred to the field. At the end of the first year of field cultivation, DefH9-iaaM grape plants were phenotypically homogeneous and did not show any morphological alterations in vegetative growth. The expression of DefH9-iaaM gene was detected in transgenic flower buds of both cultivars. CONCLUSIONS: The phenotypic homogeneity of the regenerated plants highlights the validity of this method for both propagation and genetic transformation of table grape cultivars. Expression of the DefH9-iaaM gene takes place in young flower buds of transgenic plants from both grape cultivars. BioMed Central 2002-09-27 /pmc/articles/PMC130035/ /pubmed/12354328 http://dx.doi.org/10.1186/1472-6750-2-18 Text en Copyright © 2002 Mezzetti et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Research Article
Mezzetti, Bruno
Pandolfini, Tiziana
Navacchi, Oriano
Landi, Lucia
Genetic transformation of Vitis vinifera via organogenesis
title Genetic transformation of Vitis vinifera via organogenesis
title_full Genetic transformation of Vitis vinifera via organogenesis
title_fullStr Genetic transformation of Vitis vinifera via organogenesis
title_full_unstemmed Genetic transformation of Vitis vinifera via organogenesis
title_short Genetic transformation of Vitis vinifera via organogenesis
title_sort genetic transformation of vitis vinifera via organogenesis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC130035/
https://www.ncbi.nlm.nih.gov/pubmed/12354328
http://dx.doi.org/10.1186/1472-6750-2-18
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