Factors involved in Agrobacterium tumefaciens-mediated gene transfer into Pinus nigra Arn. ssp. salzmannii (Dunal) Franco |
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Authors: | Marián López Jaime M. Humara Roberto Rodríguez Ricardo J. Ordás |
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Affiliation: | (1) Lab. Fisioloía Vegetal, Dept. Biología de Organismos y Sistemas, Ftad. Biología, C/Catedrático Rodrígo Uría s/n, Univ. Oviedo, E-33071 Oviedo Asturias, Spain;(2) Instituto Universitario de Biotecnología de Asturias-CNB (CSIC), E-33071 Oviedo Asturias, Spain |
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Abstract: | Cotyledons from dissected sterile embryos of salgareño pine (Pinus nigra Arn. ssp. salzmannii (Dunal) Franco) were inoculated with different disarmed Agrobacterium tumefaciens strains harbouring the binary vector p35SGUSint. The transient expression of a β-glucuronidase gene (uidA) was studied, using a histochemical staining procedure. Nineteen days after inoculation, the activity of β-glucuronidase was detected in epidermal and subepidermal layers of cotyledonary explants. The EHA105 strain harbouring a disarmed agropine-type Ti-plasmid (pTiBO542) was the most effective for gene transfer of the uidA gene. The effects of exudates and extracts from 0-day-old embryos on induction of vir gene expression in A. tumefaciens were also examined. The results of this study showed that salgarño pine embryo exudates contain a substance(s) that induce vir gene expression, in similar way to that observed with 100 μM acetosyringone (AS).All these findings suggest that T-DNA processing and transfer might take place when Agrobacterium infects suitable tissues of salgareño pine. |
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Keywords: | conifers salgare?o pine tissue culture transformation transient gene expression uidA expression vir gene |
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