Résumé
With the successful plant of Botany 77(8): 1168-1177) an Agrobacterium tumefaciensmediated genetic transformation has been developed (Blanc et al., 2006, PCR 24(12) 724733) and has led to genetically transformed plant expressing the gusA reporter gene driven either by the CAMV 35S or the Hev2. 1 promoters. A way to improve this procedure was to use a non-destructive visual reporter gene, the Green Fluorescent Protein (GFP). Then, a binary vector has been constructed containing the two reporter genes driven by the CAMV 35S promoter. The first results showed that the green fluorescence was strongly visible in H. brasiliensis calli and that earlier selection of transgenic calli was thus possible from the first sub-culture. Comparison of the GUS and GFP activities revealed that the two markers can be used to evaluate transformation efficiency. Using GFP visual marker, transgenic fluorescent calli were selected earlier than transgenic calli using the GUS staining. GFP visual marker consequently can be used as a selection marker, which could avoid the use of gene conferring antibiotic resistance and shorten drastically the duration of the establishment of the transgenic callus lines. This transformation system is being used for over-expressing genes of interest such those involved in stress tolerance. (Texte intégral) (Résumé d'auteur)