Efficient Agrobacterium tumefaciens-mediated transformation of embryogenic calli and regeneration of Hevea brasiliensis Müll Arg. plants

Blanc Géraldine, Baptiste Christelle, Oliver Gérald, Martin Florence, Montoro Pascal. 2006. Efficient Agrobacterium tumefaciens-mediated transformation of embryogenic calli and regeneration of Hevea brasiliensis Müll Arg. plants. Plant Cell Reports, 24 (12) : 724-733.

Résumé : An efficient procedure for producing transgenic Hevea brasiliensis callus and plant lines from clone PB 260 was established with Agrobacterium tumefaciens using strain EHA 105 harbouring the vector pCAMB1A230I . Transformation capacity and competence of the embryo-genic calli were improved after two cycles of cryopreservation. When the cocultivation temperature was reduced from 27 to 20°C, the duration of this phase could be increased up to 7 days, promoting an increase in GUS activity. These transformation conditions led to the isolation of 24 callus lines resistant to paromomycin, which is used as a selection agent. Nineteen of these lines revealed the existence of one to four copies of T-DNA by Southern-blot analysis. Nine of them were transferred for regeneration by somatic embryo-genesis. Three hundred seventy-four transgenic plants have thus been generated from six independent lines bearing 1, 2 or 3 copies of T-DNA. The efficiency and reproducibility of this method means that functional characterization of genes involved in natural rubber production can be envisaged.

Mots-clés Agrovoc : Hevea brasiliensis, Agrobacterium tumefaciens, transformation génétique, Embryogénèse somatique, antibiotique, plante transgénique, séquence nucléotidique, adn, génie génétique, cryoconservation

Mots-clés complémentaires : Bêta-glucuronidase, GUS

Classification Agris : F30 - Génétique et amélioration des plantes

Champ stratégique Cirad : Axe 1 (2005-2013) - Intensification écologique

Auteurs et affiliations

• Blanc Géraldine
• Baptiste Christelle
• Oliver Gérald, CIRAD-CP-UMR BEPC (FRA)
• Martin Florence, CIRAD-CP-UMR BEPC (FRA)
• Montoro Pascal, INRA (FRA)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/530928/)

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