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

Blanc G., Baptiste C., Oliver G., Martin F., Montoro P.. 2006. Plant Cell Reports, 24 (12) : p. 724-733.

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. (Résumé d'auteur)

Mots-clés : cryoconservation; génie génétique; adn; séquence nucléotidique; plante transgénique; antibiotique; embryogénèse somatique; transformation génétique; agrobacterium tumefaciens; hevea brasiliensis; gus; bêta-glucuronidase

Thématique : Génétique et amélioration des plantes

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