Efficient Agrobacterium tumefaciens-mediated transformation of embryogenic calli and regeneration of Hevea brasiliensis Müll Arg. plants
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.
| Main Authors: | , , , , |
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| Format: | article biblioteca |
| Language: | eng |
| Subjects: | F30 - Génétique et amélioration des plantes, Hevea brasiliensis, Agrobacterium tumefaciens, transformation génétique, Embryogénèse somatique, antibiotique, plante transgénique, séquence nucléotidique, adn, génie génétique, cryoconservation, http://aims.fao.org/aos/agrovoc/c_3589, http://aims.fao.org/aos/agrovoc/c_27444, http://aims.fao.org/aos/agrovoc/c_3220, http://aims.fao.org/aos/agrovoc/c_36911, http://aims.fao.org/aos/agrovoc/c_492, http://aims.fao.org/aos/agrovoc/c_27619, http://aims.fao.org/aos/agrovoc/c_27583, http://aims.fao.org/aos/agrovoc/c_2347, http://aims.fao.org/aos/agrovoc/c_15974, http://aims.fao.org/aos/agrovoc/c_24155, |
| Online Access: | http://agritrop.cirad.fr/530928/ http://agritrop.cirad.fr/530928/1/document_530928.pdf |
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