TY  - BOOK
AU  - Blanc Geraldine
AU  - Baptiste Christelle
AU  - Martin Florence
AU  - Montoro Pascal
AU  - Oliver Gerald
TI  - Efficient Agrobacterium tumefaciens-mediated transformation of embryogenic calli and regeneration of Hevea brasiliensis Mull Arg. plants
PY  - 2006///
PB  - Plant Cell Reports
KW  - Agrobacterium tumefaciens
KW  - Cocultivation
KW  - Cryopreservation
KW  - Genetic transformation
KW  - GUS
KW  - Temperature
N2  - An efficient procedure for producing transgenic Hevea brasiliensis callus and plants lines from clone PB 260 was established with Agrobacterium tumefaciens using strain EHA 105 harbouring the vector pCAMBIA 2301. Transformation capacity and competence of the embryogenic calli were improved after two cycles of cryopreservation. When the cocultivation temperature was reduced from 27 to 20 oC, the duration of this phase could be increased uo 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 embryogenesis. 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
ER  -