000			01804nam a2200241Ia 4500
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100			_aBlanc Geraldine
245		0	_aEfficient Agrobacterium tumefaciens-mediated transformation of embryogenic calli and regeneration of Hevea brasiliensis Mull Arg. plants
260			_bPlant Cell Reports _c2006
300			_a724-733
520			_aAn 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.
650			_aAgrobacterium tumefaciens
650			_aCocultivation
650			_aCryopreservation
650			_aGenetic transformation
650			_aGUS
650			_aTemperature
700			_aBaptiste Christelle
700			_aMartin Florence
700			_aMontoro Pascal
700			_aOliver Gerald
942			_cJS
999			_c58424 _d58424