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An improved selection strategy and the use of acetosyringone in shoot induction medium increase almond transformation efficiency by 100-fold
Authors:Milene S. Costa  Célia Miguel  M. Margarida Oliveira
Affiliation:(1) Instituto de Tecnologia Química e Biológica/Instituto de Biologia Experimental e Tecnológica (ITQB/IBET), Quinta do Marquês, 2784-505 Oeiras, Portugal;(2) Dep. Biologia Vegetal, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal
Abstract:
An improved almond transformation system was developed with a 100× increase in efficiency (12.3%) as compared to the existing transformation method (0.1%) in spite of the lower regeneration ability of the explants used. Leaf transformation was performed with Agrobacterium EHA105/p35 SGUSINT. Main modifications introduced in the transformation protocol were the use of 150 μM acetosyringone (AS) during the 21-days induction period, and a different selection strategy. Transformed shoots were assayed using PCR, GUS analyses and Southern blotting. The improved methodology is being applied for transformation of in vitro propagated cultivars and opens the possibility of using almond as model for functional studies in Prunoideae.
Keywords:adventitious regeneration   Agrobacterium-mediated transformation  kanamycin resistance   Prunus dulcis   Rosaceae
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