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.
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Fundação para a Ciência e a Tecnologia is acknowledged for funding Project POCTI/AGR/38507/2001.
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Costa, M.S., Miguel, C. & Oliveira, M.M. An improved selection strategy and the use of acetosyringone in shoot induction medium increase almond transformation efficiency by 100-fold . Plant Cell Tiss Organ Cult 85, 205–209 (2006). https://doi.org/10.1007/s11240-005-9073-z
- adventitious regeneration
- Agrobacterium-mediated transformation
- kanamycin resistance
- Prunus dulcis