An improved selection strategy and the use of acetosyringone in shoot induction medium increase almond transformation efficiency by 100-fold

被引:21
作者
Costa, MS
Miguel, C
Oliveira, MM [1 ]
机构
[1] IBET, ITQB, P-2784505 Oeiras, Portugal
[2] Univ Lisbon, Fac Ciencias, Dept Biol Vegetal, P-1749016 Lisbon, Portugal
关键词
adventitious regeneration; Agrobacterium-mediated transformation; kanamycin resistance; Prunus dulcis; Rosaceae;
D O I
10.1007/s11240-005-9073-z
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 [微生物学]; 0836 [生物工程]; 090102 [作物遗传育种]; 100705 [微生物与生化药学];
摘要
An improved almond transformation system was developed with a 100x 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 mu M 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.
引用
收藏
页码:205 / 209
页数:5
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