Arencibia Ariel D, Carmona Elva R
Genetic and Phytopathologic Department, National Institute for Sugarcane Research, Carretera CUJAE Km 2. Boyeros 19 390, Havana City, Cuba.
Methods Mol Biol. 2006;344:227-35. doi: 10.1385/1-59745-131-2:227.
We describe the procedures for recovering transgenic sugarcane from co-cultivation of both calli and in vitro plants with Agrobacterium tumefaciens. The correct tissue culture strategies and the use of super-binary vector or super-virulent strain are crucial for the successful sugarcane transformation. Both plant regeneration via calli culture and micropropagation strategies can be optimized to a wide spectrum of sugarcane genotypes, thus the procedures presented here could be applied to genetic engineering of Saccharum spp. after minor modifications. For the case of sugarcane transformation using in vitro plants, four selective micropropagation steps must be sufficient to eliminate chimera plants.
我们描述了通过愈伤组织和离体植株与根癌农杆菌共培养来获得转基因甘蔗的方法。正确的组织培养策略以及使用超二元载体或超毒力菌株对于甘蔗转化的成功至关重要。通过愈伤组织培养进行植株再生和微繁殖策略都可以针对多种甘蔗基因型进行优化,因此这里介绍的方法经过微小修改后可应用于甘蔗属的基因工程。对于使用离体植株进行甘蔗转化的情况,四个选择性微繁殖步骤必须足以消除嵌合体植株。