Van Dang Thi, Ferro Noel, Jacobsen Hans-Jörg
Fruit and Vegetable Research Institute, Hanoi, Vietnam.
GM Crops. 2010 Nov-Dec;1(5):312-21. doi: 10.4161/gmcr.1.5.14703.
The transformation of tomato (Solanum lycopersicum) through Agrobacterium tumefaciens is still far from being routine, particularly when it comes to commercial varieties. In the present paper, we present an efficient and simple protocol for leaf disc transformation of three Vietnamese tomato cultivars (DM8, MTS, FM372C) by comparing shoot regeneration media for expanding leaves and examining different parameters of inoculation, co-culture and selection conditions. The present transformation method requires neither feeder layers of cell suspension cultures nor pre-culture. The data clearly show that appropriate cytokinin- and auxin combinations and concentrations provide competent tissues for transformation. Supplementing of 8 µM trans-zeatin and 5 µM indoleacetic acid (IAA) into pre-treatment, inoculation and co-culture media resulted in higher frequency of transformation and stronger GUS-expression than that of media supplemented with 4 µM trans-zeatin and 2 µM IAA. The experiments also exhibited that tomato leaf tissues were more sensitive to glufosinate after inoculation with Agrobacteria compared to the untreated controls, so a more sophisticated scheme for the glufosinate selection had to be established.
通过根癌农杆菌对番茄（Solanum lycopersicum）进行转化仍远未成为常规操作，尤其是对于商业品种而言。在本文中，我们通过比较用于扩展叶片的芽再生培养基，并研究接种、共培养和选择条件的不同参数，提出了一种高效且简单的方法用于对三个越南番茄品种（DM8、MTS、FM372C）进行叶盘转化。目前的转化方法既不需要细胞悬浮培养物的饲养层，也不需要预培养。数据清楚地表明，适当的细胞分裂素和生长素组合及浓度可为转化提供合适的组织。在预处理、接种和共培养基中添加8 μM反式玉米素和5 μM吲哚乙酸（IAA），相比于添加4 μM反式玉米素和2 μM IAA的培养基，转化频率更高且GUS表达更强。实验还表明，与未处理的对照相比，接种农杆菌后的番茄叶片组织对草铵膦更敏感，因此必须建立更完善的草铵膦选择方案。
