Lutke W Kevin
Tissue Culture and Transformation Facility, Donald Danforth Plant Science Center, St. Louis, MO, USA.
Methods Mol Biol. 2006;344:339-49. doi: 10.1385/1-59745-131-2:339.
Petunia hybrida genetic transformation continues to be a valuable tool for genetic research into biochemical pathways and gene expression, as well as generating commercial products with varying floral colors. In this chapter, we describe a simple and reproducible genetic transformation protocol for generating transgenic petunia plants harboring a gene of interest and selectable marker. The system utilizes Agrobacterium tumefaciens for transgene integration with plant recovery via shoot organogenesis from leaf explant material. Selection for transgenic plants is achieved using the bar gene conferring resistance to glufosinate or nptII gene for resistance to kanamycin. Transformation efficiencies of around 10% are achievable with shoots being recovered about 8 wk after transgene insertion and rooted plants transferred to the greenhouse about twelve weeks after inoculation.
矮牵牛的遗传转化仍然是研究生化途径和基因表达以及培育具有不同花色商业产品的重要遗传研究工具。在本章中,我们描述了一种简单且可重复的遗传转化方案,用于培育携带感兴趣基因和选择标记的转基因矮牵牛植株。该系统利用根癌农杆菌将转基因整合到植物中,并通过叶片外植体材料的芽器官发生实现植物再生。通过使用赋予草铵膦抗性的bar基因或赋予卡那霉素抗性的nptII基因来筛选转基因植物。转基因插入后约8周可获得芽,接种后约12周将生根的植株转移到温室,转化效率可达10%左右。