Science. 1985 Mar 8;227(4691):1229-31. doi: 10.1126/science.227.4691.1229.
Transformed petunia, tobacco, and tomato plants have been produced by means of a novel leaf disk transformation-regeneration method. Surface-sterilized leaf disks were inoculated with an Agrobacterium tumefaciens strain containing a modified tumor-inducing plasmid (in which the phytohormone biosynthetic genes from transferred DNA had been deleted and replaced with a chimeric gene for kanamycin resistance) and cultured for 2 days. The leaf disks were then transferred to selective medium containing kanamycin. Shoot regeneration occurred within 2 to 4 weeks, and transformants were confirmed by their ability to form roots in medium containing kanamycin. This method for producing transformed plants combines gene transfer, plant regeneration, and effective selection for transformants into a single process and should be applicable to plant species that can be infected by Agrobacterium and regenerated from leaf explants.
利用一种新颖的叶片圆盘转化-再生方法,已经成功地培育出转化的矮牵牛、烟草和番茄植株。将表面消毒的叶片圆盘接种到含有改良的肿瘤诱导质粒的根癌农杆菌菌株中(其中已删除了来自转移 DNA 的植物激素生物合成基因,并被卡那霉素抗性的嵌合基因所取代),并培养 2 天。然后将叶片圆盘转移到含有卡那霉素的选择培养基上。在 2 至 4 周内发生芽再生,并且通过它们在含有卡那霉素的培养基中形成根的能力来确认转化体。这种生产转化植物的方法将基因转移、植物再生和转化体的有效选择结合到一个单一的过程中,应该适用于可以被根癌农杆菌感染并从叶片外植体再生的植物物种。