Friedrich Miescher-Institut, P.O.Box 2543, CH-4002 Basel, Switzerland.
EMBO J. 1984 Dec 1;3(12):2717-22. doi: 10.1002/j.1460-2075.1984.tb02201.x.
Evidence for direct, gene-mediated stable genetic transformation of plant cells of Nicotiana tabacum is presented. A selectable hybrid gene comprising the protein coding region of the Tn5 aminoglycoside phosphotransferase type II gene under control of cauliflower mosaic virus gene VI expression signals was introduced into plant protoplasts as part of an Escherichia coli plasmid. The gene was stably integrated into plant genomic DNA and constitutively expressed in selected, drug resistant, protoplast-derived cell clones. The mode of integration of the foreign gene into the plant genome resembled that observed for DNA transfection of mammalian cells. Plants regenerated from transformed cell lines were phenotypically normal and fertile, and they maintained and expressed the foreign gene throughout the development of vegetative and generative organs. Microspores, grown in anther culture, developed into resistant and sensitive haploid plantlets. Genetic crossing analysis of one of the transformed plants revealed the presence of one dominant trait for kanamycin resistance segregating in a Mendelian fashion in the F(1) generation.
本文提出了烟草细胞的直接基因介导稳定遗传转化的证据。一种可选择的杂种基因，包括 Tn5 氨基糖苷磷酸转移酶 II 基因的蛋白编码区，在花椰菜花叶病毒基因 VI 表达信号的控制下，被作为大肠杆菌质粒的一部分导入植物原生质体。该基因稳定地整合到植物基因组 DNA 中，并在选择的、抗药性的、源自原生质体的细胞克隆中组成型表达。外源基因整合到植物基因组中的方式类似于哺乳动物细胞 DNA 转染所观察到的方式。从转化细胞系再生的植物在表型上正常且可育，并且它们在营养和生殖器官的发育过程中维持并表达外源基因。在花药培养中生长的小孢子发育成具有抗性和敏感性的单倍体植物。对其中一株转化植物的遗传杂交分析表明，在 F1 代中存在一种显性的卡那霉素抗性性状，以孟德尔方式分离。
