Plant Breeding and Genetics Research Laboratory, Japan Tobacco Inc., 700 Higashibara, Toyoda, Iwata, 438, Shizuoka, Japan.
Theor Appl Genet. 1990 Aug;80(2):167-71. doi: 10.1007/BF00224381.
A leaf-disk transformation experiment was performed with tobacco (Nicotiana tabacum L.) using a binary vector and a strain of Agrobacterium tumefaciens that carried a wild-type Ti-plasmid, pTiBo542. Although the majority of kanamycin-resistant, transgenic plants was morphologically normal, one of the plants was double-flowered and had a slightly wavy stem and leaves whose edges were bent slightly upwards. The abnormal morphology was controlled by a single, dominant Mendelian gene. Young plants that carried this gene were distinguishable from normal plants at the stage of cotyledons. The homozygotes, with respect to this gene, were more seriously deformed than the heterozygotes. DNA segments derived from the binary vector and from the TL-and TR-DNA of pTiBo542 were detected in the double-flowered plant, but the T-DNA genes involved in biosynthesis of phytohormones were absent from the plant. The abnormal morphology, the resistance to kanamycin, and the segments of foreign DNA were genetically linked, and the linkage was very tight, at least between the abnormal morphology and the resistance to kanamycin; the meiotic recombination frequency was less than 0.02%, if recombination occurred at all.
采用二元载体和携带野生型 Ti 质粒 pTiBo542 的根癌农杆菌菌株,对烟草(Nicotiana tabacum L.)进行了叶盘转化实验。尽管大多数卡那霉素抗性、转基因植株在形态上是正常的,但其中一株是重瓣花,茎和叶略微波浪状,边缘略微向上弯曲。这种异常形态受一个单一的、显性的孟德尔基因控制。携带该基因的幼株在子叶阶段就可以与正常植株区分开来。在该基因方面,纯合子比杂合子的变形更严重。在重瓣花植株中检测到了来自二元载体和 pTiBo542 的 TL 和 TR-DNA 的 DNA 片段,但参与植物激素生物合成的 T-DNA 基因在植株中缺失。异常形态、对卡那霉素的抗性和外源 DNA 片段在遗传上是连锁的,至少在异常形态和对卡那霉素的抗性之间是紧密连锁的;如果发生重组,其减数分裂重组频率小于 0.02%。