Centro Nacional de Pesquisa em Recursos Genéticos e Biotecnologia, (CENARGEN/EMBRAPA) SAIN Parque Rural, P.O. Box 012372, Brasilia, DF, Brazil.
Plant Cell Rep. 1994 Sep;13(12):666-70. doi: 10.1007/BF00231620.
In search of establishing a system for genetic transformation of Brazilian potato cultivars, Agrobacterium tumefaciens carrying the plasmid pGV1040, was used to transform leaf discs of three cultivars of local importance, i.e., Aracy, Baronesa and Mantiqueira. This plasmid contains marker genes for resistance to kanamycin and phosphinothricin plus the gene for the enzyme β-glucuronidase. A two step regeneration/selection procedure produced shoots of potato cultivar Mantiqueira with in vitro resistance to kanamycin and to phosphinothricin. After transfer to the greenhouse, the potentially transgenic plants, sprayed with the herbicide Finale(®) (20% a.i.; Hoechst(®)) remained green as compared to control clones that died immediately afterwards. Southern blot analysis and histochemical and fluorimetric assay for β-glucuronidase indicated that the gene coding for the enzyme was integrated in the potato genome and could be expressed in potato tissues. No success was obtained for transformation of cultivars Aracy and Baronesa using this procedure.
为了建立巴西马铃薯品种的遗传转化体系,我们使用携带质粒 pGV1040 的根癌农杆菌来转化三种具有地方重要性的马铃薯品种(Aracy、Baronesa 和 Mantiqueira)的叶片圆盘。该质粒含有对卡那霉素和膦丝菌素的抗性标记基因以及β-葡萄糖醛酸酶的基因。两步再生/选择程序产生了马铃薯品种 Mantiqueira 的芽,这些芽在体外对卡那霉素和膦丝菌素具有抗性。转移到温室后,与立即死亡的对照克隆相比,用除草剂 Finale(®)(20%有效成分;Hoechst(®))喷洒的潜在转基因植物保持绿色。Southern blot 分析和β-葡萄糖醛酸酶的组织化学和荧光测定表明,编码该酶的基因已整合到马铃薯基因组中,并可在马铃薯组织中表达。使用该程序,未能成功转化 Aracy 和 Baronesa 这两个品种。