Tjokrokusumo D, Heinrich T, Wylie S, Potter R, McComb J
Biological Sciences and Biotechnology, Murdoch University, Murdoch 6150, Western, Australia e-mail:
Plant Cell Rep. 2000 Jul;19(8):792-797. doi: 10.1007/s002990050009.
Genetic transformation of Petunia hybrida with a reporter gene and selectable marker gene (35S-bar) was achieved in similar frequencies by pollinating flowers with pollen vacuum-infiltrated with Agrobacterium tumefaciens or applying a drop of Agrobacterium suspension to the stigma immediately prior to pollination. Nine percent of the T, and 5% of the T progeny germinated in nutrient medium with 3 mgl/l Basta. Polymerase chain reaction assays indicated that of the Basta-resistant plants, 66% of the T plants, and 61% of the T plants harboured the GUS gene. Histochemical assays showed that 10% of the putatively transformed T plants and 5% of their progeny expressed GUS in leaf tissue, pistils and young anthers. Southern hybridization confirmed genomic integration of the bar gene in one to three places in selected T and T progeny.
通过用真空渗入根癌农杆菌的花粉对矮牵牛花朵进行授粉，或在授粉前立即将一滴农杆菌悬浮液滴在柱头上，以相似的频率实现了用报告基因和选择标记基因（35S-bar）对矮牵牛进行遗传转化。9%的T₁代和5%的T₂代后代在含有3 mg/l Basta的营养培养基中发芽。聚合酶链反应分析表明，在抗Basta的植株中，66%的T₁代植株和61%的T₂代植株含有GUS基因。组织化学分析显示，10%的推定转化T₁代植株及其5%的后代在叶片组织、雌蕊和幼嫩花药中表达GUS。Southern杂交证实了bar基因在选定的T₁代和T₂代后代基因组中的一至三个位点整合。
