Chung M H, Chen M K, Pan S M
Department of Botany, National Taiwan University, Taipei, Republic of China.
Transgenic Res. 2000 Dec;9(6):471-6. doi: 10.1023/a:1026522104478.
In this study, floral spray and floral dip were used to replace the vacuum step in the Agrobacterium-mediated transformation of a superoxide dismutase (SOD) gene into Arabidopsis. The transgene was constructed by using a CaMV 35S promoter to drive a rice cytosolic CuZnSOD coding sequence in Arabidopsis. The transgene construct was developed in binary vectors and mobilized into Agrobacterium. When Arabidopsis plants started to initiate flower buds, the primary inflorescence shoots were removed and then transformed by floral spray or floral dip. More than 300 transgenic plants were generated to assess the feasibility of floral spray used in the in planta transformation. The result indicates that the floral spray method of Agrobacterium can achieve rates of in planta transformation comparable to the vacuum-infiltration and floral dip methods. The floral spray method opens up the possibility of in planta transformation of plant species which are too large for dipping or vacuum infiltration.
在本研究中，采用花喷雾法和花浸染法替代农杆菌介导的超氧化物歧化酶（SOD）基因转化拟南芥过程中的真空步骤。通过使用CaMV 35S启动子驱动水稻胞质CuZnSOD编码序列在拟南芥中构建转基因。转基因构建体在二元载体中构建并导入农杆菌。当拟南芥植株开始形成花芽时，去除主花序轴，然后通过花喷雾或花浸染进行转化。生成了300多株转基因植株以评估花喷雾法用于植物体内转化的可行性。结果表明，农杆菌花喷雾法可实现与真空渗透法和花浸染法相当的植物体内转化率。花喷雾法为因植株过大而无法进行浸染或真空渗透的植物物种的植物体内转化开辟了可能性。
