Denchev P D, Songstad D D, McDaniel J K, Conger B V
Department of Plant and Soil Science, The University of Tennessee, Knoxville, TN 37901-1071, USA, , , , , , US.
Pioneer Hi-Bred International Inc, 7300 N. W. 62nd Avenue, Johnston, IA 50131, USA, , , , , , US.
Plant Cell Rep. 1997 Oct;16(12):813-819. doi: 10.1007/s002990050326.
Young leaf tissue of orchardgrass (Dactylis glomerata L.) was placed on Schenk and Hildebrandt medium containing 30 µM dicamba. Microprojectiles coated with DNA containing the selectable bar gene (Basta® tolerance) and the reporter gene uidA coding for β-glucuronidase (GUS), both driven by the maize ubiquitin promoter (Ubi1), were propelled into the tissue with a particle inflow gun. Transient GUS expression was observed as blue spots of various sizes on leaf segments. Somatic embryos staining entirely blue were also produced, and embryos germinated on medium containing 3.0 mg 1 bialaphos. Leaves of 67 putative transformed plants were painted with 0.1% Basta. Ten showed no reaction, and 6 showed only a localized response. Cultured leaf segments from tolerant plants also produced somatic embryos that expressed GUS. The genetic transformation was confirmed by Southern blot hybridization and PCR analyses of T plants and by PCR analyses of somatic embryos produced from T plants.
将果园草（鸭茅）的幼叶组织置于含有30 µM麦草畏的 Schenk 和 Hildebrandt 培养基上。用含有由玉米泛素启动子（Ubi1）驱动的选择标记 bar 基因（抗 Basta®）和编码 β-葡萄糖醛酸酶（GUS）的报告基因 uidA 的 DNA 包被的微粒，通过粒子流入枪将其射入组织中。在叶段上观察到瞬时 GUS 表达为大小不一的蓝色斑点。还产生了完全染成蓝色的体细胞胚，并且胚在含有3.0 mg/L 双丙氨膦的培养基上萌发。用0.1% Basta 涂抹67株推定转化植株的叶片。10株无反应，6株仅表现出局部反应。来自耐受植株的培养叶段也产生了表达 GUS 的体细胞胚。通过对 T 代植株的 Southern 杂交和 PCR 分析以及对 T 代植株产生的体细胞胚的 PCR 分析证实了遗传转化。
