Molecular Biology Division, The Upjohn Co., 301 Henrietta Street, 49007, Kalamazoo, MI, USA.
Plant Cell Rep. 1990 Sep;9(5):245-8. doi: 10.1007/BF00232293.
Cotyledons of cucumber seedlings (Cucumis sativus L. cv. Poinsett 76) were co-cultivated with disarmed Agrobacterium strain C58Z707. The Agrobacterium strain contained the Agrobacterium-derived binary vector plasmid pGA482, its T-DNA region contains a plant expressible bacterial derived neomycin phosphotransferase II (NPT II) gene which upon transfer, genome integration, and expression in plant tissues confers resistance to the antibiotic kanamycin. After growth of inoculated cotyledon sections on selective medium containing 100 mg/l kanamycin, transformed embryogenic calli were obtained followed by the development of embryos and plant regeneration. Transformed R0 and R1 cucumber plants appeared normal and tested positive for NPT II enzyme activity. Genomic DNAs isolated from the NPT II positive plants all showed hybridization to the characteristic 2.0 kb (BamHI to HindIII) NPT II gene-containing fragment. These results show that the Agrobscterium-mediated gene transfer system and regeneration via somatic embryogenesis is an effective method for the transfer of genetic material into plant species belonging to the family Cucurbitaceae.
黄瓜幼苗子叶（Cucumis sativus L. cv. Poinsett 76）与去武装根瘤农杆菌菌株 C58Z707 共培养。该根瘤农杆菌菌株含有源自农杆菌的二元载体质粒 pGA482，其 T-DNA 区域包含可在植物中表达的细菌衍生新霉素磷酸转移酶 II（NPT II）基因，该基因在转移、基因组整合和植物组织表达后赋予对抗生素卡那霉素的抗性。在含有 100mg/L 卡那霉素的选择培养基上生长接种的子叶切片后，获得了转化的胚性愈伤组织，随后发育出胚胎和植物再生。转化的 R0 和 R1 黄瓜植株外观正常，并检测到 NPT II 酶活性呈阳性。从 NPT II 阳性植物中分离的基因组 DNA 均与特征性的 2.0kb（BamHI 至 HindIII）NPT II 基因内含片段发生杂交。这些结果表明，根瘤农杆菌介导的基因转移系统和通过体细胞胚胎发生的再生是将遗传物质转移到葫芦科植物物种的有效方法。
