Datta S K, Datta K, Soltanifar N, Donn G, Potrykus I
Swiss Federal Institute of Technology, ETH-Zurich, Institute of Plant Sciences, Switzerland.
Plant Mol Biol. 1992 Nov;20(4):619-29. doi: 10.1007/BF00046447.
The commercially important Indica rice cultivar Oryza sativa cv. IR72 has been transformed using direct gene transfer to protoplasts. PEG-mediated transformation was done with two plasmid constructs containing either a CaMV 35S promoter/HPH chimaeric gene conferring resistance to hygromycin (Hg) or a CaMV 35S promoter/BAR chimaeric gene conferring resistance to a commercial herbicide (Basta) containing phosphinothricin (PPT). We have obtained so far 92 Hgr and 170 PPTr IR72 plants from protoplasts through selection. 31 Hgr and 70 PPTr plants are being grown in the greenhouse to maturity. Data from Southern analysis and enzyme assays proved that the transgene was stably integrated into the host genome and expressed. Transgenic plants showed complete resistance to high doses of the commercial formulations of PPT.
商业上重要的籼稻品种水稻品种IR72已通过将基因直接导入原生质体进行了转化。使用两种质粒构建体进行聚乙二醇(PEG)介导的转化,其中一种含有赋予潮霉素(Hg)抗性的CaMV 35S启动子/HPH嵌合基因,另一种含有赋予对含草丁膦(PPT)的商业除草剂(Basta)抗性的CaMV 35S启动子/BAR嵌合基因。到目前为止,我们通过筛选从原生质体中获得了92株潮霉素抗性(Hgr)和170株草丁膦抗性(PPTr)的IR72植株。31株Hgr和70株PPTr植株正在温室中生长至成熟。Southern分析和酶分析的数据证明,转基因已稳定整合到宿主基因组中并表达。转基因植物对高剂量的PPT商业制剂表现出完全抗性。