Huang J Q, Wel Z M, An H L, Zhu Y X
National Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai.
Cell Res. 2001 Jun;11(2):149-55. doi: 10.1038/sj.cr.7290080.
Immature embryos of rice varieties "Xiushuill" and "Chunjiang 11" precultured for 4d were infected and transformed by Agrobacterium tumefaciens strain EHA101/pExT7 (containing the spider insecticidal gene). The resistant calli were transferred onto the differentiation medium and plants were regenerated. The transformation frequency reached 56% approximately 72% measured as numbers of Geneticin (G418)-resistant calli produced and 36% approximately 60% measured as numbers of transgenic plants regenerated, respectively. PCR and Southern blot analysis of transgenic plants confirmed that the T-DNA had been integrated into the rice genome. Insect bioassays using T1 transgenic plants indicated that the mortality of the leaffolder (Cnaphalocrasis medinalis) after 7d of leaf feeding reached 38% approximately 61% and the corrected mortality of the striped stem borer (Chilo suppressalis) after 7d of leaf feeding reached 16% approximately 75%. The insect bioassay results demonstrated that the transgenic plants expressing the spider insecticidal protein conferred enhanced resistance to these pests.
水稻品种“秀水 11”和“春江 11”的未成熟胚预培养 4 天后，用根癌农杆菌菌株 EHA101/pExT7（含蜘蛛杀虫基因）进行侵染和转化。将抗性愈伤组织转移到分化培养基上再生植株。转化频率分别以产生的潮霉素（G418）抗性愈伤组织数量计约为 56%（约 72%），以再生的转基因植株数量计约为 36%（约 60%）。对转基因植株进行的 PCR 和 Southern 杂交分析证实 T-DNA 已整合到水稻基因组中。使用 T1 代转基因植株进行的昆虫生物测定表明，取食叶片 7 天后，稻纵卷叶螟的死亡率达到约 38%（约 61%），二化螟取食叶片 7 天后的校正死亡率达到约 16%（约 75%）。昆虫生物测定结果表明，表达蜘蛛杀虫蛋白的转基因植株对这些害虫具有增强的抗性。
