由根癌农杆菌介导将葡萄糖氧化酶基因转入水稻

摘要:将具有广谱抗病作用的葡萄糖氧化酶（glucose oxidase , GO）基因插入具有潮霉素抗性选择标记的双元载体pCAMBIA1301，新构建了水稻高效表达载体pCAG1301。将此质粒导入根癌农杆菌（Agrobacterium tumefaciens ）菌株LBA4404后,转化粳稻(Oryza. sativa L.ssp.japonica )品种日本晴(Nipponbare)的幼胚，并由筛选出的潮霉素抗性愈伤组织分化再生植株。对所得潮霉素抗性水稻植株的Southern杂交分析表明，GO基因已整合到受体基因组。淀粉-碘化钾显色反应检测到了转基因植株产生的过氧化氢，证实GO基因表达产生的葡萄糖氧化酶已经在水稻中发挥功能。抗病性鉴定表明，所得转基因水稻对稻瘟菌(Magnaporthe grisea )具有良好的抗性。

Introducing Glucose Oxidase Gene into Rice Mediated by Agrobacterium tumefaciens

Abstract: The glucose oxidase (GO ) gene with the function of broad-spectrum pathogen defense was inserted into a binary vector pCAMBIA1301 containing selectable marker of hygromycin resistant gene to produce a new expression vector, pCAG1301. The vector was introduced into Agrobacterium tumefaciens strain LBA4404 and then transformed them to immature embryos of rice (Oryza sativa L.ssp. japonica ) cultivar Nipponbare. Putative transformed plants were regenerated from hygromycin resistant calli, and the integration of GO gene was confirmed by Southern blotting analysis on rice genome. The H2O2 generated by the transgenic plants was detected by color reaction with starch-KI and this proved that the glucose oxidase generated by the expression of GO gene had exerted its function in rice. The transgenic plants were proved to have strong resistance to rice blast disease(Magnaporthe grisea ).