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以水稻基因组测序品种日本晴(OryzasativaL.ssp.japonica)为材料,对根癌农杆菌介导的水稻转化系统进行了优化。结果表明,水稻种子经质量分数2.5%的次氯酸钠灭菌30min后,愈伤组织诱导率最高;所获得的胚性愈伤组织经过6～7d的继代培养后转化率达到95%;感染液菌浓度OD600值为0.145～0.190时,转化效率高;将抗性愈伤组织进行7d的暗培养可使分化效率提高到85%。应用本转化系统已经获得了大量的转化体,GUS表达率达到30%,并且3个月即可获得转化植株。
The Agrobacterium tumefaciens-mediated rice transformation system was optimized by using Oryza sativa L.sepjaponica as a material. The results showed that the callus induction rate was the highest after rice seeds were treated with 2.5% sodium hypochlorite for 30min. The transformation rate of embryogenic callus reached 95% after 6-7 days subculture; When the concentration of OD600 was 0.145 ~ 0.190, the transformation efficiency was high. After culturing the resistant callus for 7 days, the differentiation efficiency was increased to 85%. A large number of transformants have been obtained using this transformation system. The GUS expression rate reaches 30%, and the transformed plants can be obtained after 3 months.