反义Waxy基因共转化籼型杂交稻亲本的初步研究

A Preliminary Study on Co-transformation Parent-lines of Indica Hybrid Rice with Anti-sense Waxy Gene

以籼型杂交稻亲本II32B为供试材料,对根癌农杆菌共转化过程中影响转化效率的因素进行了研究。结果表明:在诱导培养基和分化培养基中附加NAA、KT和6-BA的激素配比可以改进愈伤组织质量,对愈伤组织的形成和苗的分化起决定作用;农杆菌OD600为0.8,侵染时间为10 min是II32B合适的转化参数;潮霉素浓度为30 mg/L是理想的筛选浓度;部分植株经PCR检测为阳性,证明外源目的基因已经整合到转基因水稻的基因组中,从而建立适合于供试水稻材料的遗传转化体系。

The indica hybrid rice parent-line II32B was taken as the tested material,the factors that affecting the transformation efficiency in the process of Agrobacterium-mediated co-transformation were studied.The results showed that the hormone ratio of NAA,KT and 6-BA supplemented to the induction medium and differentiation medium can improve quality of callus and play a decisive role in the callus formation and shoot differentiation;the appropriate parameters for II32B conversion were engineered strain EHA105 Bacilli with OD600 value of 0.8 and infection time of 10 min;the ideal screening concentration was hygromycin concentration of 30 mg/L;part of the plants were positive through PCR test to prove that the foreign gene has been integrated into the genome of transgenic rice,therefore it could establish suitable genetic transformation system for the test material of rice.