摘要
基因枪转化技术在水稻的遗传转化上已被广泛地应用并获得显著的成效。与原生质体转化法相比较,基因枪法具有不受或少受基因型限制的优点且提高了转化效率,但对籼稻,仍有不少问题需要解决,转化系统尚须进一步完善。因此有必要对影响转化频率的因素进行深入的研究。我们在以barnase基因对籼稻的遗传转化以诱发工程雄性不育的研究中。
Four factors influence on transformation of indica rice, which were high osmotic treatment; different explant as the target tissue; pressure of rupture disk and quantity of plasmid DNA, were investigated in this experiment. High osmotic treatment of target tissue prior to and after bombardment increased 3. 2-fold for Gus transient expression than control. The best treatment of high osmotic was that the target tissues were kept in the target-bed medium which contained 0. 4-0. 6 mol/L sorbitol and manitol each for 4h prior to bombardment and for 16h after bombardment. Four explants: scutellum from mature seed, young panicle, embryogenic callus and suspension cells of indica rice were tested as target explant by particle bombardment. The results ofGus transient showed that the highest expression was scutellum and for other three explants, the order from high to low was young panicle, embryogenic callus and suspension cell. Transgenic plants were obtained from all of the explants except young panicle. For the pressure of rupture disk on transformation, 1100psi or 1300psi of the pressure of rupture disk were best one for the transformation and higher than 1300psi could damage the target tissue which become black and died in the following culture duration. For the quantity of plasmid DNA, the results showed that 0. 83μg of plasmid DNA per bombardment was preferred for the transformation of indica rice.
出处
《实验生物学报》
CSCD
2000年第1期85-88,共4页
Acta Biologiae Experimentalis Sinica
基金
国家自然科学基金(39470445)
广东省自然科学基金(940496)
美国Rockefeller Foundation(1994-0001-02420)资助课题
关键词
籼稻
基因枪转化
渗透剂
外植体
DNA用量
Indica rice. Particle bombardment. Factors influencing transformation.
