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[影响籼稻粒子轰击转化的因素]

[Factors influence on transformation by particle bombardment in Indica rice].

作者信息

Tao L Z, Xu Q S, Liang C Y, Ling D H

机构信息

South China Institute of Botany, Academia Sinica, Guangzhou 510650.

出版信息

Shi Yan Sheng Wu Xue Bao. 2000 Mar;33(1):85-8.

Abstract

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 4 h prior to bombardment and for 16 h 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 of Gus 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, 1100 psi or 1300 psi of the pressure of rupture disk were best one for the transformation and higher than 1300 psi 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 0.83 microgram of plasmid DNA per bombardment was preferred for the transformation of indica rice.

摘要

本实验研究了影响籼稻转化的四个因素,即高渗处理;作为靶组织的不同外植体;破裂盘压力和质粒DNA数量。轰击前后对靶组织进行高渗处理,Gus瞬时表达比对照提高了3.2倍。最佳高渗处理是在轰击前将靶组织置于含有0.4 - 0.6 mol/L山梨醇和甘露醇的靶床培养基中4小时,轰击后放置16小时。测试了四种外植体:成熟种子的盾片、幼穗、胚性愈伤组织和籼稻悬浮细胞作为粒子轰击的靶外植体。Gus瞬时表达结果表明,最高表达的是盾片,其他三种外植体从高到低依次为幼穗、胚性愈伤组织和悬浮细胞。除幼穗外,所有外植体均获得了转基因植株。对于破裂盘压力对转化的影响,1100 psi或1300 psi的破裂盘压力最有利于转化,高于1300 psi会损伤靶组织,使其在后续培养过程中变黑死亡。对于质粒DNA数量,结果表明,每次轰击0.83微克质粒DNA最有利于籼稻的转化。

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