基因枪轰击转化棉花茎尖分生组织影响因素的探讨

以农大KB18的茎尖分生组织为受体材料,利用基因枪轰击法将植酸酶基因(PhyA)导入棉花茎尖分生组织,经过再生、筛选、分子检测等过程获得若干转基因植株。通过研究碳源、外源激素、活性炭、轰击距离和轰击次数以及茎尖的恢复培养与抗性筛选等,建立了较为完善的棉花基因枪茎尖转化体系。结果表明,植株再生率与再生培养基的成分,如激素的有无、活性炭的含量和碳源等有关,轰击距离以及筛选程序中卡那霉素的起始筛选时间、筛选浓度和浓度梯度等因素对转化周期和转化率有直接影响。

Influencing Factors of Shoot Apical Meristem Transformation via Particle Bombardment in Cotton

The phytase gene(PhyA) was transformed into shoot apical meristems of NDKB18 via microprojectile-bombardment.Some transgenic plants were obtained from these bombarded explant materials.An improved transformation system of meristem via gene-gun bombardment was established based on some influential factors investigated.The results showed that the ratios of regenerated plant were related with the ingredients of regeneration medium,such as hormones,active charcoal and different sugars.The efficiency of transgenic plant and transformation cycle were affected by the bombardment distance,the initiative time of selection,the initiative concentration and concentration gradients of kanamycin in the process of selection.