小麦幼胚基因枪转化的影响因素研究  

Effect of Factors on GUS Transient Expression of Wheat Immature Embryo Transformated by Particle Bombardment

作　　者：夏晓晖[1] 陈耀锋[1] 李春莲[1] 吕瑞华[1] 曹团武[1] 曹欣[1] 赵云祥[1] 

机构地区：[1]西北农林科技大学农学院国家小麦改良中心杨凌分中心,陕西杨凌712100

出　　处：《麦类作物学报》2006年第2期42-45,共4页Journal of Triticeae Crops

基　　金：国家转基因专项(jy03-B-23-02);陕西省科技攻关项目(2003K03-G1-04)

摘　　要：为了明确不同因素对小麦基因枪法转化效率的影响。用PDS-1000／He型基因棺将抑制叶片衰老基因Psag12-ipt转入小麦幼胚细胞，通过对报告基因GUS瞬时表达的检测，研究了金粉用量、质粒DNA用量及轰击次数对转化率的影响。结果表明，金粉用量在100-400μg范围内，时幼胚瞬时表达率影响不大，幼胚瞬时表达率在73．7%～85．0％之间，但随金粉用量的增加，细胞瞬时表迭率明显提高；质粒DNA用量以每枪2．0μg为最佳；轰击2枪幼胚和细胞瞬时表达率明显高于轰击1枪。Psag12-ipt gene was transformed into wheat immature embryo cells by particle bombardment. The effects of gold particle concentration and DNA amount per bombardment on transformation efficiency of wheat immature embryo cells were studied by GUS transient expression. The result showed that the gold particle concentration had little affect on transient expression of immature embryos, while it played a remarkable role in the cell transient expression at a relatively high gold particle amount per bombardment. It was found that the transformation frequency of wheat immature embryo cells was the superior when coated by 2.0 μg DNA and bombarded for 2 times per dish.

关 键 词：小麦幼胚 基因枪 GUS基因 瞬时表达 转化效率 

分 类 号：S512.1[农业科学—作物学] S336