以花药为受体的小麦转基因技术体系的构建

为了在转化当代获得纯合的转基因株系,本试验对以小麦花药为受体的转基因方法进行了系统研究。结果表明,金粉粒度0.6μm、金粉浓度100μg/枪、轰击距离6 cm、轰击压力1 300 psi为基因枪转化小麦花药的最佳参数组合。用含有5%DMSO的0.2%秋水仙碱溶液处理单倍体植株4 h,染色体加倍效率最高,达到100%;用含1%DMSO的溶液处理12 h加倍效率最低,为62.2%。PCR结果显示所有T0代阳性植株在T1代都无分离,说明该方法得到的转基因株系在T0代即为纯合系。本研究成功建立了以花药为受体的小麦转基因技术体系,为小麦遗传改良提供了一种新的方法。

Construction of Wheat Transgenetic Technology System using Isolated Anthers as Particle Bombardment Receptor

To obtain homozygous transgenic wheat within one generation , the parameters involved in bombardment and chromosome doubling were studied in details .The results indicated that the best parameter combination for anther bombardment were 0.6-μm particle size, 100 μg per shot, 6-cm bombardment dis-tance and 1 300-psi bombardment pressure .The highest doubling efficiency （ 100%） was obtained when haploid plants were treated with 0.2%colchicine solution containing 5%DMSO for 4 hours, and that contai-ning 1%DMSO had the lowest doubling efficiency （62.2%）.Specific PCR amplification showed that all T 0 positive plants had no segregation in T 1 generation, which indicated that the homozygous transgenic lines were obtained within one generation .This study established a successful transgenic technology system using isolated wheat anthers as bombardment recipient , which provided an efficient approach for wheat genetic improvement .