根癌农杆菌介导的水稻转化系统的优化及转反义Wx基因植株的获得

Optimization of Agrobacterium-Mediated Transformation of Rice and Regeneration of Transgenic Plants with Antisense Waxy Gene

以具不同直链淀粉含量的籼粳稻品种为受体材料 ,试验了适宜于根癌农杆菌介导转化的水稻愈伤组织诱导培养基及其诱导培养的时间。结果表明 ,一种基于 MS基本培养基的商品培养基较适合作为籼粳稻幼胚愈伤组织的诱导培养基 ;对于籼型杂交稻亲本协青早 B,转化前幼胚较适合的愈伤组织诱导培养天数为 8d左右。在此基础上 ,将反义蜡质 (Wx)基因分别导入上述受体亲本中 ,获得了一批转基因水稻植株。PCR和 Southern杂交分析表明反义 Wx基因已经整合进了转基因水稻的基因组中。遗传分析表明外源基因在转基因水稻后代中的分离符合 3∶

Several rice varieties with various amylose content in grains were used for optimization of Agrobacterium mediated transformation of rice, and the suitable medium for induction of callus and suitable initially culture time were studied. For this transformation system, the HRM medium, based on the MS basal medium, was suitable for induction of callus from immature embryos. In the case of indica rice Xieqingzao B, the suitable time of initially culture in this medium was 8 days before co culturing with Agrobacterium. Based on this transformation system, the antisense waxy gene was introduced into different rice varieties and many transgenic lines were produced. Most of these transgenic plants were confirmed by PCR and Southern blot technique, and the results indicated that the T DNA had been integrated into the genome of transgenic rice plants. Genetic analysis of some T 1 transgenic plants showed that the transgenes were segregated normally in the progenies.