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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

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【作者】 于恒秀刘巧泉陈秀花陆美芳王兴稳王宗阳顾铭洪

【Author】 YU Heng xiu 1, LIU Qiao quan 1, CHEN Xiu hua 1, LU Mei fang 1, WANG Xing wen, WANG Zong yang 2, GU Ming hong 1,* ( 1 Department of Agronomy, Agricultural College, Yangzhou University, Yangzhou 225009, China; 2Shanghai Institute of P

【机构】 扬州大学农学院农学系中国科学院上海植物生理生态研究所扬州大学农学院农学系 江苏扬州225009江苏扬州225009上海200032

【摘要】 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∶ 1的理论比例

【Abstract】 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.

【基金】 国家科技部转基因植物研究与产业化专项资助项目(J99-A-0 0 5 );国家高技术研究计划资助项目(2 0 0 1AA2 12 10 1);江苏省科技厅资助项目
  • 【文献出处】 中国水稻科学 ,Chinese Journal of Rice Science , 编辑部邮箱 ,2002年04期
  • 【分类号】S511
  • 【被引频次】51
  • 【下载频次】274
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