中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

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

于恒秀 1; 刘巧泉 1; 陈秀花 1; 陆美芳 1; 王兴稳 1; 王宗阳 2; 顾铭洪 1,*   

  1. 1扬州大学 农学院 农学系， 江苏 扬州 225009; 2中国科学院 上海植物生理生态研究所， 上海 200032; * 通讯联系人，E-mail: ricegb@yzu.edu.cn
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2002-10-10 发布日期:2002-10-10

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

YU Heng-xiu 1; LIU Qiao-quan 1; CHEN Xiu-hua 1; LU Mei-fang 1; WANG Xing-wen1; WANG Zong-yang 2; GU Ming-hong 1; *   

  1. （1Department of Agronomy; Agricultural College; Yangzhou University; Yangzhou 225009; China; 2Shanghai Institute of P;
  • Received:1900-01-01 Revised:1900-01-01 Online:2002-10-10 Published:2002-10-10
  • Contact: GU Ming-hong

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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8 d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。

关键词: 转基因, 水稻, 根癌农杆菌, 反义蜡质基因, 直链淀粉含量

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 T1 transgenic plants showed that the transgenes were segregated normally in the progenies.

Key words: transgene, rice, Agrobacterum tumefaciens, antisense waxy gene, amylose content