中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

转基因水稻经花药培养获得纯系的研究

斯华敏 1;傅亚萍 1;肖 晗 1;胡国成 1;曹军萍 1;黄大年 2;孙宗修 1   

  1. 1 中国水稻研究所 水稻生物学农业部重点开放实验室，杭州310006; 2 中国水稻研究所生物工程系，杭州 310006
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1999-01-10 发布日期:1999-01-10

Homozygous Lines of Transgenic Rice (Oryza sativa L.) Obtained via Anther Culture

Si Huamin; Fu Yaping; Xiao Han; Hu Guocheng; Cao Junping; Huang Danian; Sun Zongxiu

  

  • Received:1900-01-01 Revised:1900-01-01 Online:1999-01-10 Published:1999-01-10

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摘要： 利用基于苯乙酸的一步成苗法，获得了来源于多个品种转基因水稻当代植株的花培苗上百株。对其中两个含bar基因插入的转基因粳稻当代植株京引119-B3和京引119-B4的花培植株及其后代进行了详细的研究分析。bar基因在所获得的单倍体和二倍体花培植株中均可正常表达出对除草剂Basta的抗性。PCR、Southern分析进一步证明导入的bar基因在绝大多数花培植株中没有发生变化。即使对粳稻京引119-B4这一有多位点插入的转基因植株，花药培养也可达到使外源基因一次纯合、后代不再分离的目的。结果表明对转基因植株进行花药培养可用于快速纯合外源基因以获得纯系。并就这一技术体系的技术要点进行了讨论。

关键词: 转基因, 花药培养, 纯合

Abstract: Hundreds of anther-derived lines from transgenic rice plants with insertion of bar gene were obtained by a PAA based one-step method of anther culture. Inheritance and expression of transgene bar in 6 and 25 anther-derived lines from two primary transformants Jingyin 119-B3 and Jingyin 119-B4 respectively which were obtained with Biolistics were analyzed. bar gene expressed normally in haploid and diploid plants as in their parents. PCR and Southern blot analysis of genomic DNA from some anther-derived lines indicated that bar gene was intact after anther culture and there was no change of integrationpatterns in most lines. Our results demonstrated that anther culture could be used for rapid production of transgene-homozygous lines from transgenic rice plants, even from transgenic plants with multi-locus insertion. System of transformation-anther culture-breeding and/or genetic study was also discussed.

Key words: anther culture, gene transfer, homozygous lines