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林业科学 ›› 1995, Vol. 31 ›› Issue (2): 97-102.

• 论文及研究报告 •    下一篇

苏云金杆菌杀虫晶体蛋白基因转化美洲黑杨的研究

陈颖1,韩一凡1,田颖川2,李铃1,聂绍基3   

  1. 1. 中国林业科学研究院林业研究所 北京 100091
    2. 中国科学院微生物研究所 北京 100080
    3. 东北林业大学 哈尔滨 150040
  • 收稿日期:1993-10-21 出版日期:1995-03-25 发布日期:1995-03-25

STUDY ON THE PLANT REGENERATION FROM POPULUS DELTOIDES EXPLANT TRANSFORMED WITH BT. TOXIN GENE

Ying Chen1,Yifan Han1,Yingchuan Tian2,Ling Li1,Shaoji Nie3   

  1. 1. The Research Institute of Forestry CAF Beijing 100091
    2. Institute of Microbiology, Academia Sinica Beijing 100080
    3. Northeast Forestry University Harbin 150040
  • Received:1993-10-21 Online:1995-03-25 Published:1995-03-25

摘要:

本研究首先建立了美洲黑杨叶片外植体的再生系统, 并利用Horsch等人1985年发明的叶盘法, 将分别带有嵌合基因NPTⅡ和1.8kb或2.1kb Bt.基因的土壤杆菌LBA4404与美洲黑杨叶片共培养。采用30mg/l和50mg/l两种浓度的卡那霉素筛选转化子, 约28天左右的时间, 有不定芽从外植体切口处形成。两种卡那霉素浓度下共形成275个伸长芽, 存活了187株。这187株分别进入卡那霉素30mg/l, 50mg/l的生根培养基后, 生根率分别为9.62%和8.02%。PCR反应证明部分植株已成功地进行了转化。

关键词: 美洲黑杨, Bt.基因, 转化, PCR反应

Abstract:

The effective regenetation system for Populus deltoides was developed firstly in the experiment. Based on the system, Agrobacterium tumefaciens LBA 4404 strains hauboring the Bt. toxin gene expression vectors pB48.214 and pB48.215 respectively were uesd for transformation of poplar plants by means of the leaf disc method. After transformating, adventitious buds were formed at the cut of explant on the media with two concentrations of kanamycin 50mg/l and 30mg/l nearly one month later. Variance analysis showed that the effects of two kanamycin concentrations on the transformation rate and mean adventitious bud numbers were significant at p < 0.05. When shoots reached 1cm long, they were transferred to rooting media and supplemented with 30mg/l and 50mg/l kanamycin and the rooting percentage was 9.62% and 8.02% respectively. PCR analysis showed that Bt. genes were successfully transformed into Populus deltoides chromosomes.

Key words: Bt. gene, Populus deltoides, Transformation, PCR analysis