草地学报

›› 1996, Vol. 4 ›› Issue (2): 116-120.DOI: 10.11733/j.issn.1007-0435.1996.02.005

• 研究论文 • 上一篇    下一篇

发根农杆菌介导的百脉根的基因转化

陈燕1, 李聪1, 苏加楷1, 黄文惠1, 吕德杨2, 赵广宇2   

  1. 1. 中国农业科学院畜牧研究所, 北京100094;
    2. 中国科学院遗传研究所, 北京100012
  • 出版日期:1996-05-15 发布日期:2012-07-11
  • 基金资助:
    国家科委863计划

Agrobacterium rhizogenes Mediated Gene Transfer of the Forage Legume Lotus Corniculatus L

Chen Yan1, Li Cong1, Su Jiakai1, Huang Wenhui1, Lü Deyang2, Zhao Guangyu2   

  1. 1. Animal Science Institute of CAAS, Beijing 100094;
    2. Genetic Institute of Academia Sinica, Beijing 100012
  • Online:1996-05-15 Published:2012-07-11

PDF

摘要： 本文报道了豆科植物简单有效的转化再生实验系统。百脉根子叶外植体被含发根性Ri质粒载体的发根农杆菌感染,孩载体带有一个npt-Ⅱ基因和一个甘露碱合成酶基因。在含卡那霉素的筛选培养基上,外值体能诱导出毛根,并不断生长,毛根诱导频率为45.5%。切取毛根根段在筛选培养基上培养,毛很能膨大,并分化再生成苗和植株,其再生率为77.9%。抗性植株形态正常,浅层根系发达。DNA斑点杂交与Southern杂交证明外源基因已整合到百脉根基因组上。甘露碱测定结果表明外源甘露碱合成酶基因已在百脉根植株水平上表达。

关键词: 发根农杆菌, 基因转化, 百脉根, 再生植株

Abstract: This paper reports a simple and effective experimental system for genetranefer and regenera tion of leguminous plants.Cotyledon explants of Lotus corniculatus (Leo) was infected with Agrobacterium rhizogenes containing Ri plasmid carring a kanamycin resistant gene npt-II and a mannopine synthase gene.Trans formed hairy roots were produced and grew on kanamycin containing medium.The trans formation fre quency was 45.5 %.Trans formed Lotus corniculatus plants were recovered from A.rhizogenes-induced hairy roots segments.The regeneration frequency was 77.9%.These transgenic plants were characterized by nor mal leaf morphology and stem growth but a root system that was shallow and more extensive than normal.DNA Dot blot and Southern bIot confirmed the presence of npt-II gene from Ri plasmid in the plant genome.Mannopine was detected in transformed hairy roots and regernerated plants.

Key words: Lotus corniculatus L, Gene transformation, Plant regeneration, Agrobacterium rhizogenes