烟草叶片直接再生—基因转化体系的建立

Establishment of Tobacco Leaf Direct-Regeneration- Gene Transformation System

烟草叶片在附加激素IAA与BA的MS培养基上表现出不同的分化途径和分化频率,不同浓度的IAA、BA配比,可分别导致愈伤组织的形成,根、芽和胚状体的直接分化。在MS + 0 .1mg/LIAA + 0 .5mg/LBA的B处理培养基上,叶片外植体边缘细胞经器官发生途径高频率直接再生植株,而该分化过程受到卡那霉素的强烈抑制,因而可以做为共培养法进行烟草基因转化的良好体系。

Different differentiation way and frequency were discovered when disc leaf of tobacco cv. K326 were cultured on MS medium supplemented with different concentration of IAA and BA. High concentration of IAA/BA could induce callus and root formation, while low concentration of IAA/BA was benefit for shoot formation. Somatic embroys were discovered on MS medium with 0.1 mg/L BA and 0.1 mg/L IAA.High frequency of direct-regeneration were induced from the edge of disc leaf on MS medium with 0.1 mg/L IAA and 0.5 mg/L BA, and this differential process inhibited by 25-30 mg/L kanamycin and proved to be an effective system for tobacco gene transformation.