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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (18): 213-218.

所属专题： 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

根癌农杆菌介导ACO基因对亚洲百合的转化

赵欢蕊 刘雅莉 王跃进 徐伟荣   

  • 收稿日期:2010-07-19 修回日期:2010-07-31 出版日期:2011-09-20 发布日期:2011-09-20
  • 基金资助:
    国家自然基金资助项目

  • Received:2010-07-19 Revised:2010-07-31 Online:2011-09-20 Published:2011-09-20

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摘要： 以亚洲百合‘普利安娜’的鳞块为基因转化的受体材料，利用根癌农杆菌介导法将ACO基因导入百合，研究了影响其转化的因素。结果表明，以鳞块为受体材料，外植体预培养0天的污染率最低，有利于农杆菌的侵染；菌液浓度OD600=0.8左右时侵染30 min效果较好；重悬后菌液活化1~2 h有利于抗性芽的分化；MS重悬液和共培养基中均添加20 mg/L乙酰丁香酮（AS）可提高抗性芽分化率；去除大量元素中的NH4NO3可提高转化率；抗性植株经PCR检测部分呈现阳性，初步证明外源基因已整合到百合基因组中。

关键词: 闽北、梨、现状、对策, 闽北、梨、现状、对策

Abstract: By using bulbous tubers of ‘Pollyanna’ as the gene transformation acceptors, the ACO gene was transferred into lily with Agrobacterium tumefaciens-mediated transformation. The factors which affected transformation were studied. The results suggested that the minimum contamination rate was gotten when the bulbous tubers were used as acceptors. It was better for Agrobacterium tumefaciens instrusion with OD600=0.8 in 30 min. Agrobacterium tumefaciens which was activated for 1-2 h tended to differentiate resistant plants. The differentiation rate of resistant plants could be increased by adding into 20 mg/L AS in MS liquid and solid co-cultivation medium. NH4NO3-free medium could improve the transformation rate. PCR screening showed that some Kanamycin resistant plants were positive, which indicated preliminarily that the ACO gene had integrated into Lily’s genome.