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园艺学报 ›› 2014, Vol. 41 ›› Issue (1): 149-156.

• 研究报告 • 上一篇    下一篇

逆境诱导转录因子AtDREB2A 转化百合的研究

尚爱芹1,*,高永鹤1,段龙飞1,杨利平2   

  1. 1 河北农业大学园艺学院,河北保定 071001;2 长江师范学院生命科学与技术学院,重庆 408100
  • 出版日期:2014-01-25 发布日期:2014-01-25
  • 基金资助:

    河北省自然科学基金项目(C2008000265);2011 年度河北省博士后科研项目择优资助项目

Studies on Transformation of Lily with Dehydration Responsive Element Binding Transcription Factor AtDREB2A

SHANG Ai-qin1,*,GAO Yong-he1,DUAN Long-fei1,and YANG Li-ping2   

  1. 1College of Horticulture,Agricultural University of Hebei,Baoding,Hebei 071001,China;2College of Life Science and
    Technology,Yangtze Normal University,Chongqing 408100,China
  • Online:2014-01-25 Published:2014-01-25

摘要: 以东方百合‘西伯利亚’为试验材料,探讨影响农杆菌转化效率的因素,优化了以无菌小
鳞片为外植体的遗传转化体系,通过农杆菌介导法将逆境诱导转录因子AtDREB2A 基因转入百合基因组
中。结果表明:外植体预培养3 d,侵染菌液浓度OD600 = 0.8,侵染时间20 min,25 ℃下共培养3 d,获
得的卡那霉素阳性植株率最高。对获得的59 株抗性单株进行PCR 检测,有28 个单株的DNA 经扩增后
产生了与阳性对照相同的特异扩增带。进一步对转基因阳性植株进行Southern 杂交鉴定,结果表明外源
基因已经整合到转化植株的基因组中。生理指标测定表明转基因植株的对高温的适应性有一定程度的提
高。

关键词: 百合, 抗热性, 转录因子, AtDREB2A, 遗传转化

Abstract: In the study,by investigating the factors influencing the transformation efficiency of Lilium
‘Siberia’,a genetic transformation system of scales was optimized. By Agrobacterium-mediated,the
AtDREB2A gene was transformed into‘Siberia’. The results indicated that the highest rate of kanamycinresistant
plants was obtained through the following transformation procedure:After preculture for 3 d,the
explants were infected for 20 min with diluted OD600 = 0.8 Agrobacterium tumefaciens solution and then
co-cultivated for 3 d(under 25 ℃). Molecular detection showed 59 transformed lines of‘Siberia’were
detected by PCR analysis and 28 of them show the positive band same as objective band(AtDREB2A).
The fragment was 402 bp approximately. It preliminarily suggested that the exogenous gene was integrated
into the genome of‘Siberia’. Determination of physiological indexes showed that the transgenic plants
enhanced the tolerance to high temperature.

Key words: lily, heat resistance, transcription factor, AtDREB2A, genetic transformation

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