逆境诱导转录因子AtDREB2A转化百合的研究  

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

作　　者：尚爱芹[1] 高永鹤[1] 段龙飞[1] 杨利平[2] 

机构地区：[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　　处：《园艺学报》2014年第1期149-156,共8页Acta Horticulturae Sinica

基　　金：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

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。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 kanamycin- resistant 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 ~C ） . 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.

关 键 词：百合 抗热性 转录因子 AtDREB2A 遗传转化 

分 类 号：S682.2[农业科学—观赏园艺]