01农杆菌介导的OsDREB转化芦荟的研究(赵老师20080311).doc

农杆菌介导OsDREB基因转化芦荟（Aloe vera L）的研究1,2，何聪芬2，秦允荣2，刘蕾2，钟秦11 (1. 南京理工大学化工学院, 南京 210094；2. 北京工商大学化学与环境工程学院，北京 100037) 摘要： 本研究通过农杆菌介导法将提高植物抗寒能力的水稻OsDREB基因转入芦荟基因组中并得到转基因芦荟，共培养后的芦荟外植体在含10~15 mg L-1G418筛选剂的MS培养基上经多次筛选和分化获得了一定数量的抗性再生芽对移栽成活的抗性再生植株进行PCR检测。OsDREB基因采用电导率测定的方法检测转化DREB基因对芦荟抗逆性的影响，以低温逆境对转化DREB基因得到的芦荟再生植株进行胁迫处理，同时观察转基因植株的生长发育和抗性变化，发现对照芦荟植株的叶片显褐色干枯半透明，出现严重冻伤的迹象，而转化植株生长情况较好，且转化植株的相对电导率与对照植株相比明显较低。结果表明两个外源基因已成功整合入芦荟基因组DNA中并得到稳定表达。Study on Aloe Transformation of OsDREB Gene Mediated by Agrobacterium Tumerfaciens Zhao Hua1,2, He Cong-fen2, Qin Yun-rong2, Liu Lei2, Zhong Qin1 1. School of Chemical Engineering, Nanjing University of Science and Technology, Nanjing 210094; 2. College of Chemistry and Environment Engineering, Beijing Technology and Business University, Beijing, 100037 Abstract: Aloe has been deeply researched recently because of its attracting function on medical treatment, hairdressing, health protection and edibility. It was investigated that DREB transcription factor were able to improve crop the tolerance to abiotic and biotic stresses. In this experiment, aloe stems were used as explants for OsDREB gene transformation mediated by Agrobacterium. PCR analyses demonstrated that OsDREB gene was successfully transferred into aloe genome with a transformation efficiency of 0.5%. Relationship between relative electrical conductivity and cold resistance was studied through positive and negative plants treated with the same low-temperature.The results of relative electrical conductivity in positive plants were higher than the negative plants. And the leaves of negative plants appear severe evidence of freeze injury with brown, withered and translucent, as the positive plants appear good growing condition. 芦荟是较重要的一种亚热带经济作物，具有药用价值、美容功效和食品营养价值。联合国粮农组织（FAO）曾把芦荟誉为“21世纪人类的最佳保健品”[] 。随着芦荟产业的不断发展，芦荟产量的要求也越来越高。然而芦荟抗旱不抗寒，不利于芦荟的大规模种植。由于芦荟开花不结实或结实率极低，无法采用传统的育种方法来增强其抗逆性，因此需要基因工程的方法对其进行改良。 基因工程技术在改良作物抗性上的广泛应用，为增强植物抗逆性提供了新的手段。OsDREB基因是利用酵母单杂交技术从水稻基因组中克隆的转录因子基因，该类基因编码的蛋白能识别启动子区带