速生型刺槐遗传转化体系的建立  

ESTABLISHMENT OF GENETIC TRANSFORMATION SYSTEM OF FAST_GROWING BLACK LOCUST

作　　者：沈俊岭[1] 赵芳[1] 李云[1] 陈受宜[2] 田秀红[2] 

机构地区：[1]北京林业大学林木花卉遗传育种教育部重点实验室,北京100083 [2]中国科学院遗传与发育生物研究所,北京100101

出　　处：《核农学报》2006年第6期477-481,共5页Journal of Nuclear Agricultural Sciences

基　　金：国家林业局重点推广项目(2003-5-2)

摘　　要：为获得稳定的转基因刺槐植株，以速生型刺槐叶轴为转化受体，对转化过程中的一些影响因素进行了探究，建立了较为完善的遗传转化体系。结果表明：继代20～35d之间刺槐叶轴最适宜转化；卡那霉素的筛选浓度为30mg／L；头孢霉素的抑菌浓度为200mg／L；合适的侵染时间是20min；30mg／L的乙酰丁香酮有利于抗性芽的发生；共培养2d对转化适宜；抑菌7d后再加选择压有利于抗性芽的产生。最后得到的抗性植株经PCR检测，初步证明外源目的基因OsDREB1已整合到刺槐基因组中。The rachis of fast-growing Black Locust was used as explants in the study. The faetors that affected transformation ratio were studied, and a stable transformation system was established. The results suggested that germinated shoots which 20～35dsdd in vitro were suitable for transgenic. The optimal concentration of kanamycim to the rachis was 30mg/L and that of cefotaxine was 200mg/L. The ideal time for rachis dipping in Agrobacterium was 20min and optimal co-cultivation was 2 days. 30mg/L acetosyringone could increase the transformation ratio. The selection of the inoculated rachis was done at 7^th days of bacterial inhibiting treatment by cefotaxine. The PCR analysis showed that the target gene OsDREB 1 was integrated into the genome of black locust.

关 键 词：刺槐 叶轴 农杆菌 遗传转化 

分 类 号：S792.27[农业科学—林木遗传育种]