露地菊脚芽农杆菌真空渗透转化技术体系的建立  

Establishment of vacuum infiltration and transformation technique of Agrobacterium in Chrysanthemum morifolium foot buds

作　　者：王羽晗 李子豪 李世彪 陈驰航 王瑜麟 吴雅妮[1] 张旸[1] WANG Yu-han;LI Zi-hao;LI Shi-biao;CHEN Chi-hang;WANG Yu-lin;WU Ya-ni;ZHANG Yang(College of Life Science,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《南方农业学报》2018年第11期2236-2241,共6页Journal of Southern Agriculture

基　　金：国家科技支撑计划项目(2013AA102706);大学生创新创业训练计划项目(201710225302)

摘　　要：【目的】建立露地菊脚芽农杆菌真空渗透转化技术体系,为菊花品质改良提供参考依据。【方法】采用农杆菌真空渗透和浸染转化法对露地菊火焰的脚芽进行目的基因遗传转化,分析其PCR结果,以抗性分化苗叶片进行β-葡萄糖苷酸酶(β-glucuronidase,GUS)组织染色验证转基因植株,分析农杆菌真空渗透转化率。【结果】将含AtLFY基因和AtCO基因的表达载体分别转入露地菊根部脚芽中,通过PCR检测GUS染色鉴定后,真空渗透1次的平均转化率为11.97%,真空渗透2次的平均转化率为15.86%,菌液浸染10 min的平均转化率为3.64%,菌液浸染20 min的平均转化率为11.14%,总体嵌合率为14.58%。【结论】建立的露地菊脚芽农杆菌真空渗透转化技术体系对露地菊的遗传转化效率高,可操作性强。【Objective】The objective of this study was to establish a vacuum infiltration transformation technology system of Agrobacterium in Chrysanthemum morifolium foot buds in open field and provide reference for improving the quality of Chrysanthemum.【Method】Target genes of C.morifolium Flame foot buds were genetically transformed by Agrobacterium infiltration and transformation,and the results of PCR were analyzed.Gus-stained leaves of resistance differentiation seedlings were used to verify transgenic plants,and Agrobacterium transformation rate under vacuum infiltration was studied.【Result】The expression vectors containing AtLFY and AtCO genes were respectively transferred into the foot buds of C.morifolium.After being identified by PCR andβ-glucuronidase(GUS)staining,the transformation rate of vacuum infiltration for once was 11.97%on average,transformation rate of vacuum infiltration for twice was 15.86%on average.The average transformation rate was 3.64%after 10 min of dipping in bacteria liquid,the average transformation rate was 11.14%after dipping in bacteria liquid for 20 min,and the overall chimerism rate was 14.58%.【Conclusion】Therefore,the established Agrobacterium vacuum infiltration and transformation method for C.morifolium is with high genetic transformation efficiency and strong operability.

关 键 词：露地菊 农杆菌 真空渗透转化技术 浸染 脚芽 遗传转化 

分 类 号：S682.1[农业科学—观赏园艺]