橡胶树HbFCA启动子在转基因烟草中的功能缺失分析  

5' Deletion Analysis of HbFCA Promoter from Hevea brasiliensis in Transgenic Tobacco

作　　者：李季[1] 戴雪梅[1] 孙芳[1] 黄天带[1] 华玉伟[1] 黄华孙[1] 

机构地区：[1]中国热带农业科学院橡胶研究所/农业部橡胶树生物学重点开放实验室,海南儋州571737

出　　处：《热带农业科学》2017年第7期62-66,74,共6页Chinese Journal of Tropical Agriculture

基　　金：海南省自然科学基金"橡胶树內源组成型启动子的构建及其表达分析研究"(No.20153089);海南省重点研发计划"一种植物遗传转化新型报告基因的发掘及在橡胶树的应用研究"(No.ZDYF2016220)

摘　　要：利用5′缺失设计一系列引物,扩增HbFCA启动子质粒,获得5个启动子缺失片段,用这些片段分别替换PBI121载体中的CaMV35S启动子,得到5个HbFCA的5′缺失表达载体,即pA、pB、pC、pD和pE。利用农杆菌介导叶盘法进行烟草遗传转化,对组培瓶内生根较好的烟草叶片GUS染色,结果发现,长度为最小片段的276 bp的pE可以较好地实现HbFCA启动子的功能。对橡胶树体细胞胚瞬时表达检测发现,pE片段也能启动表达,有较好的GUS活性,可以实现HbFCA启动子的功能。Using the 5′ end serial deletion analysis method, five HbFCA promoter deletion fragments which had different distances from transcription start site were amplified by PCR, and they were used to replace the CaMV35S promoter of PBI121 carrier to obtain 5 HbFCA 5′deletion expression vectors, i.e. pA, pB, pC, pD and pE. The expression vectors were transformed into tobacco by using agrobacterium-meditated leaf disc method, and the well-rooted transformed tobacco plants in tissue culture vessels were directly detected with GUS staining. The results showed that the pE 267 bp, the shortest among the fragments, had promoter activity. The transient expression analysis of somatic embryo of rubber tree showed that the fragment pE also had better GUS staining, and could function as a Hb FCA promoter.

关 键 词：橡胶树 HbFCA 启动子 5′端缺失分析法 核心结构域 

分 类 号：S794.1[农业科学—林木遗传育种]