四川农业大学学报 ›› 2008, Vol. 26 ›› Issue (01): 15-19.doi: 10.16036/j.issn.1000-2650.2008.01.009

• 研究论文 • 上一篇    下一篇

可去除选择标记的DREB基因双T-DNA载体共转化玉米

谭登峰, 韩兆雪, 曹墨菊, 潘光堂, 荣廷昭   

  1. 四川农业大学 玉米研究所, 四川 雅安 625014
  • 收稿日期:2008-02-19 出版日期:2008-03-31 发布日期:2017-02-28
  • 基金资助:
    四川省生物技术专项

Co-Transformating Maize with Double T-DNA Vector of DREB Gene

TAN Deng-feng, HAN Zhao-xue, Cao Mo-ju, Pan Guang-tang, Rong Ting-zhao   

  1. Maize Research Institute, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2008-02-19 Online:2008-03-31 Published:2017-02-28

摘要: 通过体外重组构建双T-DNA双元载体pCDMARpWDT-Hyg,选择标记基因hpt(hygromycin phospho-transferase)和抗逆转录因子基因DREB分别位于两个独立的T-DNA。用农杆菌介导法转化玉米胚性愈伤组织,通过在抗性培养基上严格筛选,在获得的再生转化植株中,同时整合hpt基因和DREB基因的共转化率达到26.3%。该转化系统的建立为下一步得到无选择标记的转基因植株奠定基础。此外,本实验还通过gus基因的瞬时表达分析了农杆菌转化玉米的影响因素。

关键词: 玉米, DREB基因, 双T-DNA载体, 农杆菌介导法, 瞬时表达

Abstract: The double T-DNA binary vector pCDMARpWDT-Hyg with two independent T-DNAs, one containing selective marker gene hpt (hygromycin phosphotransferase) and the other containing stress-resistance transcriptional factor gene DREB, was constructed by recombination in vitro and used to transform maize embryonic calli by agrobacterium-mediated method. After strict selection on resistant medium, the co-transformation ratio of the transformed regenerated plants both by hpt and DREB genes was as high as 26.3%. The establishment of the double T-DNA vector system provided fundamental data and materials for the development of marker-free transgenic maize plants. Moreover, we discussed the influencing factors to maize transformation by Agrobacterium-mediated method by gus gene transient expression.

Key words: maize, DREB, double T-DNA vector, Agrobacterium-mediated transformation, transient expression

中图分类号: 

  • S513.035