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植物研究 ›› 2001, Vol. 21 ›› Issue (3): 437-443.

• 论文 • 上一篇    下一篇

影响根癌农杆菌介导水稻转化的因素分析

尹鸿瑛, 安韩冰, 安利佳   

  1. 大连理工大学生物工程系, 大连 116012
  • 收稿日期:2001-04-16 出版日期:2001-09-15 发布日期:2016-06-14
  • 作者简介:尹鸿瑛(1976-), 女, 博士研究生, 主要从事植物生理、分子生物学, 组织培养研究。

ANALYSIS OF FACTORS AFFECTING THE EFFICIENCY OF AGROBACTERIUM-MEDIATED RICE TRANSFORMATION

YIN Hong-ying, AN Han-bing, AN Li-jia   

  1. Bioenqineering Department, Dalian University of Technology, Dalian, 116012
  • Received:2001-04-16 Online:2001-09-15 Published:2016-06-14

摘要: 根癌农杆菌与来自水稻成熟种子盾片的愈伤组织共培养, 将GUS 基因导入水稻愈伤组织, 并获得了转基因植株。通过比较影响根癌农杆菌转化频率的各种因素, 表明激素配比为2, 4-D 1mg/L、TDZ 0.5mg/L、NAA 1mg/L 时, 可以大大促进籼稻愈伤组织的分化能力;酚类化合物的加入使农杆菌的转化频率提高8.9%~23.5%;共培养时农杆菌的稀释方式及适当调整潮霉素(hygB)的使用浓度影响到农杆菌的转化频率。

关键词: 水稻愈伤组织, 植株再生, 根癌农杆菌, 转化频率

Abstract: The rice calli were infected and transformed by A.tumefaciens strain LBA4404 harboring Ti plasmid pTOK233. After continuous hygB selection, stable hygromycin-resistant, GUS-positive calli and plants were recovered. Some putative transgenic R0 plants were conducted with molecular assays, the results indicated that foreign genes had been stably integrated in the genome of transgenic rice. The results suggested that the ingredients of holmen of 2, 4-D 1mg/L、TDZ 0.5mg/L、NAA 1mg/L were efficient for Agrobacterium-mediated transformation of rice, besides, the addition of AS increased the efficiency of transformation by 8.9%~23.5%. At the same time, we found that both the dilution of agrobacteria and the adjusting of the concentration of hygB affected the efficiency of transformation.

Key words: rice calli, transgenic rice, A.tumefaciens, frequency of transformation