中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

提高农杆菌介导转化水稻效率的因素

李双成;王世全;尹福强;邹良平;起登风;江洪;李平   

  1. 四川农业大学 水稻研究所， 四川 温江 611130； *通讯联系人， E-mail：liping@cngk.com
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-05-10 发布日期:2005-05-10

Some Factors Affecting Agrobacterium-Mediated Transformation Frequency in Rice

LI Shuang-cheng , WANG Shi-quan , YIN Fu-qiang , ZOU Liang-ping , QI Deng-feng , JIANG Hong , LI Ping
  

  • Received:1900-01-01 Revised:1900-01-01 Online:2005-05-10 Published:2005-05-10

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摘要： 以3个籼稻品种和2个粳稻品种为对象，对农杆菌转化水稻过程中影响转化效率的因素进行了研究。结果表明，菌株AGL1和EHA105按一定比例混合共转化和转化前菌体重悬对抗性愈伤率有显著影响；琼脂粉加倍和超净工作台上风干4 h的方法因能明显提高抗性愈伤率和分化率而被认为是最适合的干燥培养方式；分化培养基中加入二甲基亚砜（DMSO）或脯氨酸（Pro）可以提高分化率；壮苗时加入适量的NAA有利于壮根并提高移栽成活率。应用此农杆菌转化系统，获得了一批经PCR和点杂交鉴定的转基因植株。

关键词: 水稻, 农杆菌转化法, 共转化, 干燥培养, 分化, 生根

Abstract: Several important factors affecting rice transformation mediated by Agrobacterium were investigated with three indica and two japonica rice varieties. Co-transformation with Agrobacterium strains AGL1 and EHA 105, and by re-suspending the strains with NB culture media before transformation could improve the resistant calli rate significantly. Because of distinct influence on both the resistant calli rate and the differentiation rate, doubling the agar powder and being blew on the worktable for 4 h were considered as suitable approaches for desiccation culture. Adding DMSO or proline into the differentiation media, and NAA to the rooting media could, to some extent, improve the regeneration rate and the surviving rate, respectively. Many useful transgenic plants identified by PCR and Dot-blotting were obtained by using this protocol.

Key words: rice, Agrobacterium-mediated transformation, co-transformation, desiccation culture, differentiation, rooting