一种获得转基因芥菜的简便方法  

A Simple Method to Obtain Transgenic Mustard

作　　者：代法国[1] 胡宗利[1] 陈国平[1] 王炳琴[1] 周爽[1] 

机构地区：[1]重庆大学生物工程学院,中国重庆400030

出　　处：《生命科学研究》2011年第1期19-23,共5页Life Science Research

基　　金：重庆市攻关项目(CSTC;2009AB5174);重庆大学研究生创新团队建设项目(200909B1007)

摘　　要：随着转基因技术的不断革新,更简便高效的转基因方法逐渐发展起来.通过对花序浸染法的探索和改进,采取用移液枪点滴重悬液的方式转化芥菜(雪里蕻,Brassica juncea var.crispifolia)花蕾,最终获得了转基因芥菜.同时通过对影响转化效率的蔗糖浓度、表面活性剂浓度、菌液OD600值等进行研究,发现用5%的蔗糖(W/V)+0.03%的Silwet-77(V/V)重悬农杆菌,使重悬液的菌液浓度OD600为0.8时,可以获得较高的转化效率,转化率为2.3%.该方法避免了真空操作及浸染时间的影响,使得操作更加简便,且转化周期更短,为芥菜的遗传转化提供了一种更为简便的转化方法.With the constant innovation of transgenic technology,more simple and efficient transgenic methods developed gradually.Through the exploration and improvement of floral-dip,transgenic mustard（potherb mustard,Brassica juncea var.crispifolia） was obtained via performing direct drop-by-drop inoculation to every bud by using a pipette.Meanwhile,with the study on the factors which may affect the transformation efficiency,such as sucrose concentration,surfactant concentration,and the OD600 of the Agrobacterium inoculum,it showed that a higher transformation efficiency can be gotten by using 5% sucrose（W/V） ＋ 0.03% Silwet -77（V/V） resuspended Agrobacterium when the OD600 of the Agrobacterium inoculum was 0.8,and the transformation rate was 2.3%.The method avoids vacuum processing and influence of immerged time,which makes the manipulation simpler and the transformation period shorter,thus a more convenient method of transformation for the genetic transformation of mustard was provided.

关 键 词：芥菜 花序浸染 遗传转化 雪里蕻 根癌农杆菌 

分 类 号：Q943[生物学—植物学]