一种不依赖于无菌培养的拟南芥活体转基因种子筛选方法

摘要

拟南芥活体转基因种子的筛选仍主要采用无菌培养技术,存在操作麻烦,容易污染而造成转基因材料损失等问题.本研究在传统组织培养筛选技术的基础上,通过去除原培养基成分中对种子发芽和筛选不必需的蔗糖和有机成分,降低大量元素的浓度,缩短筛选时间,从而免除了传统转基因种子筛选中培养基和种子消毒的繁琐步骤,建立了一种不依赖于无菌培养的转基因种子筛选方法.与无菌培养筛选比较,筛选效果相同,但简化了试验步骤,提高了筛选操作的可靠性.%Sterile culture is normally employed in screening in planta transformed seeds, which is both labor-intensive and contamination-prone, and material loss events often occur because of failure in sterilization. An aseptic culture free method was developed for screening Arabidopsis transgenic seeds from in planta transformation based on the traditional method by depleting sucrose and organic components unnecessary for seed germination and screening, reducing macronutrients and shortening the screening time. The method inherits all the advantages of the traditional method, but is simpler and more reliable in operation.