农杆菌介导的生菜（Lactuca sativa）转化。

Transformation of lettuce (Lactuca sativa) mediated by Agrobacterium tumefaciens.

机构信息

Department of Vegetable Crops, University of California, 95616, Davis, CA, USA.

出版信息

Plant Cell Rep. 1987 Dec;6(6):439-42. doi: 10.1007/BF00272777.

Abstract

Lactuca sativa can be routinely transformed using Ti plasmids of Agrobacterium tumefaciens containing a chimeric kanamycin resistance gene (NOS.NPTII.NOS). Critical experimental variables were plant genotype, bacterial concentration, presence of a nurse culture and timing of transfers between tissue culture media. Transformation was confirmed by the ability to callus and root in the presence of kanamycin, nopaline production, and by hybridization in Southern blots. Transformation has been achieved with several Ti vectors. Several hundred transformed plants have been regenerated. Kanamycin resistance was inherited monogenically. Homozygotes can be selected by growing R2 seedlings on media containing G418.

摘要

莴苣可使用含有嵌合卡那霉素抗性基因（NOS.NPTII.NOS）的根瘤农杆菌 Ti 质粒进行常规转化。关键的实验变量包括植物基因型、细菌浓度、是否存在护士培养物以及组织培养基之间的转移时间。通过在卡那霉素存在下产生愈伤组织和生根、产生胭脂碱和 Southern 印迹杂交来确认转化。已经使用几种 Ti 载体实现了转化。已经再生了几百株转化植物。卡那霉素抗性是单基因遗传的。通过在含有 G418 的培养基上培养 R2 幼苗可以选择纯合子。