Gasic Ksenija, Korban Schuyler S
Department of Natural Resources & Environment Sciences, University of Illinois, Urbana, IL, USA.
Methods Mol Biol. 2006;343:281-9. doi: 10.1385/1-59745-130-4:281.
All economically important Brassica species have been successfully transformed using Agrobacterium tumefaciens. Although different tissues have been used as explants, hypocotyls remain the most desirable explants for Brassica tissue culture owing to their amenability to regeneration. Young explants excised from 3- to 4-d-old seedlings have exhibited optimal regeneration potential; the addition of adjuvants such as silver nitrate to the selection medium is necessary to achieve high efficiency of transformation. This chapter describes an Agrobacterium-mediated transformation protocol for Indian mustard based on inoculation of hypocotyls. The selectable marker gene used encodes for neomycin phosphotransferase II (nptII), and the selection agent is kanamycin.
所有具有重要经济价值的芸苔属物种都已通过根癌农杆菌成功转化。尽管不同的组织都被用作外植体，但由于下胚轴易于再生，因此仍然是芸苔属组织培养中最理想的外植体。从3至4日龄幼苗上切下的幼嫩外植体具有最佳的再生潜力；向选择培养基中添加诸如硝酸银之类的佐剂对于实现高效转化是必要的。本章描述了一种基于下胚轴接种的用于印度芥菜的农杆菌介导的转化方案。所使用的选择标记基因编码新霉素磷酸转移酶II（nptII），选择剂是卡那霉素。
