Wheat (Triticum aestivum L.)
This chapter describes a procedure for Agrobacterium-mediated wheat transformation. Freshly isolated immature embryos, precultured immature embryos, or embryogenic calli are inoculated with a disarmed A. tumefaciens strain C58 (ABI) harboring the binary vector pMON18365 containing the β-glucuronidase (GUS) gene with an intron, and a selectable marker, neomycinphosphotranferase (NPT) II gene. The inoculated explants are selected on callus induction medium with the selective agent G418 for approximately 2 wk. The resistant callus pieces that develop are then transferred onto medium with the selective agent and reduced plant growth regulators for plant regeneration and further selection. It takes approximately 2.5 to 3 mo from inoculation to the establishment of R0 plants in soil. All the transformants should be morphologically normal and set seeds. Approximately 35% of the transgenic plants have a single copy of the transgene based on data obtained from previous experiments.
Key WordsAgrobacterium tumefaciens wheat Triticum aestivum L. plant transformation monocotyledon