Protoplast Isolation, Transformation, and Regeneration for Forage and Turf Grasses

Forage and turf grasses are widely grown and contribute significantly to sustainable agriculture. This chapter describes a protocol for protoplast transformation and plant regeneration for major forage and turf grass species, including tall fescue, red fescue, meadow fescue, perennial ryegrass, and Italian ryegrass. Embryogenic calli induced from caryopsis were used to establish embryogenic cell suspension cultures. Protoplasts were isolated from embryogenic suspension cultures and used for direct gene transfer. Chimeric genes were introduced into protoplasts by polyethylene glycol treatment. Upon selection with antibiotics or herbicide, resistant calli were obtained and transgenic plants were regenerated from these calli.