アグロバクテリウム・ツメファシエンスを介した形質転換によるトランスジェニックトマト系統の作出と選抜のためのプロトコル

Here, we present a protocol for generating and selecting stable transgenic tomato lines through Agrobacterium tumefaciens-mediated transformation of cotyledon and hypocotyl explants. We describe steps for sterilizing and planting tomato seeds, cotyledon and hypocotyl excision and preculture, Agrobacterium tumefaciens preparation and co-cultivation with explants, and recovering explants and bacterial overgrowth restriction. We then detail procedures for selecting green calli; regenerating explant excision and growth in non-selective media; plant hardening, potting, and growing; and T-DNA insertion confirmation by PCR. For complete details on the use and execution of this protocol, please refer to Mountourakis et al.^1,2,3.