Protocol for generating and selecting transgenic tomato lines through Agrobacterium tumefaciens-mediated transformation

Abstract

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 ex-plants 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 Mountourak is et al.1-3

Published in

STAR Protocols
2026, volume: 7, number: 1, article number: 104322
Publisher: CELL PRESS

SLU Authors

• Moschou, Panagiotis Nikolaou

  • Department of Molecular Sciences, Swedish University of Agricultural Sciences
    
  • Foundation for Research and Technology - Hellas (FORTH)
  • University of Crete

UKÄ Subject classification

Developmental Biology

Publication identifier

• DOI: https://doi.org/10.1016/j.xpro.2025.104322

Permanent link to this page (URI)

https://res.slu.se/id/publ/146000