Transgenic Plants: Methods and Protocols pp 141-150

Part of the Methods in Molecular Biology™ book series (MIMB, volume 286) | Cite as

Organogenesis From Transformed Tomato Explants

  • Anne Frary
  • Joyce Van Eck

Summary

Tomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transformation and the technique has been useful for the isolation and identification of many genes involved in plant disease resistance, morphology and development. In this method, cotyledon explants from in vitro-grown seedlings are precultured overnight on a tobacco suspension feeder layer. The explants are then inoculated with Agrobacterium and returned to the feeder layer for a 2-d period of cocultivation. After cocultivation, the explants are transferred to an MS-based selective regeneration medium containing zeatin. Regenerated shoots are then rooted on a separate selective medium. This protocol has been used with several tomato cultivars and routinely yields transformation efficiencies of 10–15%.

Key Words

Agrobacterium tumefaciens biotechnology gene transfer genetic engineering genetic transformation Lycopersicon esculentum morphogenesis regeneration transgenic plants 

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Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • Anne Frary
    • 1
  • Joyce Van Eck
    • 2
  1. 1.Department of Molecular Biology and GeneticsIzmir Institute of TechnologyIzmirTurkey
  2. 2.The Boyce Thompson Institute for Plant ResearchIthaca

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