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In Planta Agrobacterium-Mediated Transformation by Vacuum Infiltration

  • Brian W. Tague
  • Joanna Mantis
Part of the Methods in Molecular Biology™ book series (MIMB, volume 323)

Abstract

In planta Agrobacterium-mediated transformation using vacuum infiltration results in transgenic Arabidopsis thaliana without the use of sterile conditions or plant regeneration. Plants are grown in pots, in standard potting mix. Agrobacterium tumefaciens, carrying an appropriate plant transformation vector, is suspended in an infiltration medium that contains, at a minimum, sucrose and the surfactant Silwet L-77. Flower buds are immersed in the suspension of A. tumefaciens. The application of a vacuum drives the bacteria into the intercellular air spaces. A portion of the Agrobacterium Ti plasmid known as the T-DNA region, which has been engineered to carry a selectable marker, becomes integrated into the plant genomic DNA. Plants are allowed to set seed. Seeds are germinated in selective conditions to recover transformants.

Key Words

In planta transformation Agrobacterium tumefaciens Arabidopsis thaliana vacuum infiltration 

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Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Brian W. Tague
    • 1
  • Joanna Mantis
    • 1
  1. 1.Department of BiologyWake Forest UniversityWinston-Salem

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