Floral dip: a simplified method for Agrobacterium ‐mediated transformation of Arabidopsis thaliana

Floral dip: a simplified method for Agrobacterium ‐mediated transformation of Arabidopsis thaliana Summary The Agrobacterium vacuum infiltration method has made it possible to transform Arabidopsis thaliana without plant tissue culture or regeneration. In the present study, this method was evaluated and a substantially modified transformation method was developed. The labor‐intensive vacuum infiltration process was eliminated in favor of simple dipping of developing floral tissues into a solution containing Agrobacterium tumefaciens , 5% sucrose and 500 microliters per litre of surfactant Silwet L‐77. Sucrose and surfactant were critical to the success of the floral dip method. Plants inoculated when numerous immature floral buds and few siliques were present produced transformed progeny at the highest rate. Plant tissue culture media, the hormone benzylamino purine and pH adjustment were unnecessary, and Agrobacterium could be applied to plants at a range of cell densities. Repeated application of Agrobacterium improved transformation rates and overall yield of transformants approximately twofold. Covering plants for 1 day to retain humidity after inoculation also raised transformation rates twofold. Multiple ecotypes were transformable by this method. The modified method should facilitate high‐throughput transformation of Arabidopsis for efforts such as T‐DNA gene tagging, positional cloning, or attempts at targeted gene replacement. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Plant Journal Wiley

Floral dip: a simplified method for Agrobacterium ‐mediated transformation of Arabidopsis thaliana

The Plant Journal, Volume 16 (6) – Dec 1, 1998

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Publisher
Wiley
Copyright
Copyright © 1998 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0960-7412
eISSN
1365-313X
D.O.I.
10.1046/j.1365-313x.1998.00343.x
Publisher site
See Article on Publisher Site

Abstract

Summary The Agrobacterium vacuum infiltration method has made it possible to transform Arabidopsis thaliana without plant tissue culture or regeneration. In the present study, this method was evaluated and a substantially modified transformation method was developed. The labor‐intensive vacuum infiltration process was eliminated in favor of simple dipping of developing floral tissues into a solution containing Agrobacterium tumefaciens , 5% sucrose and 500 microliters per litre of surfactant Silwet L‐77. Sucrose and surfactant were critical to the success of the floral dip method. Plants inoculated when numerous immature floral buds and few siliques were present produced transformed progeny at the highest rate. Plant tissue culture media, the hormone benzylamino purine and pH adjustment were unnecessary, and Agrobacterium could be applied to plants at a range of cell densities. Repeated application of Agrobacterium improved transformation rates and overall yield of transformants approximately twofold. Covering plants for 1 day to retain humidity after inoculation also raised transformation rates twofold. Multiple ecotypes were transformable by this method. The modified method should facilitate high‐throughput transformation of Arabidopsis for efforts such as T‐DNA gene tagging, positional cloning, or attempts at targeted gene replacement.

Journal

The Plant JournalWiley

Published: Dec 1, 1998

References

  • Stable genetic transformation of Arabidopsis thaliana by Agrobacterium inoculation
    Chang, Chang; Park, Park; Kim, Kim; Kang, Kang; Kim, Kim; Nam, Nam
  • RPS2 , an Arabidopsis disease resistance locus specifying recognition of Pseudomonas syringae strains expressing the avirulence gene
    Kunkel, Kunkel; Bent, Bent; Dahlbeck, Dahlbeck; Innes, Innes; Staskawicz, Staskawicz
  • Differences in susceptibility of Arabidopsis ecotypes to crown gall disease may result from a deficiency in T‐DNA integration.
    Nam, Nam; Matthysse, Matthysse; Gelvin, Gelvin
  • Foreign genes in plants: transfer, structure, expression and applications.
    Weising, Weising; Schell, Schell; Kahl, Kahl

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