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Plant Cell Reports

, Volume 25, Issue 12, pp 1325–1335 | Cite as

Transformation of barley (Hordeum vulgare L.) by Agrobacterium tumefaciens infection of in vitro cultured ovules

  • Inger Bæksted Holme
  • Henrik Brinch-Pedersen
  • Mette Lange
  • Preben Bach Holm
Genetic Transformation and Hybridization

Abstract

We report on a novel transformation procedure for barley by Agrobacterium infection of in vitro cultured ovules. Ovules of the cultivar Golden Promise were isolated a few hours after pollination and infected with the Agrobacterium tumefaciens strain AGL0 carrying the binary vector pVec8-GFP. The vector harboured a hygromycin resistance gene and the green fluorescence protein (GFP) gene. GFP-expressing embryos were isolated from the ovules, regenerated to plants and investigated by Southern blot analysis. Transformation frequencies amounted to 3.1% with hygromycin selection and 0.8% without selection. Mendelian inheritance and stable expression of the GFP gene was confirmed in 18 independent lines over two generations. We conclude that the described technique allows for the rapid and direct generation of high quality transgenic plants.

Keywords

Agrobacterium tumefaciens Barley ovule culture GFP-expression Transformation Zygote 

Abbreviations

BAP

Benzylaminopurine

GFP

Green fluorescence protein

GFP+ embryo

Embryo expressing GFP

GFP embryo

Embryo not expressing GFP

nos

Nopaline synthetase

Notes

Acknowledgements

The authors thank Ole Bråd Hansen and Lis Bagnkop Holte for skillful technical assistance. Michael Hansen, Department of Plant Biology, the Royal Veterinary and Agricultural University, Denmark, is thanked for performing the confocal microscopy. This work was funded by a grant from the Danish Research Council.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Inger Bæksted Holme
    • 1
  • Henrik Brinch-Pedersen
    • 1
  • Mette Lange
    • 1
  • Preben Bach Holm
    • 1
  1. 1.Department of Genetics and BiotechnologyDanish Institute of Agricultural Sciences, Research Centre FlakkebjergSlagelseDenmark

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