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

  • Genetic Transformation and Hybridization
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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.

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Abbreviations

BAP:

Benzylaminopurine

GFP:

Green fluorescence protein

GFP+ embryo:

Embryo expressing GFP

GFP− embryo:

Embryo not expressing GFP

nos:

Nopaline synthetase

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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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Authors and Affiliations

  1. Department of Genetics and Biotechnology, Danish Institute of Agricultural Sciences, Research Centre Flakkebjerg, 4200, Slagelse, Denmark

    Inger Bæksted Holme, Henrik Brinch-Pedersen, Mette Lange & Preben Bach Holm

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  1. Inger Bæksted Holme
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  2. Henrik Brinch-Pedersen
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  3. Mette Lange
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  4. Preben Bach Holm
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Correspondence to Inger Bæksted Holme.

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Communicated by W. Harwood

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Holme, I.B., Brinch-Pedersen, H., Lange, M. et al. Transformation of barley (Hordeum vulgare L.) by Agrobacterium tumefaciens infection of in vitro cultured ovules. Plant Cell Rep 25, 1325–1335 (2006). https://doi.org/10.1007/s00299-006-0188-4

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  • DOI: https://doi.org/10.1007/s00299-006-0188-4

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