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A rapid and efficient transformation protocol for the grass Brachypodium distachyon

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

A fast and efficient microprojectile bombardment-mediated transformation protocol is reported for the grass species Brachypodium distachyon, a proposed alternative model plant to Oryza sativa for functional genomics in grasses. Embryogenic calli derived from immature embryos were transformed by a construct containing the uidA (coding for β-glucuronidase) and bar (coding for phosphinothricin acetyl transferase) genes, and bialaphos, a non-selective herbicide, was used as the selection agent throughout all phases of the tissue culture. Average transformation efficiencies of 5.3% were achieved, and for single bombardments transformation efficiencies of up to 14% were observed. The time frame from the bombardment of embryogenic callus to the harvesting of transgenic T1 seeds was 29 weeks and 25 weeks for the diploid and two tetraploid accessions used, respectively. Since the seed-to-seed life cycle is 19 weeks for the diploid and 15 weeks for the tetraploid accessions, our B. distachyon transformation system allows testing of both the T0 and the T1 generation as well as production of T2 seeds within 1 year.

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Abbreviations

BAP:

Benzylaminopurine

2,4-D:

2,4-Dichlorophenoxyacetic acid

GUS:

β-Glucuronidase

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Acknowledgements

We thank Julia Kinane (Risø National Laboratory, Denmark) and Niels Roulund (DLF-Trifolium A/S, Denmark) for help on the statistical analyses. A special thanks to the group of Dr. John Draper for their kind assistance during the 4-month stay of Pernille Christiansen at the Institute of Biological Science, University of Wales, UK.

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Authors and Affiliations

  1. Risø National Laboratory, Frederiksborgvej 399, P.O. Box 49, 4000, Roskilde, Denmark

    Pernille Christiansen & Thomas Didion

  2. DLF-Trifolium A/S, Research Division, Højerupvej 31, 4660, Store Heddinge, Denmark

    Claus Henrik Andersen, Marianne Folling & Klaus Kristian Nielsen

Authors
  1. Pernille Christiansen
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  2. Claus Henrik Andersen
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  3. Thomas Didion
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  4. Marianne Folling
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  5. Klaus Kristian Nielsen
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Corresponding author

Correspondence to Pernille Christiansen.

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Communicated by J.M. Widholm

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Christiansen, P., Andersen, C.H., Didion, T. et al. A rapid and efficient transformation protocol for the grass Brachypodium distachyon. Plant Cell Rep 23, 751–758 (2005). https://doi.org/10.1007/s00299-004-0889-5

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  • DOI: https://doi.org/10.1007/s00299-004-0889-5

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