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Agrobacterium-mediated transformation and inbred line development in the model grass Brachypodium distachyon


Brachypodium distachyon (Brachypodium) has been proposed as a model temperate grass because its physical, genetic, and genome attributes (small stature, simple growth requirements, small genome size, availability of diploid ecotypes, annual lifecycle and self fertility) are suitable for a model plant system. Two additional requirements that are necessary before Brachypodium can be widely accepted as a model system are an efficient transformation system and homogeneous inbred reference genotypes. Here we describe the development of inbred lines from 27 accessions of Brachypodium. Determination of c-values indicated that five of the source accessions were diploid. These diploid lines exhibit variation for a variety of morphological traits. Conditions were identified that allow generation times as fast as two months in the diploids. An Agrobacterium-mediated transformation protocol was developed and used to successfully transform 10 of the 19 lines tested with efficiencies ranging from 0.4% to 15%. The diploid accession Bd21 was readily transformed. Segregation of transgenes in the T 1 generation indicated that most of the lines contained an insertion at a single genetic locus. The new resources and methodologies reported here will advance the development and utilization of Brachypodium as a new model system for grass genomics.

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2,4-D :

2,4-dichlorophenoxyacetic acid


Linsmaier and Skoog basal medium


Murashige and Skoog salts and vitamins


callus inducing medium


  1. Arumuganathan K, Earle ED, (1991) Estimation of nuclear DNA content of plants by flow cytometry Plant Mol. Biol. Rep. 9:229–241

  2. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K, (1996). Current Protocols in Molecular Biology New York Wiley

  3. Bablak P, Draper J, Davey MR, Lynch PT, (1995) Plant regeneration and micropropagation of Brachypodium distachyon Plant Cell Tiss Org Cult 42:97–107

  4. Bennett MD, Leitch IJ, (2005) Nuclear DNA amounts in Angiosperms: progress, problems and prospects Annals of Botany 95:45–90

  5. Cheng M, Lowe BA, Spencer TM, Ye X, Armstrong CL, Cheng M, (2004) Invited review: Factors influencing Agrobacterium-mediated transformation of monocotyledonous species In Vitro Cell Dev Biol Plant 40:31–45

  6. Christiansen P, Didion T, Andersen CH, Folling M, Nielsen KK, (2005) A rapid and efficient transformation protocol for the grass Brachypodium distachyon Plant Cell Rep. 23:751–758

  7. Draper J, Mur LAJ, Jenkins G, Ghosh-Biswas GC, Bablak P, Hasterok R, Routledge APM, (2001) Brachypodium distachyon. A new model system for functional genomics in grasses Plant Phys. 127:1539–1555

  8. Edwards, K, Johnstone C & Thompson C (1991) A simple and rapid method for the preparation of plant genomic DNA For PCR Analysis. Nucleic Acids Res. 19

  9. Feldmann KA, (1991) T-DNA insertion mutagenesis in Arabidopsis: Mutational spectrum Plant J. 1:71–82

  10. Garfinkel M. Nester EW. (1980) Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism J. Bacteriol. 144:732–746

  11. Gaut BS, (2002) Evolutionary dynamics of grass genomes New Phytol. 154:15–28

  12. Hasterok R, Draper J, Jenkins G, (2004) Laying the cytotaxonomic foundations of a new model grass, Brachypodium distachyon (L.) beauv Chromosome Res. 12:397–403

  13. Jeon J-S, Lee S, Jung K-H, Jun S-H, Jeong D-H, Lee J, Kim C, Jang S, Lee S, Yang K, Nam J, An K, Han M-J, Sung R-J, Choi H-S, Yu J-H, Choi J-H, Cho S-Y, Cha S-S, Kim S-I, An G, (2000) T-DNA insertional mutagenesis for functional genomics in rice Plant J. 22:561–570

  14. Kellogg EA, (2001) Evolutionary history of the grasses Plant Phys. 125:1198–1205

  15. Lazo GR, Stein PA, Ludwig RA, (1991) A DNA transformation-competent Arabidopsis genomic library in Agrobacterium Bio/Technology 9:963–967

  16. Linsmaier EM, Skoog F, (1965) Organic growth factor requirements of tobacco tissue cultures Plant Phys. 18:100–127

  17. Riede CR, Anderson JA, (1996) Linkage of RFLP markers to an aluminum tolerance gene in wheat Crop Sci. 36:905–909

  18. Shi Y, Draper J, Stace C, (1993) Ribosomal DNA variation and its phylogenetic implication in the genus Brachypodium (Poaceae) Plant Syst. Evol. 188:125–138

  19. Sundaresan V, Springer P, Volpe T, Haward S, Jones JDG, Dean C, Ma H, Martienssen R, (1995) Patterns of gene action in plant development revealed by enhancer trap and gene trap transposable elements Genes Dev. 9:1797–1810

  20. Svitashev SK, Somers DA, (2002) Characterization of transgene loci in plants using FISH: A picture is worth a thousand words Plant Cell Tiss. Org. Cult. 69:205–214

  21. Wolfe KH, (2001) Yesterday’s polyploids and the mystery of diploidization Nature Rev. Genet. 2:333–341

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Correspondence to John P. Vogel.

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Vogel, J.P., Garvin, D.F., Leong, O.M. et al. Agrobacterium-mediated transformation and inbred line development in the model grass Brachypodium distachyon . Plant Cell Tiss Organ Cult 84, 199–211 (2006).

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  • c-value
  • embryogenic callus
  • genome size
  • model system
  • tissue culture