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

Abstract

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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Abbreviations

BA:

benzylaminopurine

2,4-D :

2,4-dichlorophenoxyacetic acid

LS:

Linsmaier and Skoog basal medium

MS:

Murashige and Skoog salts and vitamins

CIM:

callus inducing medium

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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). https://doi.org/10.1007/s11240-005-9023-9

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Keywords

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