Plant & Animal Genomes XV Conference

January 13-17, 2007
Town & Country Convention Center
San Diego, CA

Transformation Of The Temperate Grass Brachypodium distachyon (Line Bd21) Using The Green Fluorescent Protein (GFP) As A Screenable Marker

Philippe Vain , Barbara Worland , Vera Thole , Lesley J. Fish , Michael W. Bevan , John W. Snape

  John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK.

Genetic transformation of Brachypodium distachyon is an essential step to establishing the species as a functional genomics model for temperate cereals and grasses. A modified form of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria was used to develop a facile and rapid transformation system for the diploid line Bd21 of B. distachyon using Agrobacterium-mediated transformation of compact embryogenic calli derived from immature embryos. The GFP4 gene under the control of the maize Ubiquitin1 promoter and intron sequences produced bright green fluorescence easily detectable and screenable in sectors of developing Bd21 calli two weeks after transformation. Visual screening of transformed Bd21 tissue, associated with hygromycin selection, enabled early identification of transformation events and drastically reduced the quantity of tissue to be handled during the selection process. GFP expression was observed in primary transformed Bd21 plants (T0) and their progeny (T1). The advantages of using fluorescent proteins as screenable markers in B. distachyon genetic engineering and functional genomic programmes are discussed.