title: Transformation of barley (Hordeum vulgare L.) by Agrobacterium tumefaciens infection of in vitro cultured ovules.
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.
authors.affiliation: Department of Genetics and Biotechnology, Danish Institute of Agricultural Sciences, Research Centre Flakkebjerg, 4200, Slagelse, Denmark. Inger.Holme@agrsci.dk
authors.forename: Inger Baeksted
authors.initials: IB
authors.lastname: Holme
authors.forename: Henrik
authors.initials: H
authors.lastname: Brinch-Pedersen
authors.forename: Mette
authors.initials: M
authors.lastname: Lange
authors.forename: Preben Bach
authors.initials: PB
authors.lastname: Holm
mesh_terms: D016960:Agrobacterium tumefaciens / Q000472:pathogenicity* / Q000502:physiology*; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000378:metabolism; D018744:DNA, Plant / Q000378:metabolism; D018628:Gene Dosage; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D001467:Hordeum / Q000166:cytology / Q000235:genetics* / Q000382:microbiology*; D040582:Inheritance Patterns; D008970:Molecular Weight; D012038:Regeneration; D012639:Seeds / Q000166:cytology / Q000235:genetics* / Q000382:microbiology*; D014170:Transformation, Genetic; D019076:Transgenes
chemical_list: D004269:DNA, Bacterial; D018744:DNA, Plant; C036483:T-DNA; D049452:Green Fluorescent Proteins