Transformation of barley (Hordeum vulgare L.) by Agrobacterium tumefaciens infection of in vitro cultured ovules.
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.
Holme
Inger Baeksted
IB
Department of Genetics and Biotechnology, Danish Institute of Agricultural Sciences, Research Centre Flakkebjerg, 4200, Slagelse, Denmark. Inger.Holme@agrsci.dk
Brinch-Pedersen
Henrik
H
Lange
Mette
M
Holm
Preben Bach
PB
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
D004269:DNA, Bacterial; D018744:DNA, Plant; C036483:T-DNA; D049452:Green Fluorescent Proteins