High-throughput Agrobacterium-mediated barley transformation

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Abstract
Background: Plant transformation is an invaluable tool for basic plant research, as well as a useful technique for the direct improvement of commercial crops. Barley (Hordeum vulgare) is the fourth most abundant cereal crop in the world. It also provides a useful model for the study of wheat, which has a larger and more complex genome. Most existing barley transformation methodologies are either complex or have low (Agrobacteriumstrain AGL1, carrying vectors from the pBract series that contain thehptgene (conferring hygromycin resistance) as a selectable marker. Results of large scale experiments utilising theluc(firefly luciferase) gene as a reporter are described. The method presented here has been used to produce hundreds of independent, transgenic plant lines and we show that a large proportion of these lines contain single copies of thelucgene.Conclusion: This protocol demonstrates significant improvements in both efficiency and ease of use over existing barley transformation methods. This opens up opportunities for the development of functional genomics resources in barley.