Abstract
BackgroundPlant 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 (<10%) transformation efficiencies.ResultsA robust, simple and reproducible barley transformation protocol has been developed that yields average transformation efficiencies of 25%. This protocol is based on the infection of immature barley embryos with Agrobacterium strain AGL1, carrying vectors from the pBract series that contain the hpt gene (conferring hygromycin resistance) as a selectable marker. Results of large scale experiments utilising the luc (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 the luc gene.ConclusionThis 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.
Highlights
Plant transformation is an invaluable tool for basic plant research, as well as a useful technique for the direct improvement of commercial crops
The genetic transformation of cereal crops has been widely studied as a tool for crop improvement and as a vital part of the development of functional genomics resources
Development of the improved Agrobacterium-mediated barley transformation protocol Development of the high-throughput Agrobacteriummediated barley transformation system was dependent on the availability of immature embryos from good quality plants grown under controlled conditions
Summary
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 provides a useful model for the study of wheat, which has a larger and more complex genome. The first fertile transgenic barley plants were produced by particle bombardment of immature embryos [1]. This was followed by reports of Agrobacterium-mediated transformation of barley immature embryos [2]. In a comparison of particle bombardment and Agrobacterium-mediated techniques, it was shown that Agrobacterium-mediated transformation was more efficient and led to the production of transgenic lines with lower numbers (page number not for citation purposes)
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