Agrobacterium-Mediated Transformation of Barley Immature Embryos
Barley transformation is an essential tool for a range of functional genomics studies as well as for future crop improvement. The demand for efficient crop transformation systems continues to grow, with new genome editing technologies adding to that demand. Here we describe an efficient and routine transformation protocol for the spring barley Golden Promise, based on Agrobacterium-mediated inoculation of immature embryos. This protocol has been widely used for overexpression and RNAi applications and more recently for CRISPR/Cas9 mediated genome editing. Average transformation efficiencies of 25% can be easily achieved.
Key wordsBarley transformation Agrobacterium tumefaciens Immature embryos Genome editing Genome engineering
We acknowledge support from the Biotechnology and Biological Sciences Research Council (BBSRC) via grant [BB/N019466/1] and grant [BB/P013511/1] to the John Innes Centre.
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