Abstract
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.
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References
Lawrenson T, Shorinola O, Stacey N, Liu C, Østergaard L, Patron N, Uauy C, Harwood W (2015) Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. Genome Biol 16:258
Ricroch A, Clairand P, Harwood W (2017) Use of CRISPR systems in plant genome editing: towards new opportunities in agriculture. Emerging Topics Life Sci 1(2):169–182
Wan Y, Lemaux PG (1994) Generation of large numbers of independently transformed fertile barley plants. Plant Physiol 104:37–48
Tingay S, McElroy D, Kalla R, Fieg S, Wang M, Thornton S, Brettell R (1997) Agrobacterium tumefaciens-mediated barley transformation. Plant J 11:1369–1376
Bartlett JG, Alves SC, Smedley M, Snape JW, Harwood WA (2008) High-throughput Agrobacterium-mediated barley transformation. Plant Methods 4:22
Travella S, Ross SM, Harden J, Everett C, Snape JW, Harwood WA (2005) A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques. Plant Cell Rep 23:780–789
Volpi e Silva N, Patron N (2017) CRISPR-based tools for plant genome engineering. Emerging Topics Life Sci 1:135–149
Harwood W, Smedley M (2009) Barley transformation using biolistic techniques. In: Jones H, Shewry P (eds) Transgenic wheat, barley and oats. Methods in molecular biology™ (Methods and Protocols), vol 478, Humana Press, pp 125–136
Garfinkel M, Nester EW (1980) Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J Bacteriol 144:732–743
Smedley MA, Harwood WA (2014) Gateway-compatible plant transformation vectors. Methods Mol Biol 1223:3–16
Acknowledgments
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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Hinchliffe, A., Harwood, W.A. (2019). Agrobacterium-Mediated Transformation of Barley Immature Embryos. In: Harwood, W. (eds) Barley. Methods in Molecular Biology, vol 1900. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8944-7_8
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DOI: https://doi.org/10.1007/978-1-4939-8944-7_8
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-8944-7
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