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Transformation in Brachypodium distachyon
JIC
Contact: Philippe Vain
Current status
At JIC the Brachypodium diploid genotype Bd21 can be routinely transformed at around 55% efficiency using Agrobacterium and embryogenic callus derived from immature embryos. Molecular analyses (including Flanking Sequence Tags - FSTs) of transgenic Brachypodium plants and inheritance studies are ongoing. FSTs have been submitted to public databases and the corresponding Bd21 insertion lines are available from JIC upon request.
Costs and time scales
Current work and future directions at JIC
- To improve the efficiency of Agrobacterium-mediated transformation systems enabling the production of thousands of T-DNA lines per year if possible from mature embryos. This will allow facile high-throughput T-DNA insertional mutagenesis strategies for functional genomic approaches in Brachypodium.
- To develop novel promoter and gene trap vectors for Brachypodium based on the pCLEAN dual binary vector system developed at JIC (http://www.jic.ac.uk/staff/philippe-vain/vectors.htm)
- To study the expression pattern of transcription factors controlling shoot development in Brachypodium - 12 months studentship with Mary Byrne and Philippe Vain (JIC) already funded.
Recent relevant publications
Journal Article
2008
- Vain, P., Worland, B., Thole, V., McKenzie, N., Opanowicz, M., Fish, L.J., Bevan, M.W. and Snape, J.W. (2008) "Agrobacterium-mediated transformation of the temperate grass Brachypodium distachyon (genotype Bd21) for T-DNA insertional mutagenesis", Plant Biotechnology Journal, 6, 236-245
- Opanowicz, M., Vain, P., Draper, J., Parker, D. and Doonan, J.H. (2008) "Brachypodium distachyon: making hay with a wild grass", Trends in Plant Science, 13, 172-177
2007
- Thole, V., Worland, B., Snape, J.W. and Vain, P. (2007) "The pCLEAN dual binary vector system for Agrobacterium-Mediated plant transformation.", Plant Physiology, 145, 1211-1219
- Vain, P. (2007) "Thirty years of plant transformation technology development.", Plant Biotechnology Journal,, 5, 221-229
- Vain, P. (2007) "Trends in GM crop, food and feed safety literature", Nature Biotechnology, 25, 624-626.
2006
- Lacroix, B., Tzfira, T., Vainstein, A. and Citovsky, V. (2006) "A case of promiscuity: Agrobacterium\'s endless hunt for new partners", Trends in Genetics,, 22, 29-37
- Vain, P. (2006) "Global trends in plant transgenic science and technology (1973-2003). ", Trends in Biotechnology, 24, 206-211.
2005
- Afolabi, A.S., Worland, B., Snape, J. and Vain, P. (2005) "Novel pGreen/pSoup dual-binary vector system in multiple T-DNA co-cultivation as a method of producing marker-free (clean gene) transgenic rice (Oriza sativa L) plant", African Journal of Biotechnology, 4, 531-540.
- Vain, P. (2005) "Plant transgenic science knowledge", Nature Biotechnology, 23, 1348-1349.
Brachypodium distachyon (IBERS)
Brachypodium distachyon (RRes)
Contact: Huw D Jones
Current status
We have applied published methods and confirmed that biolistics and Agrobacterium tumefaciens are suitable DNA-delivery methods for mature embryo-derived callus of Bd21. Regeneration is via somatic embryogenesis and subsequent selection using either antibiotic or herbicide.
Costs and time scales
We have not yet calculated the full economic costs of transforming and producing seed from Bd21. Time scales are 14-16 weeks from mature seeds to PCR-positive plantlets in media or soil, 6 weeks for vernalisation and a further 8-10 weeks to harvest mature T1 seeds.
Current work and future directions at RRes
We will use Brachypodium Bd21 to validate Ac/Ds transposition and the potential of this transposon for gene- and promoter-tagging.
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