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Transformation in Barley
JIC
Contact: Wendy Harwood
Current status
Agrobacterium-mediated transformation is used as the method of choice in barley. The spring genotype Golden Promise is the most responsive barley genotype and is used for all routine transformation. At JIC, large-scale experiments have produced hundreds of independent transgenic lines with average transformation efficiencies of 25%. Recent experiments have yielded efficiencies up to 50%. These efficiencies are comparable to, or better than, those reported elsewhere and good enough to allow the development of transformation-based functional genomics tools such as T-DNA insertion in this important crop. As barley is closely related to wheat, it also serves as an excellent model for this crop, thus benefiting both the wheat and barley research communities.
Time scale and costs
The current cost for barley transformation at JIC is £3,117 / construct (FEC) to deliver 20 independently transformed T0 plants. Rooted transgenic plants in tubes can be produced within 12-14 weeks. A further 4-5 months is needed to obtain T1 seed.
Current work and future directions
- Further improvement of transformation efficiency and transformation of other genotypes.
- Understanding transgene insertion and controlling transgene expression.
- The effect of introns on transgene expression.
- Further development of methods for the isolation of transgene flanking regions in GM crops.
- Analysis of Flanking Sequence Tags (FST's) in barley.
- A number of collaborative projects involving barley transformation are also in progress.
- Training in barley transformation has been provided to a number of laboratories and the methodology successfully transferred to labs within and outside the UK.
Recent relevant publications
Book or Book Chapter
2008
- Harwood W A, Bartlett J, Alves S, Perry M, Smedley M, Leyland N, Snape JW (2008) "Barley transformation using Agrobacterium-mediated techniques.", Transgenic Wheat, Barley and Oats: Production and Characterisation, The Humana Press , Methods in Molecular Biology Vol 478, Chapter 9, pp137-147
- Harwood W A, Smedley M (2008) "Barley transformation using biolistic techniques. ", Transgenic Wheat, Barley and Oats: Production and Characterisation, The Humana Press, Methods in Molecular Biology Vol 478 , Chapter 8, pp 125-136
2004
- Harwood W A, Bilham L J, Travella S, Salvo-Garrido H, Snape J W (2004) "Fluorescence in situ hybridization to localise transgenes in plant chromosomes", Methods in Molecular Biology: Transgenic Plants, (Ed. L Pena) Vol. 286: 327-340
Journal Article
2005
- Travella, S, Ross S M, Harden J, Everett C, Snape J W, Harwood W A (2005) "A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques", Plant Cell Reports, 23 pp780-789
2004
- Bourdon V, Wickham A, Lonsdale D, Harwood W A (2004) "Additional introns inserted within the luciferase reporter gene stabilize transgene expression in wheat", Plant Science , 167 pp 1143-1149
- Salvo G H, Travella S, Bilham L J, Harwood W A, Snape J W (2004) "The distribution of transgene insertion sites in barley determined by physical and genetic mapping", Genetics , 167 -pp 1371-1379
- Harwood W A, Bilham L J, Travella S, Bourdon V, Salvo H, Harden J, Perry M, Snape J W (2004) "Genetic transformation of barley: Improved technology, safety assessment and future potential", Czech Journal of Genetics and Plant Breeding , 40 pp 77
SCRI
Contact: Jennifer Stephens
Current status
Our facilities are currently being updated to establish SCRI as a centre of excellence for barley transformation in and around Scotland. We routinely use Agrobacterium tumefaciens to transform the spring cultivar Golden Promise with great success. We are actively involved in developing technologies to improve T-DNA transfer to less amenable varieties including the spring malting cultivar, Optic.
Costs and time scales
Barley transformation at SCRI will be available to Research Institutes/Universities both nationally and internationally from early 2009. We will provide help/advice in construct design and can make constructs to order. We aim to produce 10-20 independently transformed plantlets within 12 weeks. These plants will have been grown on selective antibiotics and screened by PCR to confirm the presence of the transgene. Southern analysis to determine gene copy number and generation of T1 seed can also be performed.
Current work and future directions
- Development of new vectors:
- Tissue-specific and inducible promoters
- Systematic comparison of different selectable markers
- Marker-free transformation
- Improvement of transformation efficiencies for high-throughput technologies
See also: SCRI Functional Genomics (FunGen)
News
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| 19/07/2010 NEW 20th International Triticeae Mapping Initiative (ITMI) workshop more ... |
| 24/06/2010 One voice for UK Plant and Crop Science more ... |
| 24/06/2010 Rothamsted Research - Barley Diversity Collection more ... |
| 21/05/2010 Appointments to BBSRC Strategy Panels, Pool of Experts: Peer Review more ... |
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