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An Efficient Agrobacterium-Mediated Transformation Protocol for Hexaploid and Tetraploid Wheat
Sadiye Hayta1, Mark A Smedley1, Martha Clarke1
1John Innes Centre, Department of Crop Genetics, Norwich Research Park, Norwich, Norfolk, UK.
Current Protocols
|March 3, 2021
Summary
A new Agrobacterium-mediated gene transformation system for wheat (Triticum aestivum) significantly improves efficiency. This breakthrough enhances wheat breeding by enabling precise genome editing in elite varieties.
Area of Science:
- Plant Biotechnology
- Crop Improvement
- Molecular Biology
Background:
- Wheat gene transformation lags behind other cereals, hindering crop improvement.
- Genome editing offers precise DNA manipulation but is limited by low regeneration and genotype dependence.
Purpose of the Study:
- To develop a robust and efficient Agrobacterium-mediated transformation system for wheat.
- To overcome limitations in wheat tissue culture and genotype dependency for gene editing.
Main Methods:
- Immature embryos of hexaploid spring wheat 'Fielder' were used as starting material.
- A centrifugation pretreatment was employed before Agrobacterium inoculation.
- The protocol was adapted for hexaploid wheat 'Cadenza' and durum wheat 'Kronos'.
Main Results:
- Achieved up to 33% transformation efficiency in 'Fielder' wheat.
- Demonstrated efficiencies of up to 4% in 'Cadenza' and 10% in 'Kronos'.
- Combined gene fusion GRF-GIF with the system improved transformation to 77.5% in 'Fielder'.
Conclusions:
- The developed system is high-throughput, efficient, and repeatable for wheat transformation.
- This technology facilitates the application of gene editing tools in elite wheat varieties.
- The system alleviates genotype dependence, accelerating wheat breeding advancements.
