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Background: The project
builds on the extensive expertise in wheat transformation
developed in the Australian CRC for Molecular Plant Breeding
(MPBCRC) including the highly successful biolistics-based
wheat transformation protocols at CIMMYT. The efficiency of
wheat transformation systems strongly depends on the transformation
parameters, the condition of the donor plant, and the plant
genotype chosen for the transformation process. In bread wheat
all these factors have been well examined, as well as the
ability to produce embryogenic callus, regeneration in selection
medium, and overall transformation performance. As a result
of this study, MPBCRC identified two bread wheat lines (MPB
Bobwhite-26 and MPB Bobwhite-56) as “super transformable”,
leading to a quantum leap in wheat transformation efficiency
in various laboratories throughout the world, and establishing
the basis for the use of this technology in wheat transgenic
breeding and functional genomics. The established transformation
protocol was based on biolistic transformation of immature
zygotic embryos and glufosinate selection, leading to 40-60%
transformation efficiencies. While biolistics-based methods
are effective at studying candidate genes initially, the recovery
of transgenic plants with multiple and rearranged transgene
copies is not desirable for final product development. Agrobacterium-based
methods have been shown to result in lower copy and less rearranged
transgenic events and has been used successfully in several
cereals, including wheat.
Objectives:
- Evaluate various Agrobacterium-based
transformation methods using the super transformable MPB
Bobwhite-26 and MPB Bobwhite-56 varieties
- Evaluate other wheat varieties to optimise
the transformation efficiencies
- Modify the Agrobacterium-based
method to optimise the efficiency in wheat
Target Germplasm: MPB Bobwhite-26,
MPB Bobwhite-56, other wheat varieties
Gene(s): The reporter genes
gusA and gfp
Partner(s): Department
of Primary Industries (DPI), Australia
Status: Initial transformants
are being produced.
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