Doubled Haploid Transgenic Wheat Lines by Microspore Transformation

Doubled Haploid Transgenic Wheat Lines by Microspore Transformation

Microspores are preferred explant choice for genetic transformation, as their use shortens the duration of obtaining homozygous transformants. All established gene-delivery methods of particle bombardment, electroporation, and cocultivation with Agrobacterium tumefaciens were optimized on androgenic...

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Bibliographic Details
Published in:Methods in molecular biology (Clifton, N.J.) Vol. 1679; p. 213
Main Authors: Rustgi, Sachin, Ankrah, Nii O, Brew-Appiah, Rhoda A T, Sun, Yue, Liu, Weiguo, von Wettstein, Diter
Format: Journal Article
Language:English
Published: United States 2017
Subjects:
Agrobacterium tumefaciens - genetics
Biolistics - methods
Cell Culture Techniques
Electroporation
Gene Transfer Techniques
Genetic Vectors - genetics
Haploidy
Phenotype
Pollen - genetics
Transformation, Genetic
Triticum - genetics
Triticum - growth & development
Agrobacterium tumefaciens
Electroporation
Genetic transformation
Microspore embryogenesis
Particle bombardment
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Summary:Microspores are preferred explant choice for genetic transformation, as their use shortens the duration of obtaining homozygous transformants. All established gene-delivery methods of particle bombardment, electroporation, and cocultivation with Agrobacterium tumefaciens were optimized on androgenic microspores or derived tissues. In the biolistic gene delivery method 35-40 days old haploid microspore embryoids were used for genetic transformation, whereas freshly isolated androgenic microspores were used for genetic transformation in the electroporation and Agrobacterium cocultivation-based methods. The genetic transformation methods of biolistic gene-delivery and electroporation gave rise to the chimeric plants, whereas the method involving cocultivation with Agrobacterium yielded homozygous transformants. These methods were tested on a large number of cultivars belonging to different market classes of wheat, and found to be fairly independent of the explant genotype. Other benefits of using microspores or derived tissues for transformation are: (1) a few explant donors are required to obtain desired transformants and (2) the time required for obtaining homozygous transformants is about 8 months in case of spring wheat genotypes and about a year in case of winter wheat genotypes.
ISSN:1940-6029
DOI:10.1007/978-1-4939-7337-8_13