Doubled Haploid Transgenic Wheat Lines by Microspore Transformation.
Methods Mol Biol
; 1679: 213-234, 2017.
Article
in En
| MEDLINE
| ID: mdl-28913803
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.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
Pollen
/
Transformation, Genetic
/
Triticum
/
Gene Transfer Techniques
/
Haploidy
Language:
En
Journal:
Methods Mol Biol
Year:
2017
Type:
Article
Affiliation country:
United States