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Updated: May 17, 2026
10:28
Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes
Published on: February 14, 2020
Gene targeting in maize by somatic ectopic recombination
Ayhan Ayar1, Sophie Wehrkamp-Richter, Jean-Baptiste Laffaire
1Biogemma, Centre de Recherche de Chappes, Chappes, France.
Plant Biotechnology Journal
|October 26, 2012
Summary
This study presents a novel gene targeting system for maize, enhancing transformation efficiency and reducing unwanted events. The system utilizes inducible endonuclease expression to facilitate precise homologous recombination for gene editing in plants.
Area of Science:
- Plant biotechnology
- Molecular genetics
- Crop improvement
Background:
- Gene targeting in plants is hindered by low transformation efficiency and high rates of non-targeted events.
- Developing efficient and precise gene editing tools is crucial for crop improvement.
Purpose of the Study:
- To demonstrate a gene targeting system in maize that overcomes limitations in transformation efficiency.
- To establish a method for precise homologous recombination-mediated gene insertion in maize.
Main Methods:
- Integrated donor DNA, endonuclease gene (I-SceI), and a target locus (TL) with a defective selectable marker into maize genome.
- Used a dexamethasone-inducible I-SceI::GR protein for controlled cleavage of the TL.
- Assayed homologous recombination by selecting for the repaired selectable marker (nptII).
Main Results:
- Achieved a recombination frequency of 0.085% per immature embryo, with 29% of recombinants showing successful targeting.
- Demonstrated that excision of the donor locus may be unnecessary and can lead to unwanted repair events.
- The system enables the production of numerous cells for gene targeting from each assembled transformant.
Conclusions:
- The developed system significantly improves gene targeting efficiency in maize.
- This approach facilitates gene targeting in plant species with inherently low transformation efficiencies.
- The findings suggest potential for broader application in plant genetic engineering.
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