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Related Experiment Video

Updated: Dec 7, 2025

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

Published on: February 14, 2020

An Efficient Gene Excision System in Maize.

Ning Wang¹, Maren Arling¹, George Hoerster¹

¹Crop Genome Engineering, Applied Science and Technology, Corteva Agriscience, Johnston, IA, United States.

Frontiers in Plant Science

|September 28, 2020

Summary

This summary is machine-generated.

Maize transformation is improved by excising morphogenic genes Baby Boom (Bbm) and Wuschel2 (Wus2) post-transformation. This method increases fertile T0 plant production and enhances transformation efficiency in elite maize inbreds.

Keywords:

Agrobacterium developmentally-regulated promoters heat-shock promoters marker-free events morphogenic genes rapid maize transformation

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Last Updated: Dec 7, 2025

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

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Published on: February 14, 2020

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Published on: July 23, 2014

Area of Science:

Plant Biotechnology
Maize Genetic Engineering
Molecular Biology

Background:

Morphogenic genes Baby Boom (Bbm) and Wuschel2 (Wus2) enhance maize transformation efficiency and explant range.
Overexpression of Bbm/Wus2 can lead to compromised plant quality and sterility in regenerated T0 plants.
Developing methods to remove these genes post-transformation is crucial for producing fertile transgenic maize.

Purpose of the Study:

To develop a method for excising morphogenic genes (Bbm/Wus2) after maize transformation but before plant regeneration.
To improve the production of fertile T0 transgenic maize plants.
To enhance the efficiency and applicability of maize transformation in elite inbred lines.

Main Methods:

Utilized an inducible site-specific recombinase (Cre) system for targeted excision of morphogenic genes.
Employed developmentally regulated (e.g., Ole, Glb1) and inducible heat shock promoters (e.g., Hsp17.7, Hsp26) to drive Cre expression.
Optimized tissue culture conditions and construct design for efficient transformation and excision.

Main Results:

Developmentally regulated promoters achieved 25-100% excision rates.
Excision-activated selectable markers yielded 53-68% excision but lower transformation frequencies (13-50%).
Heat shock promoters combined with optimized conditions resulted in high T0 transformation (29-69%), excision (50-97%), and usable quality events (4-15%) with minimal non-transgenic escapes.

Conclusions:

Excision of morphogenic genes post-transformation significantly increases the production of fertile T0 maize plants.
The developed Cre-mediated excision strategy, particularly with heat shock promoters, offers a robust method for elite maize transformation.
Transgenic events generated are free from both morphogenic and marker genes, ensuring improved plant quality.