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DNA- and Selectable-Marker-Free Genome-Editing System Using Zygotes from Recalcitrant Maize Inbred B73.
Hajime Yamada1,2, Norio Kato2,3,4, Masako Ichikawa1,2
1Agri-Bio Research Center, KANEKA CORPORATION, Higashibara 700, Iwata, Shizuoka, 438-0802 Japan.
Plant & Cell Physiology
|January 30, 2024
Summary
This study introduces a DNA-free genome-editing system for recalcitrant maize. The novel method efficiently produces edited maize plantlets without selectable markers, advancing molecular breeding.
Area of Science:
- Plant Science
- Molecular Biology
- Agricultural Science
Background:
- Genome-editing tools like CRISPR/Cas9 are vital for plant breeding.
- Conventional maize genome editing requires DNA cassette removal to avoid GMO concerns.
- Recalcitrant maize genotypes pose challenges for efficient DNA-free editing.
Purpose of the Study:
- To establish an efficient DNA-free genome-editing system for recalcitrant maize genotypes.
- To overcome limitations of conventional methods and improve maize molecular breeding.
Main Methods:
- Direct delivery of Cas9-gRNA ribonucleoproteins into isolated maize zygotes.
- Regeneration of edited plantlets using co-culture with phytosulfokine.
Main Results:
- Successfully generated DNA- and selectable-marker-free genome-edited maize plantlets from recalcitrant genotypes.
- Achieved high efficiency in editing maize using the developed zygote-based system.
Conclusions:
- The developed method enables efficient DNA-free genome editing in recalcitrant maize.
- This approach can significantly advance molecular breeding for maize and other cereals.
