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Transgene-free genome editing in poplar
Lennart Hoengenaert1,2, Chantal Anders1,2, Jan Van Doorsselaere3
1Department of Plant Biotechnology and Bioinformatics, Ghent University, Ghent, 9052, Belgium.
The New Phytologist
|January 22, 2025
Summary
This study presents a novel gene-editing method for poplar trees, successfully creating edited plants without foreign DNA integration. This breakthrough is crucial for agricultural applications, especially in vegetatively propagated species.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Genetics
Background:
- Stable integration of gene-editing cassettes is standard but problematic for vegetatively propagated plants.
- Existing methods hinder the removal of foreign DNA without compromising elite varieties.
Purpose of the Study:
- To develop an efficient method for generating gene-edited Populus tremula × P. alba (poplar) trees.
- To achieve gene editing without incorporating foreign DNA into the plant genome.
Main Methods:
- Utilized Agrobacterium tumefaciens for transient transformation with a base-editing construct targeting CCoAOMT1.
- Employed ALS genes for positive selection on chlorsulfuron medium.
- Investigated CodA gene for negative selection of T-DNA integrated lines.
Main Results:
- Approximately 50% of regenerated shoots were T-DNA free, resulting from transient transformation.
- 7% of chlorsulfuron-resistant shoots were confirmed T-DNA free, nonchimeric, and edited in CCoAOMT1.
- Long-read whole-genome sequencing verified the absence of foreign DNA in edited lines.
Conclusions:
- Developed an efficient, transgene-free gene-editing method for poplar trees.
- Transient transformation combined with selection markers enables the generation of edited plants without foreign DNA.
- The CodA negative selection marker may be beneficial for other plant varieties.
