[Erh-Min Lai/Chih-Horng Kuo/Chih-Hang Wu ] Floral Stage Selection and Immune Evasion Enhance Agrobacterium-Mediated Transformation and Genome Editing
POST:
Agrobacterium-mediated transformation via floral inoculation (AMT-FI) enables genetic engineering without tissue culture. It is widely used in the model plant Arabidopsis thaliana, yet its efficiency, underlying mechanisms and broader applicability remain limited.
To provide conceptual and practical advances in AMT-FI, the team developed dual-reporter systems to simultaneously track Agrobacterium infection, transformation events and ovule viability in real time. The team identified that flowers opened at 5 to 9 days post-inoculation (DPI) are optimal for high transformation efficiency, with a peak at 6 DPI, achieving nearly 100% of siliques harboring transformants and the ~26% stable transformation efficiency. However, Agrobacterium infection induced ovule abortion, whereas efr mutants lacking the EF-Tu receptor (EFR) showed reduced ovule abortion and exhibited significantly higher genome editing efficiency. The research group further engineered stealth Agrobacterium strains (AS201 and AS202) for evading recognition by EFR to enhance both transient expression and genome editing efficiency.
By integrating floral-stage selection, immune evasion, and Agrobacterium engineering, the team provides a practical platform to advance plant genome engineering and demonstrate that genome-edited T1 plants can be recovered without selection.
This work is supported by Academia Sinica Grand Challenge Project led by Project Director Erh-Min Lai, with co-PIs Chih-Horng Kuo and Chih-Hang Wu. The study was primarily conducted by co-first authors Mao-Sen Liu and Teng-Kuei Huang, with technical support from Yi-Chieh Wang and Si-Chong Wang.