Efficient Agrobacterium-Mediated Methods for Transient and Stable Transformation in Common and Tartary Buckwheat

Area of Science:

• Plant biotechnology
• Crop science
• Molecular biology

Background:

• Buckwheat is a nutrient-rich crop with available genome data.
• Developing efficient genetic transformation methods for buckwheat is crucial for research and breeding.

Purpose of the Study:

• To establish reliable Agrobacterium-mediated transient and stable gene transformation protocols for common and Tartary buckwheat.
• To demonstrate the utility of these methods for metabolic engineering and gene function analysis.

Main Methods:

• Agrobacterium-mediated vacuum infiltration of leaf and cotyledon tissues.
• Use of eGFP and GUS reporter genes for transient expression analysis.
• PCR, Western blotting, fluorescence microscopy, and LC-MS for confirmation of gene integration and protein/metabolite production.

Main Results:

• Optimized Agrobacterium-mediated transient transformation protocols were established for Fagopyrum esculentum and F. tartaricum.
• Successful engineering of the gramine biosynthetic pathway via transient expression of HvAMIS and HvNMT genes.
• Achieved in planta stable transformation using Agrobacterium-mediated vacuum infiltration with reporter and selectable marker genes.

Conclusions:

• The developed Agrobacterium-mediated transformation methods provide essential tools for buckwheat functional genomics and metabolic engineering.
• These protocols will accelerate research, gene editing, and breeding efforts in buckwheat improvement.