Establishment of an efficient Agrobacterium tumefaciens-mediated genetic transformation and regeneration system for...

Genetic transformation has been a crucial technology for plant molecular biology research and molecular design breeding, particularly in modern agriculture, and currently gene function validation is mostly conducted on model plants, which severely limits the development of non-model plants such as medicinal plants. Polygonum cuspidatum Sieb. et Zucc. plays a significant role in the treatment of diabetes, cancer, respiratory and cardiovascular diseases, however, its optimal regeneration and gene transformation system is still not available. Here, we at first establish an efficient regeneration system for P. cuspidatum as: stems are the most suitable explants for callus induction and regeneration of buds, and N1 medium (MS medium with 1.0 mg/L 1-naphthylacetic acid (NAA), 0.1 mg/L kinetin (KT)) is the most suitable regeneration medium. Then based on this regeneration system, an Agrobacterium tumefaciens-mediated genetic transformation system via callus regeneration (CR) and organ regeneration (OR) is established and optimized as: the stems are immersed in the A. tumefaciens resuspension medium (1/4 MS medium added with 0.02% Silwet L-77) and vacuum infiltrated for 10 min, the optimal co-culture time is 5 days, and the most suitable screening antibiotic is 100 mg/L hygromycin (Hyg). In addition, the practicality of our introduced transformation system is confirmed through the experimental verification of the reporter gene DsRED and two candidate genes: stilbene synthase (PcSTS) and MYB transcription factor (PcMYB62). In summary, our established system can not only efficiently regenerate and genetically transform P. cuspidatum, a typical non-model but highly valuable plant, but it could also serve as a reference for the construction of other non-model plant transformation systems.

The online version contains supplementary material available at 10.1007/s11032-026-01645-w.