Single-walled carbon nanotube-mediated plasmid DNA delivery in the model monocot Brachypodium distachyon

Background/aim: -mediated transformation and biolistics, are restricted by genotype dependency, lengthy tissue culture steps, and low efficiency, particularly in monocot species. Therefore, the development of alternative gene delivery strategies remains an ongoing research focus in plant biotechnology. Nanoparticle-based delivery systems have recently gained attention due to their physiochemical properties, biocompatibility, and potential for alternative gene delivery approaches in plant systems. Materials and methods: . SWCNTs were functionalized with polyethyleneimine (PEI) and plasmid DNA (pCAMBIA1301) was loaded via electrografting. Successful complex formation was confirmed with transmission electron microscopy (TEM), atomic force microscopy (AFM), and zeta potential measurements. Results: (GUS) reporter gene expression in both callus tissues and seeds, with ImageJ-based quantification showing that the 3:1 SWCNT:pDNA ratio yielded the highest mean signal intensity. Conclusion: Our findings show that the SWCNT-based system enabled the delivery of a large plasmid (~12 kb), supporting transient reporter gene expression in monocot tissues. Overall, the data suggest that SWCNT-mediated delivery represents a promising technique for DNA delivery in monocots, and further optimization is expected to improve its efficiency.