Integrated In Vitro and In Silico Evaluation of Natural Plasmid‐Curing Agents Against Multidrug‐Resistant Agrobacterium tumefaciens

Multidrug-resistant (MDR) Agrobacterium tumefaciens threatens healthcare and agriculture. Toxicity and environmental risks limit conventional plasmid-curing agents, prompting natural alternatives. Curcumin, piperine, and quercetin were tested for plasmid-curing against clinically isolated MDR A. tumefaciens. Morphological and 16S rRNA gene sequencing were used for identification of the isolate. Antimicrobial susceptibility and biofilm formation were measured pre- and post-treatment. Molecular docking was performed to analyze phytocompound-plasmid maintenance protein interactions. Broad-spectrum resistance and ethidium bromide insensitivity were found in the MDR isolate. Curcumin, piperine, and quercetin induced plasmid curing by restoring antibiotic susceptibility and inhibiting biofilm formation. The observed increases in antibiotic susceptibility following treatment were statistically significant (p < 0.05). Molecular docking showed strong binding affinities for quercetin and tetracycline, supporting these effects' mechanism. These results show that natural compounds can eliminate MDR A. tumefaciens biofilm formation and plasmid-mediated resistance. Curcumin, piperine, and quercetin disrupt plasmid stability through protein-ligand interactions, offering a promising alternative to traditional curing agents. Curcumin, piperine, and quercetin are eco-friendly, effective plasmid-curing agents that may combat A. tumefaciens multidrug resistance. This integrated in vitro and in silico approach supports phytocompound research and antibiotic-resistant pathogen treatment.