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The exogenous application of nanoscale biochar has great potential in agriculture to improve crop resistance to pathogens, although the mechanisms underlying this physiological response are unclear. We investigated the potential of nanoscale and conventional biochar amendment to induce resistance in Nicotiana benthamiana upon infection with the pathogen Phytophthora nicotianae. Nanoscale biochar synthesized at 350 °C (400 mg L−1) from corn straw with a particle size of ∼160 nm had a beneficial role in the active immunity of N. benthamiana through the ethylene pathway to provide resistance to P. nicotianae infection; in leaf assays, lesion diameters were reduced by 9.26% as compared to controls. Mechanistically, a 20.3% increase in reactive oxygen species (ROS) from biochar NP amendment activated the ethylene pathway, followed by SA-induced systemic acquired resistance (SAR) expression …