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Plants produce a huge array of secondary metabolites that play a key role in defense mechanisms against detrimental microorganisms and herbivores, and represent a suitable alternative to synthetic fungicides in sustainable agriculture. In this work, twelve crude hydroethanolic extracts derived from leaves of different potato cultivars were chemically characterized by LC/MS and their antioxidant properties were investigated in vitro. Furthermore, the biological activity against the fungal pathogen Rhizoctonia solani was evaluated both in vitro and in vivo. Extracts showed the ability to inhibit R. solani growth in vitro and significantly reduced damping-off incidence in in vivo experiments. Furthermore, R. solani mycelia exposed to the extracts showed an altered morphology (low translucency, irregular silhouette, and cytoplasmatic content coagulation) compared to the untreated control in light microscopy examination. Principal component analysis conducted on identified chemical compounds highlighted significant metabolic variations across the different extracts. In particular, those that inhibited most of the growth of the pathogen were found to be enriched in α-chaconine or α-solanine content, indicating that their biological activity is affected by the abundance of these metabolites. These results clearly indicated that plant-derived compounds represent a suitable alternative to chemicals and could lead to the development of new formulates for sustainable control of plant diseases. [ABSTRACT FROM AUTHOR]