Next-generation biofungicides for food safety: in vitro and in silico validation of Tetraclinis articulata essential oil against Mediterranean wheat pathogen for sustainable agriculture

Abstract

The rising resistance of plant pathogens to conventional fungicides requires the search for sustainable and environmentally friendly alternative strategies. Fusarium algeriense , a pathogen of durum wheat in the Mediterranean, is considered a major threat to food security. This study explores the antifungal potential of the essential oil from Tetraclinis articulata , using phytochemical, biological, and in silico approaches. The EO extracted by hydrodistillation was characterized by GC–MS, revealing a chemotype dominated by β -caryophyllene (24.73%), bornyl acetate (16.93%), and α -pinene (15.13%), defining an original profile rich in oxygenated sesquiterpenes. In vitro antifungal activity, evaluated by direct contact on agar Czapek Dox’s medium against the characterized isolate FusAlg12 (ITS, GenBank PX864254), showed dose-dependent inhibition of mycelial growth, reaching complete inhibition (100%) at the highest oil concentration (15 μL/mL). A bio-inspired hybrid molecule (HTA1), derived from the main compound, was designed and evaluated by molecular docking on fungal CYP51, a key target enzyme. HTA1 showed high affinity for the catalytic active site (−9.9 kcal/mol), higher than that of the reference ligand fluconazole (−9.1 kcal/mol), suggesting a competitive inhibition mechanism through hydrophobic locking of the active site. Molecular dynamics simulations indicate that the CYP51-HTA1 complex is structurally stable, with the ligand maintaining consistent interactions within the binding pocket while preserving protein compactness and conformation. ADMET evaluation indicates a favorable profile for phytosanitary application. These results demonstrate that T. articulata is a promising source of natural antifungal molecules and pave the way for the development of innovative biofungicides targeting a key enzyme in the physiology of phytopathogens.