, a soilborne pathogen responsible for root rot in olive trees, poses a significant threat to olive production. Managing this pathogen remains challenging due to its aggressive root colonization and the limited efficacy of conventional control methods. Given the concerns associated with chemical treatments, this study evaluated the resistance of eight olive varieties to -induced root rot under controlled greenhouse conditions by assessing structural and biochemical defense mechanisms. Greenhouse trials revealed that Arbequina, Koroneiki, and Haouziya exhibited strong resistance, with 0% disease severity, while Picholine Marocaine and Picholine Languedoc were highly susceptible, reaching 100% disease severity. Growth parameters varied significantly, with susceptible varieties showing severe reductions in root length (RL), root fresh weight (RFW), and root dry weight (RDW), whereas resistant varieties maintained these parameters unchanged. While shoot length (SL) remained unaffected across all varieties, shoot fresh weight (SFW) and shoot dry weight (SDW) were significantly reduced in susceptible ones. Fourier-transform infrared (FTIR) spectroscopy revealed that resistant varieties maintained stable levels of lignin, cellulose, and polysaccharides, while susceptible ones exhibited extensive cell wall degradation. Additionally, total polyphenol content (TPC) and total flavonoid content (TFC) significantly increased in resistant varieties upon infection, whereas susceptible varieties experienced a substantial decline. These findings highlight the crucial role of structural and biochemical defenses in olive resistance to and suggest that selecting resistant varieties could serve as a sustainable strategy for managing root rot in olive production.