Susceptible and resistant cultivars of cucumber (cv. Suyo and cv. Shogoin fushinari, respectively) were evaluated for induction of systemic resistance by plant growth-promoting fungal isolates Phoma sp. GS8-1 and nonsporulating fungus GU21-2 against Colletotrichum orbiculare. Reduction in anthracnose disease was assessed at regular intervals of 0, 3, 6, and 9 days after challenge inoculation. Activities of endochitinase, β-1, 3-endoglucanase, peroxidase (PO), polyphenol oxidase, and phenylalanine ammonia lyase (PAL) were assayed at the above intervals, while the activities of exochitinase and β-1,3-exogluconase were assayed at 9 days after challenge inoculation. The data showed that all the enzymes activities increased in plants treated with plant growth-promoting fungi (PGPF) compared with nontreated plants of both cultivars. The activities of endochitinase and β-1,3-endoglucanase were highly increased in PGPF-induced plants than in nontreated plants of both cultivars at different time points. Activities of PO, polyphenol oxidase, and PAL also increased between 3 and 6 days due to inoculation with PGPF and decreased thereafter. The activity of PAL at 9 days in PGPF-induced plants was at the same level as that of nontreated plants of both cultivars. Results of this investigation suggest that PGPF-induced systemic resistance is associated with not only high increases in the activities of chitinase and β-1,3-gluconases but also other plant defense-related enzymes such as PO, PPO, and PAL. Roles of other mechanisms rather than accumulation of defense-related enzymes in PGPF-mediated induced resistance in susceptible cultivars were demonstrated.