Inoculation with biocontrol agents can affect iron (Fe) uptake by plants. The objective of this research was to study the necessity of defining a Fe threshold in growth media for the inoculation with the biocontrol agent Trichoderma asperellum T34. A completely randomized experiment with cucumber (Cucumis sativus L.) was performed involving two factors: Fe rate in the growth medium in the form of ferrihydrite (0, 8, 16, 32.5, and 75 mgkg(-1) of citrate-ascorbate-extractable Fe (CA-Fe), and plant inoculation with T34. Dry matter (DM) of aerial parts of cucumber was decreased by T34. This was related to a decreased accumulation of Fe in plants, more in aerial parts than in roots. However, at the highest Fe rate (75 mgkg(-1)), differences in DM yield, plant height, and the content and concentration of Fe in shoots between inoculated and noninoculated plants were not significant. The threshold of CA-Fe in the medium for DM yield of cucumber was 37 mgkg(-1) without T34. With T34, this threshold was 65 mgkg(-1), which implies that, below this limit, additional Fe supply is required for inoculation with T34.