A peat substrate is made from peat from drained peatlands, which is a limited resource. A realistic estimate is that 50% of the world's wetlands have been lost. Peat is used in horticulture, especially for the cultivation of vegetables in greenhouses. The consequences of peatland exploitation are an increase in the greenhouse effect and a decrease in carbon stocks. Wood fiber can be used as an alternative to peat. The chemical properties of growing media interact and change continuously due to the small volume of growing media, which is limited by the growing container. This study aims to gain new knowledge on the impact of nutrient changes in the microbial degradation of carbon compounds in wood fiber and mixtures with a peat substrate on the content and uptake of nutrients required by plants. The cucumber (Cucumis sativus L.) variety 'Dirigent H' developed in the Netherlands was cultivated in growing media of a peat substrate and wood fiber: (1) peat substrate (PS); (2) wood fiber (WF); (3) wood fiber and peat substrate 50/50 v/v (WF/PS 50/50); (4) wood fiber and peat substrate 25/75 v/v (WF/PS 25/75). The rates of fertilization were the following: (1) conventional fertilization (CF); (2) 13 g N per plant (N13); (3) 23 g N per plant (N23); (4) 30 g N per plant (N30). The experiment was carried out with three replications. As the amount of wood fiber increased, the humidity and pH of the growing media increased. The fertilization of the cucumbers with different quantities of nitrogen influenced the nutrient uptake. The plants grown in the 50/50 and 25/75 growing media had the best Cu uptake when fertilized with N23. When the plants grown in the wood fiber media and the 50/50 media were fertilized with N13, N23, and N30, the Mn content in the growing media at the end of the growing season was significantly lower than the Mn content in the media with conventional fertilization. Thus, nitrogen improved the uptake of Mn by the plants grown not only in the wood fiber, but also in the combinations with a peat substrate. Growing plants in wood fiber and fertilizing them with N13 can result in the optimum uptake of micronutrients. The number and biomass of cucumber fruits per plant were influenced by the amount of wood fiber in the growing media and the application of nitrogen fertilizer. The highest number of fruits and biomass of fruits per plant obtained were significantly higher when the cucumbers were grown in WF/PS 50/50 growing media with additional N13 fertilization.