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The development of seedling production technology and methods of establishing highyielding plantations of Picea obovata Ledeb. on a scientific basis is one of the urgent problems of forestry in Mongolia. In this study, we aimed to solve the following problems: to conduct a comparative analysis of the seedling growth parameters and biomass accumulation grown on different nutrient substrates; to assess the relationship between seedling growth, biomass accumulation and soil properties; to determine the most optimal nutrient substrates for seedling production of Siberian spruce in greenhouse conditions in Northern Mongolia. Six formulations of nutrient substrates (T1, Т2, Т3, Т4, Т5, Т6) were used for the seedling production of Picea obovata Ledeb. in greenhouses equipped with a sprinkler system. Nutrient substrates were prepared using black soil, manure, compost, peat, sawdust, sand in different composition ratios. During the 4-year-observation period height, root collar diameter, root length and aboveground and belowground biomass of seedlings were measured at the end of each growing season. We divided the biomass of seedlings into several structural elements. We found that all tested nutrient substrates, except the control substrate, had a positive effect on seedling growth in height and diameter. Comparative analyses showed that different ratio and composition of black soil, compost, manure, sawdust, and sand in the nutrient substrate had different effects on seedling growth (p > 0.001) and biomass accumulation (p > 0.001). Among the proposed nutrient substrates, the treatments T2 (50 % black soil + 20 % sand + 20 % peat + 10 % compost) and T6 (60 % black soil + 20 % sand + 10 % peat + 10 % compost) were selected as the most effective soil substrate that are suitable for further seedling production of Siberian spruce under greenhouse conditions in Mongolia. There fore, it was observed that good root system development was a determinant of seedling growth in height, diameter, and aboveground biomass accumulation especially from 3–4 years of age. Spruce seedling growth was positively correlated not only with humus content (r = 0.46), but also with soil acidity (r = 0.43) and available phosphorus (r = 0.48). The results of this investigation made an important contribution to the development of production technology for growing standard and large-sized seedlings of Picea obovata in greenhouse complexes in Northern Mongolia. For citation: Jagdag D., Batsaikhan G., Baatarbileg N., Lobanov A.I., Gerelbaatar S. Effect of Nutrient Substrate on Seedling Growth and Biomass Allocation of Picea obovata Ledeb. in Northern Mongolia. Lesnoy Zhurnal = Russian Forestry Journal, 2023, no. 6, pp. 57–69. (In Russ.). https://doi.org/10.37482/0536-1036-2023-6-57-69