Nonstructural carbohydrates (NSCs) are a key factor in the physiological regulation of plants and can reflect buffering capacity of plants under diverse environmental conditions. The effects of diverse environmental conditions on plant NSCs and tissue or organ scales have been thoroughly studied, but their effects on fine root (root diameter < 2 mm) NSC concentrations are still not completely understood. Our aims were to explore the synergistic fluctuations in root traits and NSC concentrations under diverse environmental conditions. This study was conducted on two-year-old temperate seedling tree species (Juglans mandshurica Maxim., Fraxinus mandshurica Rupr., and Phellodendron amurense Rupr.) with different drought intensities and soil substrates. The specific root length (SRL) and specific root surface area (SRA) were significantly affected by drought intensities and soil substrates, while the root tissue density (RTD) and average diameter (AD) were not significantly affected by water intensities and soil substrates in all three species. The root C, N, and P concentration did not change according to drought stress but were significantly affected by the soil substrates in all three species. Similarly, the soluble sugar (SS) and starch (ST) concentrations were significantly affected by both the drought stress and the soil substrates in all three species. The AD explained 6.8% of the total variations in soluble sugar, while the SRL explains 32.1% of the total variation in starch. The root tip C, N, and P concentrations were not significantly correlated with NSCs under different treatments. The total variations in root tip morphology, chemistry, and NSC concentrations are greater among species than compared to different drought intensities and soil substrates. However, the root NSC concentrations were closely related to root morphological traits (SRL and AD) rather than chemical traits. On the basis of different soil resources, the species with thinner diameters have higher SS concentrations, while those of a thicker diameter have higher ST concentrations.Forests 2020, 11, 415 2 of 18 declines [1-3]. The IPCC (Intergovernmental Panel on Climate Change) predicted that the most serious and frequent drought events will occur in the coming decades [4]. Nonstructural carbohydrates (NSCs, mainly composed of soluble sugar and starch) are a product of plant photosynthesis and the main substrate for plant respiration, which is responsible for the carbon budget balance [5,6]. NSC also reflects the level of substance for plant survival, growth, and their buffering capacity under environmental degradation [7] and NSCs are an important factor in the regulation of plant physiological phenomena under diverse environmental conditions. In recent years, most of the studies examining NSC dynamics at the tissue or organ scale have investigated the effects of water [1,[8][9][10], CO 2 concentrations [11], shade [10,12], and other factors. However, these studies have rarely focused on root traits, especially the root tip morphology und...