Non-structural carbohydrates (NSCs) facilitate plant adaptation to drought stress, characterize tree growth and survival ability, and buffer against external disturbances. Previous studies have focused on the distribution and dynamics of NSCs among different plant organs under drought conditions. However, discussion about the NSC levels of fine roots in different root branch orders is limited, especially the relationship between fine root trait variation and NSC content. The objective of the study was to shed light on the synergistic variation in fine root traits and NSC content in different root branch orders under different drought and soil substrate conditions. The 2-year-old Fraxinus mandshurica Rupr. potted seedlings were planted in three different soil substrates (humus, loam, and sandy-loam soil) and subjected to four drought intensities (CK, mild drought, moderate drought, and severe drought) for 2 months. With increasing drought intensity, the biomass of fine roots decreased significantly. Under the same drought intensity, seedlings in sandy-loam soil had higher root biomass, and the coefficient of variation of 5th-order roots (37.4, 44.5, and 53% in humus, loam, and sandy-loam soil, respectively) was higher than that of lower-order roots. All branch order roots of seedlings in humus soil had the largest specific root length (SRL) and specific root surface area (SRA), in addition to the lowest diameter. With increasing drought intensity, the SRL and average diameter (AD) of all root branch orders increased and decreased, respectively. The fine roots in humus soil had a higher soluble sugar (SS) content and lower starch (ST) content compared to the loam and sandy-loam soil. Additionally, the SS and ST contents of fine roots showed decreasing and increasing tendencies with increasing drought intensities, respectively. SS and ST explained the highest degree of the total variation in fine root traits, which were 32 and 32.1%, respectively. With increasing root order, the explanation of the variation in root traits by ST decreased (only 6.8% for 5th-order roots). The observed response in terms of morphological traits of different fine root branch orders of F. mandshurica seedlings to resource fluctuations ensures the maintenance of a low cost-benefit ratio in the root system development.