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Understanding the seasonal variation in root biomass (root diameter less than or equal to 5 mm) of Pinus hwangshanensis would provide a theoretical basis for the sustainable development of P. hwangshanensis forestry in China. In the natural habitat of P. hwangshanensis, five investigation sites along an elevational gradient from 1000 m to 2000 masl (meters above sea level) were established at 200 m intervals. The root biomass of P. hwangshanensis, along with the diameter at breast height (DBH) and plant height (H), were measured seasonally at each elevational site. We found that the average annual biomass of small roots (d ≤ 2 mm) and medium roots (2 mm < d ≤ 5 mm) was 158.76 g/m² and 4.68 g/m² respectively. The fine-root biomass and medium-root biomass of P. hwangshanensis changed significantly along the elevational gradient. The fine-root biomass of P. hwangshanensis did not change from season to season. At each site, however, the medium-root biomass varied significantly across seasons, with higher biomass observed in winter than in all other seasons (P < 0.05). In summary, the fine-root biomass and medium-root biomass of P. hwangshanensis varied significantly along the elevational gradient; the biomass of fine and medium roots showed divergent seasonal variations, suggesting that different seasonal growth mechanisms occurred between the two root types.