Soil organic matter primarily originates from microorganisms and litter, which exhibit significant differences in carbon (C), nitrogen (N), and phosphorus (P) content as well as their stoichiometric ratios. This variation enables the use of ecological stoichiometry methods based on input source differentiation to trace soil organic matter. However, there is currently a lack of a global database on the stoichiometric ratios of microorganisms and litter to support the application of this method. Through a global integrated analysis, 4357 ecological-chemical characteristics of litter and 3107 of microorganisms were collected, covering forest, grassland, and cropland ecosystems. The results showed that the global average atomic ratio for litter was C：N：P=2880：47：1, while for microorganisms, it was C：N：P=108：7.3：1. Significant differences in the stoichiometric ratios of litter were observed across ecosystems, with the highest C：N ratio in agricultural ecosystems (77.5), the highest C：P ratio in grassland ecosystems (4148), and the highest N：P ratio in grassland ecosystems (64.8). Similarly, there were significant differences in the stoichiometric ratios of microorganisms across ecosystems, with the highest C：N ratio in forest ecosystems (19.9), the highest C：P ratio in forest ecosystems (131), and the highest N：P ratio in grassland ecosystems (8.7). Simultaneously, within the same ecosystem, significant differences exist in the stoichiometric ratios between litter and microorganisms. This provides both feasibility and scientific justification for utilizing these differences in stoichiometric ratios to trace soil organic matter.