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The glacial retreat is observed and predicted to increase in intensity especially in high-elevation areas as a result of global warming, which leaves behind a primary succession along soil chronosequences. Although soil microbes have been recognized as main drivers of ecological and evolutionary processes, our understanding of their effects on nutrient biogeochemistry during primary succession remains limited. In this study, we investigated changes in the microbial community structure, ecoenzymatic stoichiometry, and glomalin-related soil protein (GRSP) accumulation in the Hailuogou Glacier Chronosequence, located on the eastern Tibetan Plateau. We wanted to reveal the effects of nutrient limitation on soil microbes and the relative contributions of edaphic and biotic factors. The results showed that with an increasing soil age, there was a steady increase in the microbial biomass and a shift from a bacterial to fungal dominated pattern. Soil enzyme stoichiometry and analyses on threshold elemental ratios revealed that microbial activities are limited by carbon and nitrogen during the early successional stage (3-52 years), while phosphorus was the main limiting factor during later stages (80-120 years). Moreover, the redundancy analysis and structural equation modeling suggested that during early stages edaphic factors had a greater impact on microbial processes, while the vegetation factors were most influential during the last two stages. Overall, these results highlighted the importance of integrating knowledge of the microbial community structure, soil enzyme activities and GRSP to gain a holistic view of soil-plant microbe interactions during ecosystem successions.