Aridity is known to influence the structure and function of the soil microbiome and their connection with ecosystem functions, however, whether aridity leads to gradual (or abrupt) and systemic (or specific) changes in the microbiome-ecosystem functions relationships in alpine ecosystems is largely unknown. Here, we conducted a survey of 60 sites along an aridity-gradient transect across the Tibetan Plateau to investigate changes in microbial diversity and functions and to evaluate their relationship with soil functionality. We found that plant richness and biomass, soil microbial richness (e.g., bacteria, fungi, diazotrophs, saprotrophs, pathotrophs and symbiotrophs), the relative abundance of copiotrophs and soil functionality (e.g., C-, N-, and P cycling, multifunctionality) decreased along the aridity gradient, whereas the proportion of genes associated with organic matter decomposition and denitrification increased (obtained from metagenomic sequencing). Above an aridity threshold of ∼0.6, soil multifunctionality decreased slowly and supported a larger proportion of genes associated with cellulose, hemicellulose, chitin, lignin and pectin degradation as well as genes associated with denitrification (e.g., nosZ, nosZ atypical 2, norB-cNor and norB-qNor). In drier habitats (>0.6 aridity level), microbial diversity and genes involved in organic matter decomposition and N cycling were strongly correlated with soil multifunctionality, whereas plant diversity was closely associated with soil multifunctionality in wetter habitats (<0.6 aridity level). Plant biomass and soil organic C play a key role in regulating microbial compositional and functional diversity and further affect soil multifunctionality in drier habitats. These findings suggest that soil microbiomes may play a larger role in maintaining soil multifunctionality in more arid environments than in less arid environments. Our results highlight the threshold responses of microbial diversity and functions to aridity and provide novel insight into mechanisms by which climate change affects ecosystem functions.