The impact of climate change in the European Alps has been roughly twice the global average, dramatically reducing permafrost extent and thickening of its active layer. Therefore, the study of the abiotic factors (i.e. chemical/physical parameters) affecting the microbial diversity inhabiting Alpine permafrost appears to be of dramatic relevance. Within the European Alps, the Stelvio area exhibits these effects in a particularly evident way, with important consequences on microbial ecosystems. Therefore, microbial communities inhabiting a permafrost core collected in the Scorluzzo active rock glacier (Stelvio Pass, Italian Central Alps) were investigated along a depth gradient (410 to 524 cm from the surface). The taxonomic structures of bacterial and fungal communities were investigated via a next-generation sequencing (NGS) approach (Illumina MiSeq), targeting the bacterial V3-V4 regions of 16S rDNA and the fungal ITS2 region. Abiotic soil factors (grain size, electrical conductivity, ice/water content, pH, Loss-on-Ignition - LOI, total and organic carbon, nitrogen and phosphorous) were analysed. Richness and Shannon-H diversity indices were correlated to abiotic factors. Bacterial diversity was significantly (p < 0.05) correlated with LOI, while fungal diversity was significantly (p < 0.05) correlated with the depth gradient. The Constrained Analysis of Principal (CAP) coordinates were used to study the correlation between abiotic parameters and the taxonomic structure of bacterial and fungal communities. Among all tested variables, the depth gradient, water content, pH and LOI affected the taxonomic structure of bacterial communities (in particular, the abundance of bacterial amplicon sequence variants - ASVs - assigned to Afipia sp., Chloroflexi, Gaiella sp., Oryzihumus sp. and Serratia, sp.), while fungal communities (ASVs assigned to Naganishia sp., Rhodotorula sp., Sordariomycetes and Taphrinales) were affected by the depth gradient. Co-occurrences (calculated by Pearson correlation coefficient) among microbial taxa (i.e. bacteria vs bacteria, bacteria vs fungi, fungi vs fungi) were investigated: the prevalence of significant (p < 0.05) positive co-occurrences was found, suggesting that the coexistence of different microbial taxa could play a crucial role in maintaining the ecological and taxonomic balance of both bacterial and fungal communities inhabiting the Alpine permafrost ecosystem. These findings suggest that the bacterial and fungal diversity of Alpine permafrost are affected in different ways by some abiotic factors. •The structure of bacterial communities is affected by depth, % of water, pH and LOI.•The structure of fungal communities is affected by depth.•Bacterial communities are dominated by Actinobacteria and Proteobacteria.•Meyerozyma is the prevalent fungal genera found in Alpine permafrost.•The prevalence of positive co-occurrences between bacteria and fungi was found.