The distribution characteristics of soil microbial diversity and community structure with altitude in high-altitude cold regions are the theoretical basis for understanding and predicting the construction process and driving mechanism of microbial communities under the background of climate change. [Objective] To gain an in-depth understanding of the variation patterns and influencing factors of soil fungal communities in alpine meadows at different altitudes along the altitude, this paper takes the alpine meadows at an altitude of 3,300-4,500 meters in Golok Tibetan Autonomous Prefecture, Qinghai Province as the research object. [Method] Utilizes Illumina high-throughput sequencing and the response characteristics of soil fungal community diversity in alpine meadows to altitude gradient changes and its key influencing factors were explored through redundancy analysis. [Result] The results indicate that the soil fungal communities in the 0–20 cm soil layers across different altitudes in the study area are predominantly composed of seven taxonomic groups. Ascomycota, Basidiomycota, Mortierellomycota, and unclassified fungal taxa represent the dominant fungal communities with the highest relative abundances in the alpine meadows of the study area. Notably, the abundance of unclassified fungi is significantly higher at the high-altitude site (4500 m) compared to other elevations. The soil microbial communities in the alpine environment of this study area provide valuable opportunities for the in-depth discovery and exploration of unknown microorganisms. Soil bacterial diversity has different changing rules along the elevation, and Ace and Chao1 index increase first and then decrease along the elevation gradient. The composition of soil fungal community at 3300 m, 3900 m and 4200 m above sea level is relatively similar. The composition of soil fungi community was different between 3600 m and 4500 m above sea level. [Conclusion] The structure and diversity of soil microbial communities exhibit clear selectivity toward soil physicochemical environments. Soil pH and available nitrogen are the primary factors influencing fungal diversity in the study area, while altitude determines the distribution patterns of soil fungal community composition, structure, and diversity in alpine meadows.