| Summary: | (1) Background: environmental gradient strongly affects microbial biodiversity, but which factors drive the diversity of arbuscular mycorrhizal fungi (AMF) associated with roots at relatively large spatial scales requires further research; (2) Methods: an experiment on large spatial scales of Mt. Taibai was conducted to explore the biodiversity and drivers of AMF-associated with roots using high-throughput sequencing; (3) Results: a total of 287 operational taxonomic units (OTUs) belong to 62 species representing 4 identified and 1 unclassified order were identified along different altitudinal gradients. With increasing altitude, AMF colonization could be simulated by a quadratic function trend, and altitude has a significant impact on colonization. AMF alpha diversity, including the Sobs and Shannon indexes, tended to be quadratic function trends with increasing altitude. The highest diversity indices occurred at mid-altitudes, and altitude had a significant effect on them. AMF communities have different affinities with soil and root nutrient, and <i>Glomus</i> is most affected by soil and root nutrient factors through the analysis of the heatmap. <i>Glomus</i> are the most dominant, with an occurrence frequency of 91.67% and a relative abundance of 61.29% and 53.58% at the level of species and OTU, respectively. Furthermore, AMF diversity were mostly associated with soil and root nutrients; (4) Conclusions: in general, AMF molecular diversity is abundant in Mt. Taibai, and altitude and nutrient properties of soil and root are the main influencing factors on AMF diversity and distribution.
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