The karst ecosystem maintains rich microbial diversity, and the composition and structure of arbuscular mycorrhizal fungi changes with karst vegetation succession. In this experiment, we used a space-time substitution method to collect soil from tree, bush, and herb communities from Huaxi Guiyang, Zhijin Bijie, and Huajiang Guanling from a typical karst area located in Guizhou. Using Illumina HiSeq molecular sequencing technology, we performed an operation taxonomic unit (OTU) clustering analysis and compared the annotated species with those in a database to explore the soil arbuscular mycorrhizal fungi (AMF) species diversity during the different karst succession stages. There were 275 Glomeromycota OTUs in 19 species from 4 orders, 8 families, and 13 genera in the karst habitat soil. AMF abundance at the genus level showed that Rhizophagus was the dominant genus, and Huajiang had the highest AMF richness. The common species Septoglomus constrictum, Rhizophagus intraradices, Claroideoglomus sp.MIB8381,and Entrophospora infrequens were distributed at different stages of the vegetation succession at every sampling site. The Shannon and Simpson's indices of AMF changed at the different stages of succession as follows:in Huaxi, tree/bush > herb (P < 0.05); in Huajiang, bush/herb > tree (P < 0.05); and in Zhijin, tree > bush > herb, but these relationships were not significantly different. The Chao1 and abundance-based coverage estimation (ACE) indices showed that in Huajiang, bush/herb > tree (P < 0.05). The Spearman correlation analysis showed that soil total phosphorus was significantly and negatively correlated with the ACE index of AMF, and it was negatively correlated with the Chao1 index. Available phosphorus was negatively correlated with the Shannon and Simpson's indices. The canonical correlation analysis showed that soil total nitrogen, available nitrogen, organic matter, total phosphorus, and available potassium were significantly correlated with the community distribution of AMF. The results showed that although there was no uniform variation law, the diversity of soil AMF increased or decreased with the process of karst vegetation succession, which was closely related to the physicochemical properties of the soil, and the influence of phosphorus was the greatest.