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Metagenomic sequencing was used to reveal the dynamic changes in microbiota and the metabolic functions in corncob composting for preparing cultivation medium of Pleurotus ostreatus. Results showed that the changes of physicochemical properties lead to different dominant phylum at different stages of composting. Firmicutes replaced Proteobacteria as the dominant phylum at thermophilic stage. Correlation analysis indicated that the succession of microbiota was significantly affected by the C/N ratio, pH, temperature and organic matters