To clarify the diversity and differences in the intestinal flora of mason bee Osmia excavata adult females from the diapause phase to the cocoon-breaking stage,and to analyze their role in host health and regulation of host growth and development,the Illumina NovaSeq second-generation high-throughput sequencing method was used to sequence the 16S rRNA V3 to V4 regions of gut bacteria and analyze changes in gut flora diversity from diapause to cocoon-breaking stages. The results re-vealed a total of 41 phyla,96 classes,198 orders,325 families,and 637 genera in the intestinal bacterial flora of O. excavata female adults during these stages. The core phylum in the intestinal bacterial flora of O. excavata adult females during the diapause phase to cocoon-breaking stage was Proteobacteria,with relative abundance ranging from 75.98％ to 91.41％;the core genus was Sodalis,with relative abundance ranging from 59.65％ to 74.66％ . Alpha diversity analyses showed that the diversity index of the intestinal flora increased significantly from the diapause phase to the post-diapause phase,followed by a decreasing trend from the diapause phase to the post-diapause phase. The species richness and ho-mogeneity of gut flora were highest during the post-diapause period and lowest during the diapause peri-od. Beta diversity analysis showed that the structure of the gut flora remained relatively stable from the diapause phase to the cocoon-breaking phase. LEfSe analysis revealed significant differences in intesti-nal flora at the level of phylum,class,order,family,and genus from the diapause phase to the cocoon-breaking phase,with the most significant differences appearing during the post-diapause period. The predicted functions of the gut flora at different times are closely related to the physiological process from diapause to post-diapause and cocoon-breaking in adults. This study indicated that O. excavata fe-male adults maintained the same species of core gut bacteria from the diapause phase to the cocoon-breaking phase. While the community structure was relatively stable,the diversity of the community changed significantly.
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