Nts, and molecular functions), respectively. The enrichment outcomes of the three big biological functions of GO are shown in Fig. 4A,B (P worth 0.03). Probably the most dominant terms integrated the oxidationreduction approach, integral components of membranes, oxidoreductase activity, CDK2 Inhibitor drug monooxygenase activity, and iron ion binding. KEGG pathway enrichment evaluation was also conducted; 97 and 220 DEGs have been enriched, corresponding to 77 and 103 pathways, respectively. In the method of ossification of O. sinensis, “Starch and sucrose metabolism”, “Tryptophan metabolism”, “Tyrosine metabolism”, and “Sphingolipid metabolism” pathways were considerably enriched (Fig. 4C). In the FB formation stage, the degree of enrichment of “Biosynthesis of antibiotics” and “Carbon metabolism” varied tremendously (Fig. 4D). These metabolic pathways might closely relate towards the formation of sclerotia and fruiting body. All DEGs, too as GO and KEGG evaluation final results, are shown in Table S3. On the other hand, some DEGs encoded functionally unknown proteins, which might relate to O. sinensis growth and improvement; additional research will probably be required to confirm their functionalities. Evaluation of GSEA substantial enrichment. Although GO- and KEGG-based analyses have compared theDEGs from functional categories, productive statistical solutions have been not employed to analyze the overall trend of gene expression. Consequently, GSEA was adapted to analyze the enrichment of genes differentially expressed in every GO term and KEGG pathway. In this study, normalized enrichment scores were utilized to draw a cluster heat map (P value 0.05) (Fig. five). In the process of sclerotium formation, phosphorylation-related signaling (phosphorelay signal IL-6 Antagonist MedChemExpress transduction method (GO:0000160), signal transduction by protein phosphorylation (GO:0023014), phosphorelay sensor kinase activity (GO:0023014), and oxidative phosphorylation (ko00190)) and oxidation-related (response to oxidative tension (GO:0000155), cellular oxidant detoxification (GO:0098869) peroxidase activity (GO:0004601), and monooxygenase activity (GO:0004497)) had been significantly upregulated. For the duration of the fruiting body development stage, the expression trend of ribosome-related terms and pathways have been elevated significantly, which includes the nucleolus (GO:0005730), ribosome (GO:0005840, ko030100), 90S preribosome (GO:0030686), and ribosome biogenesis in eukaryotes (ko03008). Nonetheless, carbohydrate transport (GO:0008643), fatty acidScientific Reports |(2021) 11:12944 |https://doi.org/10.1038/s41598-021-91718-x3 Vol.:(0123456789)www.nature.com/scientificreports/Figure 3. Comparative analysis of differentially expressed genes (DEGs) and milRNA (DEMs) in the MC, ST, and FB stage. (A) Numbers and (B) Venn diagram of DEGs in between two samples (MC_vs_ST, ST_vs_FB, ST_vs_FB). (C) Numbers and (D) Venn diagram of DEMs in between two samples (MC_vs_ST, ST_vs_FB, ST_vs_ FB). degradation (ko00071), glyoxylate and dicarboxylate metabolism (ko00630), and carbon metabolism related to energy metabolism had been downregulated. Some prominent pathways are listed in Table 1, for instance the MAPK signaling pathway. DEGs in this pathway incorporate mitogen-activated protein kinase kinase (pbs2), glycerol-3-phosphate dehydrogenase (gld1), catalase (cat1), and also other critical genes encoding enzymes. The expression of Cat was upregulated with log2(fold change) of 2.312 and downregulated with log2(fold adjust) of 2.160, respectively. In the citrate cycle, the genes encoding the enzymes malate dehydrogenase (mdh1), phosp.