## 3 Results  
### Metabolome Analysis of Caoyuanheino-I (Agaricus sp.) during growth and Development  
A total of 449 metabolites were detected through untargeted metabolomics. These metabolites were classified into 16 super classes, and 18 metabolites were not annotated into categories. Among them, the super class with the highest number of metabolites annotated was lipids and lipid-like molecules, which contained 109 metabolites (Table S1). The results of the principal component analysis (PCA) for differentially accumulated metabolites showed differences in metabolomic data among the three developmental periods (Figure S1). Eleven differentially accumulated metabolites were found in the HY vs. HZ group, with nine metabolites significantly increasing (\(p\) < 0.05) in content during the HZ period, including DL-dopa (HMDB0000609), methylglutaric acid (HMDB0061676), D-xylitol (HMDB0002917), uracil (HMDB0000300), pyruvate acid (HMDB000243), 1,5-anhydroorbitol (HMDB0002712), S-adenosylhylomocysteine (HMDB0000939), oxoadipic acid (HMDB0000225), and 4-[(E)-2-(3,5-dimethoxyylphenyl)]phenylol (HMDB0130987). Additionally, two metabolites significantly decreased (\(p\) < 0.05) in content during the HZ period: nicotinic acid (HMDB0001488) and urocanic acid (Figure 2a,b). For the HY vs. HZ group, the results showed that the differentially accumulated metabolites were most significantly enriched (\(p\) < 0.05) for pantobinate and CoA biosynthesis, pentose and glucuronate interconversions, methane metabolism, and the cysteine pathway (Figure 2c). In the HY vs. HC group, there were 16 differential metabolites, of which 14 had a significant increase (\(p<0.05\)) in content and 2 had a significant decrease (\(p<0.05\)) in content in the HC group. The metabolites with increased levels included 5Z-dodecenoic acid (HMDB0000529),  
Figure 2: A comparative analysis of the different metabolite types in the different developmental stages of Caoyuanheimo\(-1\) (_Agaricus_ sp.). HY represents the primordium period of Caoyuanheimo\(-1\), HZ represents the young fruiting period of Caoyuanheimo\(-1\), and HC represents the ripe fruiting period of Caoyuanheimo\(-1\). Volcano plots show the differentially accumulated metabolites between the HY vs. HZ (**a**), HY vs. HC (**d**), and HZ vs. HC (**g**) groups. Each point in the volcano map represents a metabolite; the X axis represents the logarithm of the fold change of a certain metabolite in the two samples; and the Y axis represents the logarithm of the \(p\)-value. The blue dots represent downregulated differentially accumulated metabolites; the pink dots represent upregulated differentially accumulated metabolites; and gray represents detected but not significantly differentially accumulated metabolites. The size of the dots represents different VIP values. Heatmaps visualization shows the differential metabolites accumulated between the HY vs. HZ (**b**), HY vs. HC (**e**), and HZ vs. HC (**h**) groups. The relative accumulation levels of individual samples are shown; the transition from red to blue represents the level of metabolite accumulation. The differentially accumulated metabolite enrichment analysis of the different development stages are shown as bubble charts of the KEGG enrichment analyses between the HY vs. HZ (**c**), HY vs. HC (**f**), and HZ vs. HC (**i**) groups. galactaric acid (HMDB0000639), guanosine (HMDB0000133), diaminopimelic acid (HMDB0001370), 2\({}^{\prime}\)-deoxyguanosine 5\({}^{\prime}\)-monophosphate (HMDB0001044), 3-hydroxybutyric acid (HMDB0000357), dodecenoic acid (HMDB0000638), D-xylitol (HMDB0002917), methyl-succinic acid (HMDB0001844), phenylacetic acid (HMDB0000209), pyruvate acid (HMDB0000243), 4-[(E)-2-(3,5-dimethoxyyl)ethenyl]phenol (HMDB0130987), succinic acid (HMDB0000254), and D-2,3-dihydroxypropanoic acid (HMDB0031818). The metabolites with decreased levels included folinic acid (HMDB0001562) and glutathione (HMDB0000125) (Figure 2d,e).