## 2 Results
### Functional Annotation  
To more fully understand the mechanisms governing terpenoid biosynthesis in _A. argyi_, functional annotation of FLNC transcripts was conducted using a series of databases. In total, 69,178 unique isoforms were annotated with BLASTX (v 2.2.26) and the COG (Clusters of Orthologous Groups) [12], GO [13], KEGG [14], KOG [15], Pfam (Protein family) [16], Swissprot [17], and NR [18] databases (Figure 2A, Table S10). Figure 2: Functional annotation and classification of assembled _A. argyi_ unigenes. (**A**) Annotation details arranged in a Venn diagram. (**B**) KEGG pathway classifications for putative proteins. (**C**) GO classifications for _A. argyi_ unigenes, including biological process (BP), cellular component (CC), and molecular function (MF) annotations. GO functional annotations were used to assign specific biological process (BP), molecular function (MF), and cellular component (CC) terms to the unique isoforms, yielding three primary GO categories and 56 subcategories. A large proportion of these genes were annotated in the BP terms (\"cellular process\", \"metabolic process\", and \"biological regulation\"), the CC terms (\"membrane\", \"membrane part\", and \"cell\"), and the MF terms (\"catalytic activity\", \"binding\", and \"transcription regulator activity\") (Figure 3C). In total, these genes were associated with 137 KEGG pathways, many of which were related to growth and metabolite accumulation (Figure 2B). Individual Transcriptomic Analyses of A. Argyi Leaf Samples Collected at Four Different Developmental Time Points  
Next, cluster dendrogram and principal component analysis (PCA) analyses were conducted using samples from different _A. argyi_ developmental stages. Samples from these four _A. argyi_ developmental stages were clustered into four separate groups, and the samples from the same collection month were assembled into the same branches, based on the results of the clustering tree, revealing clear differences in gene expression profiles among these samples (Figure 3A). PCA results revealed that the samples from May and April were significantly different from those collected in June and July, based upon the PC1 axis (Figure 3B), which is consistent with the results observed in the clustering dendrogram. The time-specific expression of genes was further assessed to reveal differences in the transcripts from the _A. argyi_ samples from four different time points. In total, 67,117 unigenes were applied to create gene intersection profiles. A total of 52,867 unigenes were shared among _A. argyi_ from the four harvest time points, with 1006, 658, 536, and 616 unigenes being specifically expressed in April, May, June, and July, respectively (Figure 4). Identification and Enrichment Analyses of DEGs Associated with Particular A. Argyi Developmental Stages  
To further explore patterns of differential gene expression across these _A. argyi_ samples collected at the four selected harvest time points, we employed fold change (FC) and FDR values to compare the expression among samples [11], leading to the identification of 140,611 DEGs, with individual comparisons among samples being made to identify DEGs that were up- or down-regulated. Specifically, DEGs were identified by comparing samples  
Figure 3: Gene expression levels during four _A. argyi_ developmental stages. (**A**) Clustering dendrogram of gene expression patterns in different _A. argyi_ developmental stage samples. (**B**) PCA of gene expression profiles for different _A. argyi_ developmental stage samples. from May and April, April and June, April and July, May and June, May and July, and June and July. The greatest number of DEGs (33,968) was observed when comparing samples from April and July, while the smallest number (15,351) was evident when comparing samples from June and July (Figure S3), consistent with the important roles of many of these genes during the different stages of _A. argyi_ development. In a soft clustering analysis, 12 gene clusters were obtained with distinctive characteristics.