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Figure 1. 

Transcriptome information and analysis of CsWOX1-OE. (a) The principal component analysis (PCA) of differently expressed transcripts from two groups. (b) Statistics on the number of up-regulated and down-regulated DEGs. (c) The number and proportion of DEGs annotated in different databases.

Figure 2. 

The GO classification analysis results show the differences in the number and categories of annotated DEGs between the mf and CsWOX1-OE, with three main GO categories: Cellular component, Molecular function, and Biological process. (a) GO classification of DEGs in the mf sample. (b) GO classification of DEGs in the CsWOX1-OE sample. Each band represents the annotation status and trends of different GO terms and the numbers in parentheses indicate the number of genes in each category.

Figure 3. 

KEGG pathway analysis of the DEGs from mf and CsWOX1-OE shows the number and proportion of DEGs annotated in different metabolic pathways. The KEGG pathways are categorized into five groups: cellular processes, environmental information processing, genetic information processing, metabolism, and organismal systems.

Figure 4. 

Combined analysis of mf and CsWOX1-OE DEGs. (a) Identification of eight common DEGs between mf and CsWOX1-OE. (b) Heatmap of gene expression levels of 20 common DEGs with the same trend of change. (c) Heatmap of gene expression levels of 60 common DEGs with opposite trends. The colored legend positioned to the left of each map indicates fold changes (Log₂ value).

Figure 5. 

RT-qPCR analyses of (a) CsRVE6, (b) CsRVE8, (c) CsFKF1, (d) CsPin1At, (e) CsABCB, and (f) CsGI in CsWOX1-OE and mf. Significance analyses compared to WT were performed with the two-tailed Student's t-test. Different letters (a, b, c) indicate significant differences between groups (p < 0.05). The values represent the mean ± SD (n = 3).

Figure 6. 

Yeast-one hybrid validation between CsWOX1 transcription factors and candidate genes, which showed that CsWOX1 bound to the promoters of CsRVE6 and CsPin1At.

Figure 7. 

CsWOX1 actives the expression of CsRVE6 and CsPin1At. (a) Structural diagram of the reporter and effector constructs used for LUC/REN assays. LUC activity measurement after transient expression of 35S:CsWOX1 with (b) ProCsPin1At:LUC and (c) ProCsRVE6:LUC in tobacco leaves. The LUC/REN ratio from the empty vector (62-SK) combined with ProCsPin1At:LUC and ProCsRVE6:LUC were used as the calibration. Two-tailed Student's t-test was performed and statistically significant differences were indicated by ** p < 0.01. Values are mean ± SD (n = 5).