Figure 1

Bio-guided separation of the active components from Acer truncatum leaves. (A) Flow chart of bio-guided separation. (B) Fluorescent microscopy images after treatment of separated fractions. (C) Fold change of 2-NBDG fluorescence measured by ImageJ (** p < 0.01; **** p < 0.0001). The fold changes were calculated from the mean of the blank control. Green and blue are two different batches. The higher fold change indicates the better promotion of glucose uptake.

Figure 2

Myricitrin and myricetin-induced 2-NBDG uptake. (A) Chemical structures of myricitrin and myricetin. The structures were identified on the basis of HR MS/MS (Figure S2) and NMR spectra (Figure S3). (B) Fold change of 2-NBDG fluorescence measured by ImageJ (**** p < 0.001, ns, no statistical difference). The higher fold change indicates the better promotion of glucose uptake. (C) Fluorescent microscopy images after treatment of myricitrin and emodin.

Figure 3

LC-MS/MS features of leaves from different years old trees (A. truncatum). (A) PCA analysis; (B) sPLS-DA analysis with multivariate prediction. (C) Content change trend of myricitrin-like components. 1-myricitrin; 2-myricetin; 3-myricetin-3-O-pentoside; 4-myricetin-3-rutinoside. Peak areas were calibrated with an external standard myricitrin. (D) Chemical structures of myricitrin-like components.

Figure 4

Main components in leaves affected by tree ages. (A) Hot spot map of increasing and decreasing components depending on tree age. The color palette represents the content of metabolites. (B) Components counts in different structural types. (C) Compounds up-regulated annually (FC > 32, p < 0.01). (D) Representative compounds down-regulated annually (FC > 32, p < 0.01). See Table S1 for details.

Figure 5

Visual analysis of differentially expressed genes (DEGs) and differential metabolites (DMs). (A) sPLS-DA analysis of DEGs. (B,C) valcono plot of DEGs (p < 0.01). (D) Venn plots of DEG counts. a, model vs. blank; b, test vs. model. (E) sPLS-DA analysis of DMs. (F,G) valcono plot of DMs (FC > 2, p < 0.01). (H) Venn plots of DM counts. a, model vs. blank; b, test vs. model.

Figure 6

KEGG pathway analysis based on combining DEGs and DMs.

Figure 7

RT-qPCR validation of DEGs related to myricitrin’s regulation.

Acknowledgments
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