Figure 4.

Functional characterization of TgbHLH95. A) Heatmap analysis of differentially expressed TFs. B) The regulatory effect of TFs on the TgGPPS promoter, the LUC/REN value of the empty SK vector on the TgGPPS promoter was set to 1.0, and Se values were calculated using 3 biological replicates. Different letters indicate significant differences (P < 0.05) during the postharvest ripening stage by Duncan's multiple range test. C) Subcellular localization analysis of TgbHLH95. The fluorescent signals were detected in tobacco cells cotransfected with TgbHLH95-GFP and the nuclear RFP-NLS marker. Scale bar = 20 μm. D) Yeast 1-hybrid analysis of TgbHLH95 binding capacity to the TgGPPS promoter, with the empty vector pGADT7 (AD) used as a negative control. E) The TgbHLH95 expression pattern in xiangfei nuts during the postharvest ripening stage. Error bars represent Se based on 3 biological replicates. The different letters indicate significant differences (P < 0.05) during the postharvest ripening stage by Duncan's multiple range test. F) Transient overexpression of TgbHLH95 increases total monoterpene content in tobacco leaves. Error bars represent Se based on 3 biological replicates. Significant differences are indicated with asterisks using Students's test (*P < 0.05).