International Journal of Horticulture, 2025, Vol.15, No.1, 8-20 http://hortherbpublisher.com/index.php/ijh 16 The results indicate that the flavor formation in yellow-peel pitayas is influenced not only by sugar and acid metabolism but also by dynamic changes in gene expression. These findings provide a scientific foundation for optimizing fruit quality and developing high-flavor pitaya cultivars. Figure 3 Molecular characterization of HpDof1.7 and HpDof5.4. (A) Expression patterns of HpDof1.7 and HpDof5.4 during pitaya maturation. (B) Subcellular localization assay of HpDof1.7 and HpDof5.4. Bars=50 μm. (C) Transcriptional activation assay of HpDof1.7 and HpDof5.4 in yeast cells. (D) Schematic representation of reporter and effector vectors constructed for transcriptional activity analysis. (E) Transcriptional activity analysis of HpDof1.7 and HpDof5.4 in vivo. The data are normalized to a value of 1 for the 62SK-BD group. 62SK-BD-VP16 served as positive control. Each value represents means ± SE of six biological replicates (*P<0.05, **P<0.01, compared to 62SK-BD) (Adopted from Mou et al., 2022) Image caption: The figure shows that the expression levels of the transcription factors HpDof1.7 and HpDof5.4 gradually increase with fruit maturation and are primarily localized in the nucleus, indicating their role in transcriptional regulation. Yeast two-hybrid and dual-luciferase assays further validated the transcriptional activation activity of HpDof1.7 and HpDof5.4, demonstrating their ability to significantly enhance the activity of target gene promoters. Overall, the results confirm that these two factors act as positive regulators, directly activating the expression of sugar metabolism and transport genes, thereby promoting sugar accumulation in pitaya and providing crucial molecular support for its sweetness and quality formation (Adapted from Mou et al., 2022)
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