This review article focuses on the transcriptional regulation of flavonol biosynthesis in plants, a crucial process for plant growth and stress resistance. Flavonols, a class of flavonoids, play significant roles in human health due to their antioxidant properties and potential health benefits. The article highlights recent advancements in understanding the transcriptional regulation of flavonol biosynthesis, including the characterization of various transcription factors (TFs) and microRNAs (miRNAs). Direct transcriptional regulation involves TFs such as MYB, AP2/ERF, and WRKY, which directly target key genes in the flavonoid biosynthetic pathway. Cascade transcriptional regulation involves miRNAs targeting TFs, interactions between activators and repressors, and the degradation of activators or repressors induced by UV-B light or plant hormones. These complex regulatory mechanisms allow plants to fine-tune flavonol homeostasis, balancing growth and stress responses. The review also discusses the role of specific TFs, such as SG7 R2R3-MYB, SG19 R2R3-MYB, and WRKY TFs, in regulating flavonol biosynthesis. Additionally, it explores post-transcriptional regulation by miRNAs and the influence of phytohormones like jasmonates, auxins, gibberellic acid, and abscisic acid on flavonol biosynthesis. The article concludes by emphasizing the importance of further research to uncover novel TFs and regulatory mechanisms in horticultural crops, as well as the potential for molecular design strategies to enhance flavonol content in these crops.This review article focuses on the transcriptional regulation of flavonol biosynthesis in plants, a crucial process for plant growth and stress resistance. Flavonols, a class of flavonoids, play significant roles in human health due to their antioxidant properties and potential health benefits. The article highlights recent advancements in understanding the transcriptional regulation of flavonol biosynthesis, including the characterization of various transcription factors (TFs) and microRNAs (miRNAs). Direct transcriptional regulation involves TFs such as MYB, AP2/ERF, and WRKY, which directly target key genes in the flavonoid biosynthetic pathway. Cascade transcriptional regulation involves miRNAs targeting TFs, interactions between activators and repressors, and the degradation of activators or repressors induced by UV-B light or plant hormones. These complex regulatory mechanisms allow plants to fine-tune flavonol homeostasis, balancing growth and stress responses. The review also discusses the role of specific TFs, such as SG7 R2R3-MYB, SG19 R2R3-MYB, and WRKY TFs, in regulating flavonol biosynthesis. Additionally, it explores post-transcriptional regulation by miRNAs and the influence of phytohormones like jasmonates, auxins, gibberellic acid, and abscisic acid on flavonol biosynthesis. The article concludes by emphasizing the importance of further research to uncover novel TFs and regulatory mechanisms in horticultural crops, as well as the potential for molecular design strategies to enhance flavonol content in these crops.