Heat‐responsive <scp>MaHSF11</scp> transcriptional activator positively regulates flavonol biosynthesis and flavonoid B‐ring hydroxylation in banana ( <i>Musa acuminata</i> )

Plant flavonols act primarily as ultraviolet radiation absorbers, reactive oxygen species scavengers, and phytoalexins, and they contribute to biotic and abiotic stress tolerance in plants. Banana (Musa acuminata), an herbaceous monocot and important fruit crop, accumulates flavonol derivatives in different organs, including the edible fruit pulp. Although flavonol content varies greatly in different organs, the molecular mechanisms involving transcriptional regulation of flavonol synthesis in banana are not known. Here, we characterized three SG7-R2R3 MYB transcription factors (MaMYBFA1, MaMYBFA2, and MaMYBFA3) and heat shock transcription factor (MaHSF11), to elucidate the molecular mechanism involved in transcriptional regulation of flavonol biosynthesis in banana. MaMYBFA positively regulates flavonol synthase 2 (MaFLS2) and downregulates MaFLS1. We show these transcription factors to be weak regulators of flavonol synthesis. Overexpression of MaHSF11 enhances flavonol contents, particularly that of myricetin, and promotes flavonol B-ring hydroxylation, which contributes to the diversity of flavonol derivatives. MaHSF11 directly interacts with the MaFLS1 and flavonoid 3',5'-hydroxylase1 (MaF3'5'H1) promoters, both in vitro and in vivo. MaHSF11 activates the expression of MaDREB1 directly, which is known to promote cold and chilling tolerance in banana fruit. Overall, our study elucidates a regulatory mechanism for flavonol synthesis in banana and suggests possible targets for genetic optimization to enhance nutritional value and stress responses in this globally important fruit crop.