A CRY1-HY5-MYB signaling cascade fine-tunes guard cell reactive oxygen species levels and triggers stomatal opening.

Stomatal opening facilitates CO2 uptake and causes water loss via transpiration. Compared with the considerable progress made toward understanding phototropin-mediated blue light (BL) signaling in guard cells, the significance of cryptochromes (CRYs) in stomatal opening and their downstream elements remain largely unknown. Here, we show that 3 homologous MYB transcription factor genes, namely MYB11, MYB12, and MYB111, are rapidly transactivated in guard cells during the dark-to-light transition in Arabidopsis (Arabidopsis thaliana). Genetic characterization of myb mutants demonstrates that these proteins specifically mediate light-induced stomatal opening by promoting local flavonol accumulation, thereby controlling reactive oxygen species homeostasis in guard cells. In response to light, activation of the plasma membrane H+-ATPase is inhibited in the myb11 myb12 myb111 triple mutant, compromising transmembrane K+ influx in the mutant guard cells. Furthermore, we demonstrate that MYB11/12/111 expression in guard cells upon illumination is induced by a CRY1-specific signaling cascade involving ELONGATED HYPOCOTYL 5 (HY5), a direct transcriptional activator of these MYBs. Overall, our work reveals a mechanism by which the CRY1-HY5-MYB module facilitates light-induced stomatal opening, providing evidence that flavonoid metabolism in guard cells is crucial for plant stress tolerance.