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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\u2013HY5\u2013MYB module facilitates light-induced stomatal opening, providing evidence that flavonoid metabolism in guard cells is crucial for plant stress tolerance.<\/jats:p>","DOI":"10.1093\/plcell\/koaf064","type":"journal-article","created":{"date-parts":[[2025,3,31]],"date-time":"2025-03-31T02:58:21Z","timestamp":1743389901000},"source":"Crossref","is-referenced-by-count":16,"title":["A CRY1\u2013HY5\u2013MYB signaling cascade fine-tunes guard cell reactive oxygen species levels and triggers stomatal opening"],"prefix":"10.1093","volume":"37","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9998-9802","authenticated-orcid":false,"given":"Yuankai","family":"Chang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Cotton Bio-Breeding and Integrated Utilization, School of Life Sciences, Henan University , Kaifeng, Henan Province 475004 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BS8 1TD ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6642-5306","authenticated-orcid":false,"given":"Xiaobin","family":"Ou","sequence":"additional","affiliation":[{"name":"Gansu Key laboratory of Protection and Utilization for Biological Resources and Ecological Restoration, Longdong University , Qingyang, Gansu Province 745000 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8995-9095","authenticated-orcid":false,"given":"Ke","family":"Yu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Cotton Bio-Breeding and Integrated Utilization, School of Life Sciences, Henan University , Kaifeng, Henan Province 475004 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