The LoMYB26/LoJAZ4-LoCOMT module regulates anther dehiscence via Jasmonic acid-mediated endothecium lignification in lily

Timely anther dehiscence is a key step for successful sexual reproduction in plants. Secondary cell wall thickening of anther endothecium is a vital process during anther dehiscence that provides an indispensable mechanical force for successful dehiscence. Anther dehiscence depends on anther lignification, and it is a timely and sophisticated process regulated by phytohormones and transcription factors. However, whether endothecium lignification occurs during anther dehiscence in lily and underlying mechanisms are still largely unclear. Our work focuses on identifying the course of endothecium lignification during anther dehiscence and elucidating the molecular mechanisms underlying endothecium lignification-dependent anther dehiscence in lily. Lignin fluorescence analysis and ultraviolet spectrophotometry were employed to elucidate the endothecium lignification process. Target genes were isolated from the transcriptomic data of anther dehiscence and lignification process. Virus-induced gene silencing (VIGS) and transient overexpression in lily anthers were used to analyze the LoMYB26 function. Yeast one-hybrid (Y1H), electrophoretic mobility shift assay (EMSA), and dual-luciferase (LUC) assay analyzed the regulatory mechanisms. Yeast two-hybrid (Y2H), luciferase complementation imaging (LCI), and bimolecular fluorescence complementation (BiFC) assays illustrated the interaction between LoMYB26 and LoJAZ4. Our results showed that endothecium lignification occurred in S6-S7 stages when anther dehiscence had not yet occurred. The R2R3-type MYB transcription factor, LoMYB26, was found to promote endothecium lignification. LoMYB26 directly bound to the Caffeic Acid O-methyltransferase (LoCOMT) promoter and activated its transcription. Meanwhile, LoMYB26 interacted with jasmonate-ZIM domain protein 4 (LoJAZ4), which repressed the LoMYB26-mediated activation of LoCOMT transcription. Additionally, the exogenous application of methyl-jasmonate (Me-JA) induced LoMYB26 transcription and promoted endothecium lignification. Our findings demonstrate that LoMYB26 promotes endothecium lignification and anther dehiscence. LoMYB26 interacted with LoJAZ4, forming a heterodimer that participates in JA-mediated endothecium lignification and anther dehiscence. This study offers valuable insights and a theoretical foundation for the breeding of anther-indehiscent lily.