MPK4 phosphorylates MYC2 transcription factors to regulate jasmonic acid signaling and herbivory responses in maize

Abstract Regulation of responses induced by herbivory and jasmonic acid (JA) remains poorly understood in the important staple crop maize (Zea mays). MYC2 is the key transcription factor regulating many aspects of JA signaling, while mitogen-activated protein kinases (MAPKs or MPKs) play important roles in various plant physiological processes. Using a combination of reverse genetics, transcriptome analysis, and biochemical assays, we elucidated the important role of mitogen-activated protein kinase 4 (MPK4) in maize resistance to insects and in JA signaling. Silencing MPK4 increased the JA and jasmonoyl-isoleucine levels elicited by wounding or simulated herbivory but decreased maize resistance to armyworm (Mythimna separata) larvae. We showed that MPK4 is required for transcriptional regulation of many genes responsive to methyl jasmonate, indicating the important role of maize MPK4 in JA signaling. Biochemical analyses indicated that MPK4 directly phosphorylates MYC2s at Thr115 of MYC2a and Thr112 of MYC2b. Compared with nonphosphorylated MYC2s, phosphorylated MYC2s were more prone to degradation and exhibited enhanced transactivation activity against the promoters of several benzoxazinoid biosynthesis genes, which are important for maize defense against insects. This study reveals the essential role of maize MPK4 in JA signaling and provides insights into the functions of MAPKs in maize.