Abstract: Agropyron mongolicum is one of northern China’s most representative perennial forage grasses, showing strong tolerance to cold and drought. In plants, caffeic acid O-methyltransferase (COMT) is a key gene involved in the biosynthesis of lignin and melatonin, and plays an important role in regulating plant growth, biomass quality, and stress resistance. In this study, through the analysis of the full-length transcriptome data of Agropyron mongolicum, the COMT candidate gene AmCOMT1 was cloned. AmCOMT1 is highly expressed in tissues with high lignin content, such as stem and root, and its expression is induced by a variety of abiotic stresses, including drought and salt. Expression of AmCOMT1 in Arabidopsis wild type (Col-0) and mutant (omt1-2) significantly promoted the synthesis of lignin in transgenic Arabidopsis, restoring the lignin content and composition of the mutant to wildtype level and the lignin content in Col-0/35S:AmCOMT1 was increased by 11%. In addition, overexpression of AmCOMT1 increased the melatonin content in Col-0/35S:AmCOMT1 transgenic Arabidopsis. Under salt stress conditions, the average root length of this strain increased by 20.3% compared to the wildtype, showing higher stress resistance. In this study, we identified AmCOMT1 from Agropyron mongolicum as a key gene regulating both lignin biosynthesis and melatonin biosynthesis, improving the stress tolerance of transgenic Arabidopsis. Our results highlighted the application potential of AmCOMT1 in genetic improvement of forage grasses through molecular breeding.