<p>Melatonin, a hormone and signaling molecule, plays a key role in regulating plant growth, development, and stress responses. It is particularly effective in mitigating abiotic stresses, such as salinity. However, the regulatory pathways governing the expression of melatonin biosynthesis genes, particularly <i>O-methyltransferase 1</i> (<i>OMT1</i>), in monocotyledonous plants under stress conditions remain a mystery. In this study, we employed a combination of computational and experimental approaches to investigate the molecular basis of salinity stress tolerance in wheat concerning melatonin biosynthesis. Our analysis of the <i>OMT1</i> gene promoter and RNA-Seq data of wheat leaves under salinity stress led to a groundbreaking discovery. We found that a Jasmonic acid (JA)-responsive transcription factor, WRKY70, may regulate <i>OMT1</i> gene expression. This finding was further extended through molecular and physiochemical assays and advanced statistical analyses. Our results revealed a significant negative correlation in the expression levels of these genes under JA treatment, salinity stress, and their combination. We also identified a significant relationship between gene expression and melatonin content under experimental conditions. These findings provide a novel perspective on the relationship between JA signaling and the melatonin biosynthesis pathway. Furthermore, the <i>WRKY70</i>-<i>OMT1</i> regulatory module appears to enhance salinity tolerance in wheat by modulating melatonin levels, improving photosynthetic efficiency, and significantly alleviating the adverse effects of salinity stress. This study provides new insights into the intricate regulatory networks underlying wheat’s responses to salinity stress and highlights the potential of targeting melatonin biosynthesis pathways to enhance crop resilience to abiotic stresses.</p>