Overexpression of CmDUF239-1 Enhances Cold Tolerance in Melon Seedlings by Reinforcing Antioxidant Defense and Activating the ICE-CBF-COR Pathway

Low-temperature stress is a major factor that limits the productivity and geographical distribution of melon (Cucumis melo L.). This study elucidates that CmDUF239-1 is a positive regulator of cold stress, and its underlying mechanisms are investigated using root-specific overexpression lines. Seedlings overexpressing CmDUF239-1 exhibited increased biomass and reduced relative electrical conductivity under cold stress. CmDUF239-1 overexpression promoted the accumulation of soluble sugar and proline, which was accompanied by enhanced activity of the proline biosynthetic enzyme Δ1-pyrroline-5-carboxylate synthase (P5CS) and suppressed activity of the proline-degrading enzyme proline dehydrogenase (PDH). Molecular analysis revealed that CmDUF239-1 overexpression upregulated antioxidant enzyme-related genes, sugar metabolism related genes, and proline-related genes. Furthermore, it activated key genes in the ICE-CBF-COR signaling pathway, including CmCBF1, CmCBF2, and the downstream effector gene CmCOR413-2. In conclusion, the CmDUF239-1 gene enhances melon cold tolerance by modulating antioxidant defense, enhancing osmolyte (sugar and proline) metabolism and activating a core signaling pathway. This study not only characterizes a novel function for a DUF family gene but also provides a promising candidate gene for the genetic improvement of cold resilience in melon and other related crops.