Melatonin Enhances the Low-Calcium Stress Tolerance by Regulating Brassinosteroids and Auxin Signals in Wax Gourd

In plants, calcium (Ca) serves as an essential nutrient and signaling molecule. Melatonin is a biologically active and multi-functional hormone that plays an important role in improving nutrient use efficiency. However, its involvement in plant responses to Ca deficiency remains largely unexplored. This study aimed to assess the effects of melatonin on Ca absorption, the antioxidant system, and root morphology under low-Ca (LCa) stress conditions, as well as to identify key regulatory factors and signaling pathways involved in these processes using transcriptome analysis. Under LCa conditions, wax gourd seedling exhibited significant decreases in Ca accumulation, showed inhibition of root growth, and demonstrated the occurrence of oxidative damage. However, melatonin application significantly enhanced Ca content in wax gourd seedlings, and it enhanced the absorption of Ca²⁺ in roots by upregulating Ca²⁺ channels and transport genes, including <i>BhiCNGC17</i>, <i>BhiCNGC20</i>, <i>BhiECA1</i>, <i>BhiACA1</i>, and <i>BhiCAX1</i>. Furthermore, the application of exogenous melatonin mitigated the root growth inhibition and oxidative damage caused by LCa stress. This was evidenced by increases in the root branch numbers, root tips, root surface area, and root volume, as well as enhanced root vitality and antioxidant enzyme activities, as well as decreases in the reactive oxygen species content in melatonin treated plants. Transcriptome results revealed that melatonin mainly modulated the brassinosteroids (BRs) and auxin signaling pathway, which play essential roles in root differentiation, elongation, and stress adaptation. Specifically, melatonin increased the active BR levels by upregulating <i>BR6ox</i> (a BR biosynthesis gene) and downregulating <i>BAS1</i> (BR degradation genes), thereby affecting the BR signaling pathway. Additionally, melatonin reduced IAA levels but activated the auxin signaling pathway, indicating that melatonin could directly stimulate the auxin signaling pathway via an IAA-independent mechanism. This study provides new insights into the role of melatonin in nutrient stress adaptation, offering a promising and sustainable approach to improve nutrient use efficiency in wax gourd and other crops.