PubMedInternational journal of biological macromolecules2026-07-27
Chitosan-melatonin nanobiopolymer regulates miR398-mediated antioxidant defense and TaHKT1;5-SOS-NHX ion-homeostasis networks to enhance wheat salt tolerance.
Algopishi Uthman Balgith UB, Alshegaihi Rana M RM, Khan M Nasir MN, Fayad Eman E et al.
In recent years, the application of nanobiomaterials and phytohormone-based nano-delivery systems has gained considerable attention for enhancing plant tolerance to abiotic stresses. Among these approaches, melatonin (MLT) and chitosan nanoparticles (CS-NPs) have emerged as promising biostimulants due to their ability to regulate antioxidant defense systems, ion homeostasis, photosynthetic performance, and stress-responsive molecular networks. Although the individual effects of MLT and CS-NPs on stress mitigation have been investigated in different crop species, their combined application through a nanobiopolymer delivery system and its influence on microRNA-mediated antioxidant regulation and ion-homeostasis pathways under salinity stress remain insufficiently explored. Therefore, the present study was conducted to evaluate the effectiveness of melatonin-loaded chitosan nanoparticles (MLT-CSNPs) in improving salt tolerance in wheat (Triticum aestivum L.) exposed to 100 mM NaCl. T. aestivum plants were treated with free MLT, CS-NPs, and MLT-CSNPs, where T7 represented NaCl + MLT-CSNPs equivalent to 100 μM melatonin. Our results demonstrated that salt stress significantly reduced plant growth and biomass, photosynthetic pigments, gas exchange characteristics, and ionic balance in T. aestivum plants. Moreover, salinity markedly increased Na+ accumulation, the Na+/K+ ratio, oxidative stress biomarkers, and cellular damage. However, the application of MLT-CSNPs significantly improved plant growth, photosynthetic efficiency, antioxidant defense, osmolyte accumulation, and ion homeostasis under saline conditions. In particular, T7 enhanced plant height and shoot dry weight by 40.6% and 85.5%, respectively, compared with salt-stressed plants. Likewise, total chlorophyll content and photosynthetic rate increased by 79.6% and 95.1%, respectively, indicating substantial protection of the photosynthetic machinery. Furthermore, MLT-CSNP treatment decreased Na+ accumulation by 56.5% and reduced the Na+/K+ ratio to 0.38 compared with 2.01 in salt-stressed plants. In addition, T7 markedly reduced H₂O₂ and MDA contents by 59.8% and 56.6%, respectively, while significantly enhancing the activities of SOD, CAT, POD, and APX. Molecular investigations further revealed that MLT-CSNPs strongly upregulated TaHKT1;5, TaSOS1, TaNHX1, antioxidant-related genes, and miR398 expression, suggesting coordinated regulation of ion exclusion, intracellular ion compartmentalization, and microRNA-mediated antioxidant defense mechanisms. These findings provide new insights into the role of MLT-CSNPs nano-delivery systems in modulating TaHKT1;5-SOS-NHX ion-homeostasis networks and miR398-associated antioxidant responses under salinity stress. Overall, MLT-CSNPs represent a sustainable and efficient nano-biostimulant strategy for enhancing T. aestivum resilience and productivity in saline environments.