Graphene nanoparticles improve alfalfa (Medicago sativa L.) growth through multiple metabolic pathways under salinity-stressed environment

Graphene, one of the emerging carbon nanomaterials, has many advantages and applications. Salinity stress seriously affects ecology and agroforestry worldwide. The effects of graphene on alfalfa under salinity stress were investigated. The results indicated that graphene promoted alfalfa growth under non-salinity stress but caused some degree of damage to root cells and leaf parameters. Graphene used in salinity stress had a positive effect on growth parameters, chlorophyll, photosynthetic gas parameters, stomatal opening, ion balance, osmotic homeostasis, cell membrane integrity and antioxidant system, while it decreased Na+, lipid peroxidation and reactive oxygen species levels. Correlation analysis revealed that most of the parameters were significantly correlated; and principal component analysis indicated that the first two dimensions (78.1% and 4.1%) explained 82.2% of the total variability, and the majority of them exceeded the average contribution. Additionally, Gene Ontology functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes signaling pathway enrichment analysis showed that there were numerous differentially expressed genes and pathways to regulate alfalfa responding to salinity stress. Taken together, the findings reveal that graphene does not enter the plant, but improves the properties and adsorption of soil to enhance salt tolerance and seedling growth of alfalfa through morphological, physiological, biochemical, and transcriptomic aspects. Furthermore, this study provides a reference for the application of graphene to improve soil environment and agricultural production. •Graphene does not enter roots but damages roots and leaves of alfalfa.•Graphene improves photosynthesis and stomatal opening under salinity stress.•Graphene maintains ionic and osmotic homeostasis under salinity stress.•Graphene-treated alfalfa reduces ROS and lipid peroxidation under salinity stress.•Graphene improves soil property and alfalfa growth by multiple pathways.