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Abstract Salt stress severely affects the growth and productivity of plants. Tomato (Solanum lycopersicum) as a vegetable model plant is susceptible and cannot survive under high salinity conditions. Nano-materials are considered as one of the bio-stimulants, which widely explored and used in plants as inducers to decrease the adverse effect of both abiotic and biotic stresses. Therefore in our study, Chitosan-Cu nanoparticles (Chi-Cu-NPs) and Chitosan nanoparticles (Chi-NPs) were synthesized and evaluated for their ability to be used as an inducer for tomato plant under salt stress through investigating the influence of these nanoparticles on the molecular level by studying 12 miRNA families integrated into hormonal pathways (miR156, miR159, miR160, miR164, miR167, miR172, miR319, miR390, miR393, miR396, miR398) and their target genes that may be involved in salt stress response and plant development using quantitative PCR (qPCR) technique. Our results showed that there was a differential expression in the studied miRNAs in plants treated with Chi-NPs and Chi-Cu NPs under salt stress. On the other hand, we validated target genes such as (SPL3, MYB33, ARF6, AP2, EIN2, TCP4, TIR1, DELLA, ARF2, NCED1, CCS1, CSD1, CSD2, GR, et) that showed different expression patterns. The results of these genes indicated that there was a linkage in the behavior of these genes in response to different treatments. Moreover, we evaluated the effect of these nanoparticles on the biochemical level by studying their effect on the plant growth parameters, photosynthesis and antioxidants enzymes level. Our results proposed that miRNAs and their target genes may play a crucial role in hormonal plant response to environmental abiotic stresses but the molecular mechanism of plant tolerance to abiotic stresses needs further investigation to clarify and initiate miRNA-based biotechnology for enhancing plant tolerance to abiotic stresses such as salinity stress |