Cucumber (Cucumis sativus var. vita z V2 272) was used in this experiment to examine the changes in gene expression after exposure the cucumber plants to Multi-Walled carbon nanotubes (MWCNT) during a period from 2020 - 2022. The current investigation was conducted in the Ain Shams Centre of Genetic Engineering and Biotechnology (ACGEB), Faculty of Agriculture, Ain Shams University, Egypt. Experiment was conducted at two times; T0 and T1, T0 is the time after 11 day of planting and T1 was after 18 days of planting with the same protocol. The impact of MWCNT on the genes expression of cucumber seedling including ACO1, ACO2, PINI, rnaxl1, and YUC genes at different time point T0 and T1 after 3 weeks from culture in free hormone - MS medium supplemented with MWCNTs is shown by RT-PCR technique. The results of the RT-PCR technique showed that MWCNTs had a consequence on the gene expression of cucumber seedlings, particularly with genes such as ACO1, ACO2, PINI, rnax and YUC. The relative gene expression of five genes, ACO1, ACO2, PINI, rnaxl1 and YUC, were measured in two periods of germination, after 11 days (T0) and 18 days (T1) and the results demonstrated that the gene expression of all these genes in the case of exposure to MWCNTs at the second time is very significantly multiplied compared to gene expression in positive and negative control. In conclusion, at middle concentrations, MWCNTs have great effect on gene expression of the genes that control growth of cucumber seedlings.