This study aims to improve the production of secondary metabolites from (Ocimum basilicum L. var. Genovese) by using nodal segments and callus as explants. The research focused on Maximizing various factors, containing two cytokinins (benzylaminopurine (BAP) and thidiazuron (TDZ)) at three different concentrations (0, 1, or 2 mg/L) and plant growth regulators, as well as various light-emitting diode (LED) systems (white as a control, violet, and a 1:1 combination of red and blue). After 30 days of cultivation, the results showed that Murashige and Skoog control medium completely free of hormones gave superior results in terms of fresh and dry shoots weight (1.46 gm and 0.88 gm, respectively), while the Murashige and Skoog medium with 2 mg/L of (TDZ) and white and violet LED lighting, obtained highest values for fresh and dry callus weight (1.62 gm and 0.91 gm, respectively). Moreover, it is worth noting that callus formation was not observed in response to benzyl aminopurine treatments. In comparison with the control, it was found that all treatments used in general stimulated antioxidant compounds such as (phenolics, flavonoids, anthocyanins, and ascorbic acid), surprisingly, whether in the individual effects of white light or thidiazuron 2 mg/L alone or the interactions between them compared to the other treatments.