This study was conducted to evaluate the phytotoxic effects of different antibiotics and fungicides on the procallus of date palm (Phoenix dactylifera L.). The fungicides Switch® and Beltanol and the antibiotics chloramphenicol and gentamycin were used in this study. Results confirmed the lethal effects of Switch®, gentamycin, and chloramphenicol (at 100 mg/l in dicamba treatment) on date palm tissues. The treated shoot tips did not develop into procallus, and their growth was completely inhibited. Beltanol treatments significantly decreased the biochemical parameters of the procallus treated with 2, 4-D and dicamba. In the Beltanol-treated tissues, the initial period of callus induction was prolonged from 63.33 and 69.66 days to 97.66 and 117.66 days. The callus induction percentage of the plants treated with the decontamination agents was also decreased. The fresh and dry weights of the procallus and the browning percentage and intensity of the cultured tissues in Beltanol and chloramphenicol treatments (100 mg/l) increased compared with those in the control treatments. Total carbohydrates, free proline content, total phenolic compounds, free amino acid, total soluble protein, and peroxidase activity were significantly affected by Beltanol and chloramphenicol (100 mg/l). By contrast, low chloramphenicol concentrations resulted in acceptable levels of biochemical components. Therefore, we recommend the use of low chloramphenicol and Beltanol concentrations instead of Switch and gentamycin as decontamination agents to prevent or treat microbial contamination in date palm tissue cultures.