Medicinal plants provides natural product may be used as precursors for new drug for their chemical diversity. In the present study, the potential of gamma radiation as sterilizing agent for preservation and its effect on phytochemical contents, antioxidant and antibacterial activity of Teucrium polium (T. polium) was analyzed for antimicrobial and antitumor activity. The results revealed that exposing plant powder to doses of gamma irradiation (5,10,15,20 KGy) significantly increased the ethyl acetate extraction yield (dry weight) as compared to the non-irradiated plant (control). Teucrium polium phytochemical contents of total phenolic content (TPC), total flavonoids content (TFC) and tannins content were significantly increased with gamma irradiation. In the same line, plant extract antioxidant activity was significantly increased in response manner to radiation dose. Teucrium polium ethyl acetate extracts shown antibacterial activity against Staphylococcus aureus and Pseudomonas aeroginosa. Gamma radiation doses caused significant increase in the antibacterial activity against P. aeroginosa and S. aureus at dose of 20 KGy. Total count of microbial load of T. polium was significantly reduced at 5.0 kGy by 99.95% with total decontamination of fungi count.