In vitro culture and radiation techniques were used for obtaining mutants tin tall fescue. Endophyte free and friendly tall fescue cultivars Kentucky-31 and Jesup were used for induction of genetic variability through in-vitro mutagenesis. Mature seeds was used for callus induction on 6 mg/L 2,4-D. Actively growing and compact callus was treated with three different doses of gamma rays (10 Gy, 30 Gy and 50 Gy). Maximum proliferation and plantlets regeneration growth was observed in control and minimum at 10 Gy. Furthermore, the maximum number of tiller in the irradiated population was observed in 10 Gy. The treatments 30 Gy and 50 Gy exhibited negative impact on the tillering potential of the tall fescue plant. The object of this study was to develop protocols for mutation breeding in tall fescue through radiation techniques.