The American serpentine leafminer fly, Liriomyza trifolii (Burgess), is one of the most destructive polyphagous pests world wide. In this study, we determined electron beam doses for inhibition of normal development of the L. trifolii and investigated the effect of electron beam irradiation on DNA damage and p53 stability. Eggs, larvae, pupae and adults were irradiated with increasing doses of electron beam irradiation (six levels ranges from 30 to 200 Gy). At 150 Gy, the number of adults that developed from irradiated eggs, larvae and pupae was lower than those of untreated control. Fecundity and egg hatchability decreased depending on the doses applied. Reciprocal crosses between irradiated and unirradiated flies demonstrated that males were more radiotolerant than females. Adult longevity was not affected in all stages. The levels of DNA damage in L. trifolii adults were evaluated using the alkaline comet assay. Our results indicate that electron beam irradiation increased levels of DNA damage in a dose-dependent manner. Moreover, low doses of electron beam irradiation led to the rapid appearance of p53 protein with in 6 h; however, it decreased after exposure to high doses (150 Gy and 200 Gy). These results suggest that electron beam irradiation induced not only abnormal development and reproduction but also p53 stability caused by DNA damage in L. trifolii. We conclude that a minimum dose of 150 Gy should be sufficient for sterilization of L. trifolii.