We examined damages from gamma-irradiaion and determined the optimal gamma-ray dose for mutation breeding in lentil (Lens culinaris L.) bean. Four individual lines (L-C, L-2, L-8 and L-9), that have remarkable adaptability in South Korea were gamma-irradiated at doses of 50, 70, 100, 200, 300, 400, and 500 Gy. The germination rate of seed decreased as the dose increased over 50 Gy in all lines. However, $LD_{50}$ and $RD_{50}$ were different among lines. The median lethal doses($LD_{50}$) were approximately 127 (L-C), 74 (L-2), 95 (L-8), and 144 (L-9) Gy. The median reduction doses($RD_{50}$) for plant height, number of leaves, root length, and flash weight were 156, 176, 150, and 180 Gy for L-C, 253, 198, 127, and 142 Gy for L-2, 188, 175, 200, and 190 Gy for L-8, and 162, 210, 224, and 184 for L-9, respectively. The growth characteristics of the $M_1$ generation decreased as the dose increased over 70 Gy. The optimal doses of gamma irradiation for mutation breeding of lentil were determined to be 70 Gy (L-2, L-8) and 100 Gy (L-C, L-9). We performed the comet assay to observe nuclear DNA damage induced by gamma-irradiation. In comet assay, a clear difference was identified over 100 Gy treatments. With increasing doses of gamma-ray in the range of 50 to 500 Gy, the rate of head DNA was decreased significantly from 97.5% to 81.6%. Tail length was consecutively increased from $1.9{\mu}m$ to $17.4{\mu}m$. Our result provides basic information for construction of mutant pools in lentils.