A plate coloration method by auxotroph strain for screening high L-arginine production mutants quickly and efficiently was established. Coynebacterum crenatum SYPA5-5 which is an L-arginine producer was mutated by N+ ion implantation with an energy level of 10 keV. Combined with the directed screening method and fermentation in flasks, a mutant C. crenatum SYPA5-5-36 with a higher L-arginine production and stability was selected. The yield of L-arginine produced by this mutant could reach 35.85 g/L which was 19.5% higher than that of the original strain. Therefore, the plate coloration method by auxotroph strain could be applied to screening high L-arginine production mutants quickly and efficiently.