The mature seeds of rapeseed (Brassica napus L.) from variety Gaoyou 605 were treated with -rays and Ethyl Methan Sulfonate (EMS). 152 mutants (12.67% of M2 population) with mutative traits, including the mutation of leaf color, leaf shape, plant height, number and angle of branches, diameter of main stalk, color of stalk and flower, number and size of petals, pistil shape, male sterility, bud death and date of bloom were found in screened M2 progenies, which have been identified in M3. The mutants of cotyledon and root traits were also screened by hydroponics culture and their total mutant frequency were estimated at 12.78% and 7.07% in M3, respectively. Identification of M4 showed that these mutations could be inherited stably. The mutant library including the mutants of leaf, plant-type, flower, cotyledon, root and physiological traits had been built in present experiment. These mutants might be used as important germplasm for rapeseed breeding and functional genomics study.