In order to induce and explore genetic variation in agronomic and quality traits by chemical mutagens and gamma rays in canola the following experiments were conducted: 1- determination of optimum dosage of chemical mutagens 2- field evaluation of M 1 to M 3 generations 3- assessment of genetic diversity of M 2 lines using microsatellite markers and 4- screening of mutant plants via TILLING. Based on LD 50 criterion, the 0.8 percent ethyl methanesulfonate for 6 hrs, 12 mM N-nitroso-N-ethylurea, 6 mM sodium azide and 9 mM N-nitroso-N-methylurea for 8 hrs were considered as optimum treatment conditions. The highest genetic diversity was achieved in sodium azide treatment. Treatment with 1000 and 800 Gy dosages of gamma rays were considered as the best treatment conditions for improving agronomic traits along with seed quality traits in RGS003 and Sarigol, respectively. Cluster analysis according to data derived from evaluation of genetic diversity using SSR markers, grouped M 2 mutant lines in a completely separated group from control. Screening of 750 M 2 mutant plants using TILLING led to identification of 12 mutations in BnDGAT1 and 9 mutations in BnIND . The results of this study imply the potential of mutations to enhance the genetic variation available to canola breeders. Key words: mutation, chemical mutagens, gamma rays, genetic diversity, canola.