Amphidiploid Brassica juncea (AABB, 2n=36) contains the synthesized genome of the diploid ancestors of Brassica rapa (AA, 2n=20) and Brassica nigra (BB, 2n=16), proven the ‘triangle of U’ model. Varieties of the B. juncea include vegetables, oilseed crops, and medicinal plants in South Asia, China, and other regions. ‘Dolsangat’, one of the cultivars of B. juncea is widely used as the main ingredient for ‘KatKimchi’, a kind of Korean traditional food Kimchi. To develop an efficient polyploidization protocol of B. juncea, we used twenty accessions. Among them, we could induce the amphidiploid plants with 0.23% in natural. A successful of polyploidization, it is essential of chromosome doubling regent treatment of B. juncea. At first, we tried to colchicine treatment in the embryo stage and it was very harmful to the embryo and could get few plants. The second, we made the regeneration plants from embryo to rooting phase and shocked them in 0.34% colchicine contained distilled water. We could induce amphidiploid plants with a success rate of 63.4%. Also, we surveyed glucosinolate content and JB1, Alsami, and JD6 showed high total contents. These plants will use for genetic materials for breeding, genetic and molecular studies.