In this study, the effect of different substrates of agricultural by-products on the mycelial growth rate and density of Trametes versicolor (Turkeytail mushroom) was analyzed. We found that pepper stem and rice bran with a mixing ratio of 9:1(v/v) produced the best mycelial growth of 101 mm in 10 days, while a mixing ratio of 8:2 resulted in mycelial growth of 83 mm in 10 days. The control group treated with a 9:1 mixing ratio of oak sawdust and rice bran (v/v) produced mycelial growth of 74 mm in 10 days. The following results are in the order of beanstalk, sesame stem, and perilla stem. After the harvest of the mushrooms, the mycelial growth rate and the density of T. versicolor in each substrate were as follows the group with waste substrate of Pleurotus eryngii and rice bran with a mixing ratio of 9:1(v/v) produced the best result of 76 mm in days, while a mixing ratio of 8:2 produced of 61 mm in 10 days. The control group with a 9:1 ratio of oak sawdust and rice bran produced mycelia of 74 mm in 10 days, while a mixing ratio of 8:2 resulted in mycelia of 59 mm in10 days.