Attempts were made to investigate the physiological and genetic changes when two different shiitake (Lentinula edodes) strains are mixed. Mycelial growth of KFRI 180 strain and KFRI 1 strain were investigated 82 mm and 80 mm, respectively. Concerning the weight loss percentage of medium, KFRI 1 strain decreased 2.4% and KFRI 180 strain 1.6%. Plug-shaped spawn had no-problem to incubate and there were no differences among the ratios of mixture. Also, conditions of plug-shaped spawns were similar. When the isolated mycelia from plug-shaped spawns was incubated again, KFRI 1 50%-KFRI 180 50% showed decreased growth of mycelia compared with other treatments. The same results were obtained from test tubes filled with sawdust. When surface of spawn bottles were observed, KFRI 1 50%-KFRI 180 50% showed spots, but other treatments were not different from KFRI 1 and KFRI 180. Test was made to confirm the strains by confrontation culture. The mixture of two strains was proved to be KFRI 1 regardless the ratios of mixture. However, by the RAPD primer analysis, when KFRI 1 was mixed with KFRI 180, KFRI 180 was stronger. Thus, the confrontation line on PDA was different from the bands analysis by primers. Attempts were made whether the fruit-bodies were made at the generating condition of spawn bottles. The results were that KFRI 1 100%, KFRI 1 90%-KFRI 180 10%, KFRI 1 80%-KFRI 180 20%, KFRI 1 50%-KFRI 180 50% treatment showed fruit-body formation. The shape of fruit-body was deformed, but the gill was made normally.