Taking mushroom as test materials，the browning probability of inhibiting tissue isolation by selecting different sizes of block,freshness Shiitake mushroom，water content medium，controlling the culture temperature and adding activated carbon and vitamin C into the culture medium,many factors that affecting the browning during the issue isolation were studied.The results showed that the browning rate was relatively low of 16.6% when the size of block was 0.7—0.9 cm；while the rate was 0% when Shiitake mushroom was placed for 1—3 days，and the culture medium was placed for 3 days.The browning rate became 0% when adding 0.1%—0.3% activated carbon and 0.1—0.5 g/L vitamin C.The browning rate was reduced to 16.6% when the temperature was controlled at about 25℃.The browning rate was the lower，the better the mycelia growth.