Abstract: In the research, a wild Lentinula edodes strain from Yunnan Chuxiong area was isolated and characterized. Effect of different temperature, pH, carbon source, nitrogen source and inorganic salt on mycelia growth of the strain were studied. At the same time, the wild L. edodes strain was successfully cultivated by the mixed sawdust, corn cob and other agricultural waste resources as the main substrate. The results were as follows: Combined with morphological and ITS molecular identification, the strain was identified as Lentinula fungus by morphological and molecular identification; biological characteristics experiments found that the optimum temperature for the growth of wild L. edodes mycelium was 30.0 ℃, and the optimum pH was 6.0, the best carbon source and nitrogen source are corn meal and peptone, the best inorganic salt are calcium chloride and magnesium sulfate; bag substitute material cultivation results show: the optimal bag substitute composed of 60.0% sawdust and 30.0% corncob, the biotransformation rate reached 41.49%.