The laccase activity and its gene expression regulation characteristics under different temperature cultures (20℃, 25℃, 28℃and 30℃) were studied with the Lentinus edodes Xin808 to analyze the effect of its diameter of the mycelium ring and oxidation ring. The laccase activity was measured by liquid fermentation, and the relative expression level of lac was detected by qPCR technique. The results showed that, the growth rate of Lentinus edode mycelium, formation of oxidation ring, and laccase activity were all higher under 25℃ cultivation temperature. While 30℃ inhibited the growth of mycelium and the formation of oxidation ring and resulted into the lowest laccase activity. Moreover, the Lac genes exhibited differential expression profiles cultured in different temperature. Compared to the 25℃(control), the relative expression level of 9 laccase genes (i.e. Llac1-3, Llac5 and Llac7-11) were up-regulated cultured at 20℃. Besides, the expression level of genes such as Llac5, Llac7, and the rest 8 laccase genes were up-regulated at 28℃. While 6 genes (including Llac1-2, Llac4, Llac7-8 and Llac10) were down-regulated under 30℃ treatment. It can promote Llac genes expression cultured at 20℃ , while 30℃ treatment inhibit the expression of the isozyme genes.