We aimed at revealing the effect of different culture conditions on the laccase activity of Leptographium qinlingensis. Single-factor experimental design was used for different carbon sources, nitrogen sources, Cu2+ and substrate concentration, combined with the guaiacol method for determining laccase activity, and analysis of variance was used to analyze effects of the different culture conditions on the laccase activity of L. qinlingensis using SAS (V8.1). The results showed that laccase activity of L. qinlingensis was significantly increased by maltose (as carbon source) and yeast (as nitrogen source). The laccase activity was induced by 0.6 mmol·L-1 copper ions and reached 292.36 U·L-1 in the medium containing the Cu2+, however, the laccase activity was inhibited by copper ions exceeding 0.9 mmol·L-1. Furthermore, the laccase activity was induced by phloem powder of host trees ( Pinus armandii ). The peak of laccase activity occurred on the 11th d of culture. Different culture conditions had remarkable effect on laccase activity of L. qinlingensis. The maximum laccase activity was 681.82 U·L-1 when being cultured in the medium containing maltose as carbon source, yeast as nitrogen source, 0.6 mmol·L-1 Cu2+ and 6 g·L-1 phloem powder of host trees on the 11thd.