【Objective】The effects of medium components and other culture conditions on the growth and antifungal substances production of Streptomyces globisporus JK-1 were investigated in order to determine the optimum culture conditions. 【Method】Streptomyces globisporus JK-1 was incubated under different culturing conditions to measure the cell dry weight； also， using the rice blast fungus as the indicating fungus， the mycelial growth rate method was used to determine the antifungal activity on JK-1 fermentation liquid under different culturing conditions. 【Result】The results showed that amongst the seven tested media， the best medium for fungus growth was LB and the best medium for antifungal substances production was potato sucrose broth （PSB）. For the growth and antibiotic production of S. globisporus JK-1 on medium PSB， the temperature range was 10-35℃， with the optimum temperature measured at 25℃. The cell dry weight reached maximum after 5 d of incubation and the antifungal substances production reached maximum after 7 d of incubation at 25℃. The most favorable pH range for growth and antifungal substance production was 6-8； the optimum level was measured at the pH 7. JK-1 could effectively utilize several kinds of carbon and nitrogen sources for its growth and antifungal substances production； amongst the tested sources， glycerol and tyrosine were the most favorable for growth， while maltose and ammonium sulfate were the most favorable for strong antifungal activity. 【Conclusion】The optimum culturing condition for growth and antifungal substances production of Streptomyces globisporus JK-1 was incubating the fungus at 25℃， PSB medium in pH 7 for 7 days. Also， glycerol and tyrosine were the most favorable sources for improving fungus growth， while maltose and ammonium sulfate were the most favorable sources for antifungal substances production.