Abstract: Fusarium chlamydosporum strain ZC201301 showed high herbicidal activity towards Digitaria sanguinalis. In the present study, the 'single factor at a time' method was used to determine biological characteristics of ZC201301. It was found that the optimal mycelial growth media were potato dextrose agar (PDA) and crabgrass leaf juice (CAJ), and the optimal temperature for mycelia growth ranged from 25 to 30℃. The optimal initial pH for culture media was 6-8. Strain ZC201301 can sporulate in a variety of media, but the optimal sporulation medium was CAJ. Mycelial growth did not differ significantly in fluorescent light from that in the dark, but darkness was beneficial for sporulation. The optimal carbon sources for mycelium growth was glucose, and the best nitrogen source was NH₄NO₃. Oxygen is essential for mycelial growth, and mycelial growth was inhibited when the liquid/gas ratio was less than 1:1. The herbicidal activity of strain ZC201301 was sensitive to relative humidity, and was significantly decreased when humidity was less than 60%. These results indicate the strain ZC201301 is amenable to propagation, which is important for the preparation of bio-control agents and for their survival under field conditions.