Gliocladium virens, when grown on peat moss amended with sucrose and ammonium nitrate and then applied to soil, resulted in root necrosis. Herbicidal activity was correlated with fungal production of the phytotoxin viridiol. Viridiol had a wide spectrum of activity; it was particularly effective against annual composite species but was less effective in monocot control. Emergence of most weeds was reduced >90% at application rates of 8.7% (of total volume) or less. Treated seedling dry weights were drastically reduced. Applications of 4.5% reduced root and shoot weight of redroot pigweed by 93 and 98%, respectively. Crops were affected at higher treatment levels; however, the toxicity was readily avoided by applying the mycoherbicide out of the root zone of the crop, instead applying it between the seed and the soil surface. Viridiol production, which confers herbicidal activity, was detected 3 days after incorporation of the fungus-peat mixture. Viridiol production peaked on days 5 and 6 at approximately 25 μg viridiol/100 ml soil, based upon an application rate of 11%, then declined to undetectable levels by the end of 2 weeks.