Characteristics of sophorolipid and rhamnolipid were evaluated as antifungal agents against plant pathogenic fungi. Eight percent of mycelial growth of plant pathogen (Phytophthora sp. and Pythium sp.) was inhibited by 200 mg/l of rhamnolipid or 500 mg/l of sophorolipid, and zoospore motility of Phytophthora sp. decreased by $90\%$ at 50 mg/l of rhamnolipid and $80\%$ at 100 mg/l of sophorolipid. The effective concentrations for zoospore lysis were two times higher than those of zoospore motility inhibition. The highest zoospore lysis was observed with Phytophthora capsici; $80\%$ lysis at 100 mg/I of di-rhamnolipid or lactonic sophorolipid, showing the dependency of structure on the lysis. In the pot test, the damping-off disease incidence ratio decreased to $42\%;and;33\%$ of control value at 2,000 mg/l sophorolipid and rhamnolipid, respectively. These results showed the potential of microbial glycolipid biosurfactants as an effective antifungal agent against damping-off plant pathogens.