[Objective] The paper was to screen the antagonistic Bacillus spp. strains against Phytophthora infestans (Mont.) de Bary and analyze its lipopeptide substances. [Methods] Using flat isolation and culture method, the bacteria that could produce Bacillus spp. were isolated from rhizosphere soil, and carried out primary and secondary screening by confrontation culture. The lipopeptide substances of Y-3 strain with better antifungal effect were carried out HPLC analysis. [Result] Nine strains of bacteria had certain antagonistic effects against P. infestans in primary screening, Y-1 and Y-3 strains had significant inhibition effect against the growth of P. infestans in secondary screening, with inhibition rate higher than 80%; the crude extract of lipopeptide produced by Y-3 strain had better antagonistic effect against P. infestans, with inhibition rate higher than 90%. HPLC analysis showed that the crude extract of lipopeptide mainly contained 8 components. [Conclusion] The lipopeptide substances produced by Y-3 strain had significant inhibition effect, which had potential application value for biological control of potato late blight. [Objective] The paper was to screen the antagonistic Bacillus spp. Strains against Phytophthora infestans (Mont.) De Bary and analyze its lipopeptide substances. [Methods] Using flat isolation and culture method, the bacteria that could produce Bacillus spp. Were isolated from rhizosphere soil, and carried out primary and secondary screening by confrontation culture. The lipopeptide substances of Y-3 strain with better antifungal effect were carried out HPLC analysis. [Result] Nine strains of bacteria had certain antagonistic effects against P. infestans in primary screening Y-1 and Y-3 strains had a significant inhibition effect against the growth of P. infestans in secondary screening, with inhibition rate higher than 80%; the crude extract of lipopeptide produced by Y-3 strain had better antagonistic effect against P. infestans, with inhibition rate higher than 90%. HPLC analysis showed that the crude extract of lipopeptide mainly contained 8 components. [Conclusion] The lipopeptide substances produced by Y-3 strain had significant inhibition effect, which had potential application value for biological control of potato late blight.