Abstract: A new selective medium to be used to simply and quick isolate Verticillium dahliae in cotton from naturally infested soil was introduced. The medium consisted of following ingredients per liter distilled water: leaves cotton 100g, sugar 5g, agar 11~20g, 40% PCNB 0.3g. chloramphenicol 0.05g, streptomycin 0.05g and penicillin 0.05g, pH adjusted to 6.4~7.0. The medium is simple and quick than old that. The microscclerotia V. dahliae from naturally infested soil could be effectively isolated while it was rarely infected by other pathogens. The colonies of V. dahliae on the medium were large black-colored.