Abstract: Among soil-borne fungi, cosmopolitan phytopathogen, Verticillium dahliae Klebahn is responsible for high yield losses in many plant species. Except for solarization in mediterranean countries and except in the few cases where disease-resistant cultivars are available, control of Verticillium wilt in commercial crops has been highly dependent on the application of preplant soil fumigants. Research to develop alternative control measurements should focus on biological approaches aimed at shifting the composition of soil microbial communities to suppress Verticillium. The merits of role of actinomycetes in biological control of soil-borne fungal-pathogens are known, however actinomycetes microflora of the Iranian soils has not been very well explored in searching for biofungicide agents. At the present research, in vitro studies of some biological effects of two Iranian strains of actinomycetes, Streptomyces plicatus strain 101 and Frankia sp. strain 103, are presented. Both strains revealed enzymatic activity and inhibited production of microsclerotia in V. dahliae. Treating the crude extract with chloroform, denaturized enzymatic activity of both strains. Thermal inactivation point of active phases of S. plicatus was 70 and 90°C and in Frankia sp. was determined as 60°C. Antifungal active phases of S. plicatus tolerate wide range of pH (5-13) but in Frankia sp. active phase tolerates pH 7-9. These two strains may be useful candidates for involving in integrated control programs of Verticillium vascular wilting.