Streptomyces padanus PMS-702 strain produces a polyene macrolide antibiotic fungichromin and displays antagonistic activities against many phytopathogenic fungi. In the present study, experimental formulations were assessed to improve the production of fungichromin, the efficacy of PMS-702 on the suppression of sporangial germination, and the reduction of cucumber downy mildew caused by Pseudoperonospora cubensis. PMS-702 strain cultured in a soybean meal-glucose (SMG) medium led to low levels of fungichromin accumulation and sporangial germination suppression. Increasing medium compositions and adding plant oils (noticeably coconut oil) in SMG significantly increased fungichromin production from 68 to 1,999.6 μg/ml. Microscopic examination reveals that the resultant suspensions significantly reduced sporangial germination and caused cytoplasmic aggregation. Greenhouse trials reveal that the application of PMS-702 cultural suspensions reduced downy mildew severity considerably. The addition of Tween 80 into the synthetic medium while culturing PMS-702 further increased the suppressive efficacy of downy mildew severity, particularly when applied at 24 h before inoculation or co-applied with inoculum. Fungichromin at 50 μg/ml induced phytotoxicity showing minor necrosis surrounded with light yellowish halos on cucumber leaves. The concentration that leads to 90% inhibition (IC90) of sporangial germination was estimated to be around 10 μg/ml. The results provide a strong possibility of using the S. padanus PMS-702 strain as a biocontrol agent to control other plant pathogens.