BACKGROUND: Cucumber plants suffer from a serious threatening disease, downy mildew, throughout the growing seasons irrespective of the weather temperature. The causal agent, Pseudoperonospora cubensis, tends to evolve rapidly upon sequential applications of chemical fungicides and generate new progeny possessing tolerance to such fungicides. Glycoproteins represent an environmentally safe alternative for chemically synthetized fungicides and do not trigger fungicide resistance. We studied the antifungal activity of four glycoproteins namely soybean ⊎-conglycinin, chickpea vicilin, duck egg ovomucin and catfish p22 against P. cubensis. Ten commercial fungicides of different chemical groups were used as positive controls of glycoprotein treatments. RESULTS: The results revealed that soybean ⊎-conglycinin and catfish p22 glycoproteins possess significant antifungal activity against P. cubensis. The amount of disease suppression caused by ⊎-conglycinin and p22 was comparable to the highly efficient chemical fungicides containing copper oxychloride, cymoxanil and fosetyl Al as active ingredients. Vicilin and ovomucin were less efficient biocides as they gave moderate suppression of disease severity. However, all tested glycoproteins provided full protection for the newly emerged cucumber leaves. Microscopic examination of glycoprotein-treated leaves inferred the ability of catfish p22 and soybean ⊎-conglycinin to disrupt the integrity of sporangial cell walls of P. cubensis rendering them nonviable compared to untreated ones. Expression levels of total phenolic compounds and the antioxidant enzymes catalase, superoxide dismutase and peroxidase were elevated upon glycoproteins application, which infers their involvement in disease suppression. CONCLUSION: This report emphasizes the direct and indirect roles of glycoproteins in safe management of cucumber downy mildew disease