Polyoxin is a high effective and broad-spectrum antifungal peptidyl nucleoside antibiotics inhibiting fungal cell wall biosynthesis, as a result, it was used extensively to control phytopathengenic fungi in agriculture. This review summarized the research progress on the chemical structure and physical and chemical properties of this antibiotic, and particularly described recent advance of our lab on the cloning and elucidation of the polyoxin biosynthetic pathway, as well as the combinatorial biosynthesis of this antibiotic. Simultaneously, we also take a brief perspective on the studies on nucleoside antibiotics biosynthesis.