Star anise (
Illicium verum
), a valuable spice tree, faces significant threats from fungal diseases, particularly
Alternaria
leaf spot. This study investigates the potential of a soil-derived actinomycete strain, YG-5, as a biocontrol agent against
Alternaria tenuissima
, the causative pathogen on
Alternaria
leaf spot in star anise. Through comprehensive morphology, physiology, biochemistry, and genetic analyses, we identified the isolate as
Streptomyces
sp. YG-5. The strain exhibited broad-spectrum antimicrobial activity against several plant pathogens, with inhibition rates ranging between 36.47 to 80.34%. We systematically optimized the fermentation conditions for YG-5, including medium composition and cultivation parameters. The optimized process resulted in an 89.56% inhibition rate against
A. tenuissima
, a 14.72% improvement over non-optimized conditions. Notably, the antimicrobial compounds produced by YG-5 demonstrated stability across various temperatures, pH levels, and UV irradiation. In vivo efficacy trials showed promising results, with YG-5 fermentation broth reducing
Alternaria
leaf spot incidence on star anise leaves by 56.95%. These findings suggest that
Streptomyces
sp. YG-5 holds significant potential as a biocontrol agent against
Alternaria
leaf spot in star anise cultivation, offering a sustainable approach to disease management in this valuable crop.