In order to study its commercial value, antagonistic spectrum and storage application of YS-1 Paenibacillus brasilensis were investigated in this paper. YS-1 P. brasilensis showed obvious antifungal activity to 5 different fruit pathogens, which was of broad antagonistic spectrum. Effect and application of YS-1 P. brasilensis fermentation liquid on Nanfeng mandarin at different storage temperatures were also investigated with the puncture inoculation method. Results showed that lesion diameter and disease incidence at 25 °C were higher than those at 5 °C after end of the storage, and there was significant difference between them. P. brasilensis fermentation liquid was effective for control of Penicillium italicum on Nanfeng mandarin stored at 5 °C for 25 d or 25 °C for 20 d. Preharvest treatment combined with postharvest treatment significantly reduced the decay rate of Nanfeng mandarin by 5.8% more than the control, particularly in the 1st 2 mo of storage. Fruits treated with P. brasilensis fermentation liquid in preharvest and postharvest period tended to have higher total sugar content, titratable acidity, ascorbic acid (AsA) content, and soluble solids content than those in the control group, and there was significant difference between the 2 groups. A delay was observed in the drop in AsA content. In this article, strain YS-1 is reported for the 1st time as a biocontrol agent against blue mold of Nanfeng mandarin. The research will provide an application reference for preservation of citrus.