Semi-arid grassland in northern China faces degradation and desertification problems, with fertilizer application appearing to be a potential solution by improving soil fertility and plant fields or biodiversity. In this study, mineral and organic fertilizers were used in a semi-arid natural grassland in Hebei Province for three years. The plant characteristics, biomass and species diversity index were assessed and analyzed. In the years 2016 and 2017, mineral fertilizers (RC) significantly increased the total aboveground biomass and the aboveground biomass, natural height, density and coverage of Leymus chinensis compared with organic fertilizers (RO), especially at a moderate application rate (RC2). Leymus chinensis was first divided into its own group and then separated into four groups via cluster tree analysis. The importance values of Leymus chinensis showed continuous increases in mineral fertilizer treatments, but not for organic fertilizers. Margalef’s species richness indexes increased significantly (to 2.09) in the organic treatment (RO1) when compared with RC2. Thus, it was concluded that mineral fertilizers could enhance the position of Leymus chinensis in the natural grassland, while organic fertilizers could promote species biodiversity. This study also provides recommendations regarding the use of fertilizers for the purposes of increasing plant biomass and biodiversity in semi-arid grasslands of northern China.