Nitrogen, phosphorus and potassium inorganic fertilizers are commonly used when growing apples, but their excessive application has resulted in a decline in soil fertility and therefore an inability to maintain sustainable cultivation systems. It is possible to compensate for this with biofertilizers. Nitrogen-fixing, phosphorus solubilizing, and potassium solubilizing bacteria are biofertilizers with a broad range of possible uses. In this study, beneficial microorganisms were screened from the rhizosphere soil of the apple tree, Malus sieversii (Ldb.) Roem., which is rich in microbial diversity in natural environments. It was essential to investigate their effects on the growth of apple seedlings. Eight populations of organic phosphorus solubilizing bacteria (56), inorganic phosphorus solubilizing bacteria (13), nitrogen-fixing bacteria (22), and potassium solubilizing bacteria (24) were isolated from eight populations of Xinjiang wild apple rhizosphere in Ili by medium culture. Their morphological characteristics were recorded and their activity was determined. The most active strains were Pseudomonas migulae, Pseudomonas poae, and Pseudomonas extremaustralis, which was determined by physiological and biochemical properties and 16S rDNA sequence analysis. The L₁₆ (4⁵) orthogonal experiment, which used the three strains as testing materials, was created to assess the effects of the strains on apple physiological indicators, soil nutrients, leaf nutrients, and biomass, as well as to identify the ideal combination, concentration, timing, and application method. The results indicated that the peroxidase (POD) and catalase (CAT) activities of apple seedlings increased significantly under the 10 treatment, and the (SOD) activities of the 0 (control) and 1 (inorganic fertilizer only) treatments were significantly lower than the other treatments; soluble sugar, soluble protein and chlorophyll contents increased in all treatments compared to 0 and 1, while malondialdehyde and proline contents increased or decreased compared to 0 and 1; apple seedlings in treatment 10 had the highest soil N, P, and K content, leaf N, P, and K content and biomass were also all highest in treatment 10. In summary, the strains screened for the test can be used as biofertilizers and the optimum application was determined for treatment 10, meaning that the results also provide a theoretical basis for their application in artificially grown orchards.