This study was conducted at the Soil and Water Department, Faculty of Agriculture, South Valley University, Qena, Egypt . The study included two incubation experiments to test the efficiency of Bacillus megathirum, Bacillus polymyxa, and Trichoderma harzianum in their ability to release phosphorus from rock phosphate. In addition, a pot experiment was conducted to study the effectiveness of bio-enriched rock phosphate as phosphate fertilizer and its effect on some soil chemical properties and phosphorus availability as well as growth and nutrient uptake of the wheat plant during growth season (winter 2021/22022). Results showed that, liquid medium inoculation by the three different microbials strains under study improved P release from rock phosphate compared to the uninoculated treatment (control). Also, the highest values of available phosphorous were recorded with isolating Bacillus polymyxa+Trichoderma harzianum and Bacillus megathirum+ Trichoderma harzianum in most of the incubation periods. Moreover, applying rock phosphate bio-enriched by Bacillus polymyxa+Trichoderma harzianum and Bacillus megathirum+ Trichoderma harzianum resulted in enhancing available soil phosphorus, plant height, root lengths, dry matter as well as N, P and K uptake of wheat plants compared to the other treatments. Also, in most treatments, all previous measurements were enhanced by applying bio-enriched phosphate rocks at the level of 1.2 ton ha-1 compared to the same biological treatments at the level 2.4 ton ha-1 and Superphosphate treatment. Hence, it was concluded that bio-enriched rock phosphate by Bacillus polymyxa+Trichoderma harzianum or Bacillus megathirum+ Trichoderma harzianum before direct application to the soil enhanced the release of P from rock phosphate and proved to be a suitable approach to use rock phosphate.