Phosphorus in the soil accessible to plants can easily be combined with calcium ion, the content of which is high in karst rocky desertification (KRD) regions, thereby resulting in a low utilization efficiency of phosphorus. The application of phosphate-solubilizing bacteria (PSB) from the KRD region would facilitate enhanced phosphate availability in the soil. In the present study, the strains belonging to , , and with efficient phosphate-solubilizing ability were isolated from fruit tree rhizosphere soils in KRD regions. Particularly, sp. Ac-14 had a sustained and stable phosphate-solubilizing ability (439-448 mg/L, 48-120 h). Calcium carbonate decreased the phosphate-solubilizing ability in liquid medium; however, it did not affect the solubilization index in agar-solidified medium. When cocultivated with seedling, Ac-14 increased the number of lateral roots, fresh weight, and chlorophyll content of the seedlings. Metabolomics analysis revealed that Ac-14 could produce 23 types of organic acids, majorly including gluconic acid and D-(-)-quinic acid. Expression of Ac-14 glucose dehydrogenase gene () conferred sp. Ps-12 with a sustained and stable phosphate-solubilizing ability, suggesting that the production of gluconic acid is an important mechanism that confers phosphate solubilization in bacteria. Moreover, Ac-14 could also produce indole acetic acid and ammonia. Collectively, the isolated Ac-14 from KRD regions possess an efficient phosphate-solubilizing ability and plant growth-promoting effect which could be exploited for enhancing phosphorus availability in KRD regions. This study holds significance for the improvement of soil fertility and agricultural sustainable development in phosphorus-deficient KRD regions.