Efficient utilization of organic materials based on the rich resources in the karst region can promote soil fertility. Microorganisms have a crucial influence on soil phosphorus availability. phoD is considered to be the encoding phosphatase gene that can reflect the hydrolysis of organophosphorus compounds for the soil bacterial community. Molecular analysis of the phoD-harboring bacterial gene provides insight into promoting soil phosphorus availability under different fertilization managements. However, the effects of organic materials on soil phosphorus fractions associated with phoD-harboring bacterial communities are poorly understood. This study comprehensively investigated the effects of organic materials on soil phosphorus availability and explored environmental drivers of phoD-harboring bacteria in the Karst region. Here, six treatments were designed in the field as follows:non-fertilized control (CK), inorganic fertilization (NPK), inorganic fertilization combined with straw (NPKS), inorganic fertilization combined with manure (NPKM), inorganic fertilization combined with sludge (NPKL), and inorganic fertilization combined with sugarcane ash (NPKA). The phoD-harboring bacterial community in Karst region soil was analyzed using high-throughput sequencing. The results showed that the content of total P (TP), Olsen-P, and Ca2-P increased with the years after organic material application, whereas the content of CaCl2-P first decreased and then increased. Compared to that under the CK treatment, organic material application, especially NPKL treatment, significantly increased soil total nitrogen (TN), TP, Olsen-P, CaCl2-P, and Ca2-P contents, followed by those in the NPKA and NPKM treatments. Correlation analysis showed that the contents of CaCl2-P, Ca2-P, and Olsen-P were significantly positively correlated with soil exchangeable calcium (Ca-ex) content. Redundancy analysis (RDA) showed that TN, Ca-ex, soil organic carbon (SOC), and total potassium (TK) contents were the key factors affecting soil P fractions. Using high-throughput sequencing, we found that only NPKS increased the richness of phoD-harboring bacteria compared to that under the control treatment. No significant difference was observed in the phoD-harboring bacterial community among all treatments. The RDA model selected the Ca-ex, TK, Olsen-P, pH, and SOC as the key environmental predictors for the phoD-harboring bacterial community. In summary, soil phosphorus availability can be improved through the input of organic materials and inorganic fertilizer combined with manure, sludge, and ash. These additions were suitable for nutrient management and sustainable development in farmland soil in the Karst region of Guangxi.