Soil phosphorus (P) deficiency is a major factor limiting crop productivity in many tropical and subtropical soils. Due to the acidic nature of these soils, rock phosphate (RP)-based P fertilizers that are cheaper than manufactured water-soluble P fertilizers can be an attractive alternative under certain conditions. Assessment of the efficacy of these alternative P fertilizers and a rational management of local P resources for sustainable agricultural production require an understanding of the dynamics of P in the soil–plant system and the interactions of various P sources in soils and monitoring of soil available P levels. The present work was conducted to test the applicability of the 32P isotopic kinetic method to assess the soil P fertility status and evaluate the agronomic effectiveness of local rock phosphates in subtropical China. A series of experiments was carried out in the laboratory, greenhouse and field conditions with the following specific objectives: (a) to evaluate the suitability of this isotopic kinetic method in evaluating soil P fertility in 32 soil samples collected across southern China, (b) to test and further develop chemical extraction methods for routine soil P testing, (c) to monitor the dissolution kinetics of local low to medium grade rock phosphate sources and their effect on soil properties and (d) to evaluate their agronomic effectiveness in greenhouse and field experiments. Since most of the studied soils had very low concentrations of soluble P and high P-fixing capacities, the isotopic kinetic method was found unsuitable for evaluating soil P fertility and to predict plant P uptake. In contrast, the proposed chemical extraction method (NaHCO3-NH4F) predicted very well plant P uptake, suggesting that this extraction method can be routinely used to evaluate soil bioavailable P in similar soils in subtropical China. From the incubation study, it was found that although the local low to medium grade RPs were inferior to the reactive NCPR in increasing soil available P levels, they have the potential to improve soil chemical properties. Field experiments indeed demonstrated that the medium grade Jinxiang RP significantly increased crop yield, suggesting that local low to medium grade RPs could be used as P sources to provide P to plants and also to improve soil chemical properties. Overall, these results provide important information for a rational management of P resources for sustainable agriculture in subtropical China.