AimsRational intercropping is crucial for improving phosphorus (P) uptake and utilization. This study aimed to investigate the effects of intercropping on the activation of soil P fractions and available P in acid soil under different P application rates.MethodsField experiments were conducted to investigate the effects of maize intercropping with soybean at different P application rates (0, 60, 90, and 120 kg P ha-1) on soil P fractions and P pool turnover over two consecutive years.ResultsIntercropping of maize and soybean showed an advantage in increasing P uptake and reducing apparent P balance at all P application rates, and promoting soil P pool activation. Compared with monoculture maize, intercropping significantly increased maize P uptake by 43.6-74.3% and 45.5-76.8% in consecutive years, reduced apparent P balance by 17.1-33.4% and 19.9-32.4% in those years. Additionally, intercropping maize increased labile P pools by 32.5-38.4% and 14.4-82.1% over consecutive years and reduced non-labile P pools by 7.4-10.9% and 6.6-11.6% compared with monoculture maize. Moreover, intercropping depleted NaOH-Po, conc. HCl-Pi, conc. HCl-Po and Residual-P fractions, and increased Resin-P, NaHCO3-Pi, NaHCO3-Po, NaOH-Pi, and 1 M HCl-Pi fractions compared with monoculture maize. Resin-P, NaHCO3-Pi, NaHCO3-Po increased by 4.3-41.2%, 21.1-84.6%, and 9.7-98.8%, respectively. Furthermore, intercropping at different P application rates significantly increased acid phosphatase activity (ACP) by 13.8-27.1% and 9.5-13.4%, and significantly increased alkaline phosphatase activity (ALP) by 21.2-42.6% and 19.9-28.6% in those years. Structural equation modelling (SEM) showed that both ACP and ALP played a crucial role in increasing available P directly or indirectly through their effects on organic P turnover.ConclusionsThese results demonstrate that intercropping maize with soybean increases soil P bioavailability by transforming NaOH-Po and conc. HCl-Po into soluble P (Resin-P and NaHCO3-Pi) by facilitating the accumulation of soil phosphatase activity.