The utilization of organic phosphorus(P) has directly or indirectly improved after exogenous phytase was added to soil.However,the mechanism by which exogenous phytase affected the soil phosphatases(phosphomonoesterase and phosphodiesterase) activities was not clear.The present work was aimed to study red soil,brown soil and cinnamon soil phosphomonoesterase(acid and alkaline)(AcP and AlP) and phosphodiesterase(PD) activities responding to the addition of exogenous phytase(1 g phytase/50 g air dry soil sample) based on the measurements performed via a fluorescence detection method combined with 96 microplates using a TECAN Infinite 200 Multi-Mode Microplate Reader.The results indicated that the acid phosphomonoesterase activity was significantly enhanced in red soil(p≤0.01),while it was significantly reduced in cinnamon soil;alkaline phosphomonoesterase activity was significantly enhanced in cinnamon soil(p≤0.01),while it was significantly reduced in red soil;phosphodiesterase activity was increased in three soils but it was significantly increased in brown soil(p≤0.01) after the addition of exogenous phytase.The activities still remained strong after eight days in different soils,which indicated that exogenous phytase addition could be enhance soil phosphatases activities effectively.This effect was not only related to soil properties,such as pH and phosphorus forms,but might also be related to the excreted enzyme amount of the stimulating microorganism.Using fluorescence spectroscopy to study exogenous phytase addition influence on soil phosphatase activities was the first time at home and abroad.Compared with the conventional spectrophotometric method,the fluorescence microplate method is an accurate,fast and simple to use method to determine the relationships among the soil phosphatases activities.