外源植酸酶添加对土壤磷酸酶活性影响的荧光光谱法研究（英文）

Effect of Exogenous Phytase Addition on Soil Phosphatase Activities:a Fluorescence Spectroscopy Study

外源植酸酶加入土壤中,能直接或者间接提高有机磷的生物有效性。但外源植酸酶添加后,对土壤本身的磷酸酶(单酯酶、二酯酶)活性会产生何种影响尚不明确。采用荧光物质为底物,将96微孔板和荧光检测法相结合,利用多功能酶标仪测定外源植酸酶(1g/50g干土)添加条件下红壤、棕壤以及褐土中磷酸单酯酶(酸性和碱性)和二酯酶的活性响应。结果表明,外源植酸酶添加后,酸性磷酸单酯酶活性在红壤中极显著增加(p≤0.01),而在褐土中显著降低;碱性磷酸单酯酶活性在棕壤和褐土中增加,其中在褐土中增加极显著(p≤0.01),而在红壤中显著降低;磷酸二酯酶活性在三种土壤中均不同程度地增加,其中在棕壤中增加极显著(p≤0.01);不同土壤中,培养到8天时不同酶活性亦保持增加,即红壤中的酸性磷酸单酯酶,棕壤中的磷酸二酯酶以及褐土中的碱性磷酸单酯酶分别保持活性增加,说明外源植酸酶的添加能够有效地增强土壤磷酸酶活性,这种作用不仅与土壤性质如pH和磷形态相关,并且可能与刺激微生物分泌酶量有关。运用荧光光谱法研究外源植酸酶对土壤磷酸酶活性的影响在国内外尚属首次。与传统的分光光度法相比,微孔板荧光法是一种灵敏、准确、快速、简便的土壤磷酸酶活性关系的测定方法。

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（1g 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 200Multi-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.