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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。
Exogenous phytase added to the soil, can directly or indirectly improve the biological effectiveness of organic phosphorus. However, it is unclear what effect the exogenous phytase will exert on the phosphatase (monosterase, diesterase) activity of the soil itself. Using fluorescent substance as substrate, the 96 microplate and fluorescence detection method were combined. The content of exogenous phytase (1g / 50g dry soil) was determined by multifunctional microplate reader in red soil, brown soil and phosphate Esterase (acidic and basic) and di-esterase activity response. The results showed that after the addition of exogenous phytase, the activity of acid phosphatase increased significantly in red soil (p≤0.01), but decreased significantly in cinnamon soil; alkaline phosphatase activity in brown soil and brown soil (P≤0.01), but decreased significantly in red soils. Phosphodiesterase activity increased to some extent in all three soils, but increased significantly in brown soils (p ≤0.01). In different soils, the activities of different enzymes also kept increasing after 8 days of cultivation, namely, the acid phosphatase in red soil, the phosphodiesterase in brown soil and the alkaline phosphatase in cinnamon The results showed that the addition of exogenous phytase can effectively enhance the activity of soil phosphatase, which is not only related to soil properties such as pH and phosphorus forms, but also may be related to the amount of enzyme secreting microorganisms. It is the first time to study the effect of exogenous phytase on soil phosphatase activity by fluorescence spectroscopy. Compared with the traditional spectrophotometry, microplate fluorescence method is a sensitive, accurate, rapid and simple method for determination of soil phosphatase activity.