光谱学与光谱分析
光譜學與光譜分析
광보학여광보분석
SPECTROSCOPY AND SPECTRAL ANALYSIS
2015年
5期
1294-1299
,共6页
杨晓竹%陈振华%张玉兰%陈利军
楊曉竹%陳振華%張玉蘭%陳利軍
양효죽%진진화%장옥란%진리군
荧光光谱方法%外源植酸酶添加%土壤磷酸单酯酶%土壤磷酸二酯酶
熒光光譜方法%外源植痠酶添加%土壤燐痠單酯酶%土壤燐痠二酯酶
형광광보방법%외원식산매첨가%토양린산단지매%토양린산이지매
Fluorescence spectroscopy method%Exogenous phytase addition%Soil phosphomonoesterase%Soil phosphodiesterase
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(1 g/50 g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。
外源植痠酶加入土壤中,能直接或者間接提高有機燐的生物有效性。但外源植痠酶添加後,對土壤本身的燐痠酶(單酯酶、二酯酶)活性會產生何種影響尚不明確。採用熒光物質為底物,將96微孔闆和熒光檢測法相結閤,利用多功能酶標儀測定外源植痠酶(1 g/50 g榦土)添加條件下紅壤、棕壤以及褐土中燐痠單酯酶(痠性和堿性)和二酯酶的活性響應。結果錶明,外源植痠酶添加後,痠性燐痠單酯酶活性在紅壤中極顯著增加(p≤0.01),而在褐土中顯著降低;堿性燐痠單酯酶活性在棕壤和褐土中增加,其中在褐土中增加極顯著(p≤0.01),而在紅壤中顯著降低;燐痠二酯酶活性在三種土壤中均不同程度地增加,其中在棕壤中增加極顯著(p≤0.01);不同土壤中,培養到8天時不同酶活性亦保持增加,即紅壤中的痠性燐痠單酯酶,棕壤中的燐痠二酯酶以及褐土中的堿性燐痠單酯酶分彆保持活性增加,說明外源植痠酶的添加能夠有效地增彊土壤燐痠酶活性,這種作用不僅與土壤性質如pH和燐形態相關,併且可能與刺激微生物分泌酶量有關。運用熒光光譜法研究外源植痠酶對土壤燐痠酶活性的影響在國內外尚屬首次。與傳統的分光光度法相比,微孔闆熒光法是一種靈敏、準確、快速、簡便的土壤燐痠酶活性關繫的測定方法。
외원식산매가입토양중,능직접혹자간접제고유궤린적생물유효성。단외원식산매첨가후,대토양본신적린산매(단지매、이지매)활성회산생하충영향상불명학。채용형광물질위저물,장96미공판화형광검측법상결합,이용다공능매표의측정외원식산매(1 g/50 g간토)첨가조건하홍양、종양이급갈토중린산단지매(산성화감성)화이지매적활성향응。결과표명,외원식산매첨가후,산성린산단지매활성재홍양중겁현저증가(p≤0.01),이재갈토중현저강저;감성린산단지매활성재종양화갈토중증가,기중재갈토중증가겁현저(p≤0.01),이재홍양중현저강저;린산이지매활성재삼충토양중균불동정도지증가,기중재종양중증가겁현저(p≤0.01);불동토양중,배양도8천시불동매활성역보지증가,즉홍양중적산성린산단지매,종양중적린산이지매이급갈토중적감성린산단지매분별보지활성증가,설명외원식산매적첨가능구유효지증강토양린산매활성,저충작용불부여토양성질여pH화린형태상관,병차가능여자격미생물분비매량유관。운용형광광보법연구외원식산매대토양린산매활성적영향재국내외상속수차。여전통적분광광도법상비,미공판형광법시일충령민、준학、쾌속、간편적토양린산매활성관계적측정방법。
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 phosphata-ses (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 mi-croplates using a TECAN Infinite 200 Multi-Mode Microplate Reader .The results indicated that the acid phos-phomonoesterase 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 micro-organism .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 .