微池板比色法在多酚氧化酶和β葡萄糖苷酶研究中的应用  

APPLICATION OF MICROPLATE COLORIMETRIC METHOD IN DETERMINING ACTIVITIES OF POLYPHENOL OXIDASE AND β-GLUCOSIDASE IN SOIL

作　　者：王彬彬[1] 林启美[2] 陈源泉[1] 隋鹏[1] 高旺盛[1] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193 [2]中国农业大学资源与环境学院,北京100193

出　　处：《土壤学报》2012年第4期773-779,共7页Acta Pedologica Sinica

基　　金：国家"十二五"科技支撑计划项目(2011BAD16B15;2012BAD14B03);中国农业大学研究生科研创新专项(KYCX2010029)资助

摘　　要：微池板比色法具有快速批量测定土壤酶活性的潜力,将其应用在农田土壤纤维素和木质素分解有关的β-葡萄糖苷酶(BG)和多酚氧化酶(PPO)活性测定及酶的温度敏感性(EQ10)的研究,尚未见报道。本文应用微池板比色法,在获取缓冲液酸碱度、培养时间和匀浆液水土比优化参数基础上,测定不同种植制度土壤(棉花连作和苜蓿-柳枝稷轮作)的PPO和BG活性及其温度敏感性,评价微池板比色法在土壤酶学研究中的价值。结果表明:pH7的缓冲液、2 h的培养时间和匀浆液水土比(20∶1)为最优的测定参数,其既满足批量分析要求又可提高测定的准确性。棉花连作与苜蓿—柳枝稷轮作土壤的PPO活性没有显著性差异,但BG酶活性轮作土壤显著高于连作土壤,PPO和BG活性均随温度的升高而增加,同时,两种栽培制度土壤并没有引起两种酶的EQ10发生显著性的差异。本研究结果显示,微池板比色法能够比较快速地同时测定多个土壤、多种酶活性,可作为土壤酶学常规分析技术,同时需要根据土壤的特性适度调整主要测定参数。The microplate colorimetric method has the potential of fast batch-determining activity of soil enzymes. However,so far in China,nothing has been reported on use of this method in measuring activities of lignin-and-cellulose- decomposing enzymes,i, e. polyphenol oxidase （PPO） and β-glucosidase （BG） ,in the soil and in studying their sensitivity to temperature （EQ10）. In this study,based on optimized acidity of the buffer solution, duration of the incubation and buffer/sample ratio of the slurry,the microplate colorimetric method was used to measure activities of the two enzymes in soils under two different farming systems,i, e. consecutive cropping of cotton and alfalfa-switchgrass rotation, and their sen- sitivity to temperature,and furthermore,its value to the study of soil enzymes was assessed. Results demonstrate that pH 7 of the buffer solution,2 hours of the incubation and 20：1 of the ratio were the optimal parameters for the test, satisfying both the requirements for batch-analysis and accuracy of the analysis. No significant different was observed between the two cropping systems in soil PPO activity. However, BG activity was much higher in the soil under rotation than under consecutive cropping of cotton. Nevertheless,the two enzymes became more active with rising temperature. The EQ10 values of BG and PPO did not vary much with cultivation systems. It is,thus,concluded that the microplate technique can be used as a routine analytic technique to measure quite rapidly a number of soils and enzymes at the same time in the study of enzymes. Meanwhile,it is necessary to adjust some major parameters properly for use of the microplate method in light of properties of the soil samples.

关 键 词：微池板比色法 土壤 多酚氧化酶 β葡萄糖苷酶 

分 类 号：S154[农业科学—土壤学]