长期施肥条件下不同粒径土壤团聚体中Bradford反应土壤蛋白质含量的差异

主要研究了稻—稻连作方式下长期不同施肥对不同粒径土壤团聚体中Bradford反应土壤蛋白质(BRSP)含量的影响及BRSP与土壤有机碳(SOC)和全氮(TN)之间的关系,同时为简化BRSP测定过程,探讨了用过1 mm筛土壤代替1~2 mm土壤团聚体研究BRSP含量的可行性。研究结果表明,长期不同施肥显著影响土壤1~2 mm团聚体中总BRSP(T-BRSP)和易浸提BRSP(EE-BRSP)含量。有机无机肥料配合施用能显著提高土壤中BRSP含量,以NPK+稻草还田提高BRSP含量的效果最佳。经25 a施用不同肥料后,1~2 mm土壤团聚体中2种形态BRSP含量与土壤SOC和TN含量呈显著正相关。试验发现,<1 mm土壤团聚体中同样有BRSP,且处理间的变化趋势与1~2 mm团聚体间的变化趋势相似。试验结果显示,过1 mm筛土壤中T-BRSP和EE-BRSP含量与1~2 mm团聚体中含量的相关性均达到极显著水平,且土壤SOC和TN含量的相关性也均达到极显著水平。     

A comparison of two colorimetric assays,based upon Lowry and Bradford techniques,to estimate total protein in soil extracts

Soil extracts usually contain large quantities of dissolved humified organic material, typically reflected by high polyphenolic content. Since polyphenols seriously confound quantification of extracted protein, minimising this interference is important to ensure measurements are representative. Although the Bradford colorimetric assay is used routinely in soil science for rapid quantification protein in soil-extracts, it has several limitations. We therefore investigated an alternative colorimetric technique based on the Lowry assay (frequently used to measure protein and humic substances as distinct pools in microbial biofilms). The accuracies of both the Bradford assay and a modified Lowry microplate method were compared in factorial combination. Protein was quantified in soil-extracts (extracted with citrate), including standard additions of model protein (BSA) and polyphenol (Sigma H1675-2). Using the Lowry microplate assay described, no interfering effects of citrate were detected even with concentrations up to 5 times greater than are typically used to extract soil protein. Moreover, the Bradford assay was found to be highly susceptible to two simultaneous and confounding artefacts: 1) the colour development due to added protein was greatly inhibited by polyphenol concentration, and 2) substantial colour development was caused directly by the polyphenol addition. In contrast, the Lowry method enabled distinction between colour development from protein and non-protein origin, providing a more accurate quantitative analysis. These results suggest that the modified-Lowry method is a more suitable measure of extract protein (defined by standard equivalents) because it is less confounded by the high polyphenolic content which is so typical of soil extracts.