A microplate assay to measure soil microbial biomass phosphorus

Abstract

Quantification of phosphorus (P) concentrations in microbial biomass is required to better understand how P immobilization and turnover in soils are controlled by environmental and anthropogenic factors. Soil microbial biomass P (MBP) is generally extracted using the chloroform fumigation-direct extraction procedure and then analysed for P using the ammonium molybdate-ascorbic acid method on a flow injection analysis (FIA) system. Our objective was to determine whether a microscale malachite green method on a microplate system would provide as accurate MBP analysis as the ascorbic acid method on an FIA system. Twelve soils were collected from agricultural fields in southwestern Quebec, fumigated with chloroform and extracted with 0.5 M NaHCO₃ (pH 8.5). The dissolved inorganic phosphorus (DIP) concentration in fumigated soils was not affected by the method of analysis, and results from the two systems of analysis were significantly correlated (r =0.998, P <0.05). The MBP concentrations in these agricultural soils were between 0.36 and 60.05 μg P g⁻¹, consistent with other published values. Our results indicate that MBP can be assessed equally well with the malachite green method using a microplate system as with the ascorbic acid method on an FIA system. The microplate system is rapid and requires smaller volumes of samples and reagents than the FIA system, thus reducing the quantity of waste produced. We conclude that the microscale malachite green method could be applied to measure the MBP concentration in a wide range of soils with good sensitivity, reproducibility and accuracy.