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A fluorescence-based microplate assay to quantify DOM-induced catabolic activity.
1INRA, UMR Climat, Sol et Environnement, Domaine St Paul, Site Agroparc, 84914, Avignon Cedex 9, France. Yves.Dudal@avignon.inra.fr
Analytical and Bioanalytical Chemistry
|November 5, 2005
Summary
This study introduces an inverse catabolic response profiles (CRP) assay for rapid quantification of dissolved organic matter (DOM) activity in low-volume samples. The novel method efficiently measures bacterial utilization of DOM, revealing only 9% biodegradability in a soil sample.
Area of Science:
- Environmental microbiology
- Biogeochemistry
- Analytical chemistry
Background:
- Dissolved organic matter (DOM) plays a crucial role in aquatic and soil ecosystems.
- Quantifying the catabolic activity of DOM is essential for understanding carbon cycling.
- Existing methods for DOM activity assessment often require large sample volumes and are time-consuming.
Purpose of the Study:
- To develop and validate a novel, rapid assay for quantifying DOM-induced bacterial catabolic activity.
- To adapt the catabolic response profiles (CRP) assay for low-volume, high-throughput analysis.
- To determine the readily biodegradable fraction of a surface soil DOM sample.
Main Methods:
- Development of an inverse CRP assay using Pseudomonas fluorescens and a fluorogenic redox indicator.
- Incubation of bacteria with DOM samples in a 96-well microplate format.
- Measurement of fluorescence to quantify bacterial catabolic response to DOM.
- Determination of assay sensitivity using glucose standards.
Main Results:
- The assay requires only 0.8 mL of DOM sample per analysis, including controls and replicates.
- A detection limit of 55 microM glucose (0.3 mg C L(-1)) was established.
- Analysis of a surface soil DOM sample indicated that only 9% of the total carbon was readily biodegradable.
Conclusions:
- The inverse CRP assay provides a rapid and efficient method for assessing DOM-induced catabolic activity.
- This technique is suitable for analyzing small sample volumes, facilitating high-throughput screening.
- The findings highlight the limited bioavailability of carbon in the tested soil DOM, with implications for carbon sequestration and nutrient cycling.
