A novel method for the measurement of dissolved adenosine and guanosine triphosphate in aquatic habitats: applications to marine microbial ecology

Abstract

A novel method for the measurement of dissolved adenosine-5′-triphosphate and guanosine-5′-triphosphate (D-ATP and D-GTP, respectively) in marine and freshwater habitats was developed and applied to samples collected from the oligotrophic North Pacific Ocean. Both D-ATP and D-GTP are co-precipitated by authigenically formed Mg(OH) 2 and can be concentrated by factors greater than 200-fold, for subsequent measurement by the firefly luciferin–luciferase bioluminescence reaction. The detection limit for this method was 2–3 pmol ml −1 of concentrated sample (equivalent to an in situ concentration of 10 pM) with a 5% precision at concentrations of 10 pmol ml −1 or above. A significant positive correlation ( P<0.001) was observed between particulate ATP (P-ATP) and D-ATP in water samples collected from Station ALOHA (22.75°N, 158°W; depth profiles 0–1000 m). The highest concentrations of dissolved nucleotides were found in the euphotic zone (0–175 m) below which the concentrations were low and relatively invariant. The dissolved nucleotide pools generally exceeded their corresponding particulate pools. Using radioisotopic tracer techniques and the new concentration method, turnover times for both particulate and dissolved nucleotides can be determined. The ability to measure concentrations and follow nucleotide tracers accurately in a very dilute environment provides a unique opportunity to address questions on microbial community metabolism, nutrient dynamics and energy flux.