N2 fixation estimates in real-time by cavity ring-down laser absorption spectroscopy

Journal Article (Academic article)

The most common currency for estimating N(2) fixation is acetylene reduction to ethylene. Real-time estimates of nitrogen fixation are needed to close the global nitrogen budget and these remain a critical gap in both laboratory and field experiments. We present a new method for continuous real-time measurements of ethylene production: Acetylene Reduction Assays by Cavity ring-down laser Absorption Spectroscopy (ARACAS). In ARACAS, air in the headspace of an incubation chamber is circulated with a diaphragm pump through a cavity ring-down ethylene spectrometer and back to the incubation chamber. This paper describes the new approach and its benefits compared to the conventional detection of ethylene by flame ionization detector gas chromatography. First, the detection of acetylene reduction to ethylene is non-intrusive and chemically non-destructive, allowing for real-time measurements of nitrogenase activity. Second, the measurements are made instantaneously and continuously at ppb levels, allowing for observation of real-time kinetics on time intervals as short as a few seconds. Third, the instrument can be automated for long time periods of measurement. Finally, the technique will be widely accessible by the research community as it can be readily adapted to most existing acetylene reduction protocols and is based on a modestly priced, commercially available instrument. We illustrate its use for measuring N(2) fixation using two species, the diazotrophic bacterium Azotobacter vinelandii and the lichen Peltigera praetextata. We also discuss potential limitations of the approach, primarily the implications of leaks in the analyzer, as well as future improvements.

Full Text

Duke Authors

Cited Authors

  • Cassar, ; Bellenger, JP; Jackson, RB; Karr, J; Barnett, BA

Published Date

  • 2012

Published In

Volume / Issue

  • 168 / 2

Start / End Page

  • 335 - 342

PubMed ID

  • 21879367

Electronic International Standard Serial Number (EISSN)

  • 1432-1939

International Standard Serial Number (ISSN)

  • 0029-8549

Digital Object Identifier (DOI)

  • 10.1007/s00442-011-2105-y


  • eng