Optimization of the 15N2 incorporation and acetylene reduction methods for free-living nitrogen fixation
Abstract
To optimize assay conditions of two common methods for measuring potential free-living nitrogen-fixation (FLNF), acetylene reduction assay (ARA) and 15N2-incorporation (15N2), for use with soil/rhizosphere samples. We tested the impact of different carbon (C) sources, oxygen concentrations (O2), and incubation times on FLNF rates of two low-fertility Michigan soils via ARA and 15N2.FLNF rates were greatest with addition of a C cocktail, at low O2, and with 7-day incubations for both methods. FLNF via ARA was 1700x greater with a C cocktail versus glucose only and via 15N2 was 17x greater with a C cocktail compared to other C sources and no-C controls. Specific O2 optimum varied by method and site. A 7-day incubation was needed for the ARA, but a 3-day incubation was suitable for 15N2. Lastly, we confirm previously identified issues with the ARA of acetylene-independent ethylene production/consumption resulting in potential FLNF measurement error of 1.3–52.3 μg N g–1 day–1. Here, we present an optimized method for measuring potential FLNF in soil/rhizosphere samples which will allow for consistent and comparable FLNF rate measurements. Researchers should account for C source, O2, and incubation time when assessing FLNF and use the ARA method with caution.
- Authors:
-
[1];
- Michigan State Univ., East Lansing, MI (United States)
- Michigan State Univ., Hickory Corner, MI (United States)
- Washington State Univ., Pullman, WA (United States)
- Publication Date:
- Research Org.:
- Michigan State Univ., East Lansing, MI (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1637663
- Grant/Contract Number:
- SC0018409; FC02-07ER64494; SC0014108
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Plant and Soil
- Additional Journal Information:
- Journal Volume: 445; Journal Issue: 1-2; Journal ID: ISSN 0032-079X
- Publisher:
- Springer
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; free-living nitrogen-fixation; 15N2 incorporation; acetylene reduction; Rhizosphere
Citation Formats
Smercina, Darian N., Evans, Sarah E., Friesen, Maren L., and Tiemann, Lisa K.. Optimization of the 15N2 incorporation and acetylene reduction methods for free-living nitrogen fixation. United States: N. p., 2019.
Web. doi:10.1007/s11104-019-04307-3.
Smercina, Darian N., Evans, Sarah E., Friesen, Maren L., & Tiemann, Lisa K.. Optimization of the 15N2 incorporation and acetylene reduction methods for free-living nitrogen fixation. United States. https://doi.org/10.1007/s11104-019-04307-3
Smercina, Darian N., Evans, Sarah E., Friesen, Maren L., and Tiemann, Lisa K.. Fri .
"Optimization of the 15N2 incorporation and acetylene reduction methods for free-living nitrogen fixation". United States. https://doi.org/10.1007/s11104-019-04307-3. https://www.osti.gov/servlets/purl/1637663.
@article{osti_1637663,
title = {Optimization of the 15N2 incorporation and acetylene reduction methods for free-living nitrogen fixation},
author = {Smercina, Darian N. and Evans, Sarah E. and Friesen, Maren L. and Tiemann, Lisa K.},
abstractNote = {To optimize assay conditions of two common methods for measuring potential free-living nitrogen-fixation (FLNF), acetylene reduction assay (ARA) and 15N2-incorporation (15N2), for use with soil/rhizosphere samples. We tested the impact of different carbon (C) sources, oxygen concentrations (O2), and incubation times on FLNF rates of two low-fertility Michigan soils via ARA and 15N2.FLNF rates were greatest with addition of a C cocktail, at low O2, and with 7-day incubations for both methods. FLNF via ARA was 1700x greater with a C cocktail versus glucose only and via 15N2 was 17x greater with a C cocktail compared to other C sources and no-C controls. Specific O2 optimum varied by method and site. A 7-day incubation was needed for the ARA, but a 3-day incubation was suitable for 15N2. Lastly, we confirm previously identified issues with the ARA of acetylene-independent ethylene production/consumption resulting in potential FLNF measurement error of 1.3–52.3 μg N g–1 day–1. Here, we present an optimized method for measuring potential FLNF in soil/rhizosphere samples which will allow for consistent and comparable FLNF rate measurements. Researchers should account for C source, O2, and incubation time when assessing FLNF and use the ARA method with caution.},
doi = {10.1007/s11104-019-04307-3},
journal = {Plant and Soil},
number = 1-2,
volume = 445,
place = {United States},
year = {2019},
month = {10}
}
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Cited by: 12 works
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Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.