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Applied and Environmental Microbiology, May 2004, p. 2886-2891, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2886-2891.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

New Method of Denitrification Analysis of Bradyrhizobium Field Isolates by Gas Chromatographic Determination of 15N-Labeled N2

Reiko Sameshima-Saito, Kaori Chiba, and Kiwamu Minamisawa*

Graduate School of Life Sciences, Tohoku University, Katahira, Aoba-ku, Sendai 980-8577, Japan

Received 28 November 2003/ Accepted 22 January 2004

To evaluate the denitrification abilities of many Bradyrhizobium field isolates, we developed a new 15N-labeled N2 detection methodology, which is free from interference from atmospheric N2 contamination. 30N2 (15N15N) and 29N2 (15N14N) were detected as an apparent peak by a gas chromatograph equipped with a thermal conductivity detector with N2 gas having natural abundance of 15N (0.366 atom%) as a carrier gas. The detection limit was 0.04% 30N2, and the linearity extended at least to 40% 30N2. When Bradyrhizobium japonicum USDA110 was grown in cultures anaerobically with 15NO3–, denitrification product (30N2) was detected stoichiometrically. A total of 65 isolates of soybean bradyrhizobia from two field sites in Japan were assayed by this method. The denitrification abilities were partly correlated with filed sites, Bradyrhizobium species, and the hup genotype.

* Corresponding author. Mailing address: Graduate School of Life Sciences, Tohoku University, Katahira, Aoba-ku, Sendai 980-8577, Japan. Phone: 81-22-217-5684. Fax: 81-22-217-5864. E-mail: kiwamu{at}ige.tohoku.ac.jp.

Applied and Environmental Microbiology, May 2004, p. 2886-2891, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2886-2891.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Itakura, M., Tabata, K., Eda, S., Mitsui, H., Murakami, K., Yasuda, J., Minamisawa, K. (2008). Generation of Bradyrhizobium japonicum Mutants with Increased N2O Reductase Activity by Selection after Introduction of a Mutated dnaQ Gene. Appl. Environ. Microbiol. 74: 7258-7264 [Abstract] [Full Text]  
  • Sameshima-Saito, R., Chiba, K., Hirayama, J., Itakura, M., Mitsui, H., Eda, S., Minamisawa, K. (2006). Symbiotic Bradyrhizobium japonicum Reduces N2O Surrounding the Soybean Root System via Nitrous Oxide Reductase. Appl. Environ. Microbiol. 72: 2526-2532 [Abstract] [Full Text]  


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