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Applied and Environmental Microbiology, November 2008, p. 6876-6879, Vol. 74, No. 22
0099-2240/08/$08.00+0     doi:10.1128/AEM.01533-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Nitric Oxide Reductase Gene Expression and Nitrous Oxide Production in Nitrate-Grown Pseudomonas mandelii{triangledown}

Saleema Saleh-Lakha,1,2 Kelly E. Shannon,1,2 Claudia Goyer,1* Jack T. Trevors,2* Bernie J. Zebarth,1 and David L. Burton3

Potato Research Centre, Agriculture and Agri-Food Canada, Fredericton, New Brunswick, Canada E3B 4Z7,1 Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1,2 Department of Environmental Sciences, Nova Scotia Agricultural College, Truro, Nova Scotia, Canada B2N 5E33

Received 7 July 2008/ Accepted 18 September 2008

Pure cultures of Pseudomonas mandelii were incubated with or without nitrate, which acts as a substrate and an electron acceptor for denitrification. Nitric oxide reductase (cnorB) gene expression was measured using a quantitative reverse transcription-PCR, and nitrous oxide emissions were measured by gas chromatography. P. mandelii cells in either the presence or absence of nitrate demonstrated an increase in cnorB gene expression during the first 3 h of growth. The level of expression of cnorB in nitrate-amended cells remained high (average, 2.06 x 108 transcripts/µg of RNA), while in untreated cells it decreased to an average of 3.63 x 106 transcripts/µg of RNA from 4 to 6 h. Nitrous oxide accumulation in the headspace was detected at 2 h, and cumulative emissions continued to increase over a 24-h period to 101 µmol in nitrate-amended cells. P. mandelii cnorB gene expression was not detected under aerobic conditions. These results demonstrate that P. mandelii cnorB gene expression was induced 203-fold at 4 h when nitrate was present in the medium. Accumulations of N2O indicated that the cNorB enzyme was synthesized and active.

* Corresponding author. Mailing address for C. Goyer: Potato Research Centre, Agriculture and Agri-Food Canada, Fredericton, New Brunswick, Canada E3B 4Z7. Phone: (506) 452-4851. Fax: (506) 452-3316. E-mail: goyercm{at}agr.gc.ca. Mailing address for J. T. Trevors: Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1. Phone: (519) 824-4120, ext. 53367. Fax: (519) 837-0442. E-mail: jtrevors{at}uoguelph.ca

{triangledown} Published ahead of print on 26 September 2008.

Applied and Environmental Microbiology, November 2008, p. 6876-6879, Vol. 74, No. 22
0099-2240/08/$08.00+0     doi:10.1128/AEM.01533-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Saleh-Lakha, S., Shannon, K. E., Henderson, S. L., Zebarth, B. J., Burton, D. L., Goyer, C., Trevors, J. T. (2009). Effect of Nitrate and Acetylene on nirS, cnorB, and nosZ Expression and Denitrification Activity in Pseudomonas mandelii. Appl. Environ. Microbiol. 75: 5082-5087 [Abstract] [Full Text]  
  • Saleh-Lakha, S., Shannon, K. E., Henderson, S. L., Goyer, C., Trevors, J. T., Zebarth, B. J., Burton, D. L. (2009). Effect of pH and Temperature on Denitrification Gene Expression and Activity in Pseudomonas mandelii. Appl. Environ. Microbiol. 75: 3903-3911 [Abstract] [Full Text]  


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