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Applied and Environmental Microbiology, February 2001, p. 972-976, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.972-976.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Quantification of Ammonia-Oxidizing Bacteria in Arable Soil by Real-Time PCR

Anna Hermansson and Per-Eric Lindgren*

Department of Biology, Linköping University, S-58183 Linköping, Sweden

Received 1 June 2000/Accepted 14 November 2000

Real-time PCR was used to quantify populations of ammonia-oxidizing bacteria representing the beta  subdivision of the class Proteobacteria in samples of arable soil, both nitrogen fertilized and unfertilized, from Mellby, Sweden. Primers and probes targeting a 16S ribosomal DNA region of the ammonia-oxidizing bacteria were designed and used. In the fertilized soil there were ~6.2 × 107 ammonia-oxidizing bacteria per g of soil, three times more than the number of bacteria in the unfertilized soil. The lytic efficiency of bead beating in these soils was investigated by using populations of free or loosely attached bacteria, bacteria tightly bound to particles, and bacteria in nonfractionated samples. The shapes of the curves generated in these tests showed that the concentration of template DNA released at various times remained constant after 10 to 100 s of bead beating.

* Corresponding author. Mailing address: Molecular Microbiology Group, Department of Physics and Measurement Technology, Linköping University, S-58183 Linköping, Sweden. Phone: 46 13 284471. Fax: 46 13 281399. E-mail: perli{at}ifm.liu.se.

Applied and Environmental Microbiology, February 2001, p. 972-976, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.972-976.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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