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Applied and Environmental Microbiology, November 2001, p. 5233-5239, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5233-5239.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

An Altered Pseudomonas Diversity Is Recovered from Soil by Using Nutrient-Poor Pseudomonas-Selective Soil Extract Media

Nina Aagot,1,2 Ole Nybroe,2 Preben Nielsen,3 and Kaare Johnsen1,*

Geological Survey of Denmark and Greenland, DK-2400 Copenhagen NV,1 Section of Genetics and Microbiology, Department of Ecology, The Royal Veterinary and Agricultural University, DK-1871 Frederiksberg,2 and Novozymes A/S, DK-2880 Bagsvaerd,3 Denmark

Received 24 April 2001/Accepted 30 August 2001

We designed five Pseudomonas-selective soil extract NAA media containing the selective properties of trimethoprim and sodium lauroyl sarcosine and 0 to 100% of the amount of Casamino Acids used in the classical Pseudomonas-selective Gould's S1 medium. All of the isolates were confirmed to be Pseudomonas by a Pseudomonas-specific OprF antibody and a Pseudomonas-specific PCR targeting 16S ribosomal DNA. The Pseudomonas isolates were characterized by classical physiological tests, repetitive extragenic palindromic-PCR, Fourier transform infrared spectroscopy, and carbon source utilization patterns. Several of these analyses showed that the amount of Casamino Acids significantly influenced the diversity of the recovered Pseudomonas isolates. Furthermore, the data suggested that specific Pseudomonas subpopulations were represented on the nutrient-poor media. The NAA 1:100 medium, containing ca. 15 mg of organic carbon per liter, consistently gave significantly higher Pseudomonas CFU counts than Gould's S1 when tested on four Danish soils. NAA 1:100 may, therefore, be a better medium than Gould's S1 for enumeration and isolation of Pseudomonas from the low-nutrient soil environment.

* Corresponding author. Mailing address: Danish Veterinary Laboratory, Bülowsvej 27, DK-1790 Copenhagen V, Denmark. Phone: 45 35 30 01 00. Fax: 45 35 30 01 20. E-mail: kjo{at}svs.dk.

Applied and Environmental Microbiology, November 2001, p. 5233-5239, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5233-5239.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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