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Amplification of 16S ribosomal RNA genes of autotrophic ammonia-oxidising bacteria demonstrates the ubiquity of nitrosospiras in the environment

Lookup NU author(s): Professor Ian Head


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Oligonucleotide sequences selected from the 16S rRNA genes of various species of ammonia-oxidizing bacteria were evaluated as specific PCR amplification primers and probes. The specificities of primer pairs for eubacterial, Nitrosospira and Nitrosomonas rRNA genes were established with sequence databases, and the primer pairs were used to amplify DNA from laboratory cultures and environmental samples. Eubacterial rRNA genes amplified from samples of soil and activated sludge hybridized with an oligonucleotide probe specific for Nitrosospira spp., but not with a Nitrosomonas-specific probe. Lakewater and sediment samples were analysed using a nested PCR technique in which eubacterial rRNA genes were subjected to a secondary amplification with Nitrosomonas or Nitrosospira specific primers. Again, the presence of Nitrosospira DNA, but not Nitrosomonas DNA, was detected and this was confirmed by hybridization of the amplified DNA with an internal oligonucleotide probe. Enrichments of lakewater and sediment samples, incubated for two weeks in the presence of ammonium, produced nitrite and were found to contain DNA from both Nitrosospira and Nitrosomonas as determined by nested PCR amplification and probing of 16S rRNA genes. This demonstrates that Nitrosospira spp. are widespread in the environment. The implications of the detection of Nitrosomonas DNA only after enrichment culture are discussed.

Publication metadata

Author(s): Hiorns WD, Hastings RC, Head IM, McCarthy AJ, Saunders JR, Pickup RW, Hall GH

Publication type: Article

Publication status: Published

Journal: Microbiology

Year: 1995

Volume: 141

Issue: 11

Pages: 2793-2800

ISSN (print): 1350-0872

ISSN (electronic): 1465-2080

Publisher: Society for General Microbiology


DOI: 10.1099/13500872-141-11-2793


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