Effect of nutrient and selective inhibitor amendments on methane oxidation, nitrous oxide production, and key gene presence and expression in landfill cover soils: characterization of the role of methanotrophs, nitrifiers, and denitrifiers

Appl Microbiol Biotechnol. 2009 Nov;85(2):389-403. doi: 10.1007/s00253-009-2238-7. Epub 2009 Sep 29.


Methane and nitrous oxide are both potent greenhouse gasses, with global warming potentials approximately 25 and 298 times that of carbon dioxide. A matrix of soil microcosms was constructed with landfill cover soils collected from the King Highway Landfill in Kalamazoo, Michigan and exposed to geochemical parameters known to affect methane consumption by methanotrophs while also examining their impact on biogenic nitrous oxide production. It was found that relatively dry soils (5% moisture content) along with 15 mg NH (4) (+) (kg soil)(-1) and 0.1 mg phenylacetylene(kg soil)(-1) provided the greatest stimulation of methane oxidation while minimizing nitrous oxide production. Microarray analyses of pmoA showed that the methanotrophic community structure was dominated by Type II organisms, but Type I genera were more evident with the addition of ammonia. When phenylacetylene was added in conjunction with ammonia, the methanotrophic community structure was more similar to that observed in the presence of no amendments. PCR analyses showed the presence of amoA from both ammonia-oxidizing bacteria and archaea, and that the presence of key genes associated with these cells was reduced with the addition of phenylacetylene. Messenger RNA analyses found transcripts of pmoA, but not of mmoX, nirK, norB, or amoA from either ammonia-oxidizing bacteria or archaea. Pure culture analyses showed that methanotrophs could produce significant amounts of nitrous oxide, particularly when expressing the particulate methane monooxygenase (pMMO). Collectively, these data suggest that methanotrophs expressing pMMO played a role in nitrous oxide production in these microcosms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaea / genetics
  • Archaea / isolation & purification
  • Bacteria / genetics
  • Bacteria / isolation & purification
  • Base Sequence
  • DNA / genetics
  • DNA / isolation & purification
  • DNA Primers
  • DNA, Archaeal / genetics
  • DNA, Archaeal / isolation & purification
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification
  • Global Warming
  • Greenhouse Effect
  • Inorganic Chemicals / isolation & purification
  • Methane / analysis
  • Methane / metabolism*
  • Nitrites / analysis
  • Nitrogen / isolation & purification
  • Nitrogen Oxides / analysis
  • Nitrous Oxide / analysis
  • Oxidation-Reduction
  • Polymerase Chain Reaction
  • RNA, Bacterial / genetics
  • RNA, Bacterial / isolation & purification
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction
  • Soil / analysis
  • Water Pollutants, Chemical / analysis


  • DNA Primers
  • DNA, Archaeal
  • DNA, Bacterial
  • Inorganic Chemicals
  • Nitrites
  • Nitrogen Oxides
  • RNA, Bacterial
  • RNA, Messenger
  • Soil
  • Water Pollutants, Chemical
  • DNA
  • Nitrous Oxide
  • Nitrogen
  • Methane