title: Influence of freeze-thaw stress on the structure and function of microbial communities and denitrifying populations in soil.
abstract: Microbial N2O release during the course of thawing of soil was investigated in model experiment focusing on denitrification, since freeze-thaw has been shown to cause significant physical and biological changes in soil, including a surge of N2O and CO2. The origin of these is still controversially discussed. The increase in denitrification after thawing may be attributed to the diffusion of organic substrates newly available to denitrifiers from disrupted soil aggregates, leading to an increase in microbial activity. Laboratory experiments with upper soil layer of a grassland were conducted in microcosms for real-time gas measurements during the entire phase of freeze and thaw. Shifts in microbial communities were evident on resolution of 16S and 18S rRNA genes and transcripts by denaturing gradient gel electrophoresis (DGGE). Microbial expression profiles were compared by RNA-arbitrarily primed PCR technique and subsequent resolution of amplified products on acrylamide gels. Differences in expression levels of periplasmic nitrate reductase gene (napA) and cytochrome cd1 nitrite reductase (nirS) were observed by most-probable-number-reverse transcription-PCR, with higher levels of expression occurring just after thawing began, followed by a decrease. napA and nirS DGGE profiles showed no change in banding patterns with fingerprints derived from DNA, whereas those derived from cDNA showed a clear succession of denitrifying bacteria, with the most complex pattern being observed at the end of the N2O surge. This study provides insight into the structural community changes and expression dynamics of denitrifiers as a result of freeze-thaw stress. Also, the results presented here support the belief that the gas fluxes observed during thawing is a result of freezing initiated high microbial activity.
authors.affiliation: Institute of Soil Ecology, GSF-National Research Center for Environment and Health, P.O. Box 1129, D-85764 Neuherberg, Germany. shilpi.sharma@gsf.de
authors.forename: Shilpi
authors.initials: S
authors.lastname: Sharma
authors.forename: Zsofia
authors.initials: Z
authors.lastname: Szele
authors.forename: Rolf
authors.initials: R
authors.lastname: Schilling
authors.forename: Jean Charles
authors.initials: JC
authors.lastname: Munch
authors.forename: Michael
authors.initials: M
authors.lastname: Schloter
mesh_terms: D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D017753:Ecosystem; D004591:Electrophoresis, Polyacrylamide Gel; D005615:Freezing; D015964:Gene Expression Regulation, Bacterial; D018869:Heat-Shock Response; D050901:Nitrate Reductase / Q000235:genetics / Q000378:metabolism; D009566:Nitrates / Q000378:metabolism*; D009572:Nitrite Reductases / Q000235:genetics / Q000378:metabolism; D009609:Nitrous Oxide / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012988:Soil Microbiology
chemical_list: D001426:Bacterial Proteins; D009566:Nitrates; D002245:Carbon Dioxide; D009572:Nitrite Reductases; D050901:Nitrate Reductase; D009609:Nitrous Oxide