TY - JOUR A1 - Shrestha, M. A1 - Abraham, W.-R. A1 - Shrestha, P.M. A1 - Noll, Matthias A1 - Conrad, R. T1 - Activity and composition of methanotrophic bacterial communities in planted rice soil studied by flux measurements, analyses of pmoA gene and stable isotope probing of phospholipid fatty acids JF - Environmental microbiology N2 - Methanotrophs in the rhizosphere of rice field ecosystems attenuate the emissions of CH4 into the atmosphere and thus play an important role for the global cycle of this greenhouse gas. Therefore, we measured the activity and composition of the methanotrophic community in the rhizosphere of rice microcosms. Methane oxidation was determined by measuring the CH4 flux in the presence and absence of difluoromethane as a specific inhibitor for methane oxidation. Methane oxidation started on day 24 and reached the maximum on day 32 after transplantation. The total methanotrophic community was analysed by terminal restriction fragment length polymorphism (T-RFLP) and cloning/sequencing of the pmoA gene, which encodes a subunit of particulate methane monooxygenase. The metabolically active methanotrophic community was analysed by stable isotope probing of microbial phospholipid fatty acids (PLFA-SIP) using 13C-labelled CH4 directly added to the rhizospheric region. Rhizospheric soil and root samples were collected after exposure to 13CH4 for 8 and 18 days. Both T-RFLP/cloning and PLFA-SIP approaches showed that type I and type II methanotrophic populations changed over time with respect to activity and population size in the rhizospheric soil and on the rice roots. However, type I methanotrophs were more active than type II methanotrophs at both time points indicating they were of particular importance in the rhizosphere. PLFA-SIP showed that the active methanotrophic populations exhibit a pronounced spatial and temporal variation in rice microcosms. KW - Methane oxidation KW - Active methanotrophs KW - Rice microcosms KW - PLFA KW - pmoA gene PY - 2008 DO - https://doi.org/10.1111/j.1462-2920.2007.01462.x SN - 1462-2912 SN - 1462-2920 VL - 10 IS - 2 SP - 400 EP - 412 PB - Blackwell Science CY - Oxford AN - OPUS4-16493 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Qiu, Q. A1 - Noll, Matthias A1 - Abraham, W.-R. A1 - Lu, Y. A1 - Conrad, R. T1 - Applying stable isotope probing of phospholipid fatty acids and rRNA in a Chinese rice field to study activity and composition of the methanotrophic bacterial communities in situ JF - The ISME journal : multisiciplinary journal of microbial ecology N2 - Methanotrophs in the rhizosphere play an important role in global climate change since they attenuate methane emission from rice field ecosystems into the atmosphere. Most of the CH4 is emitted via transport through the plant gas vascular system. We used this transport for stable isotope probing (SIP) of the methanotrophs in the rhizosphere under field conditions and pulse-labelled rice plants in a Chinese rice field with CH4 (99% 13C) for 7 days. The rate of 13CH4 loss rate during 13C application was comparable to the CH4 oxidation rate measured by the difluoromethane inhibition technique. The methanotrophic communities on the roots and in the rhizospheric soil were analyzed by terminal-restriction fragment length polymorphism (T-RFLP), cloning and sequencing of the particulate methane monooxygenase (pmoA) gene. Populations of type I methanotrophs were larger than those of type II. Both methane oxidation rates and composition of methanotrophic communities suggested that there was little difference between urea-fertilized and unfertilized fields. SIP of phospholipid fatty acids (PLFA-SIP) and rRNA (RNA-SIP) were used to analyze the metabolically active methanotrophic community in rhizospheric soil. PLFA of type I compared with type II methanotrophs was labelled more strongly with 13C, reaching a maximum of 6.8 atom-% . T-RFLP analysis and cloning/sequencing of 16S rRNA genes showed that methanotrophs, especially of type I, were slightly enriched in the 'heavy' fractions. Our results indicate that CH4 oxidation in the rice rhizosphere under in situ conditions is mainly due to type I methanotrophs. KW - Methane-oxidizing bacteria KW - Rice field KW - Rhizosphere KW - Stable isotope probing KW - Phospholipid fatty acid KW - Particulate methane monooxygenase PY - 2008 DO - https://doi.org/10.1038/ismej.2008.34 SN - 1751-7362 SN - 1751-7370 IS - 2 SP - 602 EP - 614 PB - Nature Publishing Group CY - Basingstoke AN - OPUS4-17660 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Jakobs, Désirée A1 - Mathies, Helena A1 - Abraham, W.-R. A1 - Pritzkow, Wolfgang A1 - Stephan, Ina A1 - Noll, Matthias T1 - Biodegradation of a biocide (Cu-N-cyclohexyldiazenium dioxide) component of a wood preservative by a defined soil bacterial community JF - Applied and environmental microbiology N2 - The wood protection industry has refined their products from chrome-, copper-, and arsenate-based wood preservatives toward solely copper-based preservatives in combination with organic biocides. One of these is Cu-HDO, containing the chelation product of copper and N-cyclohexyldiazenium dioxide (HDO). In this study, the fate of isotope-labeled (13C) and nonlabeled (12C) Cu-HDO incorporated in wood sawdust mixed with soil was investigated. HDO concentration was monitored by high-pressure liquid chromatography. The total carbon and the δ13C content of respired CO2, as well as of the soil-wood-sawdust mixture, were determined with an elemental analyzer-isotopic ratio mass spectrometer. The concentration of HDO decreased significantly after 105 days of incubation, and after 24 days the 13CO2 concentration respired from soil increased steadily to a maximum after 64 days of incubation. Phospholipid fatty acid-stable isotope probing (PFA-SIP) analysis revealed that the dominant PFAs C19:0d8,9, C18:0, C18:1ω7, C18:2ω6,9, C17:1d7,8, C16:0, and C16:1ω7 were highly enriched in their δ13C content. Moreover, RNA-SIP identified members of the phylum Acidobacteria and the genera Phenylobacterium and Comamonas that were assimilating carbon from HDO exclusively. Cu-HDO as part of a wood preservative effectively decreased fungal wood decay and overall microbial respiration from soil. In turn, a defined bacterial community was stimulated that was able to metabolize HDO completely. KW - 16S rRNA KW - Biocide KW - PLFA KW - Stable isotope probing KW - Soil KW - Wood preservative PY - 2010 DO - https://doi.org/10.1128/AEM.01092-10 SN - 0099-2240 VL - 76 IS - 24 SP - 8076 EP - 8083 PB - American Society for Microbiology CY - Washington, DC [u.a.] AN - OPUS4-22688 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -