Making plant methane formation visible-Insights from application of 13C-labeled dimethyl sulfoxide.

Methane (CH4) formation by vegetation has been studied intensively over the last 15 years. However, reported CH4 emissions vary by several orders of magnitude, thus making global estimates difficult. Moreover, the mechanism(s) for CH4 formation by plants is (are) largely unknown.Here, we introduce a new approach for making CH4 formation by plants clearly visible. By application of 13C-labeled dimethyl sulfoxide (DMSO) onto the leaves of tobacco plants (Nicotiana tabacum) and Chinese silver grass (Miscanthus sinensis) the effect of light and dark conditions on CH4 formation of this pathway was examined by monitoring stable carbon isotope ratios of headspace CH4 (δ13C-CH4 values).Both plant species showed increasing headspace δ13C-CH4 values while exposed to light. Higher light intensities increased CH4 formation rates in N. tabacum but decreased rates for M. sinensis. In the dark no formation of CH4 could be detected for N. tabacum, while M. sinensis still produced ~50% of CH4 compared to that during light exposure.Our findings suggest that CH4 formation is clearly dependent on light conditions and plant species and thus indicate that DMSO is a potential precursor of vegetative CH4. The novel isotope approach has great potential to investigate, at high temporal resolution, physiological, and environmental factors that control pathway-specific CH4 emissions from plants.

Nitrous oxide effluxes from plants as a potentially important source to the atmosphere.

The global budget for nitrous oxide (N2 O), an important greenhouse gas and probably dominant ozone-depleting substance emitted in the 21st century, is far from being fully understood. Cycling of N2 O in terrestrial ecosystems has traditionally exclusively focused on gas exchange between the soil surface (nitrification-denitrification processes) and the atmosphere. Terrestrial vegetation has not been considered in the global budget so far, even though plants are known to release N2 O. Here, we report the N2 O emission rates of 32 plant species from 22 different families measured under controlled laboratory conditions. Furthermore, the first isotopocule values (δ15 N, δ18 O and δ15 Nsp ) of N2 O emitted from plants were determined. A robust relationship established between N2 O emission and CO2 respiration rates, which did not alter significantly over a broad range of changing environmental conditions, was used to quantify plant-derived emissions on an ecosystem scale. Stable isotope measurements (δ15 N, δ18 O and δ15 Nsp ) of N2 O emitted by plants clearly show that the dual isotopocule fingerprint of plant-derived N2 O differs from that of currently known microbial or chemical processes. Our work suggests that vegetation is a natural source of N2 O in the environment with a large fraction released by a hitherto unrecognized process.

Chloromethane Degradation in Soils: A Combined Microbial and Two-Dimensional Stable Isotope Approach.

Chloromethane (CHCl, methyl chloride) is the most abundant volatile halocarbon in the atmosphere and involved in stratospheric ozone depletion. The global CHCl budget, and especially the CHCl sink from microbial degradation in soil, still involves large uncertainties. These may potentially be resolved by a combination of stable isotope analysis and bacterial diversity studies. We determined the stable isotope fractionation of CHCl hydrogen and carbon and investigated bacterial diversity during CHCl degradation in three soils with different properties (forest, grassland, and agricultural soils) and at different temperatures and headspace mixing ratios of CHCl. The extent of chloromethane degradation decreased in the order forest > grassland > agricultural soil. Rates ranged from 0.7 to 2.5 µg g dry wt. d for forest soil, from 0.1 to 0.9 µg g dry wt. d for grassland soil, and from 0.1 to 0.4 µg g dry wt. d for agricultural soil and increased with increasing temperature and CHCl supplementation. The measured mean stable hydrogen enrichment factor of CHCl of -50 ± 13‰ was unaffected by temperature, mixing ratio, or soil type. In contrast, the stable carbon enrichment factor depended on CHCl degradation rates and ranged from -38 to -11‰. Bacterial community composition correlated with soil properties was independent from CHCl degradation or isotope enrichment. Nevertheless, increased abundance after CHCl incubation was observed in 21 bacterial operational taxonomical units (OTUs at the 97% 16S RNA sequence identity level). This suggests that some of these bacterial taxa, although not previously associated with CHCl degradation, may play a role in the microbial CHCl sink in soil.

Quantification of ecosystem C dynamics in a long-term FACE study on permanent grassland.

RATIONALE: Because of the wide-ranging appearance and high soil organic carbon (C) content of grasslands, their ecosystems play an important role in the global C cycle. Thus, even small changes in input or output rates lead to significant changes in the soil C content, thereby affecting atmospheric [CO2 ]. Our aim was to examine if a higher C supply provided under elevated CO2 will increase the soil C pool. Special attention was given to respirational processes, where CO2 emission rates and its sources (plant vs. soil) were considered. METHODS: The Giessen-FACE experiment started in 1998 with a moderate CO2 enrichment of +20% and +30% above ambient on an extensively managed grassland. The experiment consists of three control plots where no CO2 is applied, three plots where [CO2 ] is enriched by +20% and one plot receiving [CO2 ] +30%. To exclude initial CO2 step increase effects, a detailed examination of respirational processes over 30 months was carried out after 6 years of CO2 enrichment starting in June 2004. At that time, the δ(13) C signature of the enrichment-CO2 was switched from -25 ‰ to -48 ‰ without a concomitant change in CO2 concentration. RESULTS: After 9 years, the fraction of new C under [CO2 ] +20% was 37 ± 5.4% in the top 7.5 cm but this decreased with depth. No CO2 effect on soil carbon content was detected. Between June 2004 and December 2006, elevated [CO2 ] +20% increased the ecosystem respiration by 13%. The contribution of root respiration to soil respiration was 37 ± 13% (5 cm) and 43 ± 14% (10 cm) for [CO2 ] +20% and 35 ± 13% and 40 ± 13% for [CO2 ] +30%, respectively. CONCLUSIONS: Our findings of an increased C turnover without a net soil C sequestration suggest that the sink strength of grassland ecosystems might decrease in the future, because the additional C may quickly be released as CO2 to the atmosphere. Copyright © 2016 John Wiley & Sons, Ltd.

Nitrous oxide and methane emissions from cryptogamic covers.

Cryptogamic covers, which comprise some of the oldest forms of terrestrial life on Earth (Lenton & Huntingford, ), have recently been found to fix large amounts of nitrogen and carbon dioxide from the atmosphere (Elbert et al., ). Here we show that they are also greenhouse gas sources with large nitrous oxide (N2 O) and small methane (CH4 ) emissions. Whilst N2 O emission rates varied with temperature, humidity, and N deposition, an almost constant ratio with respect to respiratory CO2 emissions was observed for numerous lichens and bryophytes. We employed this ratio together with respiration data to calculate global and regional N2 O emissions. If our laboratory measurements are typical for lichens and bryophytes living on ground and plant surfaces and scaled on a global basis, we estimate a N2 O source strength of 0.32-0.59 Tg year(-1) for the global N2 O emissions from cryptogamic covers. Thus, our emission estimate might account for 4-9% of the global N2 O budget from natural terrestrial sources. In a wide range of arid and forested regions, cryptogamic covers appear to be the dominant source of N2 O. We suggest that greenhouse gas emissions associated with this source might increase in the course of global change due to higher temperatures and enhanced nitrogen deposition.

Evidence for methane production by saprotrophic fungi.

Methane in the biosphere is mainly produced by prokaryotic methanogenic archaea, biomass burning, coal and oil extraction, and to a lesser extent by eukaryotic plants. Here we demonstrate that saprotrophic fungi produce methane without the involvement of methanogenic archaea. Fluorescence in situ hybridization, confocal laser-scanning microscopy and quantitative real-time PCR confirm no contribution from microbial contamination or endosymbionts. Our results suggest a common methane formation pathway in fungal cells under aerobic conditions and thus identify fungi as another source of methane in the environment. Stable carbon isotope labelling experiments reveal methionine as a precursor of methane in fungi. These findings of an aerobic fungus-derived methane formation pathway open another avenue in methane research and will further assist with current efforts in the identification of the processes involved and their ecological implications.

Linking activity, composition and seasonal dynamics of atmospheric methane oxidizers in a meadow soil.

Microbial oxidation is the only biological sink for atmospheric methane. We assessed seasonal changes in atmospheric methane oxidation and the underlying methanotrophic communities in grassland near Giessen (Germany), along a soil moisture gradient. Soil samples were taken from the surface layer (0-10 cm) of three sites in August 2007, November 2007, February 2008 and May 2008. The sites showed seasonal differences in hydrological parameters. Net uptake rates varied seasonally between 0 and 70 µg CH(4) m(-2) h(-1). Greatest uptake rates coincided with lowest soil moisture in spring and summer. Over all sites and seasons, the methanotrophic communities were dominated by uncultivated methanotrophs. These formed a monophyletic cluster defined by the RA14, MHP and JR1 clades, referred to as upland soil cluster alphaproteobacteria (USCα)-like group. The copy numbers of pmoA genes ranged between 3.8 × 10(5)-1.9 × 10(6) copies g(-1) of soil. Temperature was positively correlated with CH(4) uptake rates (P<0.001), but had no effect on methanotrophic population dynamics. The soil moisture was negatively correlated with CH(4) uptake rates (P<0.001), but showed a positive correlation with changes in USCα-like diversity (P<0.001) and pmoA gene abundance (P<0.05). These were greatest at low net CH(4) uptake rates during winter times and coincided with an overall increase in bacterial 16S rRNA gene abundances (P<0.05). Taken together, soil moisture had a significant but opposed effect on CH(4) uptake rates and methanotrophic population dynamics, the latter being increasingly stimulated by soil moisture contents >50 vol% and primarily related to members of the MHP clade.

Enhanced formation of methane in plant cell cultures by inhibition of cytochrome c oxidase.

The claim of methane (CH4) formation in plants has caused much controversy and debate within the scientific community over the past 4 years. Here, using both stable isotope and concentration measurements, we demonstrate that CH4 formation occurs in plant cell cultures that were grown in the dark under sterile conditions. Under non-stress conditions the plant cell cultures produced trace amounts [0.3-0.6 ng g⁻¹ dry weight (DW) h⁻¹] of CH4 but these could be increased by one to two orders of magnitude (up to 12 ng g⁻¹ DW h⁻¹) when sodium azide, a compound known to disrupt electron transport flow at the cytochrome c oxidase (complex IV) in plant mitochondria, was added to the cell cultures. The addition of other electron transport chain (ETC) inhibitors did not result in significant CH4 formation indicating that a site-specific disturbance of the ETC at complex IV causes CH4 formation in plant cells. Our study is an important first step in providing more information on non-microbial CH4 formation from living plants particularly under abiotic stress conditions that might affect the electron transport flow at the cytochrome c oxidase in plant mitochondria.