Application of multiple stable isotopes to aid identification of the origin of regional and organic animal products in Hesse, Germany.

There is an increasing global demand for regional and organic produce. However, the growth of these markets depends on consumers' trust. Thus, novel methods must be developed to aid the verification of the origin of produce. We built on our previous study to identify the geographical origin and production method of animal-derived food products. Thirty-samples of eggs, 99 of milk, 34 of beef, and 62 of pork were collected from different regions in central Germany and analysed for their stable isotopic composition. The analysis followed a single-variate authentification approach using five isotope signatures, δ18O, δ2H, δ13C, δ15N, and δ34S. The best-performing indicators for verification of the geographical origin were δ15N and δ34S for beef; δ18O, δ2H, and δ13C for milk, and δ2H and δ13C for pork. These tracers indicated statistically significant differences among regions with the exception of pork; the results recorded for eggs were inconclusive. It was possible to distinguish between production methods by means of δ15N and δ34S (beef); all five tracers (eggs), and δ13C, δ15N, and δ34S (milk). This study demonstrated how the analysis of stable isotopes can be employed to determine the geographic region of origin and production method of animal-derived products in Germany.

Assessment of multiple stable isotopes for tracking regional and organic authenticity of plant products in Hesse, Germany.

As demand for regional and organically produced foodstuff has increased in Europe, the need has arisen to verify the products' origin and production method. For food authenticity tracking (production method and origin), we examined 286 samples of wheat (Triticum aestivum), potatoes (Solanum tuberosum), and apples (Malus domestica) from different regions in Germany for their stable isotope compositions of oxygen, hydrogen, carbon, nitrogen and sulphur. Single-variate authentication methods were used. Suitable isotope tracers to determine wheat's regional origin were δ18O and δ34S. δ13C helped to distinguish between organic and conventional wheat samples. For the separation of the production regions of potatoes, several isotope tracers were suitable (e.g. δ18O, δ2H, δ15N, δ13C and δ34S isotopes in potato protein), but only protein δ15N was suitable to differentiate between organic and conventional potato samples. For the apple samples, 2H and 18O isotopes helped to identify production regions, but no significant statistical differences could be found between organically and conventionally farmed apples. For food authenticity tracking, our study showed the need to take the various isotopes into account. There is an urgent need for a broad reference database if isotope measurements are to become a main tool for determining product's origin.

Compared to conventional, ecological intensive management promotes beneficial proteolytic soil microbial communities for agro-ecosystem functioning under climate change-induced rain regimes.

Projected climate change and rainfall variability will affect soil microbial communities, biogeochemical cycling and agriculture. Nitrogen (N) is the most limiting nutrient in agroecosystems and its cycling and availability is highly dependent on microbial driven processes. In agroecosystems, hydrolysis of organic nitrogen (N) is an important step in controlling soil N availability. We analyzed the effect of management (ecological intensive vs. conventional intensive) on N-cycling processes and involved microbial communities under climate change-induced rain regimes. Terrestrial model ecosystems originating from agroecosystems across Europe were subjected to four different rain regimes for 263 days. Using structural equation modelling we identified direct impacts of rain regimes on N-cycling processes, whereas N-related microbial communities were more resistant. In addition to rain regimes, management indirectly affected N-cycling processes via modifications of N-related microbial community composition. Ecological intensive management promoted a beneficial N-related microbial community composition involved in N-cycling processes under climate change-induced rain regimes. Exploratory analyses identified phosphorus-associated litter properties as possible drivers for the observed management effects on N-related microbial community composition. This work provides novel insights into mechanisms controlling agro-ecosystem functioning under climate change.

Agricultural intensification reduces microbial network complexity and the abundance of keystone taxa in roots.

Root-associated microbes play a key role in plant performance and productivity, making them important players in agroecosystems. So far, very few studies have assessed the impact of different farming systems on the root microbiota and it is still unclear whether agricultural intensification influences the structure and complexity of microbial communities. We investigated the impact of conventional, no-till, and organic farming on wheat root fungal communities using PacBio SMRT sequencing on samples collected from 60 farmlands in Switzerland. Organic farming harbored a much more complex fungal network with significantly higher connectivity than conventional and no-till farming systems. The abundance of keystone taxa was the highest under organic farming where agricultural intensification was the lowest. We also found a strong negative association (R2 = 0.366; P < 0.0001) between agricultural intensification and root fungal network connectivity. The occurrence of keystone taxa was best explained by soil phosphorus levels, bulk density, pH, and mycorrhizal colonization. The majority of keystone taxa are known to form arbuscular mycorrhizal associations with plants and belong to the orders Glomerales, Paraglomerales, and Diversisporales. Supporting this, the abundance of mycorrhizal fungi in roots and soils was also significantly higher under organic farming. To our knowledge, this is the first study to report mycorrhizal keystone taxa for agroecosystems, and we demonstrate that agricultural intensification reduces network complexity and the abundance of keystone taxa in the root microbiome.

The impact of long-term organic farming on soil-derived greenhouse gas emissions.

Agricultural practices contribute considerably to emissions of greenhouse gases. So far, knowledge on the impact of organic compared to non-organic farming on soil-derived nitrous oxide (N2O) and methane (CH4) emissions is limited. We investigated N2O and CH4 fluxes with manual chambers during 571 days in a grass-clover- silage maize - green manure cropping sequence in the long-term field trial "DOK" in Switzerland. We compared two organic farming systems - biodynamic (BIODYN) and bioorganic (BIOORG) - with two non-organic systems - solely mineral fertilisation (CONMIN) and mixed farming including farmyard manure (CONFYM) - all reflecting Swiss farming practices-together with an unfertilised control (NOFERT). We observed a 40.2% reduction of N2O emissions per hectare for organic compared to non-organic systems. In contrast to current knowledge, yield-scaled cumulated N2O emissions under silage maize were similar between organic and non-organic systems. Cumulated on area scale we recorded under silage maize a modest CH4 uptake for BIODYN and CONMIN and high CH4 emissions for CONFYM. We found that, in addition to N input, quality properties such as pH, soil organic carbon and microbial biomass significantly affected N2O emissions. This study showed that organic farming systems can be a viable measure contributing to greenhouse gas mitigation in the agricultural sector.

Organic farming enhances soil microbial abundance and activity-A meta-analysis and meta-regression.

Population growth and climate change challenge our food and farming systems and provide arguments for an increased intensification of agriculture. A promising option is eco-functional intensification through organic farming, an approach based on using and enhancing internal natural resources and processes to secure and improve agricultural productivity, while minimizing negative environmental impacts. In this concept an active soil microbiota plays an important role for various soil based ecosystem services such as nutrient cycling, erosion control and pest and disease regulation. Several studies have reported a positive effect of organic farming on soil health and quality including microbial community traits. However, so far no systematic quantification of whether organic farming systems comprise larger and more active soil microbial communities compared to conventional farming systems was performed on a global scale. Therefore, we conducted a meta-analysis on current literature to quantify possible differences in key indicators for soil microbial abundance and activity in organic and conventional cropping systems. All together we integrated data from 56 mainly peer-reviewed papers into our analysis, including 149 pairwise comparisons originating from different climatic zones and experimental duration ranging from 3 to more than 100 years. Overall, we found that organic systems had 32% to 84% greater microbial biomass carbon, microbial biomass nitrogen, total phospholipid fatty-acids, and dehydrogenase, urease and protease activities than conventional systems. Exclusively the metabolic quotient as an indicator for stresses on microbial communities remained unaffected by the farming systems. Categorical subgroup analysis revealed that crop rotation, the inclusion of legumes in the crop rotation and organic inputs are important farming practices affecting soil microbial community size and activity. Furthermore, we show that differences in microbial size and activity between organic and conventional farming systems vary as a function of land use (arable, orchards, and grassland), plant life cycle (annual and perennial) and climatic zone. In summary, this study shows that overall organic farming enhances total microbial abundance and activity in agricultural soils on a global scale.

Impact of reduced tillage on greenhouse gas emissions and soil carbon stocks in an organic grass-clover ley - winter wheat cropping sequence.

Organic reduced tillage aims to combine the environmental benefits of organic farming and conservation tillage to increase sustainability and soil quality. In temperate climates, there is currently no knowledge about its impact on greenhouse gas emissions and only little information about soil organic carbon (SOC) stocks in these management systems. We therefore monitored nitrous oxide (N2O) and methane (CH4) fluxes besides SOC stocks for two years in a grass-clover ley - winter wheat - cover crop sequence. The monitoring was undertaken in an organically managed long-term tillage trial on a clay rich soil in Switzerland. Reduced tillage (RT) was compared with ploughing (conventional tillage, CT) in interaction with two fertilisation systems, cattle slurry alone (SL) versus cattle manure compost and slurry (MC). Median N2O and CH4 flux rates were 13 µg N2O-N m-2 h-1 and -2 µg CH4C m-2 h-1, respectively, with no treatment effects. N2O fluxes correlated positively with nitrate contents, soil temperature, water filled pore space and dissolved organic carbon and negatively with ammonium contents in soil. Pulse emissions after tillage operations and slurry application dominated cumulative gas emissions. N2O emissions after tillage operations correlated with SOC contents and collinearly to microbial biomass. There was no tillage system impact on cumulative N2O emissions in the grass-clover (0.8-0.9 kg N2O-N ha-1, 369 days) and winter wheat (2.1-3.0 kg N2O-N ha-1, 296 days) cropping seasons, with a tendency towards higher emissions in MC than SL in winter wheat. Including a tillage induced peak after wheat harvest, a full two year data set showed increased cumulative N2O emissions in RT than CT and in MC than SL. There was no clear treatment influence on cumulative CH4 uptake. Topsoil SOC accumulation (0-0.1 m) was still ongoing. SOC stocks were more stratified in RT than CT and in MC than SL. Total SOC stocks (0-0.5 m) were higher in RT than CT in SL and similar in MC. Maximum relative SOC stock difference accounted for +8.1 Mg C ha-1 in RT-MC compared to CT-SL after 13 years which dominated over the relative increase in greenhouse gas emissions. Under these site conditions, organic reduced tillage and manure compost application seems to be a viable greenhouse gas mitigation strategy as long as SOC is sequestered.

Improving Crop Yield and Nutrient Use Efficiency via Biofertilization-A Global Meta-analysis.

The application of microbial inoculants (biofertilizers) is a promising technology for future sustainable farming systems in view of rapidly decreasing phosphorus stocks and the need to more efficiently use available nitrogen (N). Various microbial taxa are currently used as biofertilizers, based on their capacity to access nutrients from fertilizers and soil stocks, to fix atmospheric nitrogen, to improve water uptake or to act as biocontrol agents. Despite the existence of a considerable knowledge on effects of specific taxa of biofertilizers, a comprehensive quantitative assessment of the performance of biofertilizers with different traits such as phosphorus solubilization and N fixation applied to various crops at a global scale is missing. We conducted a meta-analysis to quantify benefits of biofertilizers in terms of yield increase, nitrogen and phosphorus use efficiency, based on 171 peer reviewed publications that met eligibility criteria. Major findings are: (i) the superiority of biofertilizer performance in dry climates over other climatic regions (yield response: dry climate +20.0 ± 1.7%, tropical climate +14.9 ± 1.2%, oceanic climate +10.0 ± 3.7%, continental climate +8.5 ± 2.4%); (ii) meta-regression analyses revealed that yield response due to biofertilizer application was generally small at low soil P levels; efficacy increased along higher soil P levels in the order arbuscular mycorrhizal fungi (AMF), P solubilizers, and N fixers; (iii) meta-regressions showed that the success of inoculation with AMF was greater at low organic matter content and at neutral pH. Our comprehensive analysis provides a basis and guidance for proper choice and application of biofertilizers.

Greenhouse gas fluxes from agricultural soils under organic and non-organic management--a global meta-analysis.

It is anticipated that organic farming systems provide benefits concerning soil conservation and climate protection. A literature search on measured soil-derived greenhouse gas (GHG) (nitrous oxide and methane) fluxes under organic and non-organic management from farming system comparisons was conducted and followed by a meta-analysis. Up to date only 19 studies based on field measurements could be retrieved. Based on 12 studies that cover annual measurements, it appeared with a high significance that area-scaled nitrous oxide emissions from organically managed soils are 492 ± 160 kg CO2 eq. ha(-1) a(-1) lower than from non-organically managed soils. For arable soils the difference amounts to 497 ± 162 kg CO2 eq. ha(-1) a(-1). However, yield-scaled nitrous oxide emissions are higher by 41 ± 34 kg CO2 eq. t(-1) DM under organic management (arable and use). To equalize this mean difference in yield-scaled nitrous oxide emissions between both farming systems, the yield gap has to be less than 17%. Emissions from conventionally managed soils seemed to be influenced mainly by total N inputs, whereas for organically managed soils other variables such as soil characteristics seemed to be more important. This can be explained by the higher bioavailability of the synthetic N fertilisers in non-organic farming systems while the necessary mineralisation of the N sources under organic management leads to lower and retarded availability. Furthermore, a higher methane uptake of 3.2 ± 2.5 kg CO2 eq. ha(-1) a(-1) for arable soils under organic management can be observed. Only one comparative study on rice paddies has been published up to date. All 19 retrieved studies were conducted in the Northern hemisphere under temperate climate. Further GHG flux measurements in farming system comparisons are required to confirm the results and close the existing knowledge gaps.

Enhanced top soil carbon stocks under organic farming.

It has been suggested that conversion to organic farming contributes to soil carbon sequestration, but until now a comprehensive quantitative assessment has been lacking. Therefore, datasets from 74 studies from pairwise comparisons of organic vs. nonorganic farming systems were subjected to metaanalysis to identify differences in soil organic carbon (SOC). We found significant differences and higher values for organically farmed soils of 0.18 ± 0.06% points (mean ± 95% confidence interval) for SOC concentrations, 3.50 ± 1.08 Mg C ha(-1) for stocks, and 0.45 ± 0.21 Mg C ha(-1) y(-1) for sequestration rates compared with nonorganic management. Metaregression did not deliver clear results on drivers, but differences in external C inputs and crop rotations seemed important. Restricting the analysis to zero net input organic systems and retaining only the datasets with highest data quality (measured soil bulk densities and external C and N inputs), the mean difference in SOC stocks between the farming systems was still significant (1.98 ± 1.50 Mg C ha(-1)), whereas the difference in sequestration rates became insignificant (0.07 ± 0.08 Mg C ha(-1) y(-1)). Analyzing zero net input systems for all data without this quality requirement revealed significant, positive differences in SOC concentrations and stocks (0.13 ± 0.09% points and 2.16 ± 1.65 Mg C ha(-1), respectively) and insignificant differences for sequestration rates (0.27 ± 0.37 Mg C ha(-1) y(-1)). The data mainly cover top soil and temperate zones, whereas only few data from tropical regions and subsoil horizons exist. Summarizing, this study shows that organic farming has the potential to accumulate soil carbon.

Biodegradability screening of soil amendments through coupling of wavelength-scanned cavity ring-down spectroscopy to multiple dynamic chambers.

A system was developed for the automatic measurements of ¹³CO2 efflux to determine biodegradation of extra carbon amendments to soils. The system combines wavelength-scanned cavity ring down laser spectroscopy (WS-CRDS) with the open-dynamic chamber (ODC) method. The WS-CRDS instrument and a batch of 24 ODC are coupled via microprocessor-controlled valves. Determination of the biodegradation requires a known δ¹³C value and the applied mass of the carbon compounds, and the biodegradation is calculated based on the ¹³CO2 mixing ratio (ppm) sampled from the headspace of the chambers. The WS-CRDS system provided accurate detection based on parallel samples of three standard gases (¹³CO2 of 2, 11 and 22 ppm) that were measured simultaneously by isotope ratio mass spectrometry (linear regression R² = 0.99). Repeated checking with the same standards showed that the WS-CRDS system showed no drift over seven months.The applicability of the ODC was checked against the closed static chamber (CSC) method using the rapid biodegradation of cane sugar-δ¹³C-labeled through C4 photosynthesis. There was no significant difference between the results from 7-min ODC and 120-min CSC measurements. Further, a test using samples of either cane sugar (C4) or beetroot sugar (C3) mixed into standard soil proved the target functionality of the system, which is to identify the biodegradation of carbon sources with significantly different isotopic signatures.

Estimation by PLFA of microbial community structure associated with the rhizosphere of Lygeum spartum and Piptatherum miliaceum growing in semiarid mine tailings.

The objective of this study was to compare the microbial community composition and biomass associated with the rhizosphere of a perennial gramineous species (Lygeum spartum L.) with that of an annual (Piptatherum miliaceum L.), both growing in semiarid mine tailings. We also established their relationship with the contents of potentially toxic metals as well as with indicators of soil quality. The total phospholipid fatty acid (PLFA) amount was significantly higher in the rhizosphere soil of the annual species than in the rhizosphere soil of the perennial species. The fungal/bacterial PLFA ratio was significantly greater in the perennial species compared to the annual species. The fatty acid 16:1ω5c, the fungal/bacterial PLFA ratio and monounsaturated/saturated PLFA ratio were correlated negatively with the soluble contents of toxic metals. The cyc/prec (cy17:0 + cy19:0/16:1ω7 + 18:1ω7) ratio was correlated positively with the soluble contents of Pb, Zn, Al, Ni, Cd, and Cu. The results of the PLFA analysis for profiling microbial communities and their stress status of both the plant species indicate that perennial and annual gramineous species appear equally suitable for use in programmes of revegetation of semiarid mine tailings.

DNA-based stable isotope probing enables the identification of active bacterial endophytes in potatoes.

A (13)CO2 (99 atom-%, 350 ppm) incubation experiment was performed to identify active bacterial endophytes in two cultivars of Solanum tuberosum, cultivars Desirée and Merkur. We showed that after the assimilation and photosynthetic transformation of (13)CO2 into (13)C-labeled metabolites by the plant, the most directly active, cultivar specific heterotrophic endophytic bacteria that consume these labeled metabolite scan be identified by DNA stable isotope probing (DNA-SIP).Density-resolved DNA fractions obtained from SIP were subjected to 16S rRNA gene-based community analysis using terminal restriction fragment length polymorphism analysis and sequencing of generated gene libraries.Community profiling revealed community compositions that were dominated by plant chloroplast and mitochondrial 16S rRNA genes for the 'light' fractions of (13)CO2-incubated potato cultivars and of potato cultivars not incubated with (13)CO2. In the 'heavy' fractions of the (13)CO2-incubated endophyte DNA, a bacterial 492-bp terminal restriction fragment became abundant, which could be clearly identified as Acinetobacter and Acidovorax spp. in cultivars Merkur and Desirée,respectively, indicating cultivar-dependent distinctions in (13)C-label flow. These two species represent two common potato endophytes with known plant-beneficial activities.The approach demonstrated the successful detection of active bacterial endophytes in potato. DNA-SIP therefore offers new opportunities for exploring the complex nature of plant-microbe interactions and plant-dependent microbial metabolisms within the endosphere.

Effects of cattle husbandry on abundance and activity of methanogenic archaea in upland soils.

In the present study, we tested the hypothesis that animal treading associated with a high input of organic matter would favour methanogenesis in soils used as overwintering pasture. Hence, methane emissions and methanogen populations were examined at sections with different degree of cattle impact in a Farm in South Bohemia, Czech Republic. In spring, methane emission positively corresponded to the gradient of animal impact. Applying phospholipid etherlipid analysis, the highest archaeal biomass was found in section severe impact (SI), followed by moderate impact (MI) and no impact. The same trend was observed for the methanogens as showed by real-time quantitative PCR analyses of methyl coenzyme M reductase (mcrA) genes. The detection of monounsaturated isoprenoid side chain hydrocarbons (i20:1) indicated the presence of acetoclastic methanogens in the cattle-impacted sites. This result was corroborated by the phylogenetic analysis of mcrA gene sequences obtained from section SI, which showed that 33% of the analysed clones belonged to the genus Methanosarcina. The majority of the sequenced clones (41%) showed close affiliations with uncultured rumen archaeons. This leads to the assumption that a substantial part of the methanogenic community in plot SI derived from the grazing cattle itself. Compared to the spring sampling, in autumn, a significant reduction in archaeal biomass and number of copies of mcrA genes was observed mainly for section MI. It can be concluded that after 5 months without cattle impact, the severely impact section maintained its methane production potential, whereas the methane production potential under moderate impact returned to background values.

Response of soil microbial biomass and community structures to conventional and organic farming systems under identical crop rotations.

In this study the influence of different farming systems on microbial community structure was analyzed using soil samples from the DOK long-term field experiment in Switzerland, which comprises organic (BIODYN and BIOORG) and conventional (CONFYM and CONMIN) farming systems as well as an unfertilized control (NOFERT). We examined microbial communities in winter wheat plots at two different points in the crop rotation (after potatoes and after maize). Employing extended polar lipid analysis up to 244 different phospholipid fatty acids (PLFA) and phospholipid ether lipids (PLEL) were detected. Higher concentrations of PLFA and PLEL in BIODYN and BIOORG indicated a significant influence of organic agriculture on microbial biomass. Farmyard manure (FYM) application consistently revealed the strongest, and the preceding crop the weakest, influence on domain-specific biomass, diversity indices and microbial community structures. Esterlinked PLFA from slowly growing bacteria (k-strategists) showed the strongest responses to long-term organic fertilization. Although the highest fungal biomass was found in the two organic systems of the DOK field trial, their contribution to the differentiation of community structures according to the management regime was relatively low. Prokaryotic communities responded most strongly to either conventional or organic farming management.

Traditional cattle manure application determines abundance, diversity and activity of methanogenic Archaea in arable European soil.

Based on lipid analyses, 16S rRNA/rRNA gene single-strand conformation polymorphism fingerprints and methane flux measurements, influences of the fertilization regime on abundance and diversity of archaeal communities were investigated in soil samples from the long-term (103 years) field trial in Bad Lauchstädt, Germany. The investigated plots followed a gradient of increasing fertilization beginning at no fertilization and ending at the 'cattle manure' itself. The archaeal phospholipid etherlipid (PLEL) concentration was used as an indicator for archaeal biomass and increased with the gradient of increasing fertilization, whereby the concentrations determined for organically fertilized soils were well above previously reported values. Methane emission, although at a low level, were occasionally only observed in organically fertilized soils, whereas the other treatments showed significant methane uptake. Euryarchaeotal organisms were abundant in all investigated samples but 16S rRNA analysis also demonstrated the presence of Crenarchaeota in fertilized soils. Lowest molecular archaeal diversity was found in highest fertilized treatments. Archaea phylogenetically most closely related to cultured methanogens were abundant in these fertilized soils, whereas Archaea with low relatedness to cultured microorganisms dominated in non-fertilized soils. Relatives of Methanoculleus spp. were found almost exclusively in organically fertilized soils or cattle manure. Methanosarcina-related microorganisms were detected in all soils as well as in the cattle manure, but soils with highest organic application rate were specifically dominated by a close phylogenetic relative of Methanosarcina thermophila. Our findings suggest that regular application of cattle manure increased archaeal biomass, but reduced archaeal diversity and selected for methanogenic Methanoculleus and Methanosarcina strains, leading to the circumstance that high organic fertilized soils did not function as a methane sink at the investigated site anymore.

Ochrobactrum oryzae sp. nov., an endophytic bacterial species isolated from deep-water rice in India.

A non-pigmented, motile, Gram-negative bacterium designated MTCC 4195(T) was isolated from surface-sterilized seeds and plant tissue from deep-water rice (Oryza sativa) cultivated in Suraha Tal Lake in northern India. This isolate was shown to reinfect and colonize deep-water rice endophytically. The highest level of 16S rRNA sequence similarity (96.8 %) to strain MTCC 4195(T) was shown by Ochrobactrum gallinifaecis DSM 15295(T). Strain MTCC 4195(T) utilized gamma-hydroxybutyric acid, adonitol, d-glucosaminic acid and arabinose as carbon sources, but failed to use gentiobiose or citrate. The cell-wall fatty acids of strain MTCC 4195(T) were characterized by the presence of a relatively large proportion of C(18 : 1)omega7c and a relative small proportion of C(16 : 0) in comparison with Ochrobactrum species. DNA-DNA relatedness studies showed less than 52 % binding with the DNAs of type strains of other species of the genus Ochrobactrum. On the basis of phenotypic and genotypic characteristics and the results of 16S rRNA gene sequence analysis, the novel species Ochrobactrum oryzae sp. nov. is proposed, with MTCC 4195(T) (=DSM 17471(T)) as the type strain.

Herbaspirillum hiltneri sp. nov., isolated from surface-sterilized wheat roots.

The genus Herbaspirillum of the Betaproteobacteria mainly comprises diazotrophic bacteria with a potential for endophytic and systemic colonization of a variety of plants. The plant-associated bacterial isolates N3(T), N5 and N9 were derived from surface-sterilized wheat roots. After phylogenetic analysis of 16S rRNA gene sequence data the isolates could be allocated to the genus Herbaspirillum, and 99.9 % similarity to the sequence of Herbaspirillum lusitanum P6-12(T) was found. A set of 16S rRNA gene-targeted oligonucleotide probes was developed for the identification of the three novel isolates and H. lusitanum (Hhilu446), and for the specific detection of several other Herbaspirillum species described recently. For higher phylogenetic resolution, the 23S rRNA gene sequences of all members of the genus was sequenced and used to construct a phylogenetic tree. Isolates N3(T), N5 and N9 formed a group that was distinct from all other Herbaspirillum species. In addition, isolate N3(T) and H. lusitanum P6-12(T) exhibited a DNA-DNA hybridization value of only 25 %. The value for DNA-DNA hybridization between N3(T) and other members of the genus Herbaspirillum was between 14 and 32 %; DNA-DNA hybridization between strain N3(T) and isolates N5 and N9 produced values above 95 %. This places the three isolates as representatives of a novel species within the genus Herbaspirillum. A Biolog GN2 assay supported this conclusion. The major fatty acids were C(16 : 1)omega7c, C(16 : 0) and C(18 : 1)omega7c, and the DNA G+C content ranged from 60.9 to 61.5 mol%. Therefore these three isolates should be classified within a novel species, for which the name Herbaspirillum hiltneri sp. nov. is proposed. The type strain is N3(T) (=DSM 17495(T)=LMG 23131(T)).

Methane fluxes in permafrost habitats of the Lena Delta: effects of microbial community structure and organic matter quality.

For the understanding and assessment of recent and future carbon dynamics of arctic permafrost soils the processes of CH(4) production and oxidation, the community structure and the quality of dissolved organic matter (DOM) were studied in two soils of a polygonal tundra. Activities of methanogens and methanotrophs differed significantly in their rates and distribution patterns among the two investigated profiles. Community structure analysis showed similarities between both soils for ester-linked phospholipid fatty acids (PLFAs) and differences in the fraction of unsaponifiable PLFAs and phospholipid ether lipids. Furthermore, a shift of the overall composition of the microbiota with depth at both sites was indicated by an increasing portion of iso- and anteiso-branched fatty acids related to the amount of straight-chain fatty acids. Although permafrost soils represent a large carbon pool, it was shown that the reduced quality of organic matter leads to a substrate limitation of the microbial metabolism. It can be concluded from our and previous findings first that microbial communities in the active layer of an Arctic polygon tundra are composed by members of all three domains of life, with a total biomass comparable to temperate soil ecosystems, and second that these microorganisms are well adapted to the extreme temperature gradient of their environment.