Millipedes and earthworms increase the decomposition rate of 15N-labelled winter rape litter in an arable field

Effects of millipedes and earthworms on the decomposition of 15N-labelled litter of winter oilseed rape were investigated in a microcosm field experiment over a period of 264 days on an oat field near G?ttingen managed by integrated farming. A total of 32 microcosms were filled with defaunated soil. 15N-labelled rape litter was placed either on top of the soil or buried into the soil simulating mulching and ploughing, respectively. To the microcosms nine adult individuals of Blaniulus guttulatus (Diplopoda) and two of Aporrectodea caliginosa (Lumbricidae) were added separately or in combination. In general, the presence of the animals accelerated the decomposition rate of the litter material. The effects were most pronounced in the presence of Aporrectodea caliginosa. The total amount of nitrate, ammonium and the amount of 35N leached from the microcosms was increased in the presence of earthworms or of both earthworms and millipedes. Both species proved to be important members of the detritus food web of the agricultural system studied.     

Influence of Earthworms on the Sulfur Turnover in the Soil

Abstract

The effects of earthworm activity on the concentration and isotopic composition of total sulfur in soils was investigated using batch experiments. Two ecologically different lumbricid species, the anecic Lumbricus terrestris and the endogeic Aporrectodea caliginosa, were used. The earthworms were fed birch leaves, beech leaves, cattle manure or mixed plant litter. All food sources differed isotopically (δ³⁴S) from the soil (Parabraunerde). As a reference, one experiment was carried out without additional food.

The experimental results show, that both earthworm species influence the total S-content and the δ³⁴S-values in the soil by digestion of the different food sources. The differences in the total S-content of the earthworm tissues and in the S-isotopic composition of the casts can be attributed to the ecological differences between the earthworm species.     

15N-Dynamics of Cucumber (Cucumis sativus L.) in Field Trials on Sandy Loam in Brandenburg (Germany)

Abstract

Field experiments were conducted to study the ¹⁵N-utilization of cucumbers (Cucumis sativus L.) grown on sandy loam under black mulch film. With the progress of the ontogenetic development the plants took up rising ¹⁵N-amounts, which were increasingly transferred to the fruits after the beginning of flowering. At the end of the vegetation period more than 55% of the applied ¹⁵N-labelled fertilizer was found in the plants, and from this portion more than 70% in the fruits. Up to 13% of the total plant nitrogen were derived from the fertilizer. In the top soil layer (0–30 cm) the ¹⁵N-content was strongly reduced during plant growth. Though most of the ¹⁵N was taken up by the plants, a ¹⁵N transfer to deeper soil layers (30–60 cm, 60–90 cm) was observed. Balancing the amount of applied ¹⁵N-fertilizer indicates a loss of 11% during the experimental period.     

Lysimeter Investigations on the Effect of Winter Catch Crops and Weeded Fallow on the N-Dynamics in a Sandy Treposol Soil of Northeast Germany

Abstract

Lysimeter experiments (soil: sandy treposol, from the region “Havelländisches Luch”, Brandenburg, Germany) with application of ¹⁵N labelled fertilizer (80 kg N per ha as ¹⁵NH₄ ¹⁵NO₃, 10 at.-%¹⁵N exc.; for simulating mineralization in the early autumn period) were carried out to determine to what extent the amount of mineral- N was temporary conserved by winter catch crops, taken up subsequently in the vegetation periods by following crops, taken by subsequently in the vegetation periods by following crops, or percolated in the leaching water, respectively. The results were as follows:

1) Until winter or spring respectively, the catch crop uptake rates of applied mineral-N were 32% for phacelia (Phacelia tanacetifolia BENTH.), 25% for winter rape (Brassica napus L. cv. ‘AKELA’), and 16% for white mustard (Sinapis alba L.).

2) In the year after, following maize incorporated from 2.1 to 4.5% of the fertilizerborne N. The following plant community of fallow took up from 0.2 to 0.5% N originating from the fertilizer-N.

3) In comparison with the catch crops, N-leaching losses under fallow conditions were highest and equivalent to 17% of the applied fertilizer-N amount. In contrast to 3% of white mustard, phacelia and winter rape reduced N-leaching losses to 0.2 and 0.3% of the applied fertilizer-N amount.

4) In spring of the first year after the beginning of investigations, N-leaching losses were highest under fallow conditions and white mustard cultivation. Thus, the amounts of nitrate losses would exceed the EU limit for drinking water.

5) Three years after the investigations had been started, 10% (white mustard) and 20% (fallow) of the applied fertilizer-N was still found in th lysimeter soil.     

Use of 13C and 15N Mass Spectrometry to Study the Decomposition of Calamagrostis epigeios in Soil Column Experiments with and Without Ash Additions

Abstract

The dynamics of C and N in terrestrial ecosystems are not completely understood and the use of stable isotopes may be useful to gain further insight in the pathways of CO₂ emissions and leaching of dissolved organic carbon (DOC) and nitrogen (DON) during decomposition of litter. Objectives were (i) to study the decomposition dynamics of Calamagrostis epigeios, a common grass species in forests, using ¹³C-depleted and ¹⁵N-enriched plants and (ii) to quantify the effect wood ash addition on the decomposition and leaching of DOC and DON. Decomposition was studied for 128 days under aerobic conditions at 8°C and moisture close to field capacity in a spodic dystric Cambisol with mor-moder layer. Variants included control plots and additions of (i) Calamagrostis litter and (ii) Calamagrostis litter plus 4 kg ash m^?2.

• Decomposition of Calamagrostis resulted in a CO₂ production of 76.2 g CO₂–C m^?2 (10% of added C) after 128 days and cumulative DOC production was 14.0 g C m^?2 out of which 0.9g C m^?2 was Calamagrostis-derived (0.1% of added C). The specific CO₂ formation and specific DOC production from Calamagrostis were 6 times higher (CO₂) and 4 times smaller (DOC) than those from the organic layer. The amount of Calamagrostis-derived total N (NH₄ ⁺, NO₃ ^?, DON) leached was 0.7g N m^?2 (4.8% of added N). Cumulative DON production was 0.8g N m^?2 which was slightly higher than for the control. During soil passage, much of the DOC and DON was removed due to sorption or decomposition. DOC and DON releases from the mineral soil (17cm depth) were 6.3g C m^?2 and 0.5g N m^?2.

  Addition of ash resulted in a complete fixing of CO₂ for 40 days due to carbonatisation. Afterwards, the CO₂ production rates were similar to the variant without ash addition. Production of DOC (98.6g C m^?2) and DON (2.5g N m^?2) was marked, mainly owing to humus decay. However, Calamagrostis-derived DOC and Culamagrostis-derived total N were only 3.9g C m^?2 (0.5 YO of added C) and 0.5g N m^?2 (3.4% of added N). The specific DOC production rate from the organic layer was 6 times higher than that from Cularnagrostis. The results suggest that with increasing humification from fresh plant residues to more decomposed material (O_F and O_H layers) the production ratio of DOC/CO₂-C increases. Addition of alkaline substances to the forest floor can lead to a manifold increase in DOC production.

     

Gel-Electrophoresis and Subsequent Optical Emission 15N Analysis to Identify 15N-Labelled Protein Fractions

Abstract

A combined procedure to detect of ¹⁵N/¹⁴N isotope ratios by emission spectrometric analysis after starch gel-electrophoresis was developed. ¹⁵N-labelled proteins of human serum were used to optimise this method. Electrophorised gel slices with protein fractions were directly digested for subsequent isotope analysis. This method is proposed for use in routine analysis for clinical application.     

Der [15N]Methacetin-Leberfunktionstest zum Nachweis von Arzneimittelnebenwirkungen. Antibiotika-Therapie mit Amplicillin und mit Gefotaxime bei Neugeborenen

Abstract

The effect of soil temperature and moisture on plant growth and mineralisation of organic residues was investigated using ¹⁵N-labelled soybean residues and temperature-controlled tanks in the glasshouse. Treatments were arranged in a factorial design with: three soil temperatures (20, 26 and 30°C), two soil moisture regimes (8% (–800 Kpa) or 12% (–100 Kpa)), soybean residues added (enriched at 1.82 atom % ¹⁵N excess) or no residues; and either sown with ryegrass or not sown. Pots were sampled six weeks after planting and ¹⁵N-enrichment and δ¹³C of the plant and soil fractions were determined. Soil inorganic N was also periodically measured.

Available inorganic N increased significantly with addition of residues and generally decreased with increasing temperature. Plant dry matter decreased significantly with increase in soil temperature and increased with increasing moisture. Root-to-shoot ratio declined with increased temperature and moisture. Percentage nitrogen derived from residues (%Ndfr) increased linearly with increased temperature and moisture. Δ¹³C decreased linearly with increasing temperature and decreasing moisture status. There was a significant correlation between transpiration and dry matter production, but there was no correlation between water use efficiency and Δ¹³C.

The results suggest that C: N ratio of the root material effects the root turnover and in turn the water supply capacity of the root system.     

Investigations on the Nitrogen Metabolism of Forest Trees by Mathematical Modelling of Natural Isotope Ratios

Abstract

Based on an experimental investigation of nitrogen concentrations and δ¹⁵N-values in different parts of Picea abies (L.) trees and soil samples and of their dependence on the age of plant parts and on damaging stress, mathematical models which represent main features of nitrogen turnover in the forest tree - soil system and associated isotope variations are developed. These models consider parts of 3 cycles of nitrogen: (1) transport of soluble nitrogen within trees, (2) turnover of protein nitrogen in different plant parts, and (3) the exchange of N between tree and soil via litter and the uptake of mineralized soil N by plant roots. Isotope effects of protein synthesis and mineralization of organic soil N are included. The models give a qualitative explanation of the data and lead to a better understanding of isotopic variations as indicators of forest damage.     

The Impact of Ozone on the 15N Incorporation and Nitrogen Assimilation of Wheat and Maize

Abstract

Young wheat (C3) and maize (C4) plants were exposed to near-ambient concentrations of ozone in open-top chambers in order to investigate the possible effects of ozone on nitrogen metabolism. Nitrogen was supplied to the plants by adding ¹⁵N-labelled tracer substances via the soil substrate. Enzyme activities (NADH nitrate reductase, nitrite reductase, glutamine synthetase and NADH glutamate dehydrogenase) and the incorporation of ¹⁵N were determined.

The findings show that nitrogen metabolism was affected by O₃, however, there were distinct differences between the two species. In plants treated with O₃, NADH nitrate reductase activity in maize leaves was reduced, while NR activity in wheat leaves only slightly declined. Only minor changes were observed with respect to the activities of nitrite reductase, glutamine synthetase and NADH glutamate dehydrogenase.

Feeding experiments using ¹⁵NO₃ ^? showed that the incorporation of nitrate nitrogen in wheat plants exposed to ozone remains virtually unchanged, whereas in maize plants reduced incorporation rates were observed for nitrate nitrogen. The incorporation of ammonium nitrogen was distinctly increased in wheat and maize by the impact of ozone.

When investigating pigment contents, reduced levels of chlorophyll a and b and carotenoids were observed, whereas the pigment content of wheat leaves remained unchanged. These results indicate that young maize plants are more susceptible than wheat plants to short-term ozone exposure.     

The Spectrofluorometric Monitoring of Water Purification by Natural Alternatives

Aim: Determination of efficiency of zeolite and hydrolytic enzymes isolated from earthworms in water purification.

Methods: The autofluorescence of water was investigated by fluorescence excitation–emission matrix. The purification of water was carried out with natural zeolite and hydrolytic enzymes isolated from earthworms. These methods were applied to samples that contain ciliates that were detected with atomic force spectroscopy.

Results: The zeolite removed ammonia and partly eliminated some organic compounds. The hydrolytic enzymes were efficient in biodegrading organic compounds present in water.

Conclusion: Zeolite and enzymes are applicable in water purification.     

The Fate of [15N]Ammonium and [15N]Nitrate in the Soil of a 140-Year-Old Spruce Stand (Picea Abies) in the Fichtelgebirge (NE-Bavaria)

Abstract

A ¹⁵N tracer-experiment was carried out in a 140-year-old spruce stand (Picea abies (L.) Karst.) in the Fichtelgebirge (NE-Bavaria, Germany). Highly enriched (98 at%) [¹⁵N]ammonium and [¹⁵N]nitrate were applied as tracers by simulation of a deposition of 41.3 mol N ha^?1 with 11 water m^?2. To examine seasonal variations of uptake by spruce and understorey vegetation, different plots were labelled in spring, summer and autumn 1994.

One aim of the present study was to perfect a method of preparation of soil extracts for isotope ratio mass spectrometry (IRMS) measurements. Ammonium and nitrate from soil extracts were prepared for IRMS measurements by steam distillation and subsequent freeze drying. Additionally, tracer distribution and transformations in the soil nitrogen pools were examined. Ammonium, nitrate and total nitrogen were examined in the organic layer and the upper 10 cm of the mineral soil during 3 months after the first tracer application in spring 1994.

In July 1994, three months after tracer application, 40% of the [¹⁵N]ammonium label and 29% of the [¹⁵N]nitrate label, respectively, were recovered in the total N pool of the investigated soil horizons. In the organic layer the L/Of horizon retained most of the recovered tracers. Nitrification, immobilisation and mineralisation occurred even under the conditions of high soil acidity at the study site.     

Carbon-13 Kinetic Isotope Effects in the Decarbonylation of Liquid Formic Acid Assisted with Acetic Anhydride

Abstract

We evaluated the use of δ¹⁵N- and δ¹³C-values to monitor the development of food web complexity and biodiversity in a regenerating ecosystem. Therefore a model food chain was established feeding cultivated woodlice (Porcellio dilatatus) on a cellulolytic fungus (Chaetomium globosum) grown on cellulose paper. Two diets of different quality (C:N ratios of 54 vs. 200) with different δ¹⁵N- (1.3‰ vs. 3.1‰) but identical δ¹³C-values caused low and high dietary stress in animals of treatment A and B, respectively. After an incubation time of 7 weeks amount, elemental and isotopic composition of collected faeces and exuviae as well as woodlice and remaining food were determined.

The increase of δ¹⁵N-values of woodlice relative to the diet was 5.7‰ and 2.5‰ in treatments A and B, respectively, whereas δ¹³C-shifts were 1.0‰ and 1.6‰, showing a reverse relationship. Modelling of elemental and isotopic mass balances indicated that faeces recycling explains the unexpected high ¹⁵N-enrichments. Moreover, ¹³C-enrichments were positively correlated to the degree of starvation. Considering the effects of starvation and recycling of faeces, stable isotopes represent a useful tool to elucidate trophic interactions in regenerating food webs.     

Nitrogen Isotope Ratios in Different Compartments of a Mixed Stand of Spruce,Larch and Beech Trees and of Understorey Vegetation Including Fungi

Abstract

Natural nitrogen isotope ratios were measured in different compartments (needles or leaves and twigs of different age classes and crown positions, roots and soil of different horizons) of spruce (Picea abies), larch (Larix decidua) and beech (Fagus sylvatica) trees in an 11-year-old mixed stand in the Fichtelgebirge, NE Bavaria, Germany. In addition, samples of understorey vegetation (mainly ericaceous shrubs and grass) and of ectomycorrhizal and saprophytic fungi were analyzed. The δ¹⁵N values found for all samples ranged between ?7.5 and + 4.5‰. No significant differences were found for the nitrogen isotope ratios of the three tree species despite of their evergreen versus deciduous foliage and despite of their different rooting depth. Ericaceous shrubs had the most negative and fungi and soil from the mineral horizon the most positive δ¹⁵N values. Positive δ¹⁵N values of the fungi indicate their ability to utilize organic soil nitrogen, but the data do not unequivocally show that plants forming mycorrhizas profit from this organic nitrogen source.     

Nitrogen isotope pattern in Mongolian larch stands at the southern Eurasian boreal forest boundary

ABSTRACT

In the last decades a drastic increase in air temperature but a stable precipitation regime in Mongolia has led to gradual drying conditions. Thus, we evaluated the effect of spatial and climatic characteristics on the soil–plant nitrogen dynamics in three representative larch stands (Larix sibirica) with different geographical and climatic conditions using stable nitrogen isotopes. The results showed significant differences in the soil inorganic N content among sites and consequently a different isotopic composition in the plant–soil system. Litter, bark and wood had the lowest δ¹⁵N values for all sites, slightly higher δ¹⁵N values for needles, while the highest δ¹⁵N values were observed for roots and soil. These differences could be the result of the larch stands age themselves, but were in agreement with the spatial and climatic characteristics of the sites. Based on the δ¹⁵N value a higher reliance on ectomycorrhizal fungi (ECMF) was observed in the warmest and driest site, while lower dependency was shown in the cooler northern site with higher soil inorganic N content. In both sites, the rate of air temperature increase has been similar in the last decades; however, their soil–plant N dynamics showed different characteristics.     

The Relationship of Nitrate Reductase Activity to Uptake and Assimilation of Atmospheric 15NO2-Nitrogen in Needles of Norway Spruce (Picea abies [L.] Karst.)

Abstract

Using ¹⁵NO₂, relations between nitrate reductase activity and stomatal conductance, ¹⁵N-uptake and ¹⁵N-glutamate were studied in the two youngest needles flushes of potted Norway spruce (Picea abies [L.] Karst.). There were linear correlations between the stomatal conductance and the ¹⁵N-uptake and between the ¹⁵N-uptake and nitrate reductase (E.C. 1.6.6.1/1.6.6.2) activity. The ¹⁵N labelling of free glutamate shows the assimilation of NO₂ from the atmosphere in addition to the nitrogen from the soil. The portion of glutamate originating from ¹⁵NO₂ was linearly related to nitrate reductase activity in spring experiments. This indicates that this enzyme activity reflected the rate of NO₂-assimilation.     

GC-MS-gestützte Synthese und Qualitätskontrolle von 15N-markiertem Stickstoffmonoxid mit hoher 15N-Häufigkeit für umweltrelevante Tracer-Untersuchungen

Abstract

[¹⁵N]nitric oxide for investigations on the effect of nitric oxide in biological systems can be synthesized economically from the commercially available [¹⁵N]nitric acid. Difficulties arise, however, for the preparation of highly enriched ¹⁵N-labelled nitric oxide on a lower mmol scale. These difficulties are caused by oxygen and result in both, yield depression and insufficient long-time stability of the NO-containing gas mixture due to its oxidation to NO₂. Therefore, a sensitive in situ oxygen detection during synthesis was carried out by on line coupling of the synthesis apparatus with a GC-MS. Additionally, this equipment offers the advantage that both, conversion and ¹⁵N enrichment may be measured almost continuously and simultaneously. In this way, the yield can be increased up to 85% without increasing by-products. The gas mixture prepared from this [¹⁵N]nitric oxide showed an adequate stability for several months at a concentration of about 100 ppm(v) NO.     

Isotopic discrimination during litter decomposition and delta13C and delta15N soil profiles in a young artificial stand and in an old floodplain forest

In the present study, rates of litter decomposition and microbial biomass nitrogen were monitored over an 8-month period in a young broadleaf plantation (18 y) and in an old floodplain forest. Moreover, delta13C and delta15N temporal variations within soil profiles were evaluated at both sites. Rates of litter decomposition were higher in spring and autumn than in summer, in both forests. At the end of the observation period the percentage of original litter remaining was not statistically different between the young and the old forest and accounted for 60-70% of the original amount. Microbial biomass nitrogen in the remaining litter and the percentage of litter mass lost during decomposition were positively correlated. The difference in litter quality affected the decomposition rate and also the changes in carbon isotopic composition during the decomposition process. In contrast, 15N isotopic signatures showed a similar trend in the litter of the two forests irrespective of the litter quality. Although delta13Csoil and delta15Nsoil showed considerable temporal variation they increased with depth in the soils of both sites but their seasonal changes did not reflect those of the decomposing litter. Within the same soil horizon, both delta13C and delta15N showed similar seasonal trends in the soils of the two forests, suggesting the involvement of environmental factors acting at regional level, such as soil temperature and rainfall variations, in regulating seasonal delta13C and delta15N soil variations.     

Konstanz summer school 2018 on isotope ecology

Abstract

Biodegradation of polychlorinated biphenyls (PCBs) in soil is considered to be very complex due to various physico-chemical factors involved. Isotope labelling technique is the best to trace fate of the xenobiotic in the environment. In this work, the uniformly ¹⁴C-labelled PCB congener 11 (3,3′-chlorobiphenyl) was chosen as a low chlorinated coplanar biphenyl which was assumed to be readily degraded by microorganisms. Pleurotus ostreatus and two Pseudomonas species, representing white rot fungi and soil bacteria were used separately or in a consortium. The amount of liberated ¹⁴CO₂ and radio-HPLC, HPLC, GC-MS, and radio-TLC analyses of extracts at the end of a two-month experiment showed that the mineralization of PCB 11 was < 0.4%, volatilization < 3.1%, and 30% of radioactivity was irreversibly bound to the soil matrix. The respective contents of all intermediate metabolites were 4.7 to 10.5 and 2.5 to 2.7% where Pseudomonas alcaligenes alone or in combination with P. putida was applied. 3-Chlorobenzoic acid was the major biodegradation product.     

15N Distribution in Wheat and Chemical Fractionation of Root-Borne 15N in the Soil

Abstract

Spring wheat plants were grown in split-root containers and labelled with ¹⁵N by fertilizing one compartment of the container with ¹⁵NH₄ ¹⁵NO₃ (95 at.-% ¹⁵N exc.). After the harvest, approx. 90% of the ¹⁵N incorporated by the plants were found in the shoots and 3% in the roots; approx. 7% had been released into the soil of the unlabelled compartment. The ¹⁵N in the soil of the unlabelled compartment was extracted with KCl and hydrolysed with HCl. Approx. 60% of the ¹⁵N was found in the hydrolysable organic N pool of the soil and 9% in the fraction of the soluble and exchangeable inorganic nitrogen.     

Influence of earthworms on the sulfur turnover in the soil

Abstract The effects of earthworm activity on the concentration and isotopic composition of total sulfur in soils was investigated using batch experiments. Two ecologically different lumbricid species, the anecic Lumbricus terrestris and the endogeic Aporrectodea caliginosa, were used. The earthworms were fed birch leaves, beech leaves, cattle manure or mixed plant litter. All food sources differed isotopically (δ(34)S) from the soil (Parabraunerde). As a reference, one experiment was carried out without additional food. The experimental results show, that both earthworm species influence the total S-content and the δ(34)S-values in the soil by digestion of the different food sources. The differences in the total S-content of the earthworm tissues and in the S-isotopic composition of the casts can be attributed to the ecological differences between the earthworm species.