Degradation of hemicellulose, cellulose and lignin in decomposing spruce needle litter in relation to N

Decomposing needles from a Norway spruce forest in southern Sweden were studied for 559 days under laboratory conditions. Falling needles were collected in control (Co) plots and plots that had received 100 kg N ha⁻¹ yr⁻¹ as (NH₄)₂SO₄ for 9 years under field conditions. One of the aims was to determine whether the previously documented low decomposition rate of the N fertilized (NS) needles could be explained by a lower degradation degree of lignin. The lignin content was studied using the alkaline CuO oxidation method, the Klason lignin method and CPMAS ¹³C NMR spectroscopy. The amounts of cellulose and hemicellulose were also determined.The fertilized needle litters initially decomposed faster than the unfertilized, but later this reaction reversed, so that at the end the mass loss was 45% initial C in the control and 35% initial C in NS. Klason lignin decreased with time in both treatments and overall, the change of Klason lignin mirrored the litter mass loss. No major difference as regards the decomposition of hemicellulose occurred between the treatments, whereas significantly lower concentrations of cellulose were found in NS needles throughout the incubation. The CuO derived compounds (VSC) were somewhat lower in NS needles throughout the decomposition time. Initially, VSC increased slightly in both treatments, which contradicts the Klason lignin data. There was a weak positive relationship (p>0.05) between VSC and Klason lignin. Both vanillyls compounds (V) and cinnamyl compounds (Ci) increased slightly during decomposition, whereas syringyl compounds (S) vanished entirely. The lignin degradation degree, i.e. the acid-to-aldehyde ratio of the vanillyl compounds expressed as (Ac/Al)_v, showed no significant effect of treatment. The ¹³C NMR analyses of the combined samples showed increased content of aromatic C with increasing decomposition time. The carbohydrate content (O-alkyl C) was lower in the fertilized needle litter throughout the incubation time. The alkyl C content tended to increase with decomposition time and N fertilization. The alkyl C/O-alkyl C ratios increased in both treatments during the incubation. The NMR results were not tested statistically.In conclusion, no major difference concerning lignin degradation could be found between the unfertilized and N fertilized needle litter. Thus, the study contradicts the hypothesis that higher amounts of N reduce lignin degradation. The reduced biological activity is probably due to direct N effects on the microorganisms and their decomposing ability.     

Long-term N addition effects on the C mineralization and DOC production in mor humus under spruce

This study was based on laboratory incubations of mor humus from two N fertilized stands of Norway spruce in Sweden (Skogaby and Stråsan), which had received repeated N additions (100 kg N ha⁻¹ yr⁻¹ as (NH₄)₂SO₄ at Skogaby and 35, 73 and 108 kg ha⁻¹ yr⁻¹ as NH₄NO₃ at Stråsan) during 8 and 24-29 years, respectively. The aim was to investigate long-term N effects on the mineralization of C and production of DOC. Mor humus (Oe and Oa) was incubated in columns at 20 °C for 49 days. Columns were leached once a week with artificial throughfall solution, which was analyzed for DOC, total N, NH₄⁺-N and NO₃⁻-N. Prior to each leaching event, CO₂ evolution from the columns was determined. C-to-N ratios in the N-treated Oe layers at Stråsan (21-24) and Skogaby (24) were significantly lower than those of the controls (Stråsan, 32; Skogaby, 28). The cumulative amount of CO₂-C showed a significant treatment effect in the Oe layer at Skogaby, i.e. 18 and 29 mg C g⁻¹ C in the N treatment and control, respectively. At Stråsan, the cumulative CO₂-C was significantly lower in the N3 treatment compared to the control in both layers (33 compared to 74 mg C g⁻¹ C in the Oe layer and 16 compared to 35 mg C g⁻¹ C in the Oa layer). Neither the DOC nor the DON production showed any significant treatment effects at the two sites. However, DOC was lower in the fertilized Oe layer at Skogaby throughout the incubation. The leaching of DON was highest in the Oe layers at both sites, and DON increased with time at Skogaby while there was a decreasing trend at Stråsan. The DOC-to-DON ratio tended to be lower in the fertilized Oe layers at both sites. The NH₄⁺ leaching at Skogaby decreased in the N-treated Oe and Oa layers. At Stråsan, NH₄⁺ from the Oe layer increased in N2 and control. The NO₃⁻ leaching was low and constant in both Skogaby layers. At Stråsan, NO₃⁻ increased in the Oe layer of N1. Cumulative CO₂ was positively correlated to C-to-N ratio (r2=0.71,p<0.01) and to cumulative DOC (r2=0.63,p<0.05) in the Oe layer at Stråsan. Our conclusion was that long-term N additions caused decreased C-to-N ratios and decreased CO₂ evolution rates. The correlation between CO₂ and C-to-N ratio in the Oe layers at Stråsan may be due to a changed quality of the fertilized forest floor material and presence of more N efficient microorganisms.     

Natural 15N and 13C abundance in Andisols influenced by long-term fertilization management in Japan

Two field experiments were conducted on Andisols in Japan to evaluate the changes in the natural ¹⁵N and ¹³C abundance in the soil profile and to determine whether the values of δ¹⁵N could be used as an indicator of fertilizer sources or fertilizer fate. The 6-year experiment conducted at the National Agricultural Research Center (NARC) consisted of the following treatments: application of swine compost (COMPOST), slow-release nitrogen fertilizer (SRNF), readily available nitrogen fertilizer (RANF), and absence of fertilization (CONTROL). Experimental plots located at the Nippon Agricultural Research Institute (NARI) received cattle compost at different rates for 12 years; a forest soil at this site was sampled for comparison. Swine compost application led to a considerable change in the δ¹⁵N distribution pattern in the soil profile, with the highest δ¹⁵N values recorded in the top 20 cm layers of the COMPOST plot, decreasing in the sequence of CONTROL >- RANF > SRNF, mainly due to the relatively high δ¹⁵N value of swine compost and its subsequent decomposition. In contrast, SRNF application resulted in the lowest δ¹⁵N values in soil, indicating the presence of negligible nitrogen losses relative to input and low nitrogen cycling rates. Values of δ¹⁵N increased with compost application rates at NARI. In the leachate collected at 1-m depth, the δ¹⁵N values decreased in the sequence of COMPOST > RANF ≥ CONTROL > SRNF. The δ¹³C values in soil peaked in the 40–60 cm layers for all the fertilizers. The δ¹³C value was lowest in forest soil due to the presence of plant residues in soil organic matter. These results indicated that the δ¹⁵N values in the upper soil layers or leachate may enable to detect pollution sources of organic or inorganic nitrogen qualitatively in Andisols.     

Effect of long-term fertilization on 15N uptake and retention in soil

We did a pot experiment with three different fertilized soils (no fertilizer (No-F), inorganic fertilizer nitrogen, phosphorus and potassium (NPK), manure plus inorganic fertilizer (MNPK)) from a 19-year fertilizer trial. Three N treatments, (1) no N, (2) 100 mg/kg urea-¹⁵N (N), (3) 50 mg/kg urea-¹⁵N + 50 mg/kg corn straw-N (1/2N + 1/2S), were applied to each soil. The residual soil from the same treatments was used to grow second wheat crop. The MNPK soil had significantly higher nitrogen use efficiency (NUE) in the first growing season, and lower N loss than the NPK, and No-F soils. The 1/2N + 1/2S treatment decreased NUE on each soil, even though the MNPK soil still had highest NUE and lowest N loss. The residual ¹⁵N use efficiency (RNUE) in 1/2N + 1/2S treatment of MNPK soil was higher than NPK and No-F soils. We concluded that long-term application of manure plus inorganic fertilizer increased NUE and decreased N loss.     

Effects of litter quality and microarthropods on N dynamics and retention of exogenous 15N in decomposing litter

Summary Surface additions of (¹⁵NH₄)₂SO₄ were used to measure the immobilization and subsequent movement of exogenous N added to two litter types of contrasting quality (Cornus florida and Quercus prinus). Litterbaskets were used to measure the litter mass loss and N dynamics and to follow the movement of the ¹⁵N label through litter, F layer, and soil pools. Half of the litterbaskets of each species were treated with naphthalene to reduce microarthropod densities. The faster decomposing C. florida litter maintained a higher excess atom % ¹⁵N, and a greater relative concentration of the labeled input (g ¹⁵N g^–1) than did Q. prinus litter. In both litter types the excess atom % ¹⁵N, relative concentration (g ¹⁵N g^–1), and absolute amount of label recovered in the litter declined over time. This occurred during a period of net accumulation of total litter N, implying simultaneous release of the initial input and immobilization of N from other sources. The concentration of ¹⁵N in the soil increased over time, while the F layer apparently acted as an intermediary in the transfer of ¹⁵N from litter to soil. Naphthalene effectively reduced microarthropod numbers in all horizons of the litterbaskets and significantly reduced the decay rates of Q. prinus, but not C. florida litter. Naphthalene did not appear to affect total N dynamics in the litter. However, with all horizons taken together, the naphthalene-treated litterbaskets retained more total ¹⁵N than the control litterbaskets. Naphthalene also changed the vertical distribution of ¹⁵N within litterbaskets, so that the litter retained less of the ¹⁵N-labeled input and the F layer and soil horizons retained more of the labeled input than in control litterbaskets. Our major conclusions are: (1) the N pool of decomposing litter is dynamic, with simultaneous N release and immobilization activating N turnover even during the net accumulation phase; (2) litter quality is an important determinant of immobilization and retention of exogenous N inputs and, therefore, turnover of the litter N pool; and (3) microarthropod activity can significantly affect the incorporation and retention of exogenous N inputs in decomposing litter, although these changes are apparently not reflected in net N accumulation or release during the 1st year of decomposition. However, the naphthalene may have affected microbially mediated N dynamics and this possibility needs to be considered in interpreting the results.     

Depth dynamics of soil N contents and natural abundances of 15N after 43 years of long-term fertilization and liming in sub-tropical Alfisol

The aim of this study was to understand impacts of long-term (43 years) fertilization on soil aggregation, N accumulation rates and δ¹⁵N in surface and deep layers in an Alfisol. Soil samples from seven treatments were analysed for N stocks, aggregate-associated N in 0–30 cm and the changes in δ¹⁵N in 0–90 cm depths. The treatments were: unfertilized control (control); recommended N dose (N); recommended N and phosphorus doses (NP); recommended N, P and potassium doses (NPK); 150% of recommended N, P and K doses (150% NPK); NPK + 10 Mg FYM ha^?1 (NPK + FYM) and NPK + 0.4 Mg lime ha^?1 (NPK + L). Results revealed that plots under NPK + FYM had ~39% higher total N concentrations than NPK + L in 0–30 cm soil layers. In NPK + L, macro-aggregates had 35 and 11% and microaggregates had 20 and 9% lower δ¹⁵N values than NPK + FYM in 0–15 and 15–30 cm soil layers, respectively. However, plots receiving NPK + FYM had ~39% greater deep soil (30–90 cm) N accumulation than NPK + L. These results would help understanding N supplying capacity by long-term fertilization and assist devising N management strategies in sub-tropical acidic Alfisols.     

Temporal changes in distribution and composition of N from labeled fertilizer in soil organic matter fractions

Variations in the amount and composition of immobilized nitrogen (N) in major soil organic matter fractions were investigated in a 730-day soil incubation experiment using ¹⁵N-labeled urea and ¹⁵N nuclear magnetic resonance spectroscopy with the cross polarization/magic angle spinning (¹⁵N CPMAS NMR) method. After 730 days, 24.7% of the applied N was recovered from the soil as organic N. The urea-derived N recovered from humic acids and humin decreased from 11.2 and 33.8% of the applied amount after 14 days to 1.6 and 20.4% after 730 days, respectively. When these values were corrected for the microbial biomass (MB) N, they ranged from 9.0 to 1.2% and 28 to 18%, respectively. The proportion of urea-derived N recovered from fulvic acids was low, ranging between 0.4 and 5.8% (with MB N) or 5.6% (without MB N) of the applied amount, whereas that from water-soluble nonhumic substances (WS-NHS; NHS in the fulvic acid fraction) remained high, 28–33% of the applied amount after correction for the contribution of MB N up to day 365, and decreased to 0.9% thereafter. The ¹⁵N CPMAS NMR spectra of humic acids, fulvic acids, and humin showed the largest signal at −254 to −264 ppm, corresponding to peptide/amide N. The proportions of heterocyclic, peptide/amide, guanidine/aniline, and free amino N in the urea-derived humic acid N were 3–7, 83–90, 5–7, and 2–4%, respectively. More than 80% loss of the urea-derived humic acid N did not markedly alter their composition. No time-dependent variations were also observed for the proportions of respective N functional groups in humin N, which were 3–5, 71–78, 12–17, and 6–10% in the same order as above. These results suggest the greater importance of physical stability than structural variation for the initial accumulation of organic N in soil.     

Carbon and nitrogen dynamics in a forest soil amended with purified tannins from different plant species

Tannins are purported to be an important factor controlling nitrogen cycling in forest ecosystems, and the ability of tannins to bind proteins in protein-tannin complexes is thought to be the primary mechanism responsible for these effects. In this study, we examined the influence of well-characterized tannins purified from five different plant species on C and N dynamics of a forest soil A horizon. Tannic acid, a commonly used and commercially available hydrolyzable tannin (HT), and cellulose were also included for comparison. With the exception of tannins from huckleberry (Vaccinium ovatum), the amendments increased respiration 1.4-4.0 fold, indicating that they were acting as a microbial C source. Tannic acid was significantly more labile than the five purified tannins examined in this study. All treatments decreased net N mineralization substantially, through greater N immobilization and decreased mineralization. The six tannins inhibited gross ammonification rates significantly more than cellulose. This suggests that added tannins had effects in addition to serving as an alternative C source. Tannins purified from Bishop pine (Pinus muricata) were the only tannins that significantly inhibited potential gross nitrification rates, however, rates were low even in the control soil making it difficult to detect any inhibition. Differences in tannin structure such as condensed versus HTs and the hydroxylation pattern of the condensed tannin B-ring likely explain differences observed among the tannin treatments. Contrary to other studies, we did not find that condensed tannins were more labile and less inhibitory than HTs, nor that shorter chained tannins were more labile than longer chained tannins. In addition to supporting the hypothesis that reduced N availability in the presence of tannins is caused by complexation reactions, our data suggests tannins act as a labile C source leading to increased N immobilization.     

Effects of decreasing water potential on gross ammonification and nitrification in an acid coniferous forest soil

Changes in the soil water regime, predicted as a consequence of global climate change, might influence the N cycle in temperate forest soils. We investigated the effect of decreasing soil water potentials on gross ammonification and nitrification in different soil horizons of a Norway spruce forest and tested the hypotheses that i) gross rates are more sensitive to desiccation in the Oa and EA horizon as compared to the uppermost Oi/Oe horizon and ii) that gross nitrification is more sensitive than gross ammonification. Soil samples were adjusted by air drying to water potentials from about field capacity to around −1.0 MPa, a range that is often observed under field conditions at our site. Gross rates were measured using the ¹⁵N pool dilution technique. To ensure that the addition of solute label to dry soils and the local rewetting does not affect the results by re-mineralization or preferential consumption of ¹⁵N, we compared different extraction and incubation times.T₀ times ranging from 10 to 300 min and incubation times of 48 h and 72 h did not influence the rates of gross ammonification and nitrification. Even small changes of water potential decreased gross ammonification and nitrification in the O horizon. In the EA horizon, gross nitrification was below detection limit and the response of the generally low rates of gross ammonification to decreasing water potentials was minor. In the Oi/Oe horizon gross ammonification and nitrification decreased from 37.5 to 18.3 mg N kg⁻¹ soil d⁻¹ and from 15.4 to 5.6 mg N kg⁻¹ soil d⁻¹ when the water potential decreased from field capacity to −0.8 MPa. In the Oa horizon gross ammonification decreased from 7.4 to 4.0 mg N kg⁻¹ soil d⁻¹ when the water potential reached −0.6 MPa. At such water potential nitrification almost ceased, while in the Oi/Oe horizon nitrification continued at a rather high level. Hence, only in the Oa horizon nitrification was more sensitive to desiccation than ammonification. Extended drought periods that might result from climate change will cause a reduction in gross N turnover rates in forest soils even at moderate levels of soil desiccation.     

Origin of the nitrogen assimilated by soil fauna living in decomposing beech litter

We investigated the nitrogen source for main taxa of soil fauna in two beech forests of contrasted humus type using ¹⁵N-labelled beech litter and ¹⁵N analysis of soil fauna. ¹⁵N-labelled beech litter was deposited on the topsoil in December 2000 in four stands of different ages at Leinefelde (Germany) with mull humus and in one mature stand at Sorø (Denmark) with moder humus. The fate of the tracer isotope was measured in litter and soil, as well as in the soil fauna, and for each taxa, we calculated the proportion of N in the animal derived from the labelled substrate. Of the original N contained in the litter, 20-41% was lost after 9 months at Leinefelde, and only 10% at Sorø. This loss was counterbalanced by the incorporation of 24-31% external N at Leinefelde, and 31% at Sorø, partly originating from fungal colonisation of the added litter. The proportion of N assimilated from the labelled litter by the different soil animals varied in relation to their mobility and feeding preferences. Large and mobile soil animals, especially predators, derived on average less ¹⁵N because they were also able to feed outside the labelled litter boxes. Detritivores assimilated at most 15% of their nitrogen content at Leinefelde and 11% at Sorø from the decomposing labelled litter. The most labelled taxa at Leinefelde were small fungivorous and coprophagous species, mainly isotomid Collembola such as Isotomiella and Folsomia. At Sorø, best labelled taxa were saprophagous species such as Enchytraeidae, Glomeridae and Phthiracaroidea. These low rates of ¹⁵N assimilation indicate that fresh litter is not directly the main N source for soil animals. The results obtained suggest that soil fauna fed preferentially upon microorganisms colonising the litter at Leinefelde (mull) and from litter itself at Sorø (moder).     

施氮量对嘎啦幼苗~(15)N、~(13)C分配利用特性影响

【目的】采用15N、13C同位素示踪技术,通过对不同施氮量下嘎啦幼苗生长状况及氮、碳分配、利用特性等的研究,以期为苹果生产合理施肥提供依据。【方法】将2年生盆栽嘎啦幼苗进行低、中、高三个氮水平处理,同时进行15N标记。在新梢旺长初始期、新梢旺长期、新梢缓长期分别进行整株13C标记,72小时后,整株解析为叶、梢、根三部分,进行15N、13C测定。样品全氮用凯氏定氮法测定,15N丰度用ZHT-03质谱计测定。13C丰度用DELTA V Advantage同位素比率质谱仪测定。【结果】1)中、高氮水平的施肥处理可在不同程度上提高整株及叶片干物质量和新梢长度。新梢旺长初始期和新梢缓长期嘎啦幼苗整株干物质量、新梢旺长期叶片干物质分配比率在中、高氮水平处理间差异不显著,中氮水平经济有效。新梢旺长期以后新梢长度以中氮高氮低氮,三者间差异性显著,中氮处理有利于新梢生长。2)在新梢旺长初始期,低氮处理植株叶片15N分配率达50%,比其他处理高出13个百分点左右,表明低氮处理更多的氮被叶片所利用,中氮和高氮处理间差异不显著,说明在本试验施氮条件下中氮供应水平已能满足氮素营养需求。3)新梢旺长期和新梢缓长期幼苗13C固定量均以中氮处理最高,新梢旺长初始期3个处理间根系13C分配率中氮高氮低氮,表明中氮处理有利于碳同化物在嘎啦幼苗中的分配。4)不同施氮量处理的嘎啦幼苗,15N利用率随施氮水平提高而降低,高氮处理对碳同化物分配没有显著贡献。【结论】低、中、高氮不同处理新梢缓长期碳同化物在各器官间的分配比较均衡,氮素水平不能影响碳同化物的分配。盆栽试验表明,中氮水平在保证营养供应的同时,能够促进新梢生长和树势健壮。     

Leaf litter C and N cycling from a deciduous permanent crop

Our understanding of leaf litter carbon (C) and nitrogen (N) cycling and its effects on N management of deciduous permanent crops is limited. In a 30-day laboratory incubation, we compared soil respiration and changes in mineral N [ammonium (NH₄⁺-N) + nitrate (NO₃^–-N)], microbial biomass nitrogen (MBN), total organic carbon (TOC) and total non-extractable organic nitrogen (TON) between a control soil at ¹⁵N natural abundance (δ¹⁵N = 1.08‰) without leaf litter and a treatment with the same soil, but with almond (Prunus dulcis (Mill.) D.A. Webb) leaf litter that was also enriched in ¹⁵N (δ¹⁵N = 213‰). Furthermore, a two-end member isotope mixing model was used to identify the source of N in mineral N, MBN and TON pools as either soil or leaf litter. Over 30 d, control and treatment TOC pools decreased while the TON pool increased for the treatment and decreased for the control. Greater soil respiration and significantly lower (p < 0.05) mineral N from 3 to 15 d and significantly greater MBN from 10 to 30 d were observed for the treatment compared to the control. After 30 d, soil-sourced mineral N was significantly greater for the treatment compared to the control. Combined mineral N and MBN pools derived from leaf litter followed a positive linear trend (R² = 0.75) at a rate of 1.39 μg N g^?1 soil day^?1. These results suggest early-stage decomposition of leaf litter leads to N immobilization followed by greater N mineralization during later stages of decomposition. Direct observations of leaf litter C and N cycling assists with quantifying soil N retention and availability in orchard N budgets.     

Dissipation of bacterially derived C and N through the meso- and macrofauna of a grassland soil

Feeding relationships between organisms may be determined by observations of behaviour in manipulative experiments or, as in more recent times, by the use of stable isotope labelling to trace the passage of ¹³C and ¹⁵N through food webs. Here we introduce living bacteria, labelled with both ¹³C and ¹⁵N into intact soil cores to understand further the movement of bacterially sourced C and N into the meso- and macrofauna of a grassland soil. We found that these groups showed a range of isotope levels which relate to their feeding strategies. Some had no label (e.g. dipterous larvae), whilst others were highly labelled which may indicate a preference for the added bacteria. This latter group included Collembola, generally perceived as being predominantly fungal feeders. This work describes a novel technique which has the potential to provide critical information about the dissipation of bacterially derived C and N through the soil food web.     

Savanna-derived organic matter remaining in arable soils of the South African Highveld long-term mixed cropping: Evidence from C and N natural abundance

Sustainable agriculture requires the formation of new humus from the crops. We utilized ¹³C and ¹⁵N signatures of soil organic matter to assess how rapidly wheat/maize cropping contributed to the humus formation in coarse-textured savanna soils of the South African Highveld. Composite samples were taken from the top 20 cm of soils (Plinthustalfs) cropped for lengths of time varying from 0 to 98 years, after conversion from native grassland savanna (C4). We performed natural ¹³C and ¹⁵N abundance measurements on bulk and particle-size fractions. The bulk soil δ¹³C values steadily decreased from −14.6 in (C4 dominated) grassland to −16.5‰ after 90 years of arable cropping. This δ¹³C shift was attributable to increasing replacement of savanna-derived C by wheat crop (C3) C which dominated over maize (C4) inputs. After calculating the annual C input from the crop yields and the output from literature data, by using a stepwise C replacement model, we were able to correct the soil δ¹³C data for the irregular maize inputs for a period of about one century. Within 90 years of cropping 41-89% of the remaining soil organic matter was crop-derived in the three studied agroecosystems. The surface soil C stocks after 90 years of the wheat/maize crop rotation could accurately be described with the Rothamsted Carbon Model, but modelled C inputs to the soil were very low. The coarse sand fraction reflected temporal fluctuations in ¹³C of the last C₃ or C₄ cropping and the silt fraction evidenced selective erosion loss of old savanna-derived C. Bulk soil ¹⁵N did not change with increasing cropping length. Decreasing δ¹⁵N values caused by fertilizer N inputs with prolonged arable cropping were only detected for the coarse sand fraction. This indicated that the present N fertilization was not retained in stable soil C pool. Clearly, conventional cropping practices on the South African highlands neither contribute to the preservation of old savanna C and N, nor the effective humus reformation by the crops.     

Immobilization of avian excreta-derived nutrients and reduced lignin decomposition in needle and twig litter in a temperate coniferous forest

The possible effects of excreta of the Great Cormorant Phalacrocorax carbo on decomposition processes and dynamics of nutrients (N, P, Ca, K, Mg) and organic chemical components (lignin, total carbohydrates) were investigated in a temperate evergreen coniferous forest near Lake Biwa in central Japan. Two-year decomposition processes of needles and twigs of Chamaecyparis obtusa were examined at two sites, control site never colonized by the cormorants (site C) and colonizing site (site 2). Mass loss was faster in needles than in twigs. Mass loss of these litter types was faster at site C than at site 2, which was ascribed to the decreased mass loss rate of acid-insoluble ‘lignin’ at site 2. Net immobilization of N, P, and Ca occurred in needles and twigs at site 2; whereas at site C, mass of these elements decreased without immobilization during decomposition. Duration of immobilization phase of these nutrients at site 2 was estimated to be 1.6 to 2.5 years in needles and 19.6 to 23.5 years in twigs. Immobilization potential (maximum amount of exogenous nutrient immobilized per gram initial material) was similar between needles and twigs for N and Ca but was about 10 times higher in twigs than in needles for P. δ¹³C in needles was relatively constant during the first year and then increased during the second year, whereas δ¹³C in twigs was variable during decomposition. Acid-insoluble fraction was depleted in ¹³C compared to whole needles (1.6-2.1‰) and twigs (2.0-2.5‰). δ¹⁵N of needles and twigs and their acid-insoluble fractions approached to δ¹⁵N of excreta during decomposition at site 2. This result demonstrated the immobilization of excreta-derived N into litter due to the formation of acid-insoluble lignin-like substances complexed with excreta-derived N. No immobilization occurred in K and Mg and their mass decreased during decomposition at both sites. Based on these results of nutrient immobilization during decomposition and on the data of litter fall and excreta amount at site 2, we tentatively calculated stand-level immobilization potential of litter fall and its contribution to total amount of N and P deposited as excreta. Thus, the potential maximum amount immobilized into litter fall (needles and twigs) was estimated to account for 5-7% of total excreta-derived N and P.     

Spatial variations of chemical composition, microbial functional diversity, and enzyme activities in a Mediterranean litter (Quercus ilex L.) profile

Litter decomposition on the forest floor is an essential process in soil nutrient cycles and formation. These processes are controlled by abiotic factors such as climate and chemical litter quality, and by biotic factors such as microbial community diversity and activity. The aim of this study was to investigate the importance of litter depth with respect to (i) chemical litter quality as evaluated by solid-state ¹³C NMR, (ii) enzyme activities, and (iii) microbial functional diversity in four different litter layers (OLn, OLv, OF, and OH). A Mediterranean soil profile under an evergreen oak (Quercus ilex L.) forest was used as a model. The recalcitrant OM fraction, corresponding to the deepest layer, showed low enzyme activities. Peroxidases and fluorescein diacetate hydrolases (FDA) were more active in the OLn layer and probably originated largely from plants. High cellulase activity in the OLn and the OLv layers, which are rich in polysaccharides, corresponded with the high content of O-alkyl carbon compounds. Following polysaccharide degradation, laccases and lipases were much more evident in the intermediate layers. This spatial variation in nutrient demand reflected a preferential degradation of the specific plant polymers. Phosphatases were more active along the three upper layers and probably reflected a P limitation during litter degradation. Alkaline/acid (AcPAlP/AcP) ratio increased in the deepest layer, suggesting an increased participation of bacteria AlP in phosphatase pools. Results of Biolog^TM also indicated spatial variations in microbial functionality. Indeed, FF plates showed the highest functional diversity in the uppermost layer, while ECO plate functional diversity was highest in the intermediate layers. Finally, our results indicated that microbial activity and functional diversity of micro-organisms change with litter depth on a very small scale and vary with chemical organic matter (OM) composition. Thus, the observed increases in the biological variables studied were determined by the evolution of OM chemical structures, the nature and availability in C nutrients, and they ultimately resulted in a progressive accumulation of recalcitrant compounds.     

C and N natural abundance of the soil microbial biomass

Stable isotope analysis is a powerful tool in the study of soil organic matter formation. It is often observed that more decomposed soil organic matter is ¹³C, and especially ¹⁵N-enriched relative to fresh litter and recent organic matter. We investigated whether this shift in isotope composition relates to the isotope composition of the microbial biomass, an important source for soil organic matter. We developed a new approach to determine the natural abundance C and N isotope composition of the microbial biomass across a broad range of soil types, vegetation, and climates. We found consistently that the soil microbial biomass was ¹⁵N-enriched relative to the total (3.2 ‰) and extractable N pools (3.7 ‰), and ¹³C-enriched relative to the extractable C pool (2.5 ‰). The microbial biomass was also ¹³C-enriched relative to total C for soils that exhibited a C3-plant signature (1.6 ‰), but ¹³C-depleted for soils with a C4 signature (−1.1 ‰). The latter was probably associated with an increase of annual C3 forbs in C4 grasslands after an extreme drought. These findings are in agreement with the proposed contribution of microbial products to the stabilized soil organic matter and may help explain the shift in isotope composition during soil organic matter formation.     

Effect of continuous compost application on humus composition and nitrogen fertility of soils in a field subjected to double cropping

We investigated the effect of continuous compost application on humus composition and N fertility of soils in a field subjected to double cropping (paddy rice and barley) for 25 years. Soil samples were collected from three different plots: (a) No-NF, fertilizer containing P and K but no N; (b) F, fertilizer containing N, P, and K; and (c) F+C, fertilizer plus compost. The amounts of total humus, extracted humus, and humic and fulvic acids increased in the order No-NF<F≪F+C. The amounts of humic and fulvic acids were 2.7 and 1.7 times larger in the F+C plot than in the F plot, respectively. The degree of humification of the humic acids decreased in the order No-NF<F<F+C. The absorption curves and ¹³C-NMR spectra (TOSS method) of the humic acids indicated the presence of lignin-like structure, and its degree was the strongest in the F+C plot. The ¹³C-NMR spectra showed distinct differences in the distribution of carbon species between humic and fulvic acids. In humic acids, the content of aromatic-C, ranging from 37 to 44%, was the highest among carbon species. In fulvic acids, the content of O-alkyl-C, ranging from 45 to 51%, was the highest. The amounts of phosphate buffer-extractable N (PEON) and total N (TN) increased in the order No-NF<F<F+C. The amounts of PEON and TN were 1.2 and 1.7 times larger in the F+C plot than in the F plot, respectively. Present and previous findings indicated that continuous compost application could improve various properties of soils in a field subjected to long-term double cropping.     

Plant uptake of dual-labeled organic N biased by inorganic C uptake: Results of a triple labeling study

Direct plant uptake of organic nitrogen (N) is often studied using the dual-labeling approach (¹⁵N + ¹³C or ¹⁵N + ¹⁴C). However, the method might be hampered by uptake of labeled inorganic carbon (C) produced by mineralization of labeled organic compounds. Here we report the results from a triple labeling experiment (¹⁵N + ¹³C + ¹⁴C) investigating whether root uptake of labeled inorganic C can bias the results obtained in studies of organic N uptake using dual-labeled amino acids (¹⁵N, ¹³C). In a rhizosphere tube experiment we investigated ¹³C and ¹⁴C uptake by maize either supplied with labeled glycine or , but found no differences in uptake rates between these C-sources. The uptake of inorganic C to the shoot tissue was higher for maize grown in full light compared to shading, which indicates a passive uptake of inorganic C with water. We conclude that uptake of inorganic C produced by mineralization of amino acids can significantly bias the interpretations of organic N uptake studies using dual-labeling.     

Impact of top-pruning time on the fertilizer N use efficiency of flue-cured tobacco as assessed by 15N tracing technique

To evaluate the impact of top-pruning time on fertilizer N use efficiency (NUE) of flue-cured tobacco, we adopted ¹⁵N tracing technique and conducted a 2-year experiment from 2014 to 2015 in eastern China. The experiment included three top-pruning points of time: 5th, 25th and 45th day after flowering (DAF), abbreviated, respectively, as TP5, TP25 and TP45. The amounts of plant N derived from fertilizer (Ndff) and soil (Ndfs) were observed during 0–55th DAF. Results showed that top-pruning slowed down the increase of Ndff in tobacco organs, particularly in the leaf, but accelerated the increase of Ndfs dramatically. The proportion of Ndff (%) accounted for the total N reduced dramatically after top-pruning. This reduction might attribute to the selectivity of plant to different N sources as influenced by top-pruning while had little relationship with soil N supply, according to the analysis on the soil total mineral N and mineral ¹⁵N. The average NUE for the 2 years was 32%, 41% and 47%, respectively, for TP5, TP25 and TP45, showing significant (p < 0.05) differences. We concluded that the tobacco preferred to uptake soil N rather than fertilizer N after top-pruning; thus, optimizing the top-pruning time might be one of the approaches to improve the in-season NUE of flue-cured tobacco.