A liquid chromatographic/mass spectrometric method to evaluate <Superscript>13</Superscript>C and <Superscript>15</Superscript>N incorporation into soil amino acids

Purpose  

Amino acids are highly associated with biogeochemical cycling and represent an important potential source and sink of carbon (C) and nitrogen (N) in terrestrial ecosystems. Tracing the isotope dynamics of amino acids can improve the understanding of the origin and transformation of amino acids in soil matrix at process-levels; hence, the liquid chromatographic/mass spectrometric (LC/MS) method to evaluate ¹³C or ¹⁵N enrichment in amino acids is necessary to be established.     

Effect of long-term fertilization on bacterial composition in rice paddy soil

We investigated the effect of long-term fertilization on bacterial abundance, composition, and diversity in paddy soil. The experiment started in 1990 in Taoyuan Agro-ecosystem Research Station in China (111°33′ E, 28°55′ N). The molecular approaches including real-time quantitative PCR, terminal restriction fragment length polymorphism, and clone library construction were employed using 16S rRNA gene as genetic marker. Application of inorganic fertilizers did not affect bacterial abundance, and rice straw incorporation combined with inorganic fertilizers significantly (P < 0.05) increased bacterial abundance with shifts in bacterial community composition. Among phylogenetic groups, γ-Proteobacteria was responsive to all fertilization regimes while Acidobacteria was relatively stable to fertilization practices. Inorganic fertilizer mainly affected γ-Proteobacteria and δ-Proteobacteria, while rice straw incorporation influenced β-Proteobacteria and Verrucomicrobia. Therefore, long-term fertilization can affect abundance and composition of bacterial communities in paddy soil.     

Purification and isotopic signatures (<Emphasis Type="Italic">δ</Emphasis><Superscript>13</Superscript>C, <Emphasis Type="Italic">δ</Emphasis><Superscript>15</Superscript>N, Δ<Superscript>14</Superscript>C) of soil extracellular DNA

The aim of this work was to obtain pure extracellular DNA molecules so as to estimate their longevity in soil by an isotope-based approach. Extracellular DNA molecules were extracted from all horizons of a forest soil and purified by the procedure of Davis (Purification and precipitation of genomic DNA with phenol–chloroform and ethanol. In: Davis LG, Dibner MD, Battey JF (eds) Basic methods in molecular biology. Appleton & Lange, Norwalk, 16–22, 1986) without (DNA1) or with (DNA2) a successive treatment with binding resins followed by elution. The two differently purified DNA samples were compared for their A₂₆₀/A₂₈₀ ratio, polymerase chain reaction (PCR) amplification and natural abundance of stable (¹³C and ¹⁵N) and radioactive (¹⁴C) isotopes. The purity index and the PCR amplification did not differentiate the efficiency of the two purification procedures. The isotopic signature of DNA was more sensitive and was strongly affected by the purification procedures. The isotopic measurements showed that the major contaminant of extracellular DNA1 was the soil organic matter (SOM), even if it is not possible to exclude that the similar δ ¹³C, δ ¹⁵N and Δ¹⁴C values of DNA and SOM could be due to the use of SOM-deriving C and N atoms for the microbial synthesis of DNA. For extracellular DNA2, extremely low values of Δ¹⁴C were obtained, and this was ascribed to the presence of fossil fuel-derived substances used during the purification, although in amounts not revealed by gas chromatography-mass spectrometry analysis. The fact that it is not possible to obtain contaminant-free DNA molecules and the potential use of soil native organic compounds during the microbial synthesis of DNA make it not achievable to estimate the age of soil extracellular DNA by radiocarbon dating.     

Structure of humic acids in zonal soils from <Superscript>13</Superscript>C NMR data

The structure of humic acids (HAs) in zonal soil types—soddy-podzolic soils (two samples), gray forest soil (one sample), and chernozems (two samples)—was quantitatively studied by ¹³C NMR spectros-copy. In the series considered, the content of unsubstituted carbon in the aromatic fragments of HAs increased, and the fraction of unsubstituted aliphatic structures decreased. HAs of soddy-podzolic soils were found to be enriched with carbohydrate fragments compared to HAs of chernozems and gray forest soil. The carbon skeleton of HAs from typical rich chernozem contained significantly more aliphatic and carbohydrate fragments compared to typical chernozem, which probably reflected the lower degree of HA transformation in rich chernozem.     

Soil nitrogen mineralization and fate of (<Superscript>15</Superscript>NH<Subscript>4</Subscript>)<Subscript>2</Subscript>SO<Subscript>4</Subscript> in field-incubated soil in a hardwood plantation of subtropical Australia: the effect of mulching

Background, aim, and scope  

Mulching is frequently used to overcome the drought problem in hardwood plantations that are increasingly being established in lower rainfall areas of Queensland, Australia because of increasing land values. In addition to soil water content, soil nitrogen (N) availability is another critical determinant of plantation productivity in these areas. The purpose of this study was to understand how soil mineral N dynamics, in situ N mineralization, and the fate of fertilized N would be affected by mulching during the early establishment of hardwood plantations.     

In situ mineral <Superscript>15</Superscript>N dynamics and fate of added <Superscript>15</Superscript>NH<Subscript>4</Subscript><Superscript>+</Superscript> in hoop pine plantation and adjacent native forest in subtropical Australia

Background, aim, and scope  Hoop pine (Araucaria cunninghamii) is a nitrogen (N) demanding indigenous Australia softwood species with plantations in Southeast Queensland, Australia. Soil fertility has declined with increasing rotations and comparison study of N cycling between hoop pine plantations, and adjacent native forest (NF) is required to develop effective forest management for enhancing sustainable forest production and promoting environmental benefits. Field in situ mineral ¹⁵N transformations in these two forest ecosystems have not been studied. Hence, the present study was to compare the differences in soil nutrients, N transformations, ¹⁵N fluxes, and fate between the hoop pine plantation and the adjacent native forest. Materials and methods  The study sites were in Yarraman State Forest (26°52′ S, 151°51′ E), Southeastern Queensland, Australia. The in situ core incubation method was used in the field experiments. Mineral N was determined using a LACHAT Quickchem Automated Ion Analyzer. ¹⁵N were performed using an isotope ratio mass spectrometer with a Eurovector elemental analyzer. All statistical tests were carried out by the SPSS 11.0 for Windows statistical software package. Results  Soil total C and N were significantly higher in the NF than in the 53-year-old hoop pine plantation. Concentrations of NO₃ ^– were significantly higher in the NF soil than in the plantation soil. The plantation soil had significantly higher ¹⁵N and ¹³C natural abundances than the NF soil. The NF soil had significantly lower C/N ratios than the plantation soil. NO₃ ^––N was dominated in mineral N pools in both NF and plantation soils, accounting for 91.6% and 70.3% of the total mineral N pools, respectively. Rates of net nitrification and net N mineralization were, respectively, four and three times higher in the NF soil than in the plantation soil. The ¹⁵NO₃ ^––N and mineral ¹⁵N were significantly higher in the NF soil than in the plantation soil. Significant difference in ¹⁵NH₄ ⁺–N was found in the NF soil before and after the incubation. Discussion  The NF soil had significantly higher NO₃ ^––N, mineral N, total N and C but lower δ¹⁵N, δ¹³C, and C/N ratios than the plantation soil. Moreover, the rates of soil net N mineralization and nitrification were significantly higher, but ammonification rate was lower in the NF than in the plantation. The NF soil had many more dynamic N transformations than the plantation soil due to the combination of multiple species and layers and, thus, stimulation of microbial activity and alteration of C and N pool sizes in favor of the N transformations by soil microbes. The net rate of N and ¹⁵N transformation demonstrated differences in N dynamic related to the variation in tree species between the two ecosystems. Conclusions  The change of land use and trees species had significant impacts on soil nutrients and N cycling processes. The plantation had larger losses of N than the NF. The NO₃ ^––N and ¹⁵NO₃ ^––N dominated in the mineral N and ¹⁵N pools in both forest ecosystems. Recommendations and perspectives  Native forest soil had strong N dynamic compared with the plantation soil. Composition of multiple tree species with different ecological niches in the plantation could promote the soil ecosystem sustainability. The ¹⁵N isotope dilution technique in the field can be quite useful for studying in situ mineral ¹⁵N transformations and fate to further understand actual N dynamics in natural forest soils.     

Selective effects of forest fires on the structural domains of soil humic acids as shown by dipolar dephasing <Superscript>13</Superscript>C NMR and graphical-statistical analysis of pyrolysis compounds

Purpose

Data management strategies of pyrolysis results and NMR acquisition modes were examined in humic acids (HAs) from control soils and fire-affected soils. The information supplied by dipolar dephasing (DD) ¹³C NMR spectroscopy and Curie-point pyrolysis were used to assess chemical structures hardly recognizable and measurable, or of unclear interpretation, when using ¹³C NMR under standard acquisition pulses (cross-polarization/magic angle spinning, CPMAS).

Materials and methods

The HAs were isolated from two forest soils under Pinus halepensis and Pinus sylvestris in control and burned sites affected by medium or severe-intensity wildfires. For NMR analyses, during DD acquisition conditions, a 180° ¹³C pulse was inserted to minimize phase shifts. Curie-Point pyrolysis was carried out at 510 °C for 5 s, and the pyrolysis fragments were analyzed by GC/MS. The total abundances of the major pyrolysis products were compared by an update of the classical Van Krevelen’s graphical-statistical approach, i.e., as surface density values in the space defined by the compound-specific H/C and O/C atomic ratios.

Results and discussion

The DD ¹³C NMR experiments displayed significant differences in the HA spectral profiles as regards to the standard CPMAS ¹³C NMR acquisition conditions, mainly in the chemical shift region of alkyl structures as well as for tannin- or carbohydrate-like O-alkyl structures. In fact, the comparison between DD and CPMAS solid-state NMR suggested shortening of alkyl chains and generation of carbohydrate-derived, unsaturated structures—viz. furans—which adds to the aromatic domain. Pyrolytic results showed fire-induced specific changes in HAs chemical structure and its molecular diversity. The changes were evident in the location and sizes of the different clusters of pyrolysis compounds defined by their atomic ratios.

Conclusions

The DD ¹³C NMR provided specific information on the fate of aliphatic structures and the origin of unsaturated HA structures, which could be helpful in differentiating “inherited” from “pyrogenic” aromatic structures. This is further confirmed by the analysis of the molecular assemblages of pyrolytic products, which showed accumulation of condensed polyaromatic domains in the HAs after the high-intensity fire, accompanied by a recalcitrant alkyl hydrocarbon domain. Medium-intensity fire led to aromaticity increase due to a selective accumulation of lignin-derived phenols concomitant to the depletion of aliphatic hydrocarbon constituents.

     

不同施肥管理对红壤性水稻土有机碳、氮形态的影响

A long-term experiment beginning in 1981 in Jinxian County of Jiangxi Province, subtropical China, was conducted in a paddy field under a double rice cropping system with four different fertilization regimes, including 1) no fertilizer as control (CK), 2) balanced chemical N, P, and K fertilizers (NPK), 3) organic manure using milk vetch and pig manure in the early and late rice growing season, respectively (OM), and 4) balanced chemical fertilizers combined with organic manure (NPKM). Samples (0-17 cm) of the paddy field soil, which was derived from Quaternary red clay, were collected after the late rice harvest in November 2003 for determination of total organic carbon (TOC) and total nitrogen (TN) and fractions of organic C and N. Results showed that TOC and TN in the NPKM and OM treatments were significantly higher than those in other two treatments (CK and NPK). Application of organic manure with or without chemical fertilizers significantly increased the contents of all fractions of organic C and N, whereas chemical fertilizer application only increased the contents of occluded particulate organic C (oPOC) and amino acid N. In addition, application of organic manure significantly enhanced the proportions of free particulate organic carbon (fPOC) and oPOC in total C, and those of amino sugar N and amino acid N (P < 0.01) in total N. In contrast, chemical fertilizer application only increased the proportions of oPOC and amino acid N (P < 0.05). There were no significant differences in either contents or proportions of soil organic C and organic N fractions between the NPKM and OM treatments. These indicated that organic manure application with or without chemical fertilizers played the most significant role in enhancing soil organic C and N quantity and quality in the paddy field studied.     

Application of <Superscript>13</Superscript>C NMR Spectroscopy to the Study of Soil Organic Matter: A Review of Publications

Possibilities of NMR spectroscopy with ¹³C nuclei application to the study of soil organic matter and its various fractions is considered. This is a non-destructive method, which is particularly valuable in the analysis of various fractions of soil organic matter. It is regarded as a direct method, and, unlike most of indirect methods, it allows one to obtain reliable estimates of the ratio between virtually all groups of carbon atoms in different organic molecules, including those in humus specimens. Owing to impulse technique and high sensitivity, ¹³C-NMR spectra may be obtained immediately from soil samples without any extraction operations. The modern technique of obtaining spectra, their mathematical processing (Fourier transform), and data interpretation are considered. The results of applying ¹³C-NMR to the study of humus substances, water-soluble fractions of soil organic matter, and soil litters from different natural zones are discussed.     

不同施肥制度对稻田土壤微生物的影响研究

湖南省进行了18年土壤肥力与施肥效益监测的长期定位试验,就3个定位点不同施肥制度对稻田土壤微生物的影响进行了研究。结果表明,长期施肥土壤的有机质含量与细菌总数及放线菌总数之间无直接相关,土壤有机质含量的高低并不能作为评价土壤微生物多少的指标;真菌数量可反映土壤的肥力水平,土壤真菌数量可用来评价不同施肥制度的效果。施用30%有机肥和习惯施肥处理的氮素养分和硝化细菌数量较多,硝化细菌数量与有机质含量也无相关;长期施用大量有机肥(60%)的土壤反硝化细菌数量高于其它施肥处理,这可能与长期使用大量有机肥后土壤氧化还原电位降低有关。硫化细菌和反硫化细菌数量与土壤有机质含量无直接相关性,施肥可影响土壤硫素的供应。施用单质氮肥多的土壤,氨化细菌数量较多,而有机肥施用量高的土壤,氨化细菌数量较少。无肥区的土壤微生物活度最低,60%有机肥处理的土壤微生物活度最高。土壤有机质含量高,土壤微生物活度也高,两者呈正相关。保持农业土壤的健康要从多方面来进行联合调控,而不是单纯的采用某一农业技术来进行。     

Use of a double radioactive label (<Superscript>54</Superscript>Mn and <Superscript>14</Superscript>C) in the study of organomineral manganese compounds

A fundamentally new approach to the study of natural organomineral materials was proposed. A procedure was developed for using a double carbon-metal label in the systemic study of organomineral complexes of soils and conjugated landscape objects. A significant effect of water-soluble organic ligands on the migration of manganese in soils was shown. It was found that mineral manganese compounds were transformed into organomineral ones differing in composition, solubility, and stability, and the complex of humus substances in podzolic soil was a peculiar matrix on which a complex system of organomineral compounds was developed. The input mechanism of organomineral complexes with different molecular weights (MWs) into plants was studied. Pot experiments using a root exudates sampling procedure and a double radioactive label (⁵⁴Mn and ¹⁴C) showed that the organomineral complexes of Mn with the fulvic acid fraction (MW > 10000) came intact to corn roots. The fulvic acid fraction (MW 380) and manganese ions independently passed from the soil solution into the young plants.     

Short-term uptake of <Superscript>15</Superscript>NH<Subscript>4</Subscript><Superscript>+</Superscript> into soil microbes and seedlings of pine,spruce and birch in potted soils

Short-term competition between soil microbes and seedlings of Scots pine (Pinus sylvestris L.), Norway spruce (Picea abies (L.) Karst.) and silver birch (Betula pendula Roth) for N was assessed in a pot study using (¹⁵NH₄)₂SO₄ as a tracer. Seedlings were grown in organic and mineral soil, collected from a podsol soil; 3.18 mg (¹⁵NH₄)₂SO₄ per pot were injected into the soil, corresponding to 4 µg ¹⁵N g^-1 d.m. (dry matter) mineral soil and 17 µg ¹⁵N g^-1 d.m. organic soil. The amounts of N and ¹⁵N in the seedlings and in microbial biomass derived from fumigation-extraction were measured 48 h after addition of ¹⁵N. In the mineral soil, 19–30% of the added ¹⁵N was found in the plants and 14–20% in the microbial biomass. There were no statistically significant differences between the tree species. In the organic soil, 74% of the added ¹⁵N was recovered in the microbial biomass in birch soil, compared to 26% and 17% in pine and spruce soils, respectively. Correspondingly, about 70% of the ¹⁵N was recovered in pine and spruce seedlings, and only 23% in birch seedlings. In conclusion, plants generally competed more successfully for added ¹⁵NH₄ ⁺ than soil microbes did. An exception was birch growing in organic soil, where the greater amount of available C from birch root exudates perhaps enabled micro-organisms to utilise more N.     

长期不同施肥对红壤性水稻土产量及基础地力的影响

利用长期定位施肥试验并结合盆栽试验,研究长期不同施肥模式对双季稻产量和土壤基础地力的影响,并分析水稻产量和肥料贡献率对土壤不同基础地力的响应。结果表明:长期施用氮磷钾肥(NPK)或氮磷钾肥配施稻草(NPKS)有利于双季稻产量的增加。NPK处理的早稻产量、晚稻产量和年总产量分别较对照(CK)处理增产100.7%、67.0%和81.9%,NPKS处理分别增产113.8%、77.7%和93.7%。CK处理早稻基础地力产量随试验年限的增加呈极显著下降趋势(p0.01),晚稻基础地力产量在试验的前9年随年限增加呈极显著下降(p0.01),之后基本维持稳定。长期施用氮磷钾肥或氮磷钾肥配施稻草有利于土壤基础地力的提升。土壤基础地力产量和基础地力贡献率均表现为NPKSNPKCK。NPK和NPKS处理早稻基础地力产量分别较CK提高38.5%和68.1%,晚稻分别提高25.8%和49.0%。NPK和NPKS处理早稻基础地力贡献率分别较CK提高21.4%和54.9%,晚稻分别提高12.8%和22.8%。无论施肥或不施肥,早晚稻产量均随土壤基础地力产量提高而增加;肥料对早晚稻产量贡献率随基础地力产量提高而极显著降低。土壤有机质、全氮、速效钾是影响土壤基础地力的主要养分因子,土壤全磷、全钾、碱解氮、有效磷对土壤基础地力也产生重要影响。     

Concentrations of Radionuclides (<Superscript>226</Superscript>Ra, <Superscript>232</Superscript>Th, <Superscript>40</Superscript>K,and <Superscript>137</Superscript>Cs) in Chernozems of Volgograd Oblast Sampled in Different Years

Data on the concentrations of natural (²²⁶Ra, ²³²Th and ⁴⁰K) and artificial (¹³⁷Cs) radionuclides and on the physicochemical properties of chernozems sampled in different years are presented. In 1952, upon the creation of the Penza-Kamensk state shelterbelt, three deep (up to 3 m) soil pits were examined within the former arable field under two-year-old plantations of ash and maple along the transect crossing the territory of the Beloprudskaya Experimental Station of the USSR Academy of Sciences in Volgograd oblast. The samples from these pits were included into the collection of dated soil samples of the Dokuchaev Central Soil Science Museum. Five pits were examined along the same transect in 2009: three pits under shelterbelts (analogues of the pits studied in 1952) and two pits on arable fields between the shelterbelts. In the past 57 years, certain changes took place in the soil structure, bulk density, and the content and composition of humus. The salt profile of soils changed significantly under the forests. The comparison of distribution patterns of natural soil radionuclides in 1952 and 2009 demonstrated their higher contents at the depth of 10–20 cm in 2009 (except for the western shelterbelt). Background concentrations of natural radionuclides in parent materials and relationships between their distributions and the salt profiles of soils have been determined; they are most clearly observed is the soils under shelterbelts. Insignificant contamination with ¹³⁷Cs (up to 34 Bq/kg) has been found in the samples of 2009 from the upper (0–20 cm) horizon. The activity of ¹³⁷Cs regularly decreases from the east to the west; the highest concentrations of this radionuclide are found in the topmost 10 cm. This allows us to suppose that ¹³⁷Cs was brought with aerial dust by eastern winds, and the shelterbelts served as barriers to the wind flow.     

Radioactive Contamination of Alluvial Soils in the Taiga Landscapes of Yakutia with <Superscript>137</Superscript>Cs, <Superscript>226</Superscript>Ra,and <Superscript>238</Superscript>U

The concentrations and distribution of ¹³⁷Cs in alluvial soils (Fluvisols) of the upper and middle reaches of the Markha River in the northwest of Yakutia and ²²⁶Ra and ²³⁸U in alluvial soils within the El’kon uranium ore deposit in the south of Yakutia have been studied. It is shown that the migration of radiocesium in the permafrost-affected soils of Yakutia owing to alluviation processes extends to more than 600 km from the source of the radioactive contamination. The migration of ¹³⁷Cs with water flows is accompanied by its deposition in the buried horizons of alluvial soils during extremely high floods caused by ice jams. In the technogenic landscapes of southern Yakutia, active water migration of ²³⁸U and ²²⁶Ra from radioactive dump rocks. The leaching of ²³⁸U with surface waters from the rocks is more intense than the leaching of ²²⁶Ra. The vertical distribution patterns of ²³⁸U and ²²⁶Ra in the profiles of alluvial soils are complex. Uranium tends to accumulate in the surface humus horizon and in the buried soil horizons, whereas radium does not display any definite regularities of its distribution in the soil profiles. At present, the migration of ²³⁸U and ²²⁶Ra with river water and their accumulation in the alluvial soils extend to about 30 km from the source.     

Influence of aggregate size on phosphorus changes in a soil cultivated intermittently: analysis by <Superscript>31</Superscript>P nuclear magnetic resonance

A pot trial using wet-sieved soil aggregates (>4, 4–2, 2–1, 1–0.5, 0.5–25, and remaining soil <0.25 mm) from a soil that had been cultivated out of permanent pasture and used for winter forage crops for 2 years examined changes in P forms before and after 35 weeks when resown with perennial ryegrass. Soil analyses showed that P was depleted after 35 weeks growth. Changes in P forms were analyzed by ³¹P nuclear magnetic resonance of soil NaOH-EDTA extracts, which removed 98–96% of total P (about 1,080 mg kg⁻¹ in unsieved soil before pasture growth). This indicated that aggregate size influenced the concentrations and forms of P probably via a combination of physical protection and moisture status: orthophosphate, monoesters, diesters and pyrophosphate increased with decreasing size, while phosphonates and polyphosphates were unaffected. The increase in pyrophosphate was attributed to fungal growth, while decreases in orthophosphate and labile organic P (diesters) decreased due to either leaching or mineralisation and plant uptake. The largest decrease was associated with orthophosphate, which could be replenished by fertiliser. However, given the soil’s high potential for P loss, this should only be done to meet conditions for optimal plant growth as any excess would increase the risk of loss. To further minimise P loss without affecting pasture yield, management should maintain or improve soil structure.     

Role of plants in the spatial differentiation of <Superscript>137</Superscript>Cs and <Superscript>90</Superscript>Sr statuses on the aggregate level

The increased concentration of an element in plant biomass compared to the soil mass is an essential condition for the differentiated spatial distribution and status of the element on the aggregate level. Two forms of this differentiation have been revealed for ¹³⁷Cs and ⁹⁰Sr. Transfer of ¹³⁷Cs from plant roots and concentration on the surface of soil aggregates have been established experimentally. Indirect data also point to the potential localization of ¹³⁷Cs on the surface of intraaggregate pores. The effect of radionuclide concentrating on the outer and inner surfaces of aggregates is due to the rapid and strong fixation of cesium microamounts by mineral soil components. ¹³⁷Cs from the surface of aggregates is more available for the repeated uptake by plant roots than from the intraped mass. The distortion of this spatial differentiation mainly occurs during the reaggregation of soil mass, which in turn decreases the availability of the radionuclide to plants. For ⁹⁰Sr, its elevated concentration in the form of organic residues has been revealed in the inter- and intraaggregate pore space. However, due to the high diffusion rate, ⁹⁰Sr is relatively rapidly (during several months under pot experimental conditions) redistributed throughout the entire volume of soil aggregates and its major part gradually passes into the phase of humic compounds, to which the radionuclide is bound by exchange sorption. The high level of the next root uptake (higher than for ¹³⁷Cs by one to two orders of magnitude) favors the permanent renewal of loci with increased ⁹⁰Sr concentrations in the inter- and intraaggregate pore space in the form of plant residues.     

Effect of chronic nitrogen fertilization on soil CO<Subscript>2</Subscript> flux in a temperate forest in North China: a 5-year nitrogen addition experiment

Purpose

Mounting evidence has indicated that there was dramatic increase in atmospheric nitrogen (N) deposition. The objectives of this study were to characterize how soil carbon dioxide (CO₂) flux responds to different forms and levels of N addition in 5 years. We hope to provide further understanding and detailed information of the impact of N addition on CO₂ flux in temperate forests in North China.

Materials and methods

A 5-year field experiment was conducted at the Xi Mountain Research Station of Beijing Forestry University, northern China, from 2011 to 2016. Multiple levels and forms of N addition experiment included control with no N added, NH₄NO₃, NaNO₃, and (NH₄)₂SO₄ at two levels (low N (L) 50 kg N ha^?1 year^?1 and high N (H) 150 kg N ha^?1 year^?1). Additional N was administered equally once per month during the growing season (March to October), and CO₂ flux was measured three times every month. Soil temperature, water-filled pore space, and NO₃ ^?-N and NH₄ ⁺-N concentrations were measured monthly to determine the relationships between CO₂ flux and soil physicochemical variables.

Results and discussion

Cumulative CO₂ flux increased by more than 50% under all high-level N addition and by 27% under L-NH₄NO₃ addition, while other N additions had no significant effect. H-NaNO₃ and H-NH₄NO₃ exerted stronger effects on cumulative CO₂ flux in initial years, especially the second year when maximum increases were 99 and 129%, respectively. Increasing inorganic N concentration could change soil from N-limited to N-rich, and then N-saturated, and so the promotion increased and then decreased. The effect of high-level N addition was stronger than that of low-level, and exhibited a general order: NH₄NO₃ > (NH₄)₂SO₄. Considering the amount and decrease in NH₄ ⁺-N/NO₃ ^?-N in local actual N deposition, there might be an increase in soil CO₂ flux in our study area in the future.

Conclusions

The performance of N addition on cumulative CO₂ flux depended on N forms and levels. If the experimental period had been less than 3 years, the effect of N addition on temperate forest soil would be overestimated. Our findings highlighted the importance of experimental time and multiple forms and levels of N addition with regard to the response of soil CO₂ flux to N deposition.

     

长期不同施肥条件下红壤性水稻土双季稻氮肥回收率的变化特征

在江西进贤红壤性水稻土上连续种植双季水稻26年,分析了在化肥氮用量相同条件下,不施肥（CK）, 单施氮肥（N）, 施用氮、 磷化肥（NP）, 氮、 钾化肥（NK）, 氮、 磷、 钾化肥（NPK）和氮磷钾配施有机肥（NPKM）处理的水稻氮肥回收率的演变特征及其增产效应。结果表明,不同施肥条件下的化肥氮回收率差异显著,26年的平均氮肥回收率 N 处理为 9.4%~11.6%、 NP为13.0%~18.5%、 NPK为19.8%~26.1%, NPKM为14.1%~22.9%。磷、 钾肥混施和与有机肥配施可显著提高水稻氮肥回收率,且对早稻的贡献大于晚稻,NPK和NPKM处理的早稻氮肥回收率比晚稻平均高6.3和8.8个百分点。N和NK处理的早稻和晚稻氮肥回收率均随年度增加而显著降低,平均每年下降约0.6个百分点,而NPK、 NPKM处理的氮肥回收率基本保持稳定。与NPK相比,NPKM处理早稻和晚稻籽粒产量分别增加19.0%和21.7%。因此,NPKM处理在提高化肥氮的回收率和高产稳产方面都是红壤性水稻土上可持续的施肥模式。