Automated instrumental analysis of carbon and nitrogen in plant and soil samples

Abstract

An automated CHN Analyzer was compared with the Walkley‐Black and Kjeldahl methods for organic carbon (C) and nitrogen (N). Four organic compounds, twenty nine plant materials and five soils were tested. The CHN Analyzer gave C and N values that were not significantly different (P<0.05) to the theoretical weight percents of the organic compounds. The Walkley Black method gave soil C values significantly lower (P<0.05) than those obtained with the CHN Analyzer. The Kjeldahl method gave soil N values significantly lower (P<0.05) than the CHN Analyzer on three of five soils tested. The discrepancies observed between methods appear to be due to different oxidation efficiencies. CHN Analyzer and Kjeldahl N analyses were not significantly different (P<0.05) for the plant materials except where samples contained greater than 0.7% NO3‐N. Potassium nitrate was also added as a spike to a tall fescue sample. Based on recovery of the spiked NO3‐N, the Kjeldahl method was a poor measure of total N for plant materials containing greater than 0.7% NO₃‐N. The findings suggest the CHN Analyzer can be used for the rapid, accurate and simultaneous determination of C and N in plant and soil samples.     

A rapid accurate wet oxidation diffusion procedure for determining organic and inorganic carbon in plant and soil samples

Abstract

A method is presented for determining organic carbon content of soils, extracts and plant tissue utilizing an acidic wet oxidation in modified culture tubes; samples containing carbonates are pretreated with dilute acid. Evolved CO₂ is trapped in a NaOH solution and quantified titrimetrically. The method described has a range of 0.4–11 mg. carbon and the range can be extended in either direction by modifying the CO₂ trapping/titration procedure. This method is as accurate and precise as more time consuming dry oxidation procedures.     

A rapid microwave digestion method for colorimetric measurement of soil organic carbon

Abstract

Soil organic matter is an active component of agroecosystems. Rapid and precise measurement of organic carbon (C_org) is essential to monitor changes in organic matter and soil quality. A new semi‐micro wet digestion method for the determination of C_org was developed using rapid microwave energy applied at 500 J mL^‐1 digestion mixture to enhance oxidation of C_org by K₂Cr₂O₇and conc. H₂SO₄. This proposed method and three existing methods of soil C determination were compared with the LECO dry combustion carbon analyzer. The r² value for the proposed microwave method regressed against LECO C was 0.9913. The recovery of C_org by the rapid microwave digestion method for spectrophotometric measurement was 91.7±1.2% (conversion factor 1.09) of C measured by the LECO dry combustion method. Compared to the traditional Walkley‐Black's method, the proposed spectrophotometric with microwave digestion method was rapid and more precise, used smaller reagent volumes, and produced less waste.     

氯磺隆污染对土壤生物量碳氮与总有机碳氮比值的影响

The extent and seriousness of the contamination of soils by pesticides still remain to be determined,In agricultural systems,herbicides are the largest class of pesticides used (Shea,1985),Chlorsulfuron is one of the most important sulfonylurea herbicides with important feature of very high herbicidal activity,which results in extremely low application rates of 10-40g ha^-1(Blair and Martin,1988),The sulfonylurea herbicides can persist in the soil for more than 1 year(Brown,1990),Therefore,there has been considerable interest in the side effects of these chemicals on non-target organisms,including soil microorganisms(Greaves and Malkomes,1980),It ios generally recognized that the microbial biomass is the eye of the needle through which all organic materials that enter the soil must pass(Jenkinson,1988), Changes in the microbial biomass-C(Cmic) can provide an early indication of long-term trends in the total organic-C(Corg) of soils(Carter,1986),The Cmic/Corg ratio has been found useful as an index of changes in soil organic matter resulting from land management changes(Hart et al.1989).Many studies were done on relative effect of heavy metals on the ratio of Cmic/Corg but few studies have laid particular attention to the effect of herbicides on this ratio,The present paper reports on the Cmic/Corg and microbial biomass-N/total N (Nmic/Ntotal)ratios in soil as affected by chlorsulfuron.     

A theoretical analysis of nonlinearities and feedbacks in soil carbon and nitrogen cycles

Analytical formulations of soil carbon and nitrogen cycles are used to explore the effects of model nonlinearities and feedbacks on the resulting dynamics. Two particular aspects are addressed: (i) nonlinearities in the decomposition rate of soil organic matter and (ii) nitrogen limitation feedbacks on microbial activity and plant-microbe competition. Although linear models of decomposition are more typical in the literature, nonlinear models accounting for the coupling between microbial biomass and its substrate have also been proposed. In deterministic conditions, linear models behave like exponential decay functions, while nonlinear models may also show fluctuating behavior, with dynamic bifurcations between stable-node and stable-focus equilibria as a function of the climatic parameters (e.g., soil moisture and temperature). Both data-model comparison and linear stability analysis support the conclusion that linear models are less suited to describe the soil fluctuating dynamics that arise under certain conditions. A second strong nonlinearity appears when the nitrogen-limitation feedback on decomposition is analyzed. Nitrogen limitation is often established when the substrate of the microbes is N-poor, and/or the competition with plants for mineral N is strong. Such conditions mainly occur when a large fraction of the microbial community cannot meet its nitrogen demand through organic N assimilation, so that mineral N is used. On the contrary, when the microbial community predominantly assimilates organic nitrogen, the occurrence of nitrogen-limitation is less likely and mineralization is given by microbial N surplus. The first case is traditionally modeled by the mineralization-immobilization turnover (MIT) scheme, while the second by the direct assimilation (DIR) scheme. However, since organic N assimilation and mineral N immobilization likely occur simultaneously because of soil heterogeneity and coexistence of different microbial communities, the two schemes only represent extreme cases. Thus, we combine them in a flexible model framework (parallel scheme) and explore how different efficiencies of organic nitrogen assimilation, mineral nitrogen availability and climatic factors control the outcome of plant-microbe competition. We conclude that models accounting only for the DIR pathway implicitly assume that microbes are superior competitors over plants, while models implementing only the MIT pathway might be too sensitive to N-limitation.     

Effect of the storage conditions of soil samples on carbon and nitrogen extractability

Concentrations of carbon and nitrogen extractable by 0.05 M K₂SO₄ (C_ext and N_ext, respectively) in 14 soils of different ecosystems vary from 16 to 205 and from 4 to 53 mg/kg, respectively. The portion of C_ext in soil organic matter is 0.06 to 0.38% of total carbon, and the portion of N_ext is 0.12–1.05% of total nitrogen. The storage of samples and their preparation to analysis differently affect the extractability of elements. The concentration of C_ext is less variable than the concentration of N_ext. An increase in C extractability (by 1.4–6.7 times) is a common feature of all soils under drying; at the following incubation of dried soils, the extractability of C decreases by 28–56%. The extractability of N increases not only under drying (by 1.5–7.1 times) and the following incubation of samples (by 25–60% to 2–3 times), but also under freezing of most soils and at the incubation of fresh and defrozen samples. A close direct correlation is observed between the initial water content of soil and the relative increase in C extractability under drying and N extractability under freezing and drying.     

A rapid method for nitrogen determination in plant tissue

Abstract

A method is described for determining the total N content of plant tissue by a modified Kjeldahl digestion employing a Technicon Auto‐Analyzer. It is faster than the manual AOAC Kjeldahl method. Reproducibility is not as good as that for the AOAC method, but coefficients of variability are comparable with those for the analyses of mineral elements in plant tissue by emission spectroscopy.     

A rapid chloroform-fumigation extraction method for measuring soil microbial biomass carbon and nitrogen in flooded rice soils

 A chloroform-fumigation extraction method with fumigation at atmospheric pressure (CFAP, without vacuum) was developed for measuring microbial biomass C (C_BIO) and N (N_BIO) in water-saturated rice soils. The method was tested in a series of laboratory experiments and compared with the standard chloroform-fumigation extraction (CFE, with vacuum). For both methods, there was little interference from living rice roots or changing soil water content (0.44–0.55 kg kg^–1 wet soil). A comparison of the two techniques showed a highly significant correlation for both C_BIO and N_BIO (P<0.001) suggesting that the simple and rapid CFAP is a reliable alternative to the CFE. It appeared, however, that a small and relatively constant fraction of well-protected microbial biomass may only be lysed during fumigation under vacuum. Determinations of microbial C and N were highly reproducible for both methods, but neither fumigation technique generated N_BIO values which were positively correlated with C_BIO. The range of observed microbial C:N ratios of 4–15 was unexpectedly wide for anaerobic soil conditions. Evidence that this was related to inconsistencies in the release, degradation, and extractability of N_BIO rather than C_BIO came from the observation that increasing the fumigation time from 4 h to 48 h significantly increased N_BIO but not C_BIO. The release pattern of C_BIO indicated that the standard fumigation time of 24 h is applicable to water-saturated rice soils. To correct for the incomplete recovery of C_BIO, we suggest applying the k _C factor of 2.64, commonly used for aerobic soils (Vance et al. 1987), but caution is required when correcting N_BIO data. Until differences in fumigation efficiencies among CFE and CFAP are confirmed for a wider range of rice soils, we suggest applying the same correction factor for both methods. Received: 1 June 1999     

耕作与轮作方式对黑土有机碳和全氮储量的影响

土壤有机碳(SOC)及全氮(TN)对土壤肥力、作物产量、农业可持续发展以及全球碳、氮循环等都具有重要影响。为探索不同耕作和轮作方式对耕层黑土SOC和TN储量的影响,本文以吉林省德惠市进行了8 a的田间定位试验中层黑土为研究对象,对免耕、垄作和秋翻三种耕作方式及玉米-大豆轮作和玉米连作两种轮作方式下SOC和TN在各土层的含量变化进行了分析,并采用等质量土壤有机质储量计算方法,对比分析了不同处理对0～30 cm SOC和TN储量的影响。结果表明,与试验开始前相比,玉米-大豆轮作系统中,秋翻下SOC和TN储量均有所降低;免耕显著增加了0～5 cm SOC及TN含量,但SOC在亚表层亏损,导致其储量并未增加;而垄作处理下SOC及TN含量在0～5、5～10 cm的均显著增加,0～30 cm储量亦分别增加了4.9%和10.7%。玉米连作系统的两种耕作处理(免耕和秋翻)下SOC和TN储量均有所增加,且TN储量增幅均高于玉米-大豆轮作系统,其中免耕下TN储量增幅是玉米-大豆轮作的3.2倍。所有处理下C/N均呈降低趋势,其中垄作0～5 cm C/N由12.05降至11.04,降低幅度分别是免耕和秋翻的3.2和2.8倍。综上可知,对质地黏重排水不良的中层黑土,玉米-大豆轮作系统下免耕并不是促进SOC固定的有效形式,而垄作则促进了黑土SOC和TN的积累,这不仅有利于土壤肥力的改善,而且是使农田黑土由CO2"源"变为"汇"的有效形式之一。与玉米-大豆轮作相比,玉米连作下三种耕作方式都有利于SOC和TN积累。     

长期不同管理措施下红壤剖面碳、氮储量变化特征

基于中国农业科学院国家红壤肥力长期试验平台,研究长期不同施肥、休闲和轮作措施下旱地红壤剖面(0~100 cm)土体及不同层次(20 cm/层)有机碳(SOC)和全氮(TN)储量的变化特征。结果表明,与试验初始相比,24年不施肥(CK处理)0~100 cm各层有机碳储量均有不同程度降低,但全氮储量在40~100 cm各土层表现出累积趋势。24年施化肥(N、NP、PK、NPK)和化肥配施秸秆处理(NPKS)明显增加40~100 cm有机碳和全氮储量,0~40 cm各层碳储量有所耗竭而氮储量无明显影响。24年有机无机配施(NPKM)明显增加表层0~20和40~100 cm的有机碳和全氮储量。休闲(Fallow)和轮作(NPKMR)措施大幅度提高0~100 cm各层有机碳和全氮储量,但两处理间无显著差异。对于0~100 cm整个土体而言,休闲和轮作处理大幅度增加了土壤剖面的有机碳和全氮储量,其中有机碳储量比初始分别提高64%和93%,全氮储量比初始分别提高71%和82%。不同措施下0~100 cm各层有机碳与全氮储量及其变化量间均存在显著线性相关关系(P0.01),但不同管理措施显著改变了碳氮储量间的相关关系,但储量变化量间的相互关系无统计学上的显著差异。休闲或豆科轮作措施能显著提升红壤剖面碳、氮库容,是红壤肥力恢复和培肥较优管理措施。     

Effect of added nitrogen on plant litter decomposition depends on initial soil carbon and nitrogen stoichiometry

Increasing organic carbon inputs to agricultural soils through the use of pastures or crop residues has been suggested as a means of restoring soil organic carbon lost via anthropogenic activities, such as land use change. However, the decomposition and retention of different plant residues in soil, and how these processes are affected by soil properties and nitrogen fertiliser application, is not fully understood. We evaluated the rate and extent of decomposition of ¹³C-pulse labelled plant material in response to nitrogen addition in four pasture soils of varying physico-chemical characteristics. Microbial respiration of buffel grass (Cenchrus ciliaris L.), wheat (Triticum aestivum L.) and lucerne (Medicago sativa L.) residues was monitored over 365-days. A double exponential model fitted to the data suggested that microbial respiration occurred as an early rapid and a late slow stage. A weighted three-compartment mixing model estimated the decomposition of both soluble and insoluble plant ¹³C (mg C kg⁻¹ soil). Total plant material decomposition followed the alkyl C: O-alkyl C ratio of plant material, as determined by solid-state ¹³C nuclear magnetic resonance spectroscopy. Urea-N addition increased the decomposition of insoluble plant ¹³C in some soils (≤0.1% total nitrogen) but not others (0.3% total nitrogen). Principal components regression analysis indicated that 26% of the variability of plant material decomposition was explained by soil physico-chemical characteristics (P = 0.001), which was primarily described by the C:N ratio. We conclude that plant species with increasing alkyl C: O-alkyl C ratio are better retained as soil organic matter, and that the C:N stoichiometry of soils determines whether N addition leads to increases in soil organic carbon stocks.     

氮素补充对高寒草甸土壤团聚体有机碳、全氮分布的影响

土壤结构的稳定性对高寒草甸生态系统有重要意义。为研究不同水平氮素补充对高寒草甸草地土壤团聚体、有机碳和全氮含量及分布格局的影响,于2012-2014年在青藏高原东部夏河县进行3 a的试验研究。试验为随机区组设计,包括0(对照)、50(低氮)、100(中氮)和200(高氮)kg/hm2 4个氮素补充水平。研究结果表明:低、中、高水平氮素补充处理显著提高了0~30 cm土层≥0.25 mm大团聚体质量分数(P0.05),比对照分别提高了4.74%、6.42%和1.96%;较之对照,低、中水平氮素补充处理显著增加了≥5 mm、≥2~5 mm粒级团聚体含量;低、中水平氮素补充处理显著提升了0~30 cm土壤团聚体平均质量直径,分别比对照提升了9.79%和12.63%。不同水平氮素补充处理有机碳、全氮含量大小排序分别为:中氮≈低氮对照高氮、中氮低氮≈高氮对照。不同粒级团聚体中0.25 mm微团聚体有机碳含量最高而全氮含量最低、≥0.25~2 mm粒级有机碳含量最低而全氮含量最高;低、中水平氮素补充提高了不同粒级团聚体0~30 cm土层有机碳含量而高水平氮素补充处理显著降低了有机碳含量;低、中、高水平氮素补充增加了不同粒级团聚体全氮含量,其中中水平氮素补充处理最高,低水平氮补充次之。不同粒级团聚体含量是影响团聚体养分贡献率的主要原因,≥2~5 mm粒级团聚体含量与相应粒级团聚体有机碳含量呈显著正相关关系,≥5 mm和≥2~5 mm粒级团聚体含量与相应粒级团聚体全氮含量分别呈极显著正相关、显著正相关关系。研究表明连续每年补充50~100 kg/hm2氮可以改善高寒草甸土壤结构并提高土壤肥力状况。     

Chloroform fumigation and the release of soil nitrogen: A rapid direct extraction method to measure microbial biomass nitrogen in soil

A new “direct extraction” method for measuring soil microbial biomass nitrogen (biomass N) is described. The new method (fumigation-extraction) is based on CHC1₃ fumigation, followed by immediate extraction with 0.5 M K₂SO₄ and measurement of total N released by CHC1₃ in the soil extracts. The amounts of NH₄-N and total N extracted by K₂SO₄ immediately after fumigation increased with fumigation time up to 5 days. Total N released by CHC1₃ after 1 day fumigation (1 day CHC1₃-N) and after 5 days fumigation (5 day CHC1₃-N) were positively correlated with the flush of mineral N (F_N) in 37 soils that had been fumigated, the fumigant removed and the soils incubated for 10 days (fumigation-incubation). The regression equations were 1 day CHC1₃-N = (0.79 ± 0.022) F_N and 5 day CHC1₃-N = (1.01 ± 0.027) F_N, both regressions accounting for 92% of the variance in the data.In field soils previously treated with ¹⁵N-labelled fertilizer, the amounts of labelled N, measured after fumigation-extraction, were very similar to the amounts of labelled N mineralized during fumigation-incubation; both were about 4 times as heavily labelled as the soil N as a whole. These results suggest that fumigation-extraction and fumigation-incubation both measure the same fraction of the soil organic N (probably the cytoplasmic component of the soil microbial biomass) and that measurement of the total N released by CHC1₃ fumigation for 24 h provides a rapid method for measuring biomass N.     

Preparation of plant samples for high precision nitrogen isotope ratio analysis

Abstract

Methodologies for preparation of plant samples for high precision nitrogen isotope ratio analysis are described, including detailed attention to Kjeldhal digestion, steam distillation, concentration of distillates, and generation of nitrogen gas (N₂) from ammonium sulfate [(NH₄)₂SO₄], Apparatus developed in the authors laboratory is illustrated. Using a systematic approach to the assessment of errors associated with each of the steps in preparation and analysis procedures, the methods outlined in this paper are shown capable of determining 8¹⁵N in plant material with a precision of ±0.2°/oo.     

An examination of the total analysis of major elements in soil samples

Recently conventional chemical analyses were mostly replaced by instrumental analysis. Although results from both methods were examined in details after the samples had been solubilized or extracted by appropriate techniques, the solubilization method was considerably altered so as to be convenient for the following analysis. In atomic absorption spectrophotometry (AAS), for example, fusion technique was mostly inadequate because of the higher salts concentration in the obtained solution. The use of H₂SO₄ must also be avoided in acid digestion for SO₄ ^2- precipitates with Sr²⁺ or La³⁺ which must be added to eliminate interferences in the succeeding procedure. Therefore, it is essential to consider the whole scheme including sample treatments to evaluate the suitability of instrumental analysis. In this experiment, the suitability of the methods which have been employed in our laboratory was examined by analyzing 9 rock standards.     

A rapid procedure for estimating nitrogen mineralization in manured soil

 A routine soil testing procedure for soil N mineralization is needed that is rapid and precise. Not accounting for N mineralization can result in the over-application of N, especially in soils with a history of manure application. Our objectives were to compare results from a recently proposed rapid laboratory procedure with: (1) long-term N mineralization under standard laboratory conditions, and (2) actual forage N uptake from soil receiving dairy cattle (Bos taurus) manure in a 2-year field study. The rapid procedure is based on the quantity of CO₂-C evolved during 24 h under optimum laboratory conditions following the rewetting of dried soil. Dairy cattle manure was surface applied beginning in 1992 at annual rates of 0, 112, 224, or 448 kg N ha^–1 to field plots on a Windthorst fine sandy loam soil (fine, mixed, thermic Udic Paleustalf) near Stephenville, Texas (32°N, 98°W). Results of the one-day CO₂ procedure were highly correlated with soil N mineralized from samples collected in March of 1995 (P=0.004) and 1996 (P<0.001) and with forage N uptake (P<0.001) both years of the study. Residual inorganic N in the same soil samples was poorly correlated with soil N mineralization and forage N uptake. Received: 23 February 2000     

Extraction and analysis of nitrogen,phosphorus and carbon fractions in plant material

A method is described for the extraction and analysis of various nitrogen‐, phosphorus‐ and carbon‐containing fractions from plant material. Lipids were extracted with chloroform/methanol and chloroform/methanol/water. Soluble nitrogen (nitrate, ammonia, and amino acid), phosphorus (inorganic and sugar phosphate) and carbon (sugar and tannin) fractions were extracted with cold trichloracetic acid. Hot soluble nitrogen and phosphorus (nucleic acid) and carbon (starch and tannin) fractions were extracted with hot trichloracetic acid. Protein remained in the residue. A detailed automated scheme is described for the analysis of each of the above fractions. Also included are methods for analyzing triglyceride, hydrolyzable ester phosphate and phytic acid.     

Determining the distributions of soil carbon and nitrogen in particle size fractions using near-infrared reflectance spectrum of bulk soil samples

This study aimed at assessing the potential of near-infrared reflectance spectroscopy (NIRS) for determining the distribution of soil organic matter (SOM) in particle size fractions, which has rarely been attempted. This was done on sandy soils from Burkina Faso (three sites) and Congo-Brazzaville (one site). Over the total sample set, NIRS accurately predicted carbon (C) and nitrogen (N) concentrations (g kg^?1 fraction) in the fraction <20 μm. When considering Burkina Faso only, predictions were improved in general; those of C and N amounts (g kg^?1 soil) became accurate for the fraction <20 μm but not for the coarser fractions, probably due to heterogeneous SOM repartition. However, most SOM being <20 μm in general, NIRS could be considered promising for determining SOM size distribution.