Rhizodeposition of maize: Short‐term carbon budget and composition

The aim of this study was to assess differences in rhizodeposition quantity and composition from maize cropped on soil or on 1:1 (w/w) soil–sand mixture and distribution of recently assimilated C between roots, shoots, soil, soil solution, and CO₂ from root respiration. Maize was labeled in ¹⁴CO₂ atmosphere followed by subsequent simultaneous leaching and air flushing from soil. ¹⁴C was traced after 7.5 h in roots and shoots, soil, soil solution, and soil‐borne CO₂. Rhizodeposits in the leachate of the first 2 h after labeling were identified by high‐pressure liquid chromatography (HPLC) and pyrolysis–field ionization mass spectrometry (Py‐FIMS). Leachate from soil–sand contained more ¹⁴C than from soil (0.6% vs. 0.4%) and more HPLC‐detectable carboxylates (4.36 vs. 2.69 μM), especially acetate and lactate. This is either because of root response to lower nutrient concentrations in the soil–sand mixture or decreasing structural integrity of the root cells during the leaching process, or because carboxylates were more strongly sorbed to the soil compared to carbohydrates and amino acids. In contrast, Py‐FIMS total ion intensity was more than 2 times higher in leachate from soil than from soil–sand, mainly due to signals from lignin monomers. HPLC‐measured concentrations of total amino acids (1.33 μM [soil] vs. 1.03 μM [soil–sand]) and total carbohydrates (0.73 vs. 0.34 μM) and ¹⁴CO₂ from soil agreed with this pattern. Higher leachate concentrations from soil than from soil–sand for HPLC‐measured carbohydrates and amino acids and for the sum of substances detected by Py‐FIMS overcompensated the higher sorption in soil than in sand‐soil. A parallel treatment with blow‐out of the soil air but without leaching indicated that nearly all of the rhizodeposits in the treatment with leaching face decomposition to CO₂. Simultaneous application of three methods—¹⁴C‐labeling and tracing, HPLC, and Py‐FIMS—enabled us to present the budget of rhizodeposition (¹⁴C) and to analyze individual carbohydrates, carboxylates, and amino acids (HPLC) and to scan all dissolved organic substances in soil solution (Py‐FIMS) as dependent on nutrient status.     

Compound‐specific stable‐isotope (δ13C) analysis in soil science

This review provides current state of the art of compound‐specific stable‐isotope‐ratio mass spectrometry (δ¹³C) and gives an overview on innovative applications in soil science. After a short introduction on the background of stable C isotopes and their ecological significance, different techniques for compound‐specific stable‐isotope analysis are compared. Analogous to the δ¹³C analysis in bulk samples, by means of elemental analyzer–isotope‐ratio mass spectrometry, physical fractions such as particle‐size fractions, soil microbial biomass, and water‐soluble organic C can be analyzed. The main focus of this review is, however, to discuss the isotope composition of chemical fractions (so‐called molecular markers) indicating plant‐ (pentoses, long‐chain n‐alkanes, lignin phenols) and microbial‐derived residues (phospholipid fatty acids, hexoses, amino sugars, and short‐chain n‐alkanes) as well as other interesting soil constituents such as “black carbon” and polycyclic aromatic hydrocarbons. For this purpose, innovative techniques such as pyrolysis–gas chromatography–combustion–isotope‐ratio mass spectrometry, gas chromatography–combustion–isotope‐ratio mass spectrometry, or liquid chromatography–combustion–isotope‐ratio mass spectrometry were compared. These techniques can be used in general for two purposes, (1) to quantify sequestration and turnover of specific organic compounds in the environment and (2) to trace the origin of organic substances. Turnover times of physical (sand < silt < clay) and chemical fractions (lignin < phospholipid fatty acids < amino sugars ≈ sugars) are generally shorter compared to bulk soil and increase in the order given in brackets. Tracing the origin of organic compounds such as polycyclic aromatic hydrocarbons is difficult when more than two sources are involved and isotope difference of different sources is small. Therefore, this application is preferentially used when natural (e.g., C3‐to‐C4 plant conversion) or artificial (positive or negative) ¹³C labeling is used.     

Differentiating the mineralization dynamics of the originally present and newly synthesized amino acids in soil amended with available carbon and nitrogen substrates

Newly synthesized amino acids are the principle compounds created after inorganic nitrogen (N) is rapidly immobilized into microbial tissues. However, little is known about the mineralization kinetics of these newly synthesized amino acids compared to the amino acids originally present in the soil, and how substrate availability controls their mineralization. With ¹⁵N isotope tracing, the newly synthesized (¹⁵N-labeled) amino acids can be differentiated from the amino acids originally present (unlabeled) in soil, making it possible to evaluate the mineralization of the newly synthesized amino acids in tandem with the original amino acids. As amino acids can serve as both N and carbon (C) sources for microorganisms, the mineralization dynamics of amino acids may be manipulated by the availability of extraneous C and N. In this study, an aerobic 30-week intermittent leaching experiment was conducted, using glucose as C source and (¹⁴NH₄)₂SO₄ as N source, following separate additions to soil. The newly synthesized amino acids were determined by an isotope-based high performance liquid chromatography/mass spectrometry (HPLC/MS). The newly synthesized soil amino acids mineralized faster than the original ones, which indicated more rapid cycling of N in the newly synthesized soil amino acids pool. Glucose addition significantly decreased the mineralization of both the newly synthesized and the original amino acids. However, when inorganic N was abundant, the newly synthesized amino acids decomposed rapidly, and preferentially as a C source and energy, while N addition inhibited the mineralization of the original amino acids in the soil. We conclude that the presence of readily degradable C (e.g. glucose) and inorganic N controls the mineralization of newly synthesized and original amino acid pools in soil differently, which is a crucial mechanism in adjusting the N supply and sequestration processes in soil ecosystems.     

Carbohydrate and amino acid composition of dissolved organic matter leached from soil

Low molecular weight organic substances (LMWOS) in soil and soil solution include mainly amino acids, carboxylic acids, and carbohydrates. Due to their high bioavailability they play a crucial role in the cycles of C and nutrients in soils. The variety of soil processes that involve LMWOS requires identifying their composition to elucidate reactions and transformations. In most studies, LMWOS are extracted under artificial conditions, e.g. batch experiments, which may overestimate the actual concentrations. This study measures the composition of carbohydrates and amino acids in solution of a Haplic Luvisol leached in a column experiment. A combined system for simultaneous leaching and blowout of CO₂ was used to estimate LMWOS decomposition. ¹⁴C-labeled glucose was added as a highly sensitive tracer to control the efficiency of the LMWOS extraction by leaching and to estimate LMWOS decomposition during leaching. High performance liquid chromatography (HPLC), optimized for soil extracts, was used to analyze LMWOS composition. For HPLC optimization, different preparations of leached solutions (filtration vs. centrifugation, and drying vs. no-drying) were compared. For sugar determination, drying had no influence on the solution concentrations. In contrast, amino acid concentrations significantly decreased by drying LMWOS eluted substances. Combining the HPLC identification of eluted substances with ¹⁴C tracer application revealed that about 5% of the glucose could be leached unchanged within 786 min (13.1 h), whereas about 84% remained in the soil, 9% were decomposed to CO₂, and 2% were transformed to other LMWOS and recovered in the soil solution. The total amino acid concentration (TAC) in soil solution was about 8.2 μmol l⁻¹, dominated by alanine (14.4% of TAC), glycine (13.4%), glutamic acid (9.9%), serine (9.4%), and leucine (9.3%). The total carbohydrate concentration was about 2.4 μM, dominated by glucose (29.9%), glucuronic acid (26.8%), and galacturonic acid (17.3%). Ratios of hexoses to pentoses, amino sugars glucosamine to galactosamine, and neutral sugars to uronic acids were determined. All three parameters pointed to the dominant influence of plants as the source of LMWOS in the leached soil solution. Within the small contribution of microorganisms, bacteria dominated over fungi. These used biomarker ratios as well as LMWOS concentrations differed widely from the ones obtained with conventional batch extraction. More research is necessary to evaluate the application of these biomarkers to soil solutions.     

Variability and relationships between Pb,Cu, and Zn concentrations in soil solutions and forest floor leachates at heavily polluted sites

Seasonal variability of Cu, Pb, and Zn concentrations in litter leachates and soil solutions was examined in an afforested zone surrounding a copper smelter in SW Poland. Litter leachates (with zero‐tension lysimeters) and soil solutions (with MacroRhizon suction‐cup samplers, installed at a depth of 25–30 cm) were collected monthly at three sites differing in contamination levels in the years 2009 and 2010 (total Cu: 2380, 439, and 200 mg kg^–1, respectively). Concentrations of Cu in the litter leachate were correlated with dissolved organic C (DOC), whereas Zn and Pb were mainly related to leachate pH. Metal concentrations in the soil solution were weakly influenced by their total content in soils and the monthly fluctuations reached 300, 600, and 700% for Cu, Pb, and Zn, respectively. Metal concentrations in soil solutions (Cu 110–460 μg L^–1; Zn 20–1190 μg L^–1; Pb 0.5–36 μg L^–1) were correlated with their contents in the litter leachates. Chemical speciation, using Visual Minteq 3.0, proved organically‐complexed forms even though the correlations between metal concentrations and soil solution pH and DOC were statistically insignificant. The flux of organically‐complexed metals from contaminated forest floors is believed to be a direct and crucial factor affecting the actual heavy metal concentrations and their forms in the soil solutions of the upper mineral soil horizons.     

Compound specific plant amino acid δ15N values differ with functional plant strategies in temperate grassland

The distribution and natural abundance isotopic (δ¹⁵N) content of whole tissue and individual amino acids in plants in a temperate grassland were determined using ion chromatography (IC), continuous flow‐isotope ratio mass spectrometry (CF‐IRMS), and gas chromatography‐combustion‐isotope ratio mass spectrometry (GC‐C‐IRMS). The results showed that the selected plants (Lolium perenne, Juncus effusus, and Brachythecium rutabulum) differed in their amino acid content and distribution from the parent grassland soil. Bulk and individual amino acid δ¹⁵N isotope signatures were different between the plants, which concurred with their functional strategy in relation to the relative acquisition of available N sources. The individual amino acid δ¹⁵N values of histidine and phenylalanine could be used to differentiate between the three plant species.     

Phosphorus leaching in a calcareous soil treated with plant residues and inorganic fertilizer

Column experiments were conducted over 45 d to determine the degree of P mobility. The sandy loam soil was spiked with 200 mg P kg^–1 and 5% organic residues. The treatments included: control without any water‐soluble P and plant residues, potato, wheat, water‐soluble P fertilizer, wheat + water‐soluble P, and potato + water‐soluble P. Each column was leached with distilled water, and leachates were collected and analyzed for P, K⁺, Ca²⁺, Mg²⁺, along with pH and EC. Sequential extraction was performed on soil samples at the end of leaching column experiments. The relatively high initial concentration of P in the leachates decreased to more stable values after 15 d which can be attributed to the colloid‐bound P. The P concentrations in the leachates fluctuated between 8 and 220 mg L^–1 in the water‐soluble–P fertilizer treatment, between 0.80 and 230 mg L^–1 in the potato + water‐soluble‐P treatment, and between 0.90 and 214 mg L^–1 in the wheat + water‐soluble P treatment. Leaching loss of P mainly occurred in the 15 d of leaching, accounting for 94%, 88%, and 65% of total P leached in wheat + water‐soluble‐P, potato + water‐soluble‐P, and water‐soluble‐P treatments, respectively. Maximum amount of P leached was found from an exponential kind model and was in the range 0.45 mg kg^–1 to 125.4 mg kg^–1 in control and potato + water‐soluble‐P treatments, respectively. Sequential extraction results showed that in control and amended soils the major proportion of P was associated with Ca. The leachate samples in all treatments were saturated with respect to hydroxyapatite, β‐tricalcium phosphate, and octacalcium phosphate up to 20 d of leaching, whereas they were undersaturated with respect to Mg‐P minerals through the entire leaching experiment.     

Integrated Flow-based System Displaying an In-line Mini Soil Column to Monitor Iron Species in Soils Leachates

ABSTRACT

Contamination of ground water as a consequence of soil leaching processes is an issue of major concern. In this context, a simulation of the soil leaching process was designed. A sequential injection (SI) method to monitor the soil leaching of iron complexes with in-line rain simulation for leachate production is described. The developed methodology comprises the SI determination of both iron(III) and 3-hydroxy-4-pyridinones iron(III) complexes, coupled to a mini soil column (mSC) for displaying in-line rain simulations. The described SI method enabled iron(III) determination within the range 2.0–35 µmol L^?1, with a detection limit of 0.42 µmol L^?1, and determination of iron(III) complexes in the range 1.0–45 µmol L^?1. It was successfully applied to leachates from laboratory scale soil columns (LSSC), with good precision for both iron(III) and iron complexes determinations: calculated relative standard deviation (RSD) of 5% and 6%, respectively. A step further in automation and miniaturization was attained with the incorporation of a mini soil column for the in-line leachate production. The system enabled the soil leachate production and assessment in less than 5 min, including determinations in triplicate.     

Qualitative assessment of rhizodeposits in non‐sterile soil by analytical pyrolysis

Estimation of the amount of root exudates and simultaneous identification of their composition in non‐sterile soil is a challenging objective in rhizosphere research. We coupled 3 methods: (1) labeling of corn in ¹⁴CO₂ atmosphere to separate root‐derived and soil‐derived organic substances in the rhizosphere, (2) a previously developed leaching method to collect rhizodeposits, and (3) pyrolysis field ionization mass spectrometry (Py‐FIMS) to investigate the molecular‐chemical composition of rhizodeposits. Eluted rhizodeposits accounted for 2.8 % (Loam) and 0.97 % (nutrient solution in quartz sand) of recovered ¹⁴C and showed clear differences in composition between the growth substrates. The ¹⁴CO₂ evolved mostly by root respiration accounted for 3.5–4.0 % without significant differences according to growth substrate or diurnal dynamics. Principal component analysis of the Py‐FI mass spectra of leachates showed a clear diurnal dynamics of the amount and the composition of corn rhizodeposits collected during day‐time and night‐time. Differences originated mostly from signals assigned to carbohydrates, sterols, and peptides. This approach is recommended for forthcoming studies of rhizodeposition in different soil substrates, crops grown, and time‐series of exudate sampling.     

Heavy metal binding by hydrophobic and hydrophilic dissolved organic carbon fractions in a Spodosol A and B horizon

For a one year period intact Spodosol soil columns were percolated weekly with H₂O_deion, 1.58 mmol H₂SO₄ L^?1, and 0.79 mmol H₂SO₄ L^?1+0.64 mmol HNO₃ L^?1, respectively. Decomposition rates, soil organic carbon (OC) solubilization, dissolved organic carbon (DOC) fractions, and Cr-, Cu-, and Cd-binding by dissolved hydrophobic and hydrophilic acids were studied. Acid treatment reduced significantly OC respiration as well as OC solubilization in the humic layers. The reduced OC solubility at acid addition was more pronounced for the less polar hydrophobic compounds, resulting in a decrease of the hydrophobic acids (from ca. 65 to 40–45% of DOC), and in an increase of the hydrophilic acids (from ca. 25 to 40–45% of DOC). For B horizon leachates, DOC increased at acid treatment. Generally, hydrophobic acids were retained preferentially in the B horizon. Also in the B horizon output there was an increase of the hydrophilic acids as acidity increased (from ca. 40 to 50% of DOC). Differences between the two acid treatments were negligible. The degree of metal-organic complexes decreased in the order Cr >Cu >Cd, from A to B horizon leachates, and with increasing acidity. Hydrophilic acids were found to be the dominating ligands in complexing Cr and Cu. Actual Cr- and Cu-binding by hydrophilic acids exceeded that by hydrophobic acids 2–8 times. As the hydrophilic acids represented the most mobile DOC components in the soil columns, in particular with increasing acidity, significant amounts of Cr and Cu in the B horizon leachates were organically complexed, although a great proportion of the hydrophobic acids was retained in the B horizon.     

Extraktion eines kupferbelasteten Bodens unter Verwendung eines aminosäurehaltigen Reststoffhydrolysats. 2. Elutionsdynamik und Bilanzierung der Aminosäuren nach kontinuierlichem Eintrag in Bodensäulen

Extraction of a copper contaminated soil material by the percolation of an amino acid containing residue hydrolysate. 2. Time course of amino acid elution and input/output balance of amino acids During 16 days an amino acid containing blood meal hydrolysate (amino acid concentration: 188 mMol·L^?1) was percolated through a column packed soil material (soil content per column: 4.1 kg dry weight, four parallels). The copper contaminated material (soil type: Typic Udifluvent, soil texture: sandy loam, loamy sand) was sampled from an area formerly used for cultivation of hop (Humulus lupus). Besides the investigation of the copper liberation the experiments aimed to determine the elution dynamic and input/output balance of amino acids (time span for amino acids balance 14 days). In total 11.7 L of hydrolysate, containing 2.2 Mol of amino acids, were introduced into each column. The mean amino acid output with the column effluent was 1.13 Mol. This corresponds to an elution degree of 51.2%, related to the sum of applicated amino acids, and to a mean substance specific elution degree of 48.4% reflecting the elution of 15 compounds. The substance specific elution ranged from 9.6% (serine) to 75.5% (valine). The highest concentrations of serine and threonine were determined in the effluents after two days, whereas the histidine concentration was highest at the last sampling. The differences in the percolation properties of the amino acids are discussed in terms of important retention and elimination processes (biodegradation, ad-/desorption, intercalation).     

Effect of alternative anions (Cl– vs. SO$ _4^{2-} $) on concentrations of free amino acids in young tea plants

The quality of green tea is highly dependent on the concentration of free amino acids, whose profile is dominated by the unique amino acid theanine (N⁵‐ethyl‐glutamine). A high quality is associated with a high amino acid–to–catechin ratio, but previous results indicated that excessive chloride (Cl^–) supply is detrimental for amino acid accumulation. Several experiments were conducted to investigate the effect of chloride on growth and concentrations of free amino acids in young tea plants. Soil‐grown tea plants supplied with different levels of potassium (K) as K₂SO₄ or KCl exhibited increased concentrations of free amino acid in young shoots only when supplied with K₂SO₄, and the negative effect of KCl supply was mainly due to a reduced concentration of theanine. Concentrations of other nutrients in plant tissues were not influenced. The uptake of Cl^– and its interaction with nitrogen (N) uptake were further investigated in a second experiment, in which soil‐cultivated tea plants were supplied with varying amounts of Cl^–. Chloride application reduced yield of young shoots, and severity of leaf damage was related to the concentration of Cl^– in leaves. Nitrogen uptake was reduced by Cl^– addition. To verify whether the decrease of free amino acids was simply a result of inhibited NO assimilation, a third experiment was conducted, in which tea plants were NH ‐fed in the absence or presence (equivalent to the NH concentration) of Cl^–. Again, concentrations of theanine and total free amino acids in young shoots were reduced by Cl^– supply, but changes of the free–amino acid pool did not contribute to the maintenance of charge balance. However, concentration of theanine in roots, where it is synthesized, was not influenced by Cl^–. Total N concentrations of roots and mature leaves, uptake rate of NH , and activity of glutamine synthetase in fibrous roots and young leaves were all unaffected by Cl^– as well. It is suggested that translocation of theanine from root to shoot and its catabolism in young shoots might be influenced by Cl^–.     

Long‐term field fertilization affects soil nitrogen transformations in a rice‐wheat‐rotation cropping system

Mineralization and nitrification are the key processes of the global N cycle and are primarily driven by microorganisms. However, it remains largely unknown about the consequence of intensified agricultural activity on microbial N transformation in agricultural soils. In this study, the ¹⁵N‐dilution technique was carried out to investigate the gross mineralization and nitrification in soils from a long‐term field fertilization experiment starting from 1988. Phospholipid fatty acids (PLFA) analysis was used to determine soil microbial communities, e.g., biomasses of anaerobic bacterial, bacterial, fungi, and actinobacteria. The abundance of ammonia‐oxidizing bacteria (AOB) and archaea (AOA) were measured using real‐time quantitative polymerase chain reaction. The results have demonstrated significant stimulation of gross mineralization in the chemical‐fertilizers treatment (NPK) ([6.53 ± 1.29] mg N kg^–1 d^–1) and chemical fertilizers–plus–straw treatment (NPK+S1) soils ([8.13 ± 1.68] mg N kg^–1 d^–1) but not in chemical fertilizers–plus–two times straw treatment (NPK+S2) soil when compared to the control‐treatment (CK) soil ([3.62 ± 0.86] mg N kg^–1 d^–1). The increase of anaerobic bacterial biomass is up to 6‐fold in the NPK+S2 compared to that in the CK soil ([0.7 ± 0.5] nmol g^–1), implying that exceptionally high abundance of anaerobic bacteria may inhibit gross mineralization to some extent. The gross nitrification shows upward trends in the NPK+S1 and NPK+S2 soils. However, it is only significantly higher in the NPK soil ([5.56 ± 0.51] mg N kg^–1 d^–1) compared to that in the CK soil ([3.70 ± 0.47] mg N kg^–1 d^–1) (p < 0.05). The AOB abundance increased from (0.28 ± 0.07) × 10⁶ copies (g soil)^–1 for the CK treatment to (4.79 ± 1.23) × 10⁶ copies (g soil)^–1 for the NPK treatment after the 22‐year fertilization. In contrast, the AOA abundance was not significantly different among all treatment soils. The changes of AOB were well paralleled by gross nitrification activity (gross nitrification rate = 0.263 AOB + 0.047 NH ‐N + 2.434, R² = 0.73, p < 0.05), suggesting the predominance of bacterial ammonia oxidation in the fertilized fields.     

Manganese efficiency and maganese‐uptake kinetics of raya (Brassica juncea), wheat (Triticum aestivum), and oat (Avena sativa) grown in nutrient solution and soil

Manganese (Mn) deficiency is reported worldwide and often decreases crop yield. However, plant species differ in their susceptibility to Mn deficiency. Poaceae are often inefficient, whereas Brassicaceae seem to be efficient in Mn uptake. The objective of this paper was to determine the relevance of Mn‐uptake kinetics, root‐system size, and Mn mobilization for differences in Mn efficiency of wheat, oat, and raya. To determine Mn‐uptake kinetics, wheat (Triticum aestivum L. cv. PBW 343), raya (Brassica juncea L. cv. RLM 619), and oat (Avena sativa L. cv. Aragon) were grown in a growth chamber together in complete nutrient solution having an average Mn concentration of 90, 180, 360, 910, and 2270 nmol L^–1. For determining Mn efficiency of the three species in soil, the plants were grown for 22 d in pots filled with 3 kg of a loamy soil low in Mn availability (pH (CaCl₂) 7.4; DTPA‐extractable Mn: 3.5 mg (kg soil)^–1). The soil was fertilized with 0, 1, 2, 4, and 8 mmol Mn (kg soil)^–1 resulting in Mn soil‐solution concentrations ranging from 40 to 90 nmol L^–1, hence lower than in the solution experiment. In order to determine Mn soil‐solution concentration close to the root surface, the root length density was increased by growing two plants of raya and four plants of wheat in only 250 mL soil columns for 25 d. In solution culture at high concentrations, raya showed a higher Mn uptake compared to wheat and oat. However, at low Mn supply, all three species were comparably Mn‐efficient, i.e., plant growth was similar, and also the uptake was similar. In soil, the highest yield was achieved for raya in the unfertilized treatment whereas the Poaceae needed at least a fertilization of 1 mmol Mn (kg soil)^–1. The Poaceae showed a yield reduction of about 40% in the unfertilized treatment. Manganese concentration in the shoot dry weight was always higher in raya than in wheat or oat. This was due to a higher Mn uptake whereas relative shoot‐growth rate and root‐to‐shoot ratio were similar among the species. The higher Mn uptake of raya in soil was in contradiction to the comparable Mn‐uptake kinetics of the three crops at low Mn concentration in solution. This points to plant differences in their ability to affect Mn availability in the rhizosphere. In the bulk soil, all the crops decreased Mn solution concentration, but this effect was somewhat less for raya. But in the rhizosphere, raya increased Mn soil‐solution concentration significantly to 58 nmol L^–1, as compared to 37 nmol L^–1 of the unplanted control soil. In contrast, wheat showed a Mn solution concentration of 25 nmol L^–1 which was not significantly different from the control. The results indicate that differences in Mn efficiency among the crops studied are related to their ability to affect the solubility of Mn in the rhizosphere.     

Does nitrogen concentration affect relative uptake rates of nitrate,ammonium, and glycine?

Plant roots are exposed to a variety of nitrogen forms (e.g., nitrate, ammonium, amino acids) and take up these forms at different rates. Many studies have investigated whether plants prefer nitrate, ammonium, or amino acids; but studies may not be comparable because they used substrate concentrations between 100 and 2000 μmol L^–1. This study tests the hypothesis that substrate concentrations from 10 to 1750 μmol L^–1 affect plant preference for N forms. Nitrogen uptake by the herb Ocimum basilicum and the evergreen tree Eucalyptus regnans was examined by placing roots of intact seedlings in equimolar mixtures of nitrate, ammonium, and glycine in which one of the N forms was ¹⁵N‐labelled (and ¹³C‐labelled in the case of glycine). In both species, preference for N forms was affected by substrate concentration. At 10 μmol L^–1 (O. basilicum) or 10 and 50 μmol L^–1 (E. regnans), rates of N uptake did not differ among N forms. At substrate concentrations of 50 μmol L^–1 and greater O. basilicum took up ammonium the fastest, glycine the slowest, and nitrate at an intermediate rate. At substrate concentrations from 100 to 1750 μmol L^–1, E. regnans took up ammonium the fastest with glycine and nitrate taken up at slower rates. The absence of significant differences at lower concentrations was a true biological effect rather than a function of larger relative errors. This study demonstrates that substrate concentration has a large effect on plant preference for N forms, and sounds a warning for studies of N nutrition that do not consider the concentration‐dependence of plant preference for N forms.     

Effects of fertilizer type and rate on labile soil fractions of a sandy Cambisol—long‐term and short‐term dynamics

The application of density fractionation is an established technique, but studies on short‐term dynamics of labile soil fractions are scarce. Objectives were (1) to quantify the long‐term and short‐term dynamics of soil C and N in light fraction (LFOC, LFON, ρ ≤ 2.0 g cm^–3) and microbial biomass C (C_mic) in a sandy Cambisol as affected by 28 y of different fertilization and (2) to determine the incorporation of C₄‐C into these labile fractions during one growing season of amaranth. The treatments were: straw incorporation plus application of mineral fertilizer (MSI) and application of farmyard manure (FYM) each at high (MSI_H, FYM_H, 140–150 kg N ha^–1 y^–1) and low (MSI_L, FYM_L, 50–60 kg N ha^–1 y^–1) rates at four field replicates. For all three sampling dates in 2008 (March, May, and September), stocks of LFOC, LFON and C_mic decreased in the order FYM_H > FYM_L > MSI_H, MSI_L. However, statistical significance varied markedly among the sampling dates, e.g., with LFOC being significantly different (p ≤ 0.05) in the order given above (sampling date in March), significantly different depending on the fertilizer type (May), or nonsignificant (September). The high proportion of LFOC on the stocks of soil organic C (45% to 55%) indicated the low capacity of soil‐organic‐matter stabilization on mineral surfaces in the sandy Cambisol. The incorporation of C₄‐C in the LFOC during one growing season of amaranth was small in all four treatments with C₄‐LFOC ranging from 2.1% to 3.0% of total LFOC in March 2009, and apparent turnover times of C₃‐derived LFOC ranged from 21 to 32 y for the sandy soils studied. Overall, our study indicates that stocks of LFOC and LFON in a sandy arable soil are temporarily too variable to obtain robust significant treatment effects of fertilizer type and rate at common agricultural practices within a season, despite the use of bulked six individual cores per plot, a common number of field replicates of four, and a length of treatments (28 y) in the order of the turnover time (21–32 y) of C₃‐derived LFOC.     

Co‐occurrence of organic and inorganic N influences asparagine uptake and amino acid profiles in white clover

Background : Direct plant uptake of organic nitrogen (N) may be important for plant N nutrition, but we lack knowledge of how the concentration and form of external N influence organic N uptake and plant N status. Aims : We investigated the uptake of the amino acid asparagine (Asn) in white clover in the presence of different inorganic, organic and total N concentrations. Methods : Actively N₂‐fixing white clover seedlings were for one week exposed to combinations of ${\mathrm{NO}}_3^{-}$ (3–30 µmol N kg^?1 sand DW) and Asn (3–30 µmol N kg^?1 sand DW), whereafter the Asn uptake rate was determined by addition of ¹³C₄‐Asn. Shoot and root amino acid profiles were also analyzed. Results : Increasing ${\mathrm{NO}}_3^{-}$ and total N concentrations decreased ¹³C₄‐Asn uptake rates and internal clover Asn content. In addition, total N and ${\mathrm{NO}}_3^{-}$ also affected amino acid profiles, with Asn, Asp, Glu, Gln, Cys, Gly, Pro, Ser, and Ala being more related to the low N doses, and Thr, Val, Ile, Leu, Phe, Tyr, Trp, and Met being more abundant at increasing N doses. Conclusions : Asn uptake rate in white clover is reduced by increasing inorganic N. Plant amino acid profiles are likely to be a more sensitive indicator of N supply.     

Radiogaschromatographie von Kohlenhydraten,Organischen Säuren und Aminosäuren aus Pflanzen

Radio gaschromatography of carbohydrates, organic acids and amino acids A method for the radio-gaschromatographic determination of carbohydrates, organic acids and amino acids as well as the simultaneous recording of the specific activity of radioactive labelled compounds of these fractions is described and its application to the analysis of plant extracts or protein hydrolysates demonstrated. The application of 0.05–0.1 mCi ¹⁴C/g fresh weight of plant material is enough to obtain a sufficiently strong labelling even in short-term incubation experiments (15 sec.), as for example in photosynthesis research.     

Compositional changes of selected amino acids,organic acids,and soluble sugars in the xylem sap of N,P, or K‐deficient tomato plants

Xylem sap plays a major role in long‐distance transport of water, nutrients, and metabolites. However, there is little information on the behavior of metabolites in mineral‐deficient xylem sap. For this reason, the time‐dependent changes in selected metabolites (amino acids, organic acids, and soluble sugars) from tomato xylem sap in response to nitrogen (N), phosphorus (P), or potassium (K)‐deficient condition were investigated. Tomato plants (Solanum lycopersicum L.) were grown hydroponically in liquid culture under three different mineral regimes: N‐deficient [0.5 mM Ca(NO₃)₂ and 0.5 mM KNO₃], P‐deficient (0.05 mM KH₂PO₄), and K‐deficient (0.5 mM KNO₃), respectively. Xylem sap was collected at 10:00 am after 1, 5, 15, and 30 d, and the selected metabolites were analyzed with liquid chromatography. All N, P, or K deficiencies led to a substantial increase in metabolites in the xylem sap. The predominant amino acid in the xylem sap was glutamine and, interestingly, all mineral deficiencies resulted in a substantial amount of γ‐aminobutyric acid (GABA). Additionally, organic acids (citrate and malate) and soluble sugars were strongly increased in all mineral deficiencies, and, in particular, the level of shikimate was greatly affected by N deficiency. Based on these data, it is necessary to clearly elucidate an unknown event taking place in xylem loading in a variety of environmental impacts, and we are now studying to expand our knowledge on metabolic and proteomic responses using GC‐MS and LC‐MS.