Effects of nitrogen rate and nitrogen form on glycine uptake by pakchoi (Brassica chinensis L.) under sterile culture

It is well known that plants are capable of taking up intact amino acids. However, how the nitrogen (N) rates and N forms affect amino acid uptake and amino acid nutritional contribution for plant are still uncertain. Effects of the different proportions of nitrate (NO₃^?), ammonium (NH₄⁺) and ¹⁵N-labeled glycine on pakchoi seedlings glycine uptake were investigated for 21 days hydroponics under the aseptic media. Our results showed that plant biomass and glycine uptake was positively related to glycine rate. NO₃^? and NH₄⁺, the two antagonistic N forms, both significantly inhibited plant glycine uptake. Their interactions with glycine were also negatively related to glycine uptake and glycine nutritional contribution. Glycine nutritional contribution in the treatments with high glycine rate (13.4%–35.8%) was significantly higher than that with low glycine rate (2.2%–13.2%). The high nutritional contribution indicated amino acids can serve as an important N source for plant growth under the high organic and low inorganic N input ecosystem.     

Eliminating amino acid interference during spectrophotometric NH4 analysis

Amino acids can interfere with NH₄⁺ in spectrophotometric NH₄⁺ determination hampering accurate quantification of the fate of NH₄⁺ and dissolved organic N in soils. Serious interference has been reported for soils rich in organic matter, and for soils that have been fumigated, oven-dried or fertilized where between 5 and 60% of the NH₄⁺ detected could be attributed to amino acid interference. We investigated whether a combination of gas diffusion and the classical Berthelot method can eliminate this interference, increasing the selectivity of the NH₄⁺ analysis. We tested this approach using synthetic amino acid solutions and a large set of terrestrial samples (n = 353), including pore water samples, (fumigated or dried) soil extracts, and extracts of manures, composts and crop residues. The evaluated method produced accurate (recovery > 99.7%) and reproducible (standard error = 2.2%) NH₄⁺ concentrations, eliminating any interference between amino acids and NH₄⁺ (interference < 0.3%). Interference from K₂SO₄ was also eliminated. The method is robust, broadly applicable and will improve our understanding of nitrogen cycling in various ecosystems, in particular those where high levels of amino acids occur.     

陕西省几种代表性土壤NH4+吸附、解吸动力学特征研究

采用连续液流法测定了五种土壤吸附、解吸ＮＨ＾＋４的动力学性质。研究表明：（１）ＮＨ＾＋４吸附、解吸平衡时间及反应速率，平衡时的吸附、解吸量及吸附平衡常数均随土壤粘粒和ＣＥＣ不同而变化；（２）不同动力学模型及同一模型对不同土壤的拟合性不同。     

Ammonium assimilation in rice based on the occurrence of 15N and inhibition of glutamine synthetase activity

Assimilation of ammonium (NH₄) into free amino acids and total reduced nitrogen (N) was monitored in both roots and shoots of two‐week old rice seedlings supplied with 5 mM 99% (¹⁵NH₄)₂SO₄ in aerated hydroponic culture with or without a 2 h preincubation with 1 mM methionine sulfoximine (MSX), an inhibitor of glutamine synthetase (GS) activity. ¹⁵NH₄ was not assimilated into amino acids when the GS/GOGAT (glutamate synthase) cycle was inhibited by MSX. Inhibition of glutamine synthetase (GS) activity in roots with MSX increased both the amount of NH₄ and the abundance of ¹⁵N labeled NH₄. In contrast, the amount of Gln and Glu, and their proportions as ¹⁵N, decreased in roots when GS activity was inhibited. This research confirms the importance of GS/GOGAT in NH₄ assimilation in rice roots.

¹⁵N‐labeled studies indicate that NH₄ ions incorporated by roots of rice are transformed primarily into glutamine (Gln) and glutamic acid (Glu) before being converted to other amino acids through transamination (15). The formation of amino acids such as aspartic acid (Asp) and alanine (Ala) directly from free NH₄ in roots also has been reported (4,15). Translocation of free NH₄ to plant shoots, based on the concentration of free NH₄ in xylem exudate, has been reported in tomato (13), although NH₄ in shoots primarily originates from nitrate reduction in the shoot. Photorespiration also can contribute to the accumulation of NH₄ in leaves (7).

The GS/GOGAT cycle appears to be primarily responsible for the assimilation of exogenously supplied NH₄ and NH₄ derived from nitrate reduction in leaves, as well as NH₄ derived from photorespiration (2,3,6,8). Genetic evidence cited to support this conclusion includes the lethal effect of photorespiratory conditions on plant mutants deficient in chloroplast‐localized GS and GOGAT activities (2,3,9), and the rapid accumulation of free NH₄ in GS‐deficient mutants under photorespiratory conditions (2,3,5).

The present study was initiated to quantify the in vivo amino acid synthesis in rice roots and shoots by analysis of ¹⁵N labeling, and should provide a more complete understanding of this important system for NH₄ utilization.     

A GC/MS method for the assessment of N and C incorporation into soil amino acid enantiomers

Identifying the transformation process of amino acid enantiomers was essential to probe into the fate, turnover and aging of soil nitrogen due to their important roles in the biogeochemical cycling. If this can be achieved by differentiating between the newly biosynthesized and the inherent compounds in soil, then the isotope tracer method can be considered most valid. We thereby developed a gas chromatography/mass spectrometry (GC/MS) method to trace the ¹⁵N or ¹³C isotope incorporation into soil amino acid enantiomers after being incubated with ¹⁵NH₄⁺ or U-¹³C-glucose substrates. The most significant fragments (F) as well as the related minor ions were monitored by the full scan mode and the isotope enrichment in amino acids was estimated by calculating the atom percentage excess (APE). ¹⁵NH₄⁺ incorporation was evaluated according to the relative abundance increase of m/z F+1 to F for neutral and acidic amino acids and F+2 to F (mass 439) for lysine. The assessment of ¹³C enrichment in soil amino acids was more complicated than that of ¹⁵N due to multi-carbon atoms in amino acid molecules. The abundance ratio increment of m/z F+n to F (n is the original skeleton carbon number in each fragment) indicated the direct conversion from the added glucose to amino acids, but the total isotope incorporation from the added ¹³C can only be calculated according to all target isotope fragments, i.e. the abundance ratio increment summation from m/z (Fa+1) through m/z (Fa+T) represented the total incorporation of the added ¹³C (Fa is the fragment containing all original skeleton carbons and T is the carbon number in the amino acid molecule). This method has a great advantage especially for the evaluation of high-abundance isotope enrichment in organic compounds compared with GC/C/IRMS. And in principle, this technique is also valid for amino acids besides enantiomers if stereoisomers are not concerned. Our assessment approach could shine a light on investigating the biochemical mechanism of microbial transformation of N and C in soils of terrestrial ecosystem.     

Amino acid,peptide and protein mineralization dynamics in a taiga forest soil

The availability of inorganic N has been shown to be one of the major factors limiting primary productivity in high latitude ecosystems. The factors regulating the rate of transformation of organic N to nitrate and ammonium, however, remain poorly understood. The aim of this study was to investigate the nature of the soluble N pool in forest soils and to determine the relative rate of inorganic N production from high and low molecular weight (MW) dissolved organic nitrogen (DON) compounds in black spruce forest soils. DON was found to be the dominant N form in soil solution, however, most of this DON was of high MW of which >75% remained unidentified. Free amino acids constituted less than 5% of the total DON pool. The concentration of NO₃⁻ and NH₄⁺ was low in all soils but significantly greater than the concentration of free amino acids. Incubations of low MW DON with soil indicated a rapid processing of amino acids, di- and tri-peptides to NH₄⁺ followed by a slower transformation of the NH₄⁺ pool to NO₃⁻. The rate of protein transformation to NH₄⁺ was slower than for amino acids and peptides suggesting that the block in N mineralization in taiga forest soils is the transformation of high MW DON to low MW DON and not low MW DON to NH₄⁺ or NH₄⁺ to NO₃⁻. Calculated turnover rates of amino acid-derived C and N immobilized in the soil microbial biomass were similar with a half-life of approximately 30 d indicating congruent C and N mineralization.     

氮素不同形态配比对菠菜体内游离氨基酸含量和相关酶活性的影响

采用溶液培养试验,研究了氮素不同形态配比对菠菜茎叶中游离氨基酸含量及3种主要氮代谢酶活性的影响。结果表明:1)随着营养液中铵硝比(NH4+-N/NO3--N)的降低,菠菜茎叶中游离氨基酸的总量呈下降趋势。在全硝营养下(NH4+-N/NO3--N=0∶100)下,菠菜茎叶中游离氨基酸的总量只有全铵营养(NH4+-N/NO3--N=100∶0)的34.4%。2)在全铵营养下,菠菜茎叶中游离氨基酸的主要组分是谷氨酰胺、精氨酸和谷氨酸,三者占游离氨基酸总量的百分比依次为39.8%、20.2%和8.9%;在全硝营养下,菠菜茎叶中游离氨基酸以谷氨酸、天冬氨酸和丝氨酸为主,三者占游离氨基酸总量的百分比分别为30.3%1、8.6%和8.5%。3)提高营养液中硝态氮的比例,可以显着提高菠菜茎叶中硝酸还原酶(NR)的活性,同时降低了谷氨酸脱氢酶(GDH)的活性,谷氨酰胺合成酶(GS)活性则呈现先升后降的抛物线状变化规律。4)菠菜茎叶中NR活性与谷胺酰胺含量之间存在着显著负相关关系(r=-0.968)。     

Improving the Accuracy of Diffusion for Inorganic 15N Analyses of Soil Extracts and Water

Sequential diffusion techniques used to speciate inorganic nitrogen-15 (¹⁵N) during soil or water analysis are complicated by incomplete recovery of ammonium (NH₄⁺)-N, introducing error in the subsequent determination of nitrate (NO₃^–)-N. Based on studies to evaluate different strategies for minimizing cross-contamination error in Mason-jar diffusions, a simple cleaning technique was developed that involves an additional 6-h diffusion using 0.6 M boric acid (H₃BO₃) at room temperature following the recovery of NH₄⁺-N. This technique was 60–87% effective for reducing cross-contamination of unlabeled NO₃^–-N by labeled NH₄⁺-N and became more effective for controlling analytical error with decreasing sample volumes, lower NH₄⁺-N enrichment, and larger quantities of NO₃^–-N. When used with the cleaning technique described, sequential diffusions were far superior for ¹⁵N analysis of NO₃^–-N, as compared to the nonsequential approach that involves an isotope dilution calculation after separate diffusions to determine NH₄⁺-N and total mineral N.     

农田土壤中氮同位素分级与硝化能力的关系研究

A laboratory-based aerobic incubation was conducted to investigate nitrogen(N) isotopic fractionation related to nitrification in five agricultural soils after application of ammonium sulfate((NH4)2SO4). The soil samples were collected from a subtropical barren land soil derived from granite(RGB),three subtropical upland soils derived from granite(RQU),Quaternary red earth(RGU),Quaternary Xiashu loess(YQU) and a temperate upland soil generated from alluvial deposit(FAU). The five soils varied in nitrification potential,being in the order of FAU YQU RGU RQU RGB. Significant N isotopic fractionation accompanied nitrification of NH+4. δ15N values of NH+4 increased with enhanced nitrification over time in the four upland soils with NH+4 addition,while those of NO-3 decreased consistently to the minimum and thereafter increased. δ15N values of NH+4 showed a significantly negative linear relationship with NH+4-N concentration,but a positive linear relationship with NO-3-N concentration. The apparent isotopic fractionation factor calculated based on the loss of NH+4 was 1.036 for RQU,1.022 for RGU,1.016 for YQU,and 1.020 for FAU,respectively. Zero- and first-order reaction kinetics seemed to have their limitations in describing the nitrification process affected by NH+4 input in the studied soils. In contrast,N kinetic isotope fractionation was closely related to the nitrifying activity,and might serve as an alternative tool for estimating the nitrification capacity of agricultural soils.     

Changes in amino acid and organic acid composition in tomato and cucumber plants in relation to salinity and nitrogen nutrition

Under conditions of salt stress, plants show qualitative and quantitative alterations in various organic compounds, such as nitrogen (N) compounds and organic acids. In this work, the effect of different saline levels as well as various N levels, supplied as nitrate (NO₃) or as ammonium (NH₄)+NO₃ on the concentration of amino acids and organic acids in the leaves of tomato and cucumber plants has been studied. The effect of the source of N on individual amino acid contents varied with plant species. Most of the amino acids increased when the concentration of N in the nutrient solution was increased, except when N was added as NH₄+NO₃ for tomato. The effect of salt stress depended on which amino acid was considered. The data also indicate that the effect of salinity on each particular amino acid was greatly dependent on the plant species and N source. Organic acids were differently affected by salinity and by the N source, depending on the plant species. In tomato, the concentrations of short‐chain organic acids were 2–3 times higher in NO₃‐supplied plants than in those grown with NH₄+NO₃. Finally, in cucumber, malic acid concentration increased as a function of the saline level in the medium.     

Kinetics of 15NH4 + assimilation in tomato plants: evidence for 15NH4 + assimilation via GDH in tomato roots 1

The kinetics of ¹⁵NH₄ ⁺ assimilation into free amino acids and total reduced nitrogen were monitored in both roots and shoots of two week old tomato seedlings supplied with 5 mM 99% (¹⁵NH₄)₂SO₄ via the aerated root medium in hydroponic culture, in the presence and absence of a 2 h pre‐incubation with 1 mM methionine sulfoximine (MSX). The labeling kinetics of amino acids in roots of tomato plants in the presence of MSX show that continued assimilation of ¹⁵NH₄ ⁺ can occur when the GS/GOGAT cycle is inhibited. In the presence of MSX, three amino acids [glutamate, alanine, and y‐amino butyrate (GABA)] of the root tissue continue to become labeled with ¹⁵N under conditions where labeling of the amino‐N moiety of glutamine is completely inhibited. This indicates primary ammonia assimilation via GDH, or alternatively, assimilation of ammonia into alanine via alanine dehydrogenase. Free ammonia accumulates rapidly in both shoots and roots of tomato in response to MSX. The labeled ammonia accumulated in the roots in the presence of MSX must be largely derived from the medium whereas in shoots this ammonia appears to be derived from catabolism of unlabeled amino acids and proteins. The pools of glutamine, glutamate and alanine after 24 h exposure to ¹⁵NH₄ ⁺ were, on the average, 5‐ to 10‐fold lower in the MSX‐treated than in the control (‐MSX) shoots and roots. In contrast, the pools of valine, leucine, isoleucine, proline, threonine, phenylalanine, lysine, and tyrosine increased 5‐ to 10‐fold above the control values in the shoots of MSX‐treated plants, and 2‐ to 4‐fold above control values in the roots of MSX‐treated tomato plants after 24 h. The latter amino acids all exhibited low isotope abundance, and presumably were derived from protein turnover.     

Microbial immobilization of ammonium and nitrate in relation to ammonification and nitrification rates in organic and conventional cropping systems

Agricultural systems that receive high or low organic matter (OM) inputs would be expected to differ in soil nitrogen (N) transformation rates and fates of ammonium (NH₄⁺) and nitrate (NO₃⁻). To compare NH₄⁺ availability, competition between nitrifiers and heterotrophic microorganisms for NH₄⁺, and microbial NO₃⁻ assimilation in an organic vs. a conventional irrigated cropping system in the California Central Valley, chemical and biological soil assays, ¹⁵N isotope pool dilution and ¹⁵N tracer techniques were used. Potentially mineralizable N (PMN) and hot minus cold KCl-extracted NH₄⁺ as indicators of soil N supplying capacity were measured five times during the tomato growing season. At mid-season, rates of gross ammonification and gross nitrification after rewetting dry soil were measured in microcosms. Microbial immobilization of NO₃⁻ and NH₄⁺ was estimated based on the uptake of ¹⁵N and gross consumption rates. Gross ammonification, PMN, and hot minus cold KCl-extracted NH₄⁺ were approximately twice as high in the organically than the conventionally managed soil. Net estimated microbial NO₃⁻ assimilation rates were between 32 and 35% of gross nitrification rates in the conventional and between 37 and 46% in the organic system. In both soils, microbes assimilated more NO₃⁻ than NH₄⁺. Heterotrophic microbes assimilated less NH₄⁺ than NO₃⁻ probably because NH₄⁺ concentrations were low and competition by nitrifiers was apparently strong. The high OM input organic system released NH₄⁺ in a gradual manner and, compared to the low OM input conventional system, supported a more active microbial biomass with greater N demand that was met mainly by NO₃⁻ immobilization.     

Arbuscular mycorrhizae affect the N and C economy of nodulated Phaseolus vulgaris (L.) during NH4 nutrition

In the symbiosis between nodulated legume roots and arbuscular mycorrhizal (AM) fungi, the C and N economy can be influenced by the source of N-supply from either AM-derived NH₄⁺ uptake or nodule-derived biological nitrogen fixation (BNF). This relationship was investigated in terms of NH₄⁺ supply and BNF by the two symbionts. Nodulated Phaseolus vulgaris seedlings with and without AM, were hydroponically grown with either 0 N or 1 mM NH₄⁺ supply. Plants were harvested at 30 days after emergence and measurements were taken for biomass, N₂ fixation, photosynthesis, CO₂ and O₂ root respiration, calculated C and N economy. AM roots had higher NH₄⁺ uptake and this was associated with the suppression of BNF and nodule growth. The higher NH₄⁺ uptake in AM roots occurred with lower root maintenance respiration, compared to when N was derived from BNF. There was also an increase in the below-ground sink strength of NH₄⁺ fed AM roots compared to NH₄⁺ fed non-AM roots, as evidenced by the increases in root CO₂ and O₂ respiration and photosynthetic stimulation. These results indicate that although the AM root had higher total below-ground respiratory costs during NH₄⁺ nutrition, there were lower respiratory C costs associated with N derived from AM symbionts in comparison to N from BNF.     

Determination of nitrogen by microdiffusion in mason Jars. I. inorganic nitrogen in soil extracts

Abstract

Simple microdiffusion methods are described for determination of NH₄ ⁺, NO₃ ^‐, and NO₂ ^‐ in soil extracts. These methods involve diffusion of NH₃ in a 473‐mL (1‐pint) wide‐mouth Mason jar, the diffused NH₃‐N being collected in 3 mL of boric acid‐indicator solution in a 60 mm (dia.) Petri dish suspended from the Mason jar lid, for quantitative determination by titrimetry (0.0025 M H₂SO₄). Magnesium oxide is used to liberate NH₄ ⁺; Devarda's alloy is used to reduce NO₃‐ and NO₂ ^‐ to NH₄ ⁺; and sulfamic acid is used to eliminate NO₂ ^‐. Depending upon the volume of soil extract (10–50 mL), diffusion at room temperature (a20°C) was complete in 18–72 h with orbital shaking, and in 24–86 h without shaking. The methods gave quantitative recovery of NH₄ ⁺, NO₃ ^‐, and NO₂ ^‐added to soil extracts. A potential source of interference in the methods involving use of Devarda's alloy is the liberation of NH₄ ⁺‐N from alkali‐labile organic‐N compounds.     

Root Carbon Enrichment Alleviates Ammonium Toxicity in Cucumber Plants

ABSTRACT

The addition of carbonates to a nutrient solution to alleviate ammonium (NH₄ ⁺) toxicity in hydroponically-grown cucumber (Cucumis sativus L.) plants was investigated. Stable isotopes [nitrogen (¹⁵N) and carbon (¹³C)] were used to assess the uptake of nitrogen [NH₄ ⁺ or nitrate (NO₃ ^?)] as well as carbon [bicarbonate (HCO₃ ^?)/carbonate (CO₃ ^2?)] by the roots. Ammonium as the sole N source at 5 mM decreased plant fresh weights compared to NO₃ ^?. However, at lower concentrations of NH₄ ⁺ (25% of 5 mM total N), growth was increased compared to NO₃ ^? alone. Inorganic C enrichment [calcium carbonate (CaCO₃)] of the nutrient solution increased the fresh weight of NH₄ ⁺ grown plants with up to 150% relative to control plants receiving calcium hydroxide [Ca(OH)₂] for pH regulation. Root ¹⁵N enrichment was lower in ¹⁵NH₄ ⁺ supplied plants compared to ¹⁵NO₃ ^?, while the ¹³C enrichment in leaves was increased by NH₄ ⁺ nutrition compared to NO₃ ^? or NH₄NO_3. The enhanced C capture was associated with high PEPCase activity in the roots. It is concluded that inorganic carbon enrichment of the root medium may alleviate NH₄ ⁺ toxicity via increased synthesis of C skeletons and, accordingly, increased capacity for NH₄ ⁺ assimilation and N export to the shoots.     

A method to assess whether ‘preferential use’ occurs after N ammonium addition; implication for the N isotope dilution technique

The robustness of the assumption of equilibrium between native and added N during ¹⁵N isotope dilution has recently been questioned by Watson et al. (Soil Biol Biochem 32 (2000) 2019-2030). We re-analyzed their raw data using equations that consider the added and native NH₄⁺ and NO₃⁻ pools as separate state variables. Gross mineralization rates and first-order rate constants for NH₄⁺ and NO₃⁻ consumption were obtained by combining analytical integration of the differential equations with a non-linear fitting procedure. The first-order rate constants for NH₄⁺ consumption and NO₃⁻ immobilization for the added NH₄⁺ and NO₃⁻ pool were used to estimate gross mineralization rates and first-order rate constants for nitrification of native NH₄⁺. The latter were 2-4 times lower than the first-order rate constants derived from the added N pool. This discrepancy between first-order rate constants for nitrification implies that one or more process rates estimated for the added N pools cannot be applied to the native N pools. Preferential use of the added N resulted in an overestimation of the gross mineralization by 1.5-2.5-fold, emphasizing the need for critical evaluation of the assumption of equilibrium before gross mineralization rates are calculated.     

Evidence of carbon stimulated N transformations in grassland soil after slurry application

High nitrification rates which convert ammonium (NH₄⁺) to the mobile ions NO₂⁻ and NO₃⁻ are of high ecological significance because they increase the potential for N losses via leaching and denitrification. Nitrification can be performed by chemoautotrophic or heterotrophic organisms and heterotrophic nitrifiers can oxidise either mineral (NH₄⁺) or organic N. Selective nitrification inhibitors and ¹⁵N tracer studies have been used in an attempt to separate heterotrophic and autotrophic nitrification. In a laboratory study we determined the effect of cattle slurry on the oxidation of mineral NH₄⁺-N and organic-N by labelling the NH₄⁺ or NO₃⁻ pools separately or both together with ¹⁵N. The size and enrichment of the mineral N pools were determined at intervals. To calculate gross N transformation rates a ¹⁵N tracing model was developed. This model consists of the three N-pools NH₄⁺, NO₃⁻ and organic N. Sub-models for decomposition of degradable carbon in the soil and the slurry were added to the model and linked to the N transformation rates. The model was set up in the software ModelMaker which contains non-linear optimization routines to determine model parameters. The application of cattle slurry increased the rate of nitrifcation by a factor of 20 compared with the control. The size and enrichment of the mineral N pools provided evidence that nitrification was due to the conversion of NH₄⁺ to NO₃⁻ and not the conversion of organic N to NO₃⁻. There was evidence that slurry-enhanced oxidation of NH₄⁺ to NO₃⁻ was due to a combination of autotrophic and heterotrophic transformations. Slurry application increased the mineralisation rate by approximately a factor of two compared with the control and the rate of immobilisation of NH₄⁺ by approximately a factor of three.     

不同铵钾比对高铵下拟南芥地上部和根系生长的影响

钾在缓解植物铵毒害的过程中起着重要的作用。本文研究了高铵(30 mmol/L)条件下,不同铵钾比(7.5︰1和150︰1)对拟南芥(Col-0)主根、侧根以及地上部生长的影响。结果表明:30 mmol/L NH4+条件下,高铵钾比(150)处理显著加重了拟南芥铵毒害现象,地上部和根系生长所受的抑制作用更为明显并导致更严重的氧化胁迫。相比低铵钾比水平,在高铵处理下,高铵钾比使得拟南芥主根伸长量降低57.4%,侧根数量减少33.3%,而地上部鲜重减轻69.9%。DAB(3,3￠-二氨基联苯胺,3,3￠-diaminobenzidine)叶片染色结果表明,不加铵处理下,外源不同钾水平(0.2和4.0 mmol/L)对拟南芥叶片的氧化胁迫作用没有显著差异;而高铵处理下,相比低铵钾比处理,高铵钾比显著增加了叶片中过氧化氢的含量,加重了其氧化胁迫。伊文思蓝(Evans blue,EB)染色结果表明,不加铵处理下,外源不同钾水平对拟南芥地上部和根部的膜透性没有显著差异,而高铵处理下,高铵钾比显著增强了拟南芥地上部和根部的膜透性,表明其对细胞的伤害程度加重。可见,高铵抑制拟南芥根系和地上部生长,高铵钾比则会加重这种抑制,其原因除了高浓度钾能减少植物对铵的吸收外,可能与高铵钾比条件加剧了植物的氧化胁迫有关。因此,适宜的铵钾比在植物应对铵毒害的过程中发挥重要作用。     

Ammonium immobilisation during chloroform fumigation

The assumption in using the chloroform fumigation technique for microbial biomass determination is that microbes are killed or at least inactivated by the treatment. Problems associated with transformations of the N released on or during fumigation have so far only been associated with the fumigation-incubation method. A laboratory and a field study were carried out to investigate the possible N transformations during biomass determination by the fumigation-extraction method. Labelled NH₄NO₃ (either the NO₃⁻, NH₄⁺ or both pools were ¹⁵N enriched) was applied to the soil and biomass determinations made at intervals subsequently. The size and enrichment of the ammonium (NH₄⁺), and nitrate (NO₃⁻) pools were determined before and after chloroform fumigation. The ¹⁵N enrichment of the NH₄⁺ pool after fumigation could only be explained if immobilisation of ammonium occurred at some time during the 24 h fumigation period. The extent of this immobilisation was calculated. In addition, there was evidence that nitrification occurred during the fumigation procedure at the start of the laboratory study and throughout the field study. The laboratory and field study differed mainly in the dynamics related to NO₃⁻ uptake and release. There was evidence for uptake of NO₃⁻ by the microbial biomass with and without utilization. We conclude that the ¹⁵N enrichment in the microbial biomass cannot be accurately determined when N transformations and release of non-utilized N occurs during fumigation. The possible immobilisation of mineral N during fumigation will affect the magnitude of the factor used to convert measured microbial biomass N to actual microbial biomass N in soil.     

Comparison of NH4 pool dilution techniques to measure gross N fluxes in a coarse textured soil

We compared gross N fluxes by ¹⁵N pool dilution in a coarse-textured agricultural soil when ¹⁵N was applied to the soil NH₄⁺ pool by either: (i) mixing a ¹⁵NH₄NO₃ solution into disturbed soil or (ii) injection of ¹⁵NH₃ gas into intact soil cores. The two techniques produced similar results for gross N mineralization rates indicating that NH₄⁺ production in soil was not altered by soil disturbance, method of application (gas vs. solution), or amount of N applied. This was not the case for immobilization rates, which were twofold higher when ¹⁵N label was applied to the soil NH₄⁺ pool with the mixing technique compared to the injection technique. This was attributed to the fact that more NH₄⁺ was applied with the mixing technique. Estimates of gross nitrification were accompanied by large error terms meaning differences between ¹⁵N labeling methods could not be accurately assessed for this process rate.