The automated determination of glucosamine,galactosamine, muramic acid,and mannosamine in soil and root hydrolysates by HPLC

Amino sugars make a significant contribution to soil organic N and are mainly of microbial origin. The most important amino sugars in soil are glucosamine, galactosamine, muramic acid, and mannosamine. A method was developed for the simultaneous determination of these four amino sugars by high‐performance reverse‐phase liquid chromatography in standard solutions, soil and root hydrolysates. Pre‐column derivatization with o‐phthaldialdehyde (OPA) was used in an automated sample injector with thermostatic regulation of the reagent at 4 °C. The separation of the four amino sugars was fully satisfactory and was not disturbed by other fluorescent components in the soil and root hydrolysates.     

Amino sugars and muramic acid—biomarkers for soil microbial community structure analysis

Characterizing functional and phylogenetic microbial community structure in soil is important for understanding the fate of microbially-derived compounds during the decomposition and turn-over of soil organic matter. This study was conducted to test whether amino sugars and muramic acid are suitable biomarkers to trace bacterial, fungal, and actinomycetal residues in soil. For this aim, we investigated the pattern, amounts, and dynamics of three amino sugars (glucosamine, mannosamine and galactosamine) and muramic acid in the total microbial biomass and selectively cultivated bacteria, fungi, and actinomycetes of five different soils amended with and without glucose. Our results revealed that total amino sugar and muramic acid concentrations in microbial biomass, extracted from soil after chloroform fumigation varied between 1 and 27 mg kg⁻¹ soil. In all soils investigated, glucose addition resulted in a 50-360% increase of these values. In reference to soil microbial biomass-C, the total amino sugar- and muramic acid-C concentrations ranged from 1-71 g C kg⁻¹ biomass-C. After an initial lag phase, the cultivated microbes revealed similar amino sugar concentrations of about 35, 27 and 17 g glucosamine-C kg⁻¹ TOC in bacteria, fungi, and actinomycetes, respectively. Mannosamine and galactosamine concentrations were lower than those for glucosamine. Mannosamine was not found in actinomycete cultures. The highest muramic acid concentrations were found in bacteria, but small amounts were also found in actinomycete cultures. The concentrations of the three amino sugars studied and muramic acid differed significantly between bacteria and the other phylogenetic microbial groups under investigation (fungi and actinomycetes). Comparison between the amino sugar and muramic acid concentrations in soil microbial biomass, extracted after chloroform fumigation, and total concentrations in the soil showed that living microbial biomass contributed negligible amounts to total amino sugar contents in the soil, being at least two orders of magnitude greater in the soils than in the soil inherent microbial biomass. Thus, amino sugars are significantly stabilized in soil.     

Land-use effects on amino sugars in particle size fractions of an Argiudoll

Identifying amino sugar pools from different land-use systems may advance our knowledge of land-use effects on the fate of microbially-derived substances. Surface soils (0–10 cm) from (1) native pasture, (2) a >80-years-arable site, and (3) a >80-years-afforested site were fractionated into clay, silt, fine-, and coarse-sand fractions. Then, soil organic carbon, N, glucosamine, galactosamine, mannosamine, and muramic acid were analyzed.Afforestation did not influence the amino sugar content in bulk soil, whereas cultivation reduced the content by 54%. The concentrations of amino sugars in g kg⁻¹ SOM declined after both long-term cropping and afforestation by 6% and 13%, respectively, relative to that in the grassland. The amino sugar depletion at the forest site occurred mainly from the silt fraction (by 25%), while that in the cultivated site was mainly due to preferential loss of amino sugars from clay (by 19% compared with the grassland). Both ratios of glucosamine to galactosamine and glucosamine to muramic acid increased when the prairie was converted to forest or cultivated land, suggesting that bacterial N especially is better preserved than fungal N under prairie conditions.     

Land use effects on amino sugar signature of chromic Luvisol in the semi-arid part of northern Tanzania

 Characterizing amino sugar signature in particle size separates of tropical soils is important for further understanding the fate of microbial-derived compounds during the decomposition of soil organic matter (SOM) in tropical agroecosystems. We investigated the impact of land-use changes on the nature, amount and dynamics of amino sugars in soil of the semi-arid northern Tanzania. Samples were collected from the uppermost 10 cm of native woodland, degraded woodland, fields cultivated for 3 and 15 years and homestead fields fertilized with animal manure. The amount of glucosamine, galactosamine, mannosamine and muramic acid were determined in bulk soil and size separates. Compared to the native woodland, a 68% and 72% reduction in total amino sugar contents were found in the 3- and 15-year cultivated fields, respectively. Moreover, 39% of the total amino sugar was lost from the degraded woodland. This may be attributed to accelerated decomposition of amino sugars and/or decreasing microbial biomass input under the semi-arid environment following clear-cutting and cultivation. In contrast, only a 20% decline was found from the fields where animal manure had been applied. Most of the amino sugar depletion occurred from the coarse and fine sand-associated SOM. The decline from the silt and clay-bound amino sugar was relatively small, indicating the importance of organo-mineral associations in the stabilization of microbial-derived sugars in this tropical soil. After 15 years of continuous cultivation, the ratio of glucosamine:galactosamine increased from 1.44 to 2.23, while the ratio of glucosamine:muramic acid increased from 14.5 to 26.5 (P<0.05). These results suggest that cultivation may have led to preferential depletion of bacterial-derived amino sugars (muramic acid and galactosamine) compared with fungal-derived glucosamine. Received: 22 June 2000     

Amino sugars in soils of the North American cultivated prairie

Characterizing amino sugar dynamics in cultivated soils helps to further understand the influence of cultivation on soil organic matter turnover. This study was designed to evaluate accumulations and patterns of four amino sugars in 17 surface (0–10 cm) soil samples along a climosequence in the North American long-term cultivated prairie from Saskatoon, Candada, to Texas, USA. Mean annual temperature (MAT) ranged from 0.9 to 22.2°C and mean annual precipitation (MAP) from 300 to 1308 mm. Samples were analyzed for glucosamine, mannosamine, galactosamine, and muramic acid. Amino sugar contents (mg kg^?1 soil) varied markedly among the 17 sites and were controlled by mean annual temperature (MAT) and clay and silt contents, mainly. The relationship between amino sugar-N proportions to total N (%) and MAT followed parabolic regression models. Compared with native sites, amino sugars were depleted by 53% and the amino sugar-N by 18% of the total, on average, after long-term cropping. The intensity of amino sugar-N depletion correlated positively with MAT (r = 0.77^***). Bacterially-derived galactosamine and muramic acid declined preferentially to mainly chitin-derived glucosamine after long-term cropping. The glucosamine-to-galactosamine and glucosamine-to-muramic acid ratios can be used, therefore, as indicators for the identification of land use effects on microbially-derived SOM residues.     

A novel GC/MS technique to assess N and C incorporation into soil amino sugars

Amino sugars have been used as biomarker to indicate microorganism contribution to soil organic matter turnover and sequestration. However, there is no direct gas chromatograph mass spectrometry (GC/MS) approach to assess microbial synthesis of amino sugars in soil. We developed a novel method which combines laboratory incubation of substrate containing ¹⁵N or ¹³C and a GC/MS technique to trace ¹⁵N or ¹³C isotope changes in three amino sugars, glucosamine, galactosamine, and muramic acid. Sample preparation followed the procedure of Zhang and Amelung (1996) [Zhang, X., Amelung, W., 1996. Gas chromatographic determination of muramic acid, glucosamine, galactosamine, and mannosamine in soils. Soil Biology and Biochemistry 28, 1201-1206.]. The GC/MS determination was conducted using a full scan mode with both electronic ionization (EI) and chemical ionization (CI) sources. The CI source was suitable for all of the three amino sugars, while the EI source was not applicable to muramic acid due to its low sensitivity in the determination as well as low concentration of muramic acid in soil. The enrichment of ¹⁵N or ¹³C in amino sugars during incubation was estimated by calculating the atom percentage excess (APE). ¹⁵N incorporation was evaluated according to fragment (F) abundance ratio of mass F+1 to F, whilst ¹³C incorporation was estimated according to the ratio of mass F+n to F (n is skeleton carbon number in the fragment). This novel method was assessed by using two soil samples (a Kandiudult and a Udoll) incubated with either ¹⁵N-amonium or U-¹³C-glucose. The results indicate that the GC/MS determination is reproducible, thus this technique is useful in detecting the microbial synthesis of amino sugars in soil, and especially it should be possible when looking at the position or how much labeled carbon and nitrogen atoms have been incorporated.     

Shifts in amino sugar and ergosterol contents after addition of sucrose and cellulose to soil

An incubation experiment with organic soil amendments was carried out with the aim to determine whether formation and use of microbial tissue (biomass and residues) could be monitored by measuring glucosamine and muramic acid. Living fungal tissue was additionally determined by the cell-membrane component ergosterol. The organic amendments were fibrous maize cellulose and sugarcane sucrose adjusted to the same C/N ratio of 15. In a subsequent step, spherical cellulose was added without N to determine whether the microbial residues formed initially were preferentially decomposed. In the non-amended control treatment, ergosterol remained constant at 0.44 μg g⁻¹ soil throughout the 67-day incubation. It increased to a highest value of 1.9 μg g⁻¹ soil at day 5 in the sucrose treatment and to 5.0 μg g⁻¹ soil at day 33 in the fibrous cellulose treatment. Then, the ergosterol content declined again. The addition of spherical cellulose had no further significant effects on the ergosterol content in these two treatments. The non-amended control treatment contained 48 μg muramic acid and 650 μg glucosamine g⁻¹ soil at day 5. During incubation, these contents decreased by 17% and 19%, respectively. A 33% increase in muramic acid and an 8% increase in glucosamine were observed after adding sucrose. Consequently, the ratio of fungal C to bacterial C based on bacterial muramic acid and fungal glucosamine was lowered in comparison with the other two treatments. No effect on the two amino sugars was observed after adding cellulose initially or subsequently during the second incubation period. This indicates that the differences in quality between sucrose and cellulose had a strong impact on the formation of microbial residues. However, the amino sugars did not indicate a preferential decomposition of microbial residues as N sources.     

Temporal responses of soil microorganisms to substrate addition as indicated by amino sugar differentiation

Amino sugars are one of the important microbial residue biomarkers which are associated with soil organic matter cycling. However, little is known about their transformation kinetics in response to available substrates because living biomass only contributes a negligible portion to the total mass of amino sugars. By using ¹⁵N tracing technique, the newly synthesized (labeled) amino sugars can be differentiated from the native portions in soil matrix, making it possible to evaluate, in quantitative manner, the transformation pattern of amino sugars and to interpret the past and ongoing changes of microbial communities during the assimilation of extraneous ¹⁵N. In this study, laboratory incubations of soil samples were conducted by using ¹⁵NH₄⁺ as nitrogen source with or without glucose addition. Both the ¹⁵N enrichment (expressed as atom percentage excess, APE) and the contents of amino sugars were determined by an isotope-based gas chromatography-mass spectrometry. The significant ¹⁵N incorporation into amino sugars was only observed in glucose plus ¹⁵NH₄⁺ amendment with the APE arranged as: muramic acid (MurN) > glucosamine (GlcN) > galactosamine (GalN). The dynamics of ¹⁵N enrichment in bacterial-derived MurN and fungal-derived GlcN were fitted to the hyperbolic equations and indicative for the temporal responses of different soil microorganisms. The APE plateau of MurN and fungal-derived GlcN represented the maximal extent of bacterial and fungal populations, respectively, becoming active in response to the available substrates. The different dynamics of the ¹⁵N enrichment between MurN and GlcN indicated that bacteria reacted faster than fungi to assimilate the labile substrates initially, but fungus growth was dominant afterward, leading to integrated microbial community structure over time. Furthermore, the dynamics of labeled and unlabeled portions of amino sugars were compound-specific and substrate-dependent, suggesting their different stability in soil. GlcN tended to accumulate in soil while MurN was more likely degraded as a carbon source when nitrogen supply was excessive.     

Amino sugars as specific indices for fungal and bacterial residues in soil

Amino sugars are important indices for the contribution of soil microorganisms to soil organic matter. Consequently, the past decade has seen a great increase in the number of studies measuring amino sugars. However, some uncertainties remain in the interpretation of amino sugar data. The objective of the current opinion paper is to summarize current knowledge on amino sugars in soils, to give some advice for future research objectives, and to make a plea for the correct use of information. The study gives an overview on the origin of muramic acid (MurN), glucosamine (GlcN), galactosamine (GalN), and mannosamine (ManN). Information is also provided on measuring total amino sugars in soil but also on compound-specific δ¹³C and δ¹⁵N determination. Special attention is given to the turnover of microbial cell-wall residues, to the interpretation of the GlcN/GalN ratio, and to the reasons for converting fungal GlcN and MurN to microbial residue C. There is no evidence to suggest that the turnover of fungal residues generally differs from that of bacterial residues. On average, MurN contributes 7% to total amino sugars in soil, GlcN 60%, GalN 30%, and ManN 4%. MurN is highly specific for bacteria, GlcN for fungi if corrected for the contribution of bacterial GlcN, whereas GalN and ManN are unspecific microbial markers.     

Microbial colonisation of roots as a function of plant species

Fifteen plants species were grown in the greenhouse on the same soil and sampled at flowering to obtain rhizosphere soil and root material. In both fractions, the data on fungal and bacterial tissue obtained by amino sugar analysis were compared with the total microbial biomass based on fumigation-extraction and ergosterol data. The available literature on glucosamine concentrations in fungi and on muramic acid concentrations in bacteria was reviewed to prove the possibility of generating conversion values for general use in root material. All microbial properties analysed revealed strong species-specific differences in microbial colonisation of plant roots. The root material contained considerable amounts of microbial biomass C and biomass N, reaching mean levels of 10.9 and 1.4 mg g⁻¹ dry weight, respectively. However, the majority of CHCl₃ labile C and N, i.e. 89 and 55% was root derived. The average amount of ergosterol was 13 μg g⁻¹ dry weight and varied between 0.0 for Phacelia roots and 45.5 μg g⁻¹ dry weight for Vicia roots. The ergosterol content in root material of mycorrhizal and non-mycorrhizal plant species did not differ significantly. Fungal glucosamine was converted to fungal C by multiplication by 9 giving a range of 7.1-25.9 mg g⁻¹ dry weight in the root material. Fungal C and ergosterol were significantly correlated. Bacterial C was calculated by multiplying muramic acid by 45 giving a range from 1.7 to 21.6 mg g⁻¹ dry weight in the root material. In the root material of the 15 plant species, the ratio of fungal C-to-bacterial C ranged from 1.0 in mycorrhizal Trifolium roots to 9.5 in non-mycorrhizal Lupinus roots and it was on average 3.1. These figures mean that the microbial tissue in the root material consists on average of 76% fungal C and 24% bacterial C. The differences in microbial colonisation of the roots were reflected by differences in microbial indices found in the rhizosphere soil, most strongly for microbial biomass C and ergosterol, but to some extent also for glucosamine and muramic acid.     

Kinetics of soil microbial uptake of free amino acids

 Amino acids and proteins typically form the biggest input of organic-N into most soils and provide a readily available source of C and N for soil microorganisms. Amino acids can also be taken up directly by plant roots, providing an alternative source of available soil N. However, the degree to which plants can compete against the soil microbial population for amino acids in soil solution remains poorly understood. The aim of this study was to measure the rate of microbial uptake of three contrastingly charged ¹⁴C-labelled amino acids (glutamate^1–, glycine⁰, lysine^0.9+) over a wide concentration range (0.1–5 mM) and in two contrastingly managed soils varying in their degree of erosion, organic-C content and microbial biomass. Amino acid uptake was concentration dependent and conformed to a single Michaelis-Menten equation. The mean maximum amino acid uptake rate (V _max) for the non-eroded (control) soil (high organic-C, high biomass) was 0.13±0.02 mmol kg^–1 h^–1, while half maximal uptake occurred at a concentration (K _m) of 2.63±0.07 mM. Typically, V _max was fourfold lower and K _m twofold lower in the eroded soil (low available organic-C, low biomass) compared to the non-eroded (control) soil. Amino acid substrate concentration had little effect on the proportion of amino acid utilized in catabolic versus anabolic metabolism and was similar for both. While the results obtained here represent the summation of kinetics for a mixed soil population, they indicate that amino acid uptake is saturated at concentrations within the millimolar range. Because the affinity constants also were similar to those described for plant roots, we hypothesized that competition for amino acids between plants and microbes will be strong in soil but highly dependent upon the spatial distribution of roots and microbes in soil. Received: 2 March 2000     

尿素向氨基糖的转化以及对土壤氨基糖库动态的影响

采用13CO(NH2)2为底物进行黑土培养实验,利用气相色谱/质谱技术测定土壤中三种氨基糖含量以及同位素富集比例,根据其微生物标识物作用探讨土壤中不同微生物群落对于尿素碳的同化利用特征及黑土氨基糖库对于尿素添加的响应。研究结果表明,尿素碳可以被土壤微生物同化利用,但是可利用性显著低于葡萄糖。氨基葡萄糖中13C富集比例显著高于胞壁酸,表明真菌对尿素碳的同化能力高于细菌。尿素添加使土壤有机碳含量有所下降,同时土壤氨基糖总量及其与有机碳的相对比例也显著降低,说明在碳源严重受限条件下,氨基糖可被优先分解利用以补充碳源供给。胞壁酸含量虽低,但其调节并平衡碳氮元素供给与需求的能力较强;氨基葡萄糖稳定性高于胞壁酸,但在碳源缺乏时也可部分分解。土壤氨基糖的动态与土壤碳氮的可利用性及其耦合作用密切相关,在平衡土壤碳氮需求方面具有一定的调节作用。     

Net microbial amino sugar accumulation process in soil as influenced by different plant material inputs

Identifying the impact of plant material inputs on soil amino sugar synthesis may advance our knowledge of microbial transformation processes in soils. In a 12-week laboratory microcosm incubation, 1, 2, 4, and 6% (w/w) soybean leaf or maize stalk were initially added to soil, respectively, whereas soil without plant addition was used as a control. The results showed that adding organic materials to the soil led to a net accumulation of amino sugars, because of greater microbial synthesis. The ratios of glucosamine to galactosamine and of glucosamine to muramic acid, two indicators differentiating the relative contribution to soil organic matter of fungi and bacteria, showed substantial variance across the gradient of substrate addition. Our results suggest that the amount of nutrients in a given substrate is the primary attribute determining microbial net accumulation of soil amino sugars, especially in the relatively short term, whereas the composition of nutrients might be more important in the relatively long term when nutrients are not sufficient. The use of the two ratios (glucosamine to galactosamine and glucosamine to muramic acid) reflects different dynamics of galactosamine and muramic acid during the decomposition of organic substrates in soils. Muramic acid, compared with galactosamine, is more likely to accumulate in the soil active organic fraction under abundant nutrient conditions, whereas it would be decomposed along with active organic matter when the nutrients are scarce and remain in minor quantities in the clay fraction without being attacked by microbes.     

Influence of cow manure vermicompost on the growth, metabolite contents, and antioxidant activities of Chinese cabbage (Brassica campestris ssp. chinensis)

The effects of cow manure vermicompost on plant growth, metabolite contents, and antioxidant activities of Chinese cabbage were investigated in pot cultures. Five treatments were designed by mixing vermicompost and soil at ratio of 0:7, 1:7, 2:7, 4:7, 7:0 (w/w). Marketable weight of Chinese cabbage was significantly (p < 0.05) higher in the 2:1 treatment than in the other treatments, while plants grown in the full soil treatment (0:7) showed the lowest marketable weight (Fig. 1a). Vermicompost application significantly increased the nutrient content of Chinese cabbage leaves (p < 0.05), especially in the 4:7 treatment, with increases in the contents of soluble sugar (Fig. 2a), soluble protein (Fig. 2b), vitamin C (Fig. 3a), total phenols (Fig. 3b), and total flavonoids (Fig. 3c) by 62%, 18%, 200%, 25%, and 17% compared to the full soil treatment, respectively. The antioxidant activities expressed by 2, 2-Dipenyl-1-picrylhydrazyl-scavenging activity (Fig. 4a), hydroxyl (OH)-scavenging activity (Fig. 4b), and iron (Fe²⁺)-chelating activity (Fig. 4c) were higher by 92%, 40%, and 36% in the 4:7 than 0:7 the treatment, respectively. Vermicompost application significantly increased (p < 0.05) the plant contents of 16 essential amino acids (Table 1); the total amino acid content showed the greatest increase in the 4:7 treatment, 90% compared to the full soil treatment.     

Development of a capillary electrophoresis–mass spectrometry method for small peptides in the soil solution

Small peptides are being investigated for their role in ecosystem cycling and plant uptake of organic N, but little is known of molecular forms in the soil solution. The aim of this study was to develop a capillary electrophoresis–tandem mass spectrometry procedure for profiling small peptides in the soil solution. Capillary electrophoresis–mass spectrometry was capable of separating and detecting a range of small peptide standards. Adequate recovery (>90%) of standard peptides spiked into samples of soil solution indicated that separation and detection were robust and not significantly affected by the sample matrix. The method was applied to samples of soil solution from grassland mesocosms filled with clay-loam soil from an abruptic lixisol. Soil solution (ultrafiltered <3 kDa) contained at least 298 putatively identified peptides with most being smaller than 600 Da. Less than 5% of small peptides contained basic amino acids, which may reflect their preferential adsorption to the soil stationary phase versus peptides comprised of acid or neutral amino acids. Capillary electrophoresis–mass spectrometry of small peptides is robust and has already yielded novel findings with its first proof of concept measurements on the soil solution.     

玉米不同残体添加对棕壤团聚体中氨基糖分配的影响

[目的]秸秆残体还田能引起土壤微生物残留物氨基糖的变化,然而不同部位秸秆残体因含碳氮化学组分差异,还田到不同肥力土壤后对氨基糖在团聚体中分配的影响尚不明析.因此,研究添加玉米不同残体对不同肥力棕壤团聚体中氨基糖分配的影响,并利用微生物标识物氨基葡萄糖与胞壁酸比值变化指示棕壤团聚体真菌和细菌群落组成动态变化,对深入阐明秸...     

Salt-tolerant rhizobacteria-mediated induced tolerance in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>) and chemical diversity in rhizosphere enhance plant growth

Salt-tolerant isolates Bacillus pumilus, Pseudomonas mendocina, Arthrobacter sp., Halomonas sp., and Nitrinicola lacisaponensis isolated from high saline habitats exhibited plant growth-promoting traits like P solubilization and indole acetic acid (IAA), siderophore, and ammonia production. These isolates were inoculated in wheat to assess microbe-mediated responses and plant growth promotion in salt affected soil. Maximum shoot and root length (33.8 and 13.6 cm) and shoot and root biomass (2.73 and 4.48 g dry weight) was recorded in plants inoculated with B. pumilus after 30 days. Total chlorophyll content was maximum in the leaves of the plants treated with Halomonas sp. (24.22 mg g⁻¹ dry weight) followed by B. pumilus (23.41 mg g⁻¹ dry weight) as compared to control (18.21 mg g⁻¹ dry weight) after 30 days. Total protein content was maximum in Arthrobacter sp. inoculated plant leaves (3.19 mg g⁻¹ dry weight) followed by B. pumilus (2.47 mg g⁻¹ dry weight) as compared to control (2.15 mg g⁻¹ dry weight) after 30 days. Total carotenoid content was maximum in plants inoculated with Halomonas sp. (1,075.45 and 1,113.29 μg g⁻¹ dry weight) in comparison to control (837.32 and 885.85 μg g⁻¹ dry weight) after 15 and 30 days. Inoculation of bacterial isolates increased presence of individual phenolics (gallic, caffeic, syringic, vanillic, ferulic, and cinnamic acids) and flavonoid quercetin in the rhizosphere soil. The concentration of IAA in rhizosphere soil and root exudates was also higher in all treatments than in control. Accumulation of phenolics and quercetin in the plants played a cumulative synergistic role that supported enhanced plant growth promotion of wheat in the stressed soil.     

Plant Availability and Uptake of Lead, Zinc, and Cadmium in Soils Contaminated with Anti-corrosion Paint from Pylons in Comparison to Heavy Metal Contaminated Urban Soils

Red lead (Pb₃O₄) has been used extensively in the past as an anti-corrosion paint for the protection of steel constructions. Prominent examples being some of the 200,000 high-voltage pylons in Germany which have been treated with red lead anti-corrosion paints until about 1970. Through weathering and maintenance work, paint compounds and particles are deposited on the soils beneath these constructions. In the present study, six such “pylon soils” were investigated in order to characterize the plant availability and plant uptake of Pb, Cd, and Zn. For comparison, three urban soils with similar levels of heavy metal contamination were included. One phase extractions with 1 M NH₄NO₃, sequential extractions (seven steps), and extractions at different soil pH were used to evaluate the heavy metal binding forms in the soil and availability to plants. Greenhouse experiments were conducted to determine heavy metal uptake by Lolium multiflorum and Lactuca sativa var. crispa in untreated and limed red lead paint contaminated soils. Concentrations of Pb and Zn in the pylon soils were elevated with maximum values of 783 mg Pb kg⁻¹ and 635 Zn mg kg⁻¹ while the soil Cd content was similar to nearby reference soils. The pylon soils were characterized by exceptionally high proportions of NH₄NO₃-extractable Pb reaching up to 17% of total Pb. Even if the relatively low pH of the soils is considered (pH 4.3–4.9), this appears to be a specific feature of the red lead contamination since similarly contaminated urban soils have to be acidified to pH 2.5 to achieve a similarly high Pb extractability. The Pb content in L. multiflorum shoots reached maximum values of 73 mg kg⁻¹ after a cultivation time of 4 weeks in pylon soil. Lime amendment reduced the plant uptake of Pb and Zn significantly by up to 91%. But L. sativa var. crispa cultivated on soils limed to neutral pH still contained critical Pb concentrations (up to 0.6 mg kg⁻¹ fresh weight). Possible mechanisms for the exceptionally high plant availability of soil Pb derived from red lead paint are discussed.     

Regulation of amino acid biodegradation in soil as affected by depth

Dissolved organic nitrogen (DON) and in particular free amino acids represent a key pool in the terrestrial soil C and N cycle. The factors controlling the rate of turnover of this pool in soil, however, remain poorly understood. We investigated the factors regulating the rate of amino acid turnover at different depths (up to 1.2 m) in five low-input, acid soil profiles. Within the root zone (0–60 cm), amino acids constituted 8% of the DON and represented only a small fraction of plant available N. In all the soil profiles, the rate of amino acid mineralisation decreased progressively with depth. The average half-life of the exogenously added amino acids in the soil was 5.8 h in topsoils (0–10 cm), falling to 20 h at a depth of 50 cm and to 33 h at 100 cm. Generally, the rate of amino acid mineralisation correlated positively with total soil C and N, soil microbial activity (basal soil respiration rate) and soil content. The relatively rapid rates of microbial amino acid assimilation in subsoils below the root zone (>60 cm) indicate that long-term transport of amino acids (e.g. from soil to freshwaters) will be low. Based upon the C-to-N ratio of the amino acid substrate and the microbial C assimilation efficiency, we estimate that approximately 40–60% of the amino acid-N will be excreted as . In conclusion, the rapid rate of free amino acid turnover and their low concentration in soil solution indicate that the formation of inorganic N ( and ) in soil is limited primarily by the rate of free amino acid appearance in soil and not by the rate of amino acid mineralisation.     

Determination of amino acid enantiomers in soils

Determination of amino acid enantiomers in environmental samples is difficult, because metals and organic impurities interfere with the analyses. We developed a new gas chromatographic method to assess amino acid enantiomer concentrations in complex soil matrices following hot HCl hydrolysis (6 M, 12 h, 105°C). The crucial focus was the establishment of a simple, reliable sample clean-up procedure. The presented method involved the adsorption of the enantiomers on a Dowex 50 W X8 cation exchange resin and the removal of interfering compounds with 0.1 M oxalic acid prior to amino acid elution with 2.5 M NH₄OH. After conversion to N-pentafluoropropionyl-amino acid iso-propyl esters, the diastereomers were separated by a Chirasil l-Val capillary column and quantified by a flame ionization or mass selective detector. The lower limit of quantification tested here was ≤1 pg injection amount. The recovery of amino acid enantiomers averaged 99±11% for pure standards and 97±11% for spiked soil hydrolysates. The general applicability of the method was demonstrated by determination of amino acid enantiomers in taxonomically different soils from different geographic regions. The coefficients of variation presented by d/l ratios were 12% for alanine and <10% for the other amino acids.