Effects of deforestation on phosphorus pools in mountain soils of the Alay Range, Khyrgyzia

 The amount, quality and turnover of soil P is heavily influenced by changes in soil management. The objective of this study was to investigate the effects of deforestation and pasture establishment on the concentrations, forms and turnover rate of soil P in mountain soils of the Alay Range, Khyrgyzia. A sequential extraction was applied to distinguish soil P pools. We used particle-size fractionation to follow the dynamics of different P pools in soils under forest and pasture and ³¹P-NMR spectroscopy to investigate the structure of alkali-soluble P forms. In the A horizons of the forest soils, total soil P concentration was 1093 mg kg^–1, organic P (P_o) representing 46% of the total P. Deforestation followed by pasture establishment not only increased significantly (P<0.01) the total P concentration (1560 mg kg^–1) but also the contribution of P_o to total P was increased by 17%. Pasture soils had significantly higher P pools than forest soils except highly labile inorganic P (P_i NaHCO₃) and primary P_i (P_i HCl_dil). Both in forest and pasture soils stable P increased with decreasing particle size (coarse sand 50%, clay 80% of total P) and primary P decreased with decreasing particle size. Phosphate monoesters and diesters represented 80% of P identified by ³¹P NMR. Low monoester to diester ratios in the alkali extracts of forest and pasture soils indicate low microbial activity. This is consistent with high C/P_o ratios and high stable P_o concentrations in the fine earth of forest and pasture. Received: 10 March 1999     

Organic phosphorus speciation and pedogenesis: analysis by solution 31P nuclear magnetic resonance spectroscopy

Changes in phosphorus (P) during soil development are central to the understanding of labile P for plant productivity and soil P management. We used NaOH‐EDTA extraction with ³¹P nuclear magnetic resonance spectroscopy (³¹P NMR), sequential P fractionation, and general soil chemical characterization to better our understanding of P dynamics within two chronosequences (Manawatu and Reefton) and one Basalt maturity sequence under original native vegetation. With time, orthophosphate and orthophosphate monoesters tended to increase with organic C to a maximum of about two‐thirds of NaOH‐EDTA‐extractable P in young soils (16 000 years in the Reefton chronosequence), but gradually declined thereafter to about one‐third of NaOH‐EDTA‐extractable P in the oldest soils (130 000 years old). This coincided with a depletion of P from primary minerals (e.g. apatite) and readily available P for plant production. This depletion of inorganic P resulted in a greater reliance on organic P cycling via mineralization, hence the depletion of the normally recalcitrant monoester‐P pool. Concomitantly, the build‐up of labile P species (diesters and pyrophosphate) and scyllo‐ over myo‐inositol hexakisphosphate occurred as soils developed, and might be attributed to microbial activity, including scavenging for P. This work highlights the importance of organic P cycling during pedogenesis.     

Characterization of phosphorus in sub-alpine forest and adjacent grassland soils by chemical extraction and phosphorus-31 nuclear magnetic resonance spectroscopy

We used chemical extraction methods and ³¹P-nuclear magnetic resonance (NMR) to investigate the effects of vegetation on the amount and structural composition of phosphorous (P) in the sub-alpine soils of central Taiwan. Chemical extraction methods were used to measure inorganic P (Pi) and organic P (Po) in main soil horizons. The soil P composition was assessed by ³¹P-NMR spectroscopy on alkaline EDTA–NaOH extracts. According to the results of chemical extractions, the forest soil had a higher amount of Pi than the grassland soil, which might be a result of the mineralization of Po. ³¹P-NMR spectra showed inorganic orthophosphate (up to 67%) and orthophosphate monoesters (up to 75%) as the major forms of P extracted in forest and grassland soils, respectively. Smaller proportions of orthophosphate diesters and trace amounts of phosphonates and pyrophosphate were found. With possible hydrolysis of P compounds during chemical extraction and slight systemic error in the processes of extraction with NMR, the results from NMR analysis are, in general, consistent with those of chemical extraction.     

A 31P nuclear magnetic resonance study of the phosphorus species in alkali extracts of soils from long-term field experiments

The different forms of phosphorus (P) in 0.5 m sodium hydroxide extracts of soils from long-term field experiments at Rothamsted were characterized by ³¹P-nuclear magnetic resonance spectroscopy (NMR). The extract from an old grassland soil (pH 4.6) from a plot of the Park Grass Continuous Hay Experiment that had received no fertilizer or lime for at least 125 years contained the following forms of P: inorganic orthophosphate (22% of the extracted P), orthophosphate monoesters (49%), orthophosphate diesters (14%), phosphonates (3%), pyrophosphate (4%) and two unidentified forms of P (7%). The soil extract from a Park Grass plot given inorganic phosphate fertilizer (35 kg P ha^?1) annually for 121 years contained the same forms of P and, in addition, a small amount of polyphosphate. There was also evidence of an increase in the orthophosphate monoester fraction. Another old grassland soil, of pH 6.1, contained more total and organic P than Park Grass but the extract contained fewer forms of P: inorganic orthophosphate (14% of the extracted P), orthophosphate monoesters (39%), orthophosphate diesters (34%) and an unidentified form (13%). An area of this grassland that had been ploughed up 20 years previously, and kept bare since, contained less organic P. The extract contained less of the phosphate diesters but the more stable monoesters remained relatively unchanged.     

Phosphorus fractions of a climosequence of soils in New zealand tussock grassland

The different forms of phosphorus in alkaline extracts of eight New Zealand topsoils, which are members of a climosequence, were characterized by ³¹P-nuclear magnetic resonance (NMR). A further two topsoils were used in an experiment to demonstrate that the NMR technique detected all of the P in the extracts. This direct method of estimating organic P in soil extracts enabled the different types and the relative amounts of P compounds to be estimated.Inorganic orthophosphate and orthophosphate monoesters were the major P components of the extracts from all soils, while all but the two driest soils also contained orthophosphate diesters. Only the high country and alpine soils, developed in a moist cool environment, contained phosphonates, a recently-discovered form of soil P of probable microbial origin.Across the climosequence of soils, the amount of orthophosphate diesters in the extracts was strongly and positively correlated with annual precipitation. This organic P fraction, together with phosphonates, could provide through mineralization a ready supply of “available” P in these mainly undisturbed tussock grassland ecosystems.     

Soil organic phosphorus in tropical forests: an assessment of the NaOH–EDTA extraction procedure for quantitative analysis by solution 31P NMR spectroscopy

The extraction of soil organic phosphorus by the NaOH–EDTA procedure was assessed in detail for a tropical forest soil (clay‐loam, pH 4.3, total carbon 2.7%). Optimum conditions for the quantification of soil organic phosphorus and characterization of its composition by solution ³¹P NMR spectroscopy were extraction in a solution containing 0.25 m NaOH and 50 mm Na₂EDTA in a 1:20 solid to solution ratio for 4 hours at ambient laboratory temperature. Replicate analyses yielded a coefficient of variation of 3% for organic phosphorus as a proportion of the spectral area. There was no significant difference in total phosphorus extraction from fresh and air‐dried soil, although slightly more organic phosphorus and less paramagnetic ions were extracted from dried soil. The procedure was not improved by changing the concentration of NaOH or EDTA, extraction time, or solid to solution ratio. Pre‐extraction with HCl or Na₂EDTA did not increase subsequent organic phosphorus extraction in NaOH–EDTA or improve spectral resolution in solution ³¹P NMR spectroscopy. Post‐extraction treatment with Chelex resin did not improve spectral resolution, but removed small concentrations of phosphorus from the extracts. Increasing the pH of NaOH–EDTA extracts (up to 1.0 m NaOH) increased the concentration of phosphate monoesters, but decreased DNA to an undetectable level, indicating its hydrolysis in strong alkali. The standardized NaOH–EDTA extraction procedure is therefore recommended for the analysis of organic phosphorus in tropical forest soils.     

Organic phosphorus species in humic acids of mountain soils along a toposequence in the Northern Caucasus

Organic P was investigated in humic acids extracted from mountain soils developed in the subalpine, upper subalpine and alpine zones of the Northern Caucasus. P contents of humic acids varied between 3.4 and 14.2 g P kg^?1, depending on P contents of the parent vegetation and on site conditions. Organic P was accumulated at sites where microbial activity is restrained due to soil acidity, low soil temperature and hydromorphy.³¹ P NMR spectroscopy revealed that orthophosphate monoesters were the dominent P species (72–85% of extract- able P), orthophosphate diesters amounted to 12–21%, and phospho- nates ranged between 0 and 9%. Humic acids of soils under cold and wet climatic conditions showed highest concentrations in phospho- nates and orthophosphate diesters. Hence, the accumulation of organo-P in the Caucasian mountain soils was partly due to increasing proportions of potentially available organic P species.     

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

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

Soil phosphorus characterisation by 31p nuclear magnetic resonance

Abstract

Five New Zealand topsoils, including three sampled under introduced pasture and two in native tussock grasslands, were extracted with 0.5 M NaOH, and the different classes of phosphorus compounds in the extracts distinguished by ³¹p nuclear magnetic resonance (n.m.r.).

Inorganic orthophosphate and orthophosphate monoesters were the major forms of phosphorus in all soil extracts. The tussock‐grassland soil extracts showed the greatest diversity of phosphorus forms, and included compounds with n.m.r. signals ascribed to phosphonates, a previously unreported form of soil phosphorus.     

Phosphorus forms and dynamics as influenced by land use changes in the sub-humid Ethiopian highlands

In undisturbed tropical forest ecosystems, the phosphorus (P) cycle is essentially “closed” with minimal short-term losses or gains of P. The forms and dynamics of soil P, however, can be greatly affected by land use changes, which often involve changes in vegetation cover, biomass production and nutrient cycling in the ecosystem. Sequential extraction and ³¹P nuclear magnetic resonance (NMR) spectroscopy were used to investigate the influence of land use changes on the amount and structural composition of P in the sub-humid highlands of southern Ethiopia. Samples were collected from surface soils (0–10 cm) of natural forest, tea plantations and cultivated fields (25 years) at Wushwush and from Podocarpus dominated natural forest, Cupressus plantations and cultivated fields (30 years) at Munesa sites. Significantly lower (P<0.05) amounts of total P were found following clear-cutting and long-term cultivation (31% and 39%), and establishment of plantations (21% and 22%) at Wushwush and Munesa, respectively. The largest depletion of sodium bicarbonate-extractable organic P (NaHCO₃–P_o) (74% and 77%) and sodium hydroxide-extractable P_o (NaOH–P_o) (67% and 67%) due to cultivation occurred in the sand, followed by the silt (52% and 56%, NaHCO₃–P_o and 32% and 53%, NaOH–P_o) and the clay (33% and 42%, NaHCO₃–P_o and 28% and 35%, NaOH–P_o) size separates from the two sites, respectively. ³¹P NMR spectroscopy revealed that orthophosphate monoesters were the major organic P compounds (27–66%) followed by orthophosphate diesters (9–27%) and teichoic acids (7–11%). Unknown organic P species accounted for 3–8% (unknown A) and 3–5% (unknown B), whereas phosphonates made up 2–3%. The proportion of diester-P decreased in the order: natural forests (24% and 27%)>plantations (15% and 13%)>cultivated fields (10% and 9%) at Wushwush and Munesa, respectively. The percentages of teichoic acid, unknown A, unknown B and phosphonates also decreased, whereas the proportions of orthophosphate monoesters increased following land use changes. Greater decline in proportions of diester-P and teichoic acids were found in the silt than in the clay size separates, which may be attributed to stabilization of microbially derived organic P structures by closer association with clay minerals and/or sesquioxides. The results of sequential extraction and ³¹P NMR spectroscopy indicate that continuous cultivation with little or no P input not only decreased the amount, but also influenced the structural composition and bioavailability of P in these tropical soils. Thus measures have to be designed for replenishment and subsequent maintenance of soil P stocks, to ensure sustainable crop production in sub-humid highland agroecosystems of southern Ethiopia.     

The chemical nature of organic phosphorus that accumulates in fertilized soils of a temperate pasture as determined by solution 31P NMR spectroscopy

Inefficiency of fertilizer phosphorus (P) use in grazing systems is often associated with the accumulation of inorganic and organic P in fertilized soil. However, the chemical nature of the accumulated organic P remains poorly understood. The aim of this study was to use solution ³¹P nuclear magnetic resonance (NMR) spectroscopy on sodium hydroxide–ethylenediaminetetraacetic acid (NaOH‐EDTA) extracts to identify the chemical nature of organic P in soils from a medium‐term (13 years) permanent pasture field experiment. This included an unfertilized pasture (P0), and treatments designed to maintain soil P fertility at near ‘optimum' (P1) and ‘supra‐optimum' (P2) levels for pasture growth; pastures at all levels of soil P fertility were continuously grazed with either a moderate or high stocking rate (SR09 and SR18). Approximately 20% of the fertilizer P added to pastures was recovered as organic P in NaOH‐EDTA extracts at the P1 level of soil P fertility in the 0–10 cm soil layer, and the majority (≈ 65%) of this was detected as the broad phosphomonoester signal. In addition, several specific forms of phosphomonoesters (myo‐ and scyllo‐inositol hexakisphosphate, α‐ and β‐glycerophosphate, and RNA mononucleotides) and phosphodiesters were detected across all soils but at low concentrations. This study shows that phosphate fertilization of pastures primarily results in the accumulation of complex forms of phosphomonoesters rather than that of specific forms of recognizable biomolecules (e.g ., myo‐inositol hexakisphosphate).     

31Phosphorus‐Nuclear magnetic resonance analysis in extracts of a phosphorus‐enriched volcanic soil of Chile

Abstract

The nature of organic and inorganic phosphorus (P) in fertilized and unfertilized samples of Vilcun soil, a Chilean medial mesic typic Dystrandept which was studied through ³¹phosphorus‐nuclear magnetic resonance (³¹P‐NMR) analysis carried out on a single alkaline extraction. The total P contents ranged from 1,506 mg P kg^‐1 (B horizon, unfertilized soil) to 7,541 mg P kg^‐1 (A horizon, fertilized soil). The magnitude of the P signal (SEM‐EDAX results) appears to be related to that of the iron (Fe) signal. Signals of ³¹P‐NMR spectra are attributable to inorganic orthophosphate, and orthophosphate monoesters and diesters. No important differences between horizons were observed. Apparently in this soil, P are mainly associated with iron oxides and organic matter. The organic P forms were not readily subjected to mineralization.     

Phosphorus forms in forest soil colloids as revealed by liquid‐state 31P‐NMR

Nanoparticles and colloids affect the storage and hence the availability of P in forest ecosystems. We investigated the fine colloids present in forest soils and their association with inorganic and organic P. To differentiate between the different P forms, we performed liquid‐state ³¹P‐nucelar magnetic resonance (³¹P‐NMR) measurements on forest bulk soil extracts, on colloid extracts and on the electrolyte phase of their soil suspensions. The ³¹P‐NMR spectra indicated that soil nanoparticles and colloids were more enriched with organic than with inorganic P forms compared to the electrolyte phase. The P concentration was enriched in the colloidal fraction in comparison to the bulk soil and the phosphate diesters were more dominant in the colloidal fraction when compared to the bulk soil. The colloidal P‐diester to P‐monoester ratios were 2 to 3 times higher in the colloidal fraction than in the bulk soil. In contrast, relatively large percentages of inorganic P were found in the electrolyte phase. Supplementary (not shown) Data are available at the JuSER Server (juser.fz‐juelich.de, reference number: FZJ‐2016‐01739) https://juser.fz‐juelich.de/record/283057 .     

Sequential extractions and 31P-NMR spectroscopy of phosphorus forms in animal manures, whole soils and particle-size separates from a densely populated livestock area in northwest Germany

The solubility and forms of phosphorus (P) were investigated in manures from chicken and pigs, eight whole soil samples and clay-, silt-, and sand-size separates from an arable and a grassland soil. Total P (P_t) in liquid pig manure (16.2 g kg^–1) and dry chicken manure (26.2 g kg^–1) was distributed between residual P (39–41% P_t), H₂SO₄–P (17–27% P_t), labile resin- and NaHCO₃–P (24–39% P_t), and NaOH-P (3–10% P_t). Most soils had larger proportions of NaOH-P and residual P, indicating reactions of manure-derived P compounds with pedogenic oxides and humic substances. Clay-size separates had the highest P-concentrations in all fractions and were particularly enriched in exchangeable and labile P forms. Solution ³¹P-nuclear magnetic resonance (NMR) spectra of 0.5 M NaOH extracts from manures and some soil samples showed greater signal intensities for orthophosphate and monoester P than 0.1 M NaOH extracts. This can be explained by alkaline hydrolysis phosphate diesters at higher NaOH concentrations and/or by preferential extraction of diesters at lower concentrations. The ³¹P-NMR spectra showed differences between the two manures and confirmed that increasing proportions of ester-P can be expected if they are spread to soils. The NaOH extracts of soil samples were characterized by large proportions of orthophosphate-P (mean 77% of assigned P compounds), which seemed to be slightly enriched in clay fractions whereas the extracts from silt contained more ester-P. Sequential extractions and ³¹P-NMR spectroscopy both showed that these excessively manured soils are likely to lose large amounts of P. Received: 15 July 1996     

Characterization of organic phosphorus in leachate from a grassland soil

The degree of eutrophication in fresh water ecosystems may be influenced by the forms of phosphorus (P) leached from agricultural systems. Physico-chemical fractionation of P in leachate from a grassland soil carried out over a two year period indicated that the majority of the P loss from the Lismore soil occurred in unreactive particulate (55-76%) P forms. ³¹P nuclear magnetic resonance analysis of a selected leachate sample indicated that unreactive P was mainly comprised of monoester and diester forms of organic P. The presence of phosphomonoesterase (20-200 μg p nitrophenol l⁻¹ h⁻¹) and phosphodiesterase (68 μg bis-p nitrophenol l⁻¹ h⁻¹) activity in leachate resulted in hydrolysis of 10-21% of total unreactive P (TUP), indicating that some of the monoesters and diesters can be eventually hydrolyzed into inorganic P forms during P transport. Enzyme hydrolysis showed that 23% of the TUP was present as labile monoester P (LMP), followed by 20% as inositol hexakisphosphate (IHP) and 14% as diesters (phospholipids and nucleic acids). The findings of this study suggest that LMP, IHP and diesters are an important component of organic P leaching from the grassland soil.     

Characterization of Phosphorus in Sewage Sludge from Different Sources by 31Phosphorus Nuclear Magnetic Resonance Spectroscopy

To investigate the distribution and dynamics of phosphorus (P) in soils for environmental protection and agronomical usage, ³¹P nuclear magnetic resonance spectroscopy (³¹P NMR) was used to characterize the contents and chemical properties of P in sewage sludge from 13 wastewater treatment plants in Shanghai. The samples were extracted with 0.25 M sodium hydroxide (NaOH) / 0.05 M sodium ethylenediamietetraacetic acid (Na₂EDTA) in ratio of 1:20 (w/v). Total P recovery in the extract ranged from 91 to 116% when compared to traditional chemical methods. The dominant forms of P in all samples were inorganic orthophosphates and orthophosphates monoesters. Orthophosphate diesters and pyrophosphates were present in only two and four samples, respectively. This study provides detailed information on the distribution, contents, and chemical properties of P in sewage sludge that may be of value in the utilization of sewage sludge for agronomic purposes.     

Identification of soil organic phosphorus by 31P nuclear magnetic resonance spectroscopy

Abstract

Soil samples from different land use systems were collected before cropping (in spring) and after harvest (in fall) for organic phosphorus (P) extractions by 0.4M sodium hydroxide (NaOH) and characterization by ³¹P nuclear magnetic resonance spectroscopy. To prevent hydrolysis of organic P compounds prior to sample concentration, NaOH was removed from the NaOH soil extracts using a G‐25 Sephadex column. The ³¹P NMR spectra in the NaOH soil extracts showed the presence of glucose‐6 phosphate (up to 64%), glycerophosphate (up to 45%), nucleoside monophosphates (up to 91%), and polynucleotides (up to 58%) as the major forms of organic P in soils. The relative concentration of nucleoside monophosphates and polynucleotides decreased in some of the soils after harvest. The ³¹P NMR spectra of the extracts also revealed the presence of phosphoenolpyruvates, a previously unreported form of soil organic P.     

Organophosphates in agrogray soils with periodic water logging according to the data of <Superscript>31</Superscript>P NMR spectroscopy

The composition of organic phosphorus compounds was studied using the ³¹P NMR spectroscopy method in agrogray soils with periodic water logging. The phosphorus content was determined by the specific difference between the hydrological and the redox regimes of these soils. The phosphorus of the organic compounds in the agrogray soils with contrasting water regimes is composed mostly of phosphoric monoesters and diesters, including nucleic and teichoic acids. The relative distribution of the monoesters and inorganic orthophosphates is shown depending on the climate and the soil’s position in the relief. The area of the monoester peaks increases by two times and that of the mineral orthophosphate decreases by six times in the agro-gray soils of Bryansk opolie with an optimal regime of moistening and evaporation in comparison with the agro-gray gleyed soils of Kolomna opolie. As the degree of the soil hydromorphism in the sequence of deeply gleyed soils and gleyic soils increased, the portion of monoesters decreased. Favorable conditions for the microbial activity are formed in soils with a contrasting redox regime, and this is expressed in the accumulation of labile diesters. Inverse relationships were found between the distributions of the mono- and diesters in iron-manganic nodules and in the soils enclosing them; this was caused by the different mechanisms of the stabilization of the stable and labile phosphorus containing compounds. A high percentage of mineral orthophosphate in the nodules allows assuming the presence of chemisorbed orthophosphate ions in organomineral phosphate-metal-humus complexes. The transformation of iron-manganic nodules under the influence of drying demonstrates the more direct participation of microorganisms in the nodules’ formation than the contribution of the physicochemical processes.     

Characterization of phosphorus fractions in mountain soils of the Bavarian Alps by 31P NMR spectroscopy

NaOH-extracts from different soils of the alpine and shrub zones at the Schneibstein (Nationalpark Berchtesgaden) between 1790–2275 m were studied using ³¹P NMR spectroscopy. NaOH (0.5 M) dissolved 29–49% of P_t. Orthophosphate monoesters were the dominant fraction (41–79%). Orthophosphate diesters comprised 10–20%. A higher diester content (36%) was found in a hydromorphic soil. Inorganic phosphate comprised 5–27%, depending on soil pH. So, in acid horizons (pH 4.45 and 5.05) 27 and 23% were detected, whereas in horizons with higher pH (> 6) only small amounts (5–8%) were found. Pyrophosphate and polyphosphates were found in traces. Phosphonates (ca. 6%) occurred mainly in acid soils.     

The nature and origins of diester phosphates in soils: a 31P-NMR study

Soils of two climosequences in Russia were investigated by ³¹P-NMR spectroscopy. They comprised Dystric Podzoluvisols, Haplic Greyzems, Calcic Chernozems, and Gypsic Kastanozems, which are located along temperature and precipitation gradients of the Russian Plain. Another sequence of soils included forest Humic Cambisols and Umbric Leptosols of subalpine and alpine meadows, which are formed in different climatic conditions along a climosequence of the Mt. Malaya Khatipara (northern Caucasus). The results showed that accumulation of DNA was high in the cold, wet, and acid soils (Dystric Podzoluvisol, alpine Umbric Leptosol), while phospholipids and teichoic acids mainly accumulated in the more microbially active soils. We performed a laboratory incubation experiment to test the relationship between microbial biomass P and P species identified in soil extracts. The proportions of P compounds resonating at 0.5-3.0 ppm in the NaHCO₃ and H₂SO₄ extracts from the incubated Humic Cambisol increased. The amounts of phosphate diesters resonating at 0 ppm in the same extracts and in the subsequent NaOH extracts decreased after incubation. Based on the results of ³¹P-NMR spectroscopy of native soils and of the laboratory incubation experiment we concluded that signals at 0 ppm in spectra of soil alkaline extracts belong to DNA P which is mainly stabilised in soil organic matter outside microbial cells (at least in soils with relatively low microbial activity). Phospholipids-teichoic acids P extracted with 0.5 M NaHCO₃ seems to be derived from soil microbial biomass, and its proportion can reflect the microbial activity in the soil.