Fungal biomass production and turnover in soil estimated using the acetate-in-ergosterol technique

We report the first attempt to estimate fungal biomass production in soil by correlating relative fungal growth rates (i.e., acetate incorporation into ergosterol) with fungal biomass increase (i.e., ergosterol) following amendments with dried alfalfa or barley straw in soil. The conversion factor obtained was then used in unamended soil, resulting in fungal biomass productions of 10-12 μg C g⁻¹ soil, yielding fungal turnover times between 130 and 150 days. Using a conversion factor from alfalfa-treated soil only resulted in two times higher estimates for biomass production and consequently lower turnover times. Comparing fungal biomass production with basal respiration indicated that these calculations overestimated the former. Still, the turnover times of fungal biomass in soil were in the same range as turnover times estimated in aquatic systems. The slow turnover of fungal biomass contrasts with the short turnover times found for bacteria. The study thus presents empirical data substantiating the theoretical division of bacteria and fungi into a fast and a slow energy channel, respectively, in the soil food web.     

Spatial changes of soil fungal and bacterial biomass from a sub-alpine coniferous forest to grassland in a humid, sub-tropical region

 Fungal and bacterial biomass were determined across a gradient from a forest to grassland in a sub-alpine region in central Taiwan. The respiration-inhibition and ergosterol methods for the evaluation of the microbial biomass were compared. Soil fungal and bacterial biomass both significantly decreased (P<0.05) with the shift of vegetation from forest to grassland. Fungal and bacterial respiration rates (evolved CO₂) were, respectively, 89.1 μl CO₂ g^–1 soil h^–1 and 55.1 μl CO₂ g^–1 soil h^–1 in the forest and 36.7 μl CO₂ g^–1 soil h^–1 and 35.7 μl CO₂ g^–1 soil h^–1 in the grassland surface soils (0–10 cm). The fungal ergosterol content in the surface soil decreased from the forest zone (108 μg g^–1) to the grassland zone (15.9 μg g^–1). A good correlation (R ²=0.90) was exhibited between the soil fungal ergosterol content and soil fungal CO₂ production (respiration) for all sampling sites. For the forest and grassland soil profiles, microbial biomass (respiration and ergosterol) declined dramatically with depth, ten- to 100-fold from the surface organic horizon to the deepest mineral horizon. With respect to fungal to bacterial ratios for the surface soil (0–10 cm), the forest zone had a significantly (P<0.05) higher ratio (1.65) than the grassland zone (1.05). However, there was no fungal to bacterial ratio trend from the surface horizon to the deeper mineral horizons of the soil profiles. Received: 30 March 2000     

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.     

Long-term nitrogen additions increased surface soil carbon concentration in a forest plantation despite elevated decomposition

Forests cover one-third of the Earth’s land surface and account for 30-40% of soil carbon (C). Despite numerous studies, questions still remain about the factors controlling forest soil C turnover. Present understanding of global C cycle is limited by considerable uncertainty over the potential response of soil C dynamics to rapid nitrogen (N) enrichment of ecosystems, mainly from fuel combustion and fertilizer application. Here, we present a 15-year-long field study and show an average increase of 14.6% in soil C concentration in the 0-5 cm mineral soil layer in N fertilized (defined as N+ hereafter) sub-plots of a second-rotation Pinus radiata plantation in New Zealand compared to control sub-plots. The results of ¹⁴C and lignin analyses of soil C indicate that N additions significantly accelerate decomposition of labile and recalcitrant soil C. Using an annual-time step model, we estimated the soil C turnover time. In the N+ sub-plots, soil C in the light (a density < 1.70 g cm⁻³) and heavy fractions had the mean residence times of 23 and 67 yr, respectively, which are lower than those in the control sub-plots (36 and 133 yr in the light and heavy fractions, respectively). The commonly used lignin oxidation indices (vanillic acid to vanillin and syringic acid to syringaldehyde ratios) were significantly greater in the N+ sub-plots than in the control sub-plots, suggesting increased lignin decomposition due to fertilization. The estimation of C inputs to forest floor and δ¹³C analysis of soil C fractions indicate that the observed buildup of surface soil C concentrations in the N+ sub-plots can be attributed to increased inputs of C mass from forest debris. We conclude that long-term N additions in productive forests may increase C storage in both living tree biomass and soils despite elevated decomposition of soil organic matter.     

Bacterial and fungal growth in soil heated at different temperatures to simulate a range of fire intensities

The intensity of a fire is an important factor determining the recovery of soil microorganisms after a forest fire, since it can alter the quality and quantity of carbon sources. Recovery of the microbial community was studied in a Mediterranean pine forest soil subjected to different temperatures to simulate the short-term effects of fire intensity on bacterial and fungal growth, estimated using leucine incorporation for bacteria and acetate incorporation into ergosterol for fungi. Soil samples were heated for 15 min at 50, 80, 120, 200, 300, 400 and 500 °C. After inoculation with fresh soil, and adding water to achieve 60% WHC, the soils were incubated at 20 °C for 21 days. Bacterial growth was initially inhibited in the samples heated above 50 °C (totally inhibited ≥ 200 °C), but recovered within days to levels much higher than the control, except for the samples heated at 500 °C, where growth remained low throughout the incubation period due to the destruction of most of the organic matter. After the first week of incubation, the bacterial response decreased to values close to, but still above, that of the control. Samples heated at 200 °C showed the highest cumulative bacterial growth. Fungal growth was initially lower than in the control in all the heated samples (totally inhibited ≥ 200 °C). Fungal growth recovered slowly during incubation in soils heated at ≤ 300 °C, but the cumulative growth in heated soils did not exceed that in the control. No fungal growth was observed in samples heated at the two highest temperatures. Soil respiration was initially totally inhibited in soil heated at ≥ 200 °C, but recovered rapidly in all soils; the highest respiration being observed already 1 day after inoculation. This is the first time both fungal and bacterial growth has been directly estimated in heated soils. High soil pH favouring bacteria can explain these results, but the differences in fungal and bacterial responses suggest a competitive interaction between these groups.     

Production of root-derived material and associated microbial growth in soil at different nutrient levels

Summary Maize plants were grown for 42 days in a sandy soil at two different mineral nutrient levels, in an atmosphere containing ¹⁴CO₂. The ¹⁴C and total carbon contents of shoots, roots, soil and soil microbial biomass were measured 28, 35 and 42 days after germination. Relative growth rates of shoots and roots decreased after 35 days at the lower nutrient level, but were relatively constant at the higher nutrient level. In the former treatment, 2% of the total ¹⁴C fixed was retained as a residue in soil at all harvests while at the higher nutrient level up to 4% was retained after 42 days. Incorporation of ¹⁴C into the soil microbial biomass was close to its maximum after 35 days at the lower nutrient level, but continued to increase at the higher level. Generally a good agreement existed between microbial biomass, ¹⁴C contents and numbers of fluorescent pseudomonads in the rhizosphere. Numbers of fluorescent pseudomonads in the rhizosphere were maximal after 35 days at the lower nutrient level and continued to increase at the higher nutrient level. The proportions of the residual ¹⁴C in soil, incorporated in the soil microbial biomass, were 28% to 41% at the lower nutrient level and 20%6 – 30% at the higher nutrient level. From the lower nutrient soil 18%6 – 52%6 of the residual soil ¹⁴C could be extracted with 0.5 N K₂SO₄, versus 14%6 – 16% from the higher nutrient soil.Microbial growth in the rhizosphere seemed directly affected by the depletion of mineral nutrients while plant growth and the related production of root-derived materials continued.     

Influence of different temperatures on metal tolerance measurements and growth response in bacterial communities from unpolluted and polluted soils

The effects of temperature on the growth rate and metal toxicity in soil bacterial communities extracted from unpolluted and polluted soils were investigated using the thymidine and leucine incorporation techniques. An agricultural soil, which was contaminated in the laboratory with Cu, Cd, Zn, Ni or Pb, and an uncontaminated forest soil were used. Measurements were made at 0°C and 20°C. Leucine incorporation was found to be as sensitive to heavy metals as thymidine incorporation in the short-term trial used to indicate heavy metal tolerance. Similar IC₅₀ values (the log of the metal concentration that reduced incorporation to 50%) were also obtained at 0 and 20°C, independently of the technique used. Metal tolerance could thus be measured using both techniques at any temperature in the range 0–20°C. In the long-term experiment different temperature-growth relationships were obtained on the basis of the rate of thymidine or leucine incorporation into bacterial assemblages from unpolluted and polluted soils, as judged from the minimum temperature values. This could not be attributed to the metal addition alone since different patterns were observed when different metals were added to the soil. Thus, the minimum temperature for thymidine incorporation was similar in Cu-polluted and unpolluted soil, while in soils polluted with Cd and Zn the minimum temperature increased by 2°C, and Ni and Pb additions increased the minimum temperature by 4°C compared to the unpolluted soil. This suggested that heavy metal pollution led to bacterial communities showing different temperature characteristics to those in the corresponding unpolluted soil. Similar observations were deduced from the minimum temperatures required for leucine incorporation. Three groups of bacterial communities were distinguished according to the growth response to temperature in polluted soils, one group in Cu-polluted soil, a second group in soil polluted with Zn and Cd, and a third group in soils polluted with Ni and Pb.     

Comparison of factors limiting bacterial growth in different soils

Lack of carbon has been assumed to be the most common limiting factor for bacterial growth in soil, although there are reports of limitation by other nutrients, e.g. nitrogen and phosphorus. We have studied which nutrient(s) limited instantaneous growth rates of bacteria in 28 Swedish soils using the thymidine or leucine incorporation technique to measure increased growth rate after adding different combinations of organic carbon (glucose), nitrogen and phosphorus. The soils ranged in pH between 3.1 and 8.9, in organic matter content between 1% and 91% and in soil C/N ratio between 10 and 28. We also tested the effect of adding different amounts of carbon on the bacterial change in growth rate for two soils with different organic matter content. We found that bacterial growth in most of the 28 soils was limited by a lack of carbon, indicated by an increased bacterial growth rate 48 h after adding glucose. In some soils, adding carbon together with nitrogen increased the bacterial growth rates even further. In three soils no effects were seen upon adding nutrients separately, but adding carbon and nitrogen together increased bacterial growth rates. Nitrogen addition tended to decrease bacterial growth rates, while phosphorus addition had little effect in most soils. No correlations were found between the soil C/N ratio, ammonium or nitrate content in soil and bacterial growth limitation, indicating that even soils with a C/N ratio of 28 could be carbon limited. Although the interpretation of the effects of a single limiting nutrient was in most cases straightforward, an interaction between the amount of carbon added and the organic matter content of the soil confounded the interpretation of the extent of a second limiting nutrient.     

Carbon and nitrogen dynamics in a forest soil amended with purified tannins from different plant species

Tannins are purported to be an important factor controlling nitrogen cycling in forest ecosystems, and the ability of tannins to bind proteins in protein-tannin complexes is thought to be the primary mechanism responsible for these effects. In this study, we examined the influence of well-characterized tannins purified from five different plant species on C and N dynamics of a forest soil A horizon. Tannic acid, a commonly used and commercially available hydrolyzable tannin (HT), and cellulose were also included for comparison. With the exception of tannins from huckleberry (Vaccinium ovatum), the amendments increased respiration 1.4-4.0 fold, indicating that they were acting as a microbial C source. Tannic acid was significantly more labile than the five purified tannins examined in this study. All treatments decreased net N mineralization substantially, through greater N immobilization and decreased mineralization. The six tannins inhibited gross ammonification rates significantly more than cellulose. This suggests that added tannins had effects in addition to serving as an alternative C source. Tannins purified from Bishop pine (Pinus muricata) were the only tannins that significantly inhibited potential gross nitrification rates, however, rates were low even in the control soil making it difficult to detect any inhibition. Differences in tannin structure such as condensed versus HTs and the hydroxylation pattern of the condensed tannin B-ring likely explain differences observed among the tannin treatments. Contrary to other studies, we did not find that condensed tannins were more labile and less inhibitory than HTs, nor that shorter chained tannins were more labile than longer chained tannins. In addition to supporting the hypothesis that reduced N availability in the presence of tannins is caused by complexation reactions, our data suggests tannins act as a labile C source leading to increased N immobilization.     

Dynamics of maize (Zea mays L.) leaf straw mineralization as affected by the presence of soil and the availability of nitrogen

An incubation experiment was carried out with maize (Zea mays L.) leaf straw to analyze the effects of mixing the residues with soil and N amendment on the decomposition process. In order to distinguish between soil effects and nitrogen effects for both the phyllospheric microorganisms already present on the surface of maize straw and soil microorganisms the N amendment was applied in two different placements: directly to the straw or to the soil. The experiment was performed in dynamic, automated microcosms for 22 days at 15 °C with 7 treatments: (1) untreated soil, (2) non-amended maize leaf straw without soil, (3) N amended maize leaf straw without soil, (4) soil mixed with maize leaf straw, (5) N amended soil, (6) N amended soil mixed with maize leaf straw, and (7) soil mixed with N amended maize leaf straw. ¹⁵NH₄¹⁵NO₃ (5 at%) was added. Gas emissions (CO₂, ¹³CO₂ and N₂O) were continuously recorded throughout the experiment. Microbial biomass C, biomass N, ergosterol, δ¹³C of soil organic C and of microbial biomass C as well as ¹⁵N in soil total N, mineral N and microbial biomass N were determined in soil samples at the end of the incubation. The CO₂ evolution rate showed a lag-phase of two days in the non-amended maize leaf straw treatment without soil, which was completely eliminated when mineral N was added. The addition of N generally increased the CO₂ evolution rate during the initial stages of maize leaf straw decomposition, but not the cumulative CO₂ production. The presence of soil caused roughly a 50% increase in cumulative CO₂ production within 22 days in the maize straw treatments due to a slower decrease of CO₂ evolution after the initial activity peak. Since there are no limitations of water or N, we suggest that soil provides a microbial community ensuring an effective succession of straw decomposing microorganisms. In the treatments where maize and soil was mixed, 75% of microbial biomass C was derived from maize. We concluded that this high contribution of maize using microbiota indicates a strong influence of organisms of phyllospheric origin to the microbial community in the soil after plant residues enter the soil.     

Land use effects on carbon quality and soil biological properties in Eutric Cambisol

Abstract

The choice of prospective type of farming requires knowledge about the specific relationships that exist between farm management practices and base environmental conditions. Nowadays the protection of soil organic carbon is one of the main tasks, because organic carbon in addition to soil fertility can act in elimination of soil contamination and carbon sequestration. Field experiments were focused on the effect of intensive farming without organic inputs versus grassland on organic carbon content. Organic carbon content (C_ox) and humic substance fractions (C-humic acids and fulvic acid fractions), hot water extractable carbon and selected microbial characteristics in Eutric Cambisol were monitored during the period 1999–2010. A priming effect of soil cultivation was detected immediately after tillage. Arable soil with ‘intensive’ crop sequences (exclusively cash crops, cereals, oil plants) and with an optimal level of chemical inputs (mineral fertilizers, pesticides), but without organic farmyard manure had lower content of all carbon forms compared with grassland. ¹³C NMR spectroscopy and thermal analysis (TGA) were applied to characterize humic acid (HA) structure and stability. More carbon, less oxygen and more aromatic compounds were detected in grassland HA. Slight differences were found in HA thermo-oxidative stability and degradability, which was probably caused by changes in elemental composition and structure. Even the land use had no significant effect on basic microbiological characteristics (basal respiration, microbial biomass and qCO₂); the physiology of the microbial community of grassland was altered by a higher ability to utilize L- and D-glutamic acid. The L/D ratio of glutamic acid mineralization indicated no occurrence of stress in soil for both types of farming. It has been demonstrated that although losses of carbon as a result of land-use conversions are generally more rapid, gains of carbon in grassland followed by changes in management practices can also occur.     

Measurement of protein synthesis by soil bacterial assemblages with the leucine incorporation technique

The incorporation of [¹⁴C]-leucine into protein by soil organisms was measured both in soil slurries and for bacteria extracted from soil by homogenization-centrifugation. The result was compared with thymidine incorporation. Using a soil slurry, 9.1×10^-10 mol leucine h^-1g^-1 dry weight of soil was incorporated into protein, with a calculated leucine: thymidine ratio (mol:mol) of about 34. Non-specific labelling of macromolecules other than protein was observed with both the soil slurry and the homogenization-centrifugation method. With the latter, 46.5% of the total incorporation was found in the protein fraction (hot-acid insoluble). The incorporation of leucine was linear with time for at least 4 h for extracted bacteria. Even at 2000 nM, [¹⁴C]-leucine did not saturate incorporation into protein. Isotope dilution plots indicated that with 750 nM leucine, the degree of participation of the labelled substance in protein synthesis was 0.59. With this value, the ratio of leucine:thymidine incorporation into total macromolecules was calculated as 41 for extracted bacteria. On the basis of incorporation into protein (leucine) and incorporation into DNA (thymidine) only, the leucine:thymidine ratio was calculated as 117. The mean turnover time of bacteria at 22°C, calculated using conversion factors from published studies and leucine incorporation into protein of extracted bacteria, was 4.3 days.     

Microbial community utilization of added carbon substrates in response to long-term carbon input manipulation

The chemical composition and quantity of plant inputs to soil are primary factors controlling the size and structure of the soil microbial community. Little is known about how changes in the composition of the soil microbial community affect decomposition rates and other ecosystem functions. This study examined the degradation of universally ¹³C-labeled glucose, glutamate, oxalate, and phenol in soil from an old-growth Douglas-fir (Pseudotsuga menziesii)—western hemlock (Tsuga heterophylla) forest in the Oregon Cascades that has experienced 7 y of chronic C input manipulation. The soils used in this experiment were part of a larger Detritus Input and Removal Treatment experiment and have received normal C inputs (control), doubled wood inputs, or root and litter input exclusion (no inputs). Soil from the doubled wood treatment had a higher fungal:bacterial ratio, and soil from the no inputs treatment had a lower fungal:bacterial ratio, than the control soil. Differences in the utilization of the compounds added to the field-manipulated soils were assessed by following the ¹³C tracer into microbial biomass and respiration. In addition, ¹³C-phospholipid fatty acids (PLFA) analysis was used to examine differential microbial utilization of the added substrates. Glucose and glutamate were metabolized similarly in soils of all three litter treatments. In contrast, the microbial community in the double wood soil respired more added phenol and oxalate, whereas microbes in the no inputs soil respired less added phenol and oxalate, than the control soil. Phenol was incorporated primarily into fungal PLFA, especially in soil of the double wood treatment. The addition of all four substrates led to enhanced degradation of soil organic matter (priming) in soils of all three litter treatments, and was greater following the addition of phenol and oxalate as compared to glucose and glutamate. Priming was greater in the no inputs soil as compared to the control or doubled wood soils. These results demonstrate that altering plant inputs to soil can lead to changes in microbial utilization of C compounds. It appears that many of these changes are the result of alteration in the size and composition of the microbial community.     

土壤C/N对苹果植株生长及氮素利用的影响

土壤C/N是土壤氮素循环的重要影响因素。本研究以2年生"富士"/平邑甜茶为试验材料, 应用¹⁵N示踪技术研究了不同土壤C/N[6.21(CK)、10、15、20、25、30、35和40]对苹果植株生长及氮素利用和损失的影响。结果表明: 随着土壤C/N比值的逐渐增大, 苹果新梢长度和植株鲜重均呈先升高后降低的变化趋势, C/N=15、20和25的3个处理苹果新梢长度和植株鲜重最大, 三者间无显著差异, 但均显著高于其他处理。不同C/N处理间植株15N利用率存在差异, 土壤C/N=25时, 植株¹⁵N利用率最大, 为22.87%, 与C/N=20的处理间无显著差异, 但两者均显著高于其他处理; 土壤C/N=40时, 植株¹⁵N利用率最低, 仅为15.43%, 低于CK处理的16.65%。土壤C/N处于15~25时, 植株吸收的氮素来自于肥料氮的比例较高; 而土壤C/N较低(<15)或太高(>25)时, 植株吸收的氮素来自于土壤氮的比例较高。土壤氮素残留量随土壤C/N的增大逐渐增加, C/N=40处理的土壤氮素残留量是CK的1.32倍。随着土壤C/N比值的逐渐增大, 肥料氮损失量呈先减少后增加的变化趋势, 以C/N=25时最少, 仅为施氮量的49.87%, 而对照最大, 为61.54%。因此, 综合土壤C/N对苹果植株生长及氮素平衡状况来看, 土壤C/N为15~25时, 能促进植株的生长发育, 降低氮肥损失, 提高肥料利用率。     

Interactive effects of elevated CO2 and nitrogen fertilization levels on photosynthesized carbon allocation in a temperate spring wheat and soil system

Increasing atmospheric CO2 concentration impacts the terrestrial carbon(C) cycle by affecting plant photosynthesis, the flow of photosynthetically fixed C belowground, and soil C pool turnover. For managed agroecosystems, how and to what extent the interactions between elevated CO2 and N fertilization levels influence the accumulation of photosynthesized C in crops and the incorporation of photosynthesized C into arable soil are in urgent need of exploration.We conducted an experiment simulating elevated CO2 with spring wheat(Triticum aestivum L.) planted in growth chambers.13C-enriched CO2 with an identical 13C abundance was continuously supplied at ambient and elevated CO2 concentrations(350 and 600 μmol mol-1, respectively) until wheat harvest.Three levels of N fertilizer application(equivalent to 80, 120, and 180 kg N ha-1 soil) were supplied for wheat growth at both CO2 concentrations. During the continuous 62-d 13CO2 labeling period, elevated CO2 and increased N fertilizer application increased photosynthesized C accumulation in wheat by 14%–24% and 11%–20%, respectively, as indicated by increased biomass production, whereas the C/N ratio in the roots increased under elevated CO2 but declined with increasing N fertilizer application levels. Wheat root deposition induced 1%–2.5% renewal of soil C after 62 d of 13CO2 labeling. Compared to ambient CO2, elevated CO2 increased the amount of photosynthesized C incorporated into soil by 20%–44%. However, higher application rates of N fertilizer reduced the net input of root-derived C in soil by approximately 8% under elevated CO2. For the wheat-soil system, elevated CO2 and increased N fertilizer application levels synergistically increased the amount of photosynthesized C. The pivotal role of plants in photosynthesized C accumulation under elevated CO2 was thereby enhanced in the short term by the increased N application. Therefore, robust N management could mediate C cycling and sequestration by influencing the interactions between plants and soil in agroecosystems under elevated CO2.     

添加不同外源氮对长期秸秆还田土壤中氮素转化的影响

【目的】秸秆还田能够改变土壤中各活性氮库的含量与比例,进而影响土壤氮素供应能力。本文研究了长期秸秆还田条件下添加不同外源氮对土壤中不同形态氮素的影响,旨在明确长期秸秆还田土壤活性氮库的含量差异。【方法】长期定位施肥试验点位于湖南省望城县(112°80′N、28°37′E,海拔高度100 m)。试验开始于1981年,供试土壤为第四纪红色黏土发育的水稻土,轮作制度为稻—稻—冬闲。2014年晚稻收获后,采集单施化肥和长期秸秆还田配施化肥两个处理的耕层土壤样品,开展室内培养试验。每个土壤样品设置灭菌和不灭菌两组主处理,在主处理下设:对照(CK)、添加尿素(N 150 kg/hm^2,U)、添加秸秆(N 150 kg/hm^2,S)和添加尿素和秸秆(N 300 kg/hm^2,U+S)四个副处理,4次重复。在25℃下恒温培养5、10、20、30、50、90、130天时,分析土壤铵态氮、硝态氮、微生物氮和可溶性有机氮含量。【结果】1) U、S和U+S处理均显著提高土壤铵态氮和硝态氮含量,高低顺序为U> U+S> S> CK。非灭菌条件下,U处理的土壤铵态氮含量较其他处理高出90.8%~288%。2)灭菌后土壤铵态氮长期维持在较高水平,其向硝态氮转化过程受阻。在培养90天内,土壤硝态氮、微生物氮和可溶性有机氮含量均处于较低水平。3)而不灭菌条件下,各处理土壤硝态氮均在培养50天后迅速增加,至培养结束土壤硝态氮达最大值(117.43~243.17 mg/kg)。4)土壤微生物氮和可溶性有机氮分别于培养20天(106.72~244.01 mg/kg)和30天(95.76~140.63 mg/kg)时达到最大值。5)至培养结束,灭菌条件下长期NPKS土壤中U+S处理可溶性有机氮显著高于其他处理,较U和S处理分别提高51.55%和29.96%。【结论】添加不同外源氮有利于提高长期秸秆还田土壤中活性有机氮的含量,尤其是添加秸秆和尿素处理,能够显著提高土壤氮素的供应能力。     

Induced N-limitation of bacterial growth in soil: Effect of carbon loading and N status in soil

Application of C-rich plant residues can change the soil system from C-limitation for microbial growth to limitation by other nutrients. However, the initial nutrient status of the soil may interact with the added amount of residues in determining limitation. We studied this interactive effect in soils from the Harvard Forest LTER, where annual addition of N since 1988 has resulted in soils with different N-status: No N (Unfertilized), 50 (Low N) and 150 (High N) kg N ha⁻¹. We hypothesized that adding C-rich substrate would change the soil from being C- to being N-limited for bacterial growth and that the extent of N-limitation would be higher with increasing substrate additions, while becoming less evident in soil with increasing N-status. We compared the effect of adding two C-rich substrates, starch (0, 10, 20, 40 mg g⁻¹ soil) and straw (0, 20, 40, 80 mg g⁻¹), incubating the soils for up to 3 and 4 weeks for starch and straw, respectively. Nutrient limitations were studied by measuring bacterial growth 3 days after adding C as glucose and N as NH₄NO₃ in a full factorial design. Initially bacterial growth in all soils was C-limited. As hypothesized, adding C-rich substrates removed the C-limitation, with lower amounts of starch and straw needed in the unfertilized and Low N soils than in the High N soil. Combinations of different N-status of the soil and amendment levels of starch and straw could be found, where bacterial growth appeared close to co-limited both by available C and N. However, at even higher amendment levels, presumable resulting in N-limitation, bacterial growth still responded less by adding N then C-limited soils by adding C. Thus, in a C-limited soil there appeared to be N available immediate for growth, while in an N-limited soil, easily available C was not immediately available.     

The initial rate of C substrate utilization and longer-term soil C storage

The initial reaction of microbial transformation and turnover of soil carbon inputs may influence the magnitude of longer-term net soil C storage. The objective of this study was to test the merit of the hypothesis that the more rapid substrates are initially utilized, the longer the residual products remain in the soil. We used simple model C compounds to determine their decomposition rates and persistence over time. Pure ¹⁴C compounds of glucose, acetate, arginine, oxalate, phenylalanine, and urea were incubated in soil for 125 days at 24°C. Total respired CO₂ and ¹⁴CO₂ was quantitatively measured every day for 15 days and residual soil ¹⁴C after 125 days. The percent ¹⁴C remaining in the soil after 125 days of incubation was positively and significantly correlated with the percent substrate utilized in the first day of incubation. The ¹⁴C in the microbial biomass ranged from 4–15% after 15 days and declined through day 125, contributing significantly to the ¹⁴C that evolved over the longer time period. Priming of ¹²C soil organic matter (SOM) was negative at day 3 but became positive, reaching a maximum on day 12; the total increase in soil C from added substrates was greater than the primed C. The primed C came from ¹²C SOM rather than the microbial biomass. This data supports the concept that the more rapidly a substrate is initially mineralized, the more persistent it will be in the soil over time.     

不同碳源添加对钙质土高累积硝态氮向土壤有机氮库转化的研究

Excessive amounts of nitrate have accumulated in many soils on the North China Plain due to the large amounts of chemical N fertilizers or manures used in combination with low carbon inputs. We investigated the potential of different carbon substrates added to transform soil nitrate into soil organic N (SON). A 56-d laboratory incubation experiment using the 15 N tracer (K15 NO3 ) technique was carried out to elucidate the proportion of SON derived from accumulated soil nitrate following amendment with glucose or maize straw at controlled soil temperature and moisture. The dynamics and isotopic abundance of mineral N (NO3 and NH+4 ) and SON and greenhouse gas (N2O and CO2 ) emissions during the incubation were investigated. Although carbon amendments markedly stimulated transformation of nitrate to newly formed SON, this was only a substitution effect of the newly formed SON with native SON because SON at the end of the incubation period was not significantly different (P > 0.05) from that in control soil without added C. At the end of the incubation period, amendment with glucose, a readily available C source, increased nitrate immobilization by 2.65 times and total N2O-N emission by 33.7 times, as compared with maize straw amendment. Moreover, the differences in SON and total N2O-N emission between the treatments with glucose and maize straw were significant (P < 0.05). However, the total N2O-N emission in the straw treatment was not significantly (P > 0.05) greater than that in the control. Straw amendment may be a potential option in agricultural practice for transformation of nitrate N to SON and minimization of N2O emitted as well as restriction of NO3-N leaching.     

Effect of biochar amendment on soil carbon balance and soil microbial activity

We investigated the behavior of biochars in arable and forest soil in a greenhouse experiment in order to prove that these amendments can increase carbon storage in soils. Two qualities of biochar were produced by hydrothermal pyrolysis from ¹³C labeled glucose (0% N) and yeast (5% N), respectively. We quantified respiratory losses of soil and biochar carbon and calculated mean residence times of the biochars using the isotopic label. Extraction of phospholipid fatty acids from soil at the beginning and after 4 months of incubation was used to quantify changes in microbial biomass and to identify microbial groups utilizing the biochars. Mean residence times varied between 4 and 29 years, depending on soil type and quality of biochar. Yeast-derived biochar promoted fungi in the soil, while glucose-derived biochar was utilized by Gram-negative bacteria. Our results suggest that residence times of biochar in soils can be manipulated with the aim to “design” the best possible biochar for a given soil type.