Biological pretreatment of sugarcane trash for its conversion to fermentable sugars

Pretreatment of lignocellulosic biomasses, the first step in their conversion to utilizable molecules requires very high energy (steam and electricity), corrosion resistant high-pressure reactors and high temperatures. These severe conditions not only add to the cost component of the entire process but also lead to the loss of sugars to the side reactions. Microbial pretreatments have been reported to be associated with reducing the cost factors as well as the severities of the reactions. Eight bioagents, including fungi and bacteria, were screened for their pretreatment effects on sugarcane trash. They narrowed down the C:N ratio of trash from 108:1 to a varying range of approximately 42:1 to 60:1.The maximum drop in C:N ratio of 61% was observed using Aspergillus terreus followed by Cellulomonas uda (52%) and Trichoderma reesei and Zymomonas mobilis (49%). The bioagents helped in degradation of sugarcane trash by production of cellulases, the maximum being produced by A. terreus, (12 fold) followed by C. uda (10 fold), Cellulomonas cartae (9 fold) and Bacillus macerans (8 fold). The microbial pretreatment of trash rendered the easy accessibility of sugars for enzymatic hydrolysis, which can be directed for production of alcohol.     

Ammonium carboxylate production from sugarcane trash using long-term air-lime pretreatment followed by mixed-culture fermentation

Sugarcane trash (ST) was converted to ammonium carboxylates using a novel bioprocessing strategy known as long-term air-lime pretreatment/mixed-culture fermentation. At mild conditions (50 °C, 5 weeks, 1-atm air, and excess lime loading of 0.4 g Ca(OH)₂/(g dry biomass)), air-lime pretreatment of ST had moderate delignification (64.4%) with little loss in polysaccharides. Without employing detoxification, sterility, expensive nutrients, or costly enzymes, the feedstock (80% treated ST/20% chicken manure) was fermented to primarily ammonium acetate (>75%) and butyrate by a mixed culture of marine microorganisms at 55 °C. In the best four-stage countercurrent fermentation, the product yield was 0.36 g total acids/(g VS fed) and the substrate conversion was 64%. Model predictions indicate both high acid concentrations (>47.5 g/L) and high substrate conversions (>70%) are possible at industrial scale.     

Long-term Effects of Pre-harvest Burning and Nitrogen and Vinasse Applications on Yield of Sugar Cane and Soil Carbon and Nitrogen Stocks on a Plantation in Pernambuco, N.E. Brazil

Since the 1970s the area under sugarcane in Brazil has increased from 2 million to over 5 million ha (M ha), and it is expected to pass the 7 M ha mark in 2007. More than half of the cane is harvested to produce bioethanol as a fuel for light vehicles. The distilleries produce approximately 13 L of distillery waste (vinasse) for each litre of ethanol produced. In the 1980s there was considerable concern over the long-term effects of the disposal of this material (containing about 1% carbon and high in K) on cane yields if it was applied to the field. At the same time there was a growing movement to abandon the practice of pre-harvest burning and some research was showing that some Brazilian varieties of sugar cane were able to obtain significant contributions of N from plant-associated biological nitrogen fixation (BNF). For these reasons an experiment was installed on a cane plantation in the state of Pernambuco, NE Brazil to investigate the long-term effects of vinasse and N fertiliser additions and the practice of pre-harvest burning on crop and sugar yield, soil fertility parameters, N balance and soil C stocks. The results showed that over a 16-year period, trash conservation (abandonment of burning) increased cane yields by 25% from a mean of 46 to 58 Mg ha⁻¹. Vinasse applications (80 m³ ha⁻¹ crop⁻¹) increased mean cane and sugar yield by 12 to 13% and the application of 80 kg N ha⁻¹ as urea increased cane yields by 9%, but total sugar yield by less than 6% (from 7.0 to 7.4 Mg ha⁻¹ crop⁻¹). The total N balance for the soil/plant system when only the surface 20 cm of the soil was considered was positive in plots where no N fertiliser was added. However, the data indicated that during the 16 years of the study considerable quantities of soil organic matter were accumulated below 20 cm depth such that the N balance considering the soil to 60 cm depth was strongly positive, except where N fertiliser was added. The data indicated that there were considerable BNF inputs to the system, which was consistent with its low response to N fertiliser and low N fertiliser-use-efficiency. There were no significant effects of vinasse or urea addition, or trash conservation on soil C stocks, although the higher yields proportioned by trash conservation had potentially significant benefits for increased mitigation of CO₂ emissions where the main use of the cane was for bioethanol production.     

Soil nitrogen turnover in proximal and distal stem areas of European beech trees

In European beech (Fagus sylvatica L.) forests, a large proportion of the water and ion input to the soil results from stemflow which creates a soil microsite of high element fluxes proximal to the tree trunk. The soil proximal to the stem is considered to have different rates of nitrogen turnover which might influence the estimation of N-turnover rates at the stand scale. In a previous study we reported high nitrate fluxes with seepage proximal to the stems in a forest dominated by European beech in Steigerwald, Germany. Here, we investigated the soil nitrogen turnover in the top 15 cm soil in proximal (defined as 1 m² around beech stems) and distal stem areas. Laboratory incubations and in situ sequential coring incubations were used to determine the net rates of ammonification, nitrification, and root uptake of mineral nitrogen. In the laboratory incubations higher rates of net nitrogen mineralization and nitrification were found in the forest floor proximal to the stem as compared to distal stem areas. No stem related differences were observed in case of mineral soil samples. In contrast, the in situ incubations revealed higher rates of nitrification in the mineral soil in proximal stem areas, while net nitrogen mineralization was equal in proximal and distal areas. In the in situ incubations the average ratio of nitrification/ammonification was 0.85 in proximal and 0.34 in distal stem areas. The net nitrogen mineralization was 4.4 g N m^-2 90 day^-1 in both areas. Mineralized nitrogen was almost completely taken up by tree roots with ammonium as the dominant nitrogen species. The average ratio of nitrate/ammonium uptake was 0.69 in proximal and 0.20 in distal areas. The higher water content of the soil in proximal stem areas is considered to be the major reason for the increased rates of nitrification. Different nitrogen turnover rates in proximal stem areas had no influence on the nitrogen turnover rates in soil at the stand scale. Consequently, the observed high nitrate fluxes with seepage proximal to stems are attributed to the high nitrogen input by stemflow rather than to soil nitrogen turnover. This revised version was published online in August 2006 with corrections to the Cover Date.     

甘蔗/大豆间作减量施氮对甘蔗产量、品质及经济效益的影响

在大田栽培条件下,研究了减量施氮对广州地区不同甘蔗/大豆间作模式下甘蔗产量、品质、生物量动态变化及甘蔗群体经济效益的影响.结果表明:氮肥和种植模式对甘蔗产量、品质均无显著影响,各处理甘蔗产量、品质均无显著差异.不同施氮水平的甘蔗/大豆间作系统较单作甘蔗提高了土地利用率,其土地当量比(LER)为1.36～2.12.各处理甘蔗总生物量的动态积累随生育期的推进均符合"S"型生长曲线,低氮水平下问作甘蔗的生物量动态积累特征参数最协调,高氮水平下,甘蔗生长高峰期提前,快速生长最大持续期缩短,甘蔗产量下降,因此,可以通过调整氮素的投入量来改善生物量的增长参数,从而获得高产.低氮处理甘蔗/大豆间作系统经济效益比高氮处理高3.2%～26.3%,说明减量施氮可有效提高群体经济效益.甘蔗/大豆1∶2间作模式的经济效益最好.     

Reassessing the nitrogen relations of Arctic plants: a mini-review

The Arctic is often assumed to be an NH₄⁺-dominated ecosystem. This review assesses the validity of this assumption. It also addresses the question of whether Arctic plant growth is limited by the ability to use the forms of nitrogen that are available. The review demonstrates that several sources of soil nitrogen are available to Arctic plants, including soluble organic nitrogen (e.g. glycine, aspartic acid and glutamic acid), NH₄⁺ and NO^?₃. In mesic Arctic soils, soluble organic nitrogen is potentially more important than either NH⁺₄ or NO^?₃. Many Arctic species are capable of taking up soluble organic nitrogen (either directly and/or in association with ectomycorrhizae), with the greatest potential for soluble organic nitrogen uptake being exhibited by deciduous species. The ability to take up soluble organic nitrogen may enable some Arctic plants to avoid nitrogen limitations imposed by the slow rate of organic matter decomposition. NO^?₃ is also present in many Arctic soils, especially calcareous soils and soils near flowing water, animal burrows and bird cliffs. Arctic species characteristic of mesic and xeric habitats are capable of taking up and assimilating NO^?₃. Even when present in lower concentrations in soils than NH⁺₄, NO^?₃ is still an important source of nitrogen for some Arctic plants. Arctic-plants therefore have a variety of nitrogen sources available to them, and are capable of using those nitrogen sources. Taken together, these findings demonstrate that the Arctic is not an NH⁺₄dominated ecosystem. Symbiotic fixation of atmospheric N₂ does not appear to be an important source of nitrogen for Arctic plants. The reliance of Arctic plants on internal recycling of nitrogen substantially reduces their dependence on soil nitrogen uptake (this is particularly the case for slow-growing evergreens). Despite the high level of internal nitrogen recycling, Arctic plant growth remains limited by the low levels of available soil nitrogen. However, Arctic plant growth is not limited by an inability to utilize any of the available forms of nitrogen. The potential effects of climatic warming on nitrogen availability and use are discussed. The question of whether the Arctic ecosystem is uniquely different from temperate nitrogen-deficient ecosystems is also assessed.     

Plant response to variations in nitrogen availability in a desert shrubland community

Spatial variations in nitrogen availability were studied in a desert community codominated byLarrea tridentata (DC.) Cov. andProsopis glandulosa Torr. Measurements of natural ¹⁵N values in tissues suggested thatProsopis obtains approximately half of its nitrogen through direct symbiotic fixation. Soils were collected under 1)Prosopis shrubs, 2)Larrea shrubs 2 m fromProsopis (LP), and 3)Larrea 2 m from otherLarrea but> 5 m from the nearestProsopis (LL).Prosopis soils showed significantly higher rates of nitrogen mineralization than LL soils in both A and B horizons. Rates of mineralization in LP soils were significantly higher than rates in LL soils only in the B horizon and were not significantly different from rates inProsopis soils. Leaf nitrogen concentrations were significantly higher in LP shrubs (2.06%) than in LL shrubs (1.78%), although ¹⁵N values did not differ between the two shrub types. Nitrogen concentrations inPerezia nana Gray, a perennial herb, were greater in plants underProsopis shrubs (2.09%) than under LP shrubs (1.93%) or LL shrubs (1.67%). Despite apparent differences in nitrogen availability, biomass ofLarrea and density ofPerezia did not differ significantly among these sites.     

Characterization of a proton translocating ATPase and sucrose uptake in a tonoplast-enriched vesicle fraction from sugarcane

A sucrose gradient fraction was used to characterize the tonoplast ATPase from storage tissue of the sugarcane plant ( Saccharum sp. var. H57–5175). Marker enzyme analyses and characterization of low-density vesicles isolated on a sucrose gradient were consistent with a highly enriched tonoplast fraction. ATPase and proton transport activities were both substantially inhibited by nitrate (80%), but very little by vanadate (10%), indicating a high titer of tonoplast compared to plasma-membrane vesicles in the fraction. Sensitivity toward other inhibitors, as well as ion effects, correlated closely among ATPase and proton translocation activities. Although the vesicles in this fraction showed good proton translocating activity there was no indication that ATP stimulated sucrose uptake in this tonoplast population.     

Seasonal flooding,nitrogen mineralization and nitrogen utilization in a prairie marsh

Flooding can be an important control of nitrogen (N) biogeochemistry in wetland ecosystems. In North American prairie marshes, spring flooding is a dominant feature of the physical environment that increases emergent plant production and could influence N cycling. I investigated how spring flooding affects N availability and plant N utilization in whitetop (Scolochloa festucacea) marshes in Manitoba, Canada by comparing experimentally spring-flooded marsh inside an impoundment with adjacent nonflooded marsh. The spring-flooded marsh had net N mineralization rates up to 4 times greater than nonflooded marsh. Total growing season net N mineralization was 124 kg N ha^–1 in the spring-flooded marsh compared with 62 kg N ha^–1 in the nonflooded marsh. Summer water level drawdown in the spring-flooded marsh decreased net N mineralization rates. Net nitrification rates increased in the nonflooded marsh following a lowering of the water table during mid summer. Growing season net nitrification was 33 kg N ha^–1 in the nonflooded marsh but < 1 kg N ha^–1 in the spring-flooded marsh. Added NO₃^–1 induced nitrate reductase (NRA) activity in whitetop grown in pot culture. Field-collected plants showed higher NRA in the nonflooded marsh. Nitrate comprised 40% of total plant N uptake in the nonflooded marsh but <1% of total N uptake in the spring-flooded marsh. Higher plant N demand caused by higher whitetop production in the spring-flooded marsh approximately balanced greater net N mineralization. A close association between the presence of spring flooding and net N mineralization and net nitrification rates indicated that modifications to prairie marshes that change the pattern of spring inundation will lead to rapid and significant changes in marsh N cycling patterns.     

Free amino acid, ammonium and nitrate concentrations in soil solutions of a grazed coastal marsh in relation to plant growth

Soluble free amino acids, ammonium and nitrate ions as sources of nitrogen for plant growth were measured in soils of a coastal marsh grazed by snow geese in Manitoba, Canada. Amounts of nitrogen, primarily ammonium ions, increased in the latter half of the growing season and over winter, but fell to low values early in the growing season. Free amino acid concentrations relative to ammonium concentrations were highest during the period of rapid plant growth in early summer, especially in soils in the intertidal zone, where the median ratio of amino acid nitrogen to ammonium nitrogen was 0·36 and amino acid concentrations exceeded those of ammonium ions in 24% of samples. Amino acid profiles, which were dominated by alanine, proline and glutamic acid, were similar to goose faecal profiles. In a continuous flow hydroponic experiment conducted in the field, growth of the salt‐marsh grass, Puccinellia phryganodes, on glycine was similar to growth on ammonium ions at an equivalent concentration of nitrogen. When supplies of soil inorganic nitrogen are low, amino acids represent a potentially important source of nitrogen for the re‐growth of plants grazed by geese and amino acid uptake may be as high as 57% that of ammonium ions.     

Ammonium uptake and mineralization in prairie streams: chamber incubation and short-term nutrient addition experiments

1. We used two separate approaches to estimate ambient ammonium cycling in the north and south branches of Kings Creek, a prairie stream. Chamber experiments were conducted to determine ammonium uptake and mineralization rates associated with epilithic biofilms and filamentous algae collected from the streams. A series of short-term whole-stream ammonium addition experiments were also conducted to estimate the rate of uptake at ambient stream concentrations, based on the relationship between ammonium concentrations and uptake rates.
2. Chamber experiments were scaled up to whole-stream levels, resulting in ambient gross uptake estimates of 0.08  μ g⁻² s⁻¹ for the north branch and 0.16  μ g⁻² s⁻¹ for the south branch. The substrata-specific estimates of mineralization were higher than uptake in both streams.
3. Substrata-specific measurements indicated that ammonium uptake is higher in riffle habitats than in pools habitats. The results of the short-term ammonium addition experiments support these findings.
4. Short-term ammonium addition experiments show that uptake rates saturate with increasing ammonium concentrations. The observed saturation of uptake rates is consistent with a Michaelis–Menten relationship.
5. Scaled estimates of uptake from the chamber experiments were similar to estimates of ambient ammonium uptake based on the whole-stream experiments, and were comparable with previous estimates of ammonium uptake and mineralization made by using stable isotope tracer methods in Kings Creek.     

Microcosm investigation of growth and phytoremediation potential of Azolla japonica along nitrogen gradients

Although Azolla species are among the most promising plants for use in phytoremediation, more studies on their growth and nitrogen (N) uptake along the N gradients of growing media are required. In this study, N concentration-dependent growth in growing media and phosphorus (P) and N accumulation by Azolla japonica were studied by estimating direct N uptake from media by molybdenum-iron proteins. The doubling time of A. japonica was less than a week, regardless of the N concentration (0, 5, and 25 mg N/L) present in the growth media, indicating that this plant is suitable for remediation. Plants showed a high uptake of P, probably via plant-bacteria symbiosis, indicating their potential for effective P remediation. A. japonica also showed more than 4% N content regardless of the treatment and accumulated more than 40 mg of N per microcosm in 3 weeks. iron and molybdenum levels in plants were strongly associated with N fixation, and N uptake from media was estimated to be more than 25 mg per microcosm in 3 weeks, indicating that A. japonica has N remediation potential. As A. japonica is a rapidly growing plant, capable of efficient P and N remediation, it has great potential for use in phytoremediation of nutrient-enriched waters such as agricultural or urban wastewater and eutrophicated aquatic ecosystems.     

Effect of specific leaf nitrogen content on photosynthesis of sugarcane

In the laboratory, rate of photosynthesis (P) of the youngest fully emerged leaf lamina on a shoot, whether taken from a 4 or 8 month old sugarcane plant, increased approximately linearly with specific leaf nitrogen content (SLN) from < 1.0 to c. 1.7 g m^-2; the rate of increase was greater the higher the incident photosynthetically active radiation (PAR) flux density, over the range 80 to 1600 μmol m^-2s^-1. In a field crop receiving standard fertiliser, including 150 kg N ha^-1, SLN of the green leaf canopy decreased more or less linearly with depth, from > 1.7 g m^-2at the top to c. 1.0 g m^-2at the bottom at 15 wk of age, and progressively with time to 1.2 to 0.85 g m^-2from top to bottom of the canopy by week 45. Where an extra 300 N ha^-1were applied in top-dressings, the decrease in SLN with time was somewhat smaller, to 1.3 to 0.9 g m^-2from top to bottom of the canopy by week 45. Calculations with a model, using the data from laboratory and field, suggest a decrease of about 25% in canopy gross P and about 30% in canopy net P from wk 15 to wk 45 in the crop receiving standard fertiliser only; the calculated decreases where the crop received additional nitrogen were about 17% and 20% respectively. There would seem to be a need for a change in type of cultivar to make profitable use of augmented nitrogen fertiliser.     

Successional changes in soil nitrogen availability,non-symbiotic nitrogen fixation and carbon/nitrogen ratios in southern Chilean forest ecosystems

Vast areas of southern Chile are now covered by second-growth forests because of fire and logging. To study successional patterns after moderate-intensity, anthropogenic fire disturbance, we assessed differences in soil properties and N fluxes across a chronosequence of seven successional stands (2–130 years old). We examined current predictions of successional theory concerning changes in the N cycle in forest ecosystems. Seasonal fluctuations of net N mineralization (N_min) in surface soil and N availability (N_a; N_a=NH ₄ ⁺ –N+NO ₃ ^– –N) in upper and deep soil horizons were positively correlated with monthly precipitation. In accordance with theoretical predictions, stand age was positively, but weakly related to both N_a (r ²=0.282, P<0.001) and total N (N_tot; r ²=0.192, P<0.01), and negatively related to soil C/N ratios (r ²=0.187, P<0.01) in surface soils. A weak linear increase in soil N_min (upper plus deep soil horizons) was found across the chronosequence (r ²=0.124, P<0.022). N_min occurred at modest rates in early successional stands, suggesting that soil disturbance did not impair microbial processes. The relationship between N fixation (N_fix) in the litter layer and stand age best fitted a quadratic model (r ²=0.228, P<0.01). In contrast to documented successional trends for most temperate, tropical and Mediterranean forests, non-symbiotic N_fix in the litter layer is a steady N input to unpolluted southern temperate forests during mid and late succession, which may compensate for hydrological losses of organic N from old-growth ecosystems.     

Effect of liming on mineralization of soil nitrogen as measured by plant uptake and nitrogen released during incubation

The effect of lime rates on oat yield and N uptake was measured in a 6-years pot experiment, using 12 acid surface soils (pH 4.7 to 6.0). Mineralization of nitrogen was measured by incubation of soil samples taken after harvest each year from the different lime treatments.Nitrogen uptake was significantly correlated with total N in the soils. Averaged over all 12 soils liming only to pH 7 or above, increased the oat yield significantly. Liming increased the N concentration of grain and the N uptake significantly during a 4-years period, indicating the effect of lime on N mineralization.The mineralization of organic N measured by incubation in the non-limed samples was highly correlated with the total N concentration, but it was not significantly related to the original pH of the soils. The amounts of N released as well as the duration of the lime effect on mineralization varied among soils. When pH was raised to 7 or above, considerable increases in N mineralization occurred in some soils. Based on average values, liming increased N mineralization significantly during a 3-years period. After 3 years, the lime treatments differed only slightly from the non-limed treatments.     

Temperature-dependent nitrogen transformations in acid oak-beech forest litter in the Netherlands

Laboratory incubation experiments have been carried out to quantify net nitrogen mineralization and nitrification in oak-beech litter at temperatures ranging from 0 to 30°C. Net mineralization was linearly proportional to temperature. Nitrification was inhibited at 0,5 and 30°C. As compared with soils under cultivation, there is only restricted knowledge of nitrification kinetics in acid forest litters, especially when temperature is considered. With these litter types, one should be cautious applying high incubation temperatures, which seldomly occur under field conditions.     

Interactions of light, temperature and inorganic nitrogen in controlling planktonic nitrogen utilisation in the Tagus estuary

The effect of light, temperature and ammonium on inorganic nitrogen uptake by phytoplankton was investigated from June 1994 through December 1995 at three sites in the Tagus estuary (Portugal), during high tide of neap tides. Ammonium concentrations higher than 10 M reduced nitrate uptake down to 24% but never prevented it. Below this threshold concentration, nitrate uptake was neither inhibited nor changed. Uptake of both nitrate and ammonium as a function of light intensity exhibited a saturation response. Uptake reduction occurred in the near bottom phytoplankton populations, particularly for nitrate. The ammonium uptake system was less limited by light than the nitrate uptake system, indicating the importance of ammonium as a nitrogen source for the phytoplankton which is likely to experience high changes in light in the well-mixed water column of this estuarine environment. Ammonium uptake was exponentially related to temperature in the upper estuary whereas in the mid and lower estuary this relationship was linear. The effect of temperature on nitrate uptake was linear but far less marked than for ammonium uptake.     

A putative new endophytic nitrogen-fixing bacterium Pantoea sp. from sugarcane

AIMS: To isolate and identify endophytic nitrogen-fixing bacteria in sugarcane growing in Cuba without chemical fertilizers. METHODS AND RESULTS: Two N2-fixing isolates, 9C and T2, were obtained from surface-sterilized stems and roots, respectively, of sugarcane variety ML3-18. Both isolates showed acetylene reduction and H2 production in nitrogen-free media. Nitrogenase activity measured by H2 production was about 15 times higher for isolate 9C than for T2 or for Gluconoacetobacter diazotrophicus (PAL-5 standard strain, ATCC 49037). The nifH gene segment was amplified from both isolates using specific primers. Classification of both T2 and 9C was made on the basis of morphological, biochemical, PCR tests and 16S rDNA sequence analysis. CONCLUSIONS: Isolate 9C was identified as a Pantoea species from its 16S rDNA, but showed considerable differences in physiological properties from previously reported species of this genus. For example, 9C can be cultured over a wide range of temperature, pH and salt concentration, and showed high H2 production (up to 67.7 nmol H2 h(-1) 10(10) cell(-1)). Isolate T2 was a strain of Gluconacetobacter diazotrophicus. SIGNIFICANCE AND IMPACT OF THE STUDY: A new N2-fixing endophyte, i.e. Pantoea, able to produce H2 and to grow in a wide range of conditions, was isolated from sugarcane stem tissue and characterized. The strain with these attributes may well be valuable for agriculture.     

Timing of nitrogen uptake affects winter storage and spring remobilisation of nitrogen in nectarine (Prunus persica var. nectarina) trees

Two-year old nectarine trees (Prunus persica, Batsch, var. nectarina, cv. Starkredgold on GF305 rootstock) planted in pots each received five applications of 1.0 g ¹⁵N labelled urea either from mid May to mid July (early uptake) or from mid August to the beginning of October (late uptake). All trees were supplied with a corresponding amount of unlabelled urea when they did not receive the labelled N. In autumn, all abscised leaves were collected and during winter randomly selected trees were harvested and divided into main organs. The remaining trees were transplanted into similar pots filled with sand; they received no N fertiliser and were harvested in May to evaluate the remobilisation of N. Total N and ¹⁵N abundance were determined in each organ. Nectarine trees took up similar amounts of N in the 'early' and in the 'late' period; however, more labelled nitrogen was recovered in the perennial organs during the winter when trees received the labelled N in the 'late' than in the 'early' period. Some 73–80% of the N present in the dormant trees was stored in the roots, which contained almost twice the amount of labelled N taken up 'late' than that absorbed 'early'. Nitrogen for spring growth was remobilised predominantly from the roots and accounted for some 43–49% of the labelled N recovered in the tree during winter. Results suggest that the nitrogen taken up 'late' in the season is preferentially stored in roots and used by peach trees to sustain new growth the following spring. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nitrogen transformations in two nitrogen saturated forest ecosystems subjected to an experimental decrease in nitrogen deposition

Nitrogen transformations were studied in the forest floor and mineral soil (0–5 cm) of a Douglas fir forest (Pseudotsuga menziesii (Mirb.) Franco.) and a Scots pine forest (Pinus sylvestris L.) in the Netherlands. Curren nitrogen depositions (40 and 56 kg N ha^-1 yr^-1, respectively) were reduced to natural background levels (1–2 kg N ha^-1 yr^-1) by a roof construction. The study concentrated on rates and dynamic properties of nitrogen transformations and their link with the leaching pattern and nitrogen uptake of the vegetation under high and reduced nitrogen deposition levels. Results of an in situ field incubation experiment and laboratory incubations were compared. No effect of the reduced N deposition on nitrogen transformations was found in the Douglas fir forest. In the Scots pine forest, however, during some periods of the year nitrogen transformations were significantly decreased under the low nitrogen deposition level. At low nitrogen inputs a net immobilization occurred during most of the year leading to a very small net mineralization for the whole year. In laboratory and in individual field plots nitrogen transformations were negatively correlated with initial inorganic nitrogen concentrations. Nitrogen budget estimates showed that nitrogen transformations were probably underestimated by the in situ incubation technique. Nevertheless less nitrogen was available for plant uptake and leaching at the low deposition plots.