Comparative Evidence that Salt Marshes and Mangroves May Protect Seagrass Meadows from Land-derived Nitrogen Loads

Seagrass meadows within estuaries are highly sensitive to increased supplies of nitrogen (N). The urbanization of coastal watersheds increases the delivery of N to estuaries, threatening seagrass habitats; both seagrass production per unit area and the area of seagrass meadows diminish as land-derived N loads increase. The damaging effects of land-derived N loads may be lessened where there are fringes of coastal wetlands interposed between land and seagrass meadows. Data compiled from the literature showed that production per unit area by seagrasses increased and losses of seagrass habitat were lower in estuaries with relatively larger areas of fringing wetlands. Denitrification and the burial of land-derived N within fringe wetlands may be sufficient to protect N-sensitive seagrass habitats from the detrimental effects of land-derived N. The protection furnished by fringing wetlands may be overwhelmed by increases in anthropogenic N loads in excess of 20–100 kg N ha⁻¹ y⁻¹. The relationships of land-derived N loadings, fringing coastal wetlands, and seagrass meadows demonstrate that different units of the landscape mosaic found in coastal zones do not exist as separate units, but instead are coupled and uncoupled by biogeochemical transformations and transport among environments. Received 12 December 2000; accepted 15 August 2001.     

Long-Term Nitrogen Addition Does Not Increase Soil Carbon Storage or Cycling Across Eight Temperate Forest and Grassland Sites on a Sandy Outwash Plain

Ecosystems - Experimental nitrogen (N) deposition generally inhibits decomposition and promotes carbon (C) accumulation in soils, but with substantial variation among studies. Differences in...     

Nitrogen Fixation (Acetylene Reduction) in a Salt Marsh Amended with Sewage Sludge and Organic Carbon and Nitrogen Compounds

Seasonal distribution of nitrogen fixation by Spartina alterniflora epiphytes and in surface and soil samples was investigated in a Georgia salt marsh which was amended with sewage sludge or with glucose and/or ammonium nitrate. There was no significant difference between the rates of fixation in the unamended and sewage sludge plots. Additional perturbation experiments suggested that nitrogen addition indirectly stimulates nitrogen fixation by enhancing Spartina production and root exudation. Glucose additions, on the other hand, suppressed nitrogen fixation on a long-term basis. It is suggested that the microbial population in the soil out-competed the plants for the available nitrogen and in turn suppressed plant production and possibly root exudation. A comparison of nitrogen fixation in clipped and unclipped Spartina plots substantiated the suggestion that root exudation probably supports nitrogen fixation. Fixation in the clipped plots was significantly lower (P < 0.05) than the rates in the unclipped plots.     

Non-Additive Effects of Water and Nitrogen Addition on Ecosystem Carbon Exchange in a Temperate Steppe

Changes in precipitation and nitrogen (N) deposition can influence ecosystem carbon (C) cycling and budget in terrestrial biomes, with consequent feedbacks to climate change. However, little is known about the main and interactive effects of water and N additions on net ecosystem C exchange (NEE). In a temperate steppe of northern China, a field-manipulated experiment was conducted to evaluate the responses of NEE and its components to improve N and water availability from 2005 to 2008. The results showed that both water and N additions stimulated gross ecosystem productivity (GEP), ecosystem respiration (ER), and NEE. Water addition increased GEP by 17%, ER by 24%, and NEE by 11% during the experimental period, whereas N addition increased GEP by 17%, ER by 16%, and NEE by 19%. The main effects of both water and N additions changed with time, with the strongest water stimulation in the dry year and a diminishing N stimulation over time. When water and N were added in combination, there were non-additive effects of water and N on ecosystem C fluxes, which could be explained by the changes in species composition and the shifts of limiting resources from belowground (water or N) to aboveground (light). The positive water and N additions effects indicate that increasing precipitation and N deposition in the future will favor C sequestration in the temperate steppe. The non-additive effects of water and N on ecosystem C fluxes suggest that multifactor experiments are better able to capture complex interactive processes, thus improving model simulations and projections.     

Identification of General Patterns of Nutrient and Labile Carbon Control on Soil Carbon Dynamics Across a Successional Gradient

Carbon (C) inputs and nutrient availability are known to affect soil organic carbon (SOC) stocks. However, general rules regarding the operation of these factors across a range of soil nutrient availabilities and substrate qualities are unidentified. “Priming” (stimulated decomposition by labile C inputs) and ‘preferential substrate utilization’ (retarded decomposition due to shifts in community composition towards microbes that do not mineralize SOC) are two hypotheses to explain effects of labile C additions on SOC dynamics. For effects of nutrient additions (nitrogen and phosphorus) on SOC dynamics, the stoichiometric (faster decomposition of materials of low carbon-to-nutrient ratios) and ‘microbial mining’ (that is, reduced breakdown of recalcitrant C forms for nutrients under fertile conditions) hypotheses have been proposed. Using the natural gradient of soil nutrient availability and substrate quality of a chronosequence, combined with labile C and nutrient amendments, we explored the support for these contrasting hypotheses. Additions of labile C, nitrogen (N), phosphorus (P), and combinations of C and N and C and P were applied to three sites: 2-year fallow grassland, mature grassland and forest, and the effects of site and nutrient additions on litter decomposition and soil C dynamics were assessed. The response to C addition supported the preferential substrate hypothesis for easily degradable litter C and the priming hypothesis for SOC, but only in nitrogen-enriched soils of the forest site. Responses to N addition supported the microbial mining hypothesis irrespective of C substrate (litter or SOC), but only in the forest site. Further, P addition effects on SOC support the stoichiometric hypothesis; P availability appeared key to soil C release (priming) in the forest site if labile C and N is available. These results clearly link previously contrasting hypotheses of the factors controlling SOC with the natural gradient in litter quality and nutrient availability that exists in ecosystems at different successional stages. A holistic theory that incorporates this variability of responses, due to different mechanisms, depending on nutrient availability and substrate quality is essential for devising management strategies to safeguard soil C stocks.     

Carbon Sequestration Efficiency of Organic Amendments in a Long-Term Experiment on a Vertisol in Huang-Huai-Hai Plain,China

Soil organic carbon (SOC) sequestration is important for improving soil fertility of cropland and for the mitigation of greenhouse gas emissions to the atmosphere. The efficiency of SOC sequestration depends on the quantity and quality of the organic matter, soil type, and climate. Little is known about the SOC sequestration efficiency of organic amendments in Vertisols. Thus, we conducted the research based on 29 years (1982–2011) of long-term fertilization experiment with a no fertilizer control and five fertilization regimes: CK (control, no fertilizer), NPK (mineral NPK fertilizers alone), NPK+1/2W (mineral NPK fertilizers combined with half the amount of wheat straw), NPK+W (mineral NPK fertilizers combined with full the amount of wheat straw), NPK+PM (mineral NPK fertilizers combined with pig manure) and NPK+CM (mineral NPK fertilizers combined cattle manure). Total mean annual C inputs were 0.45, 1.55, 2.66, 3.71, 4.68 and 6.56 ton/ha/yr for CK, NPK, NPKW1/2, NPKW, NPKPM and NPKCM, respectively. Mean SOC sequestration rate was 0.20 ton/ha/yr in the NPK treatment, and 0.39, 0.50, 0.51 and 0.97 ton/ha/yr in the NPKW1/2, NPKW, NPKPM, and NPKCM treatments, respectively. A linear relationship was observed between annual C input and SOC sequestration rate (SOCsequestration rate  = 0.16 Cinput –0.10, R = 0.95, P<0.01), suggesting a C sequestration efficiency of 16%. The Vertisol required an annual C input of 0.63 ton/ha/yr to maintain the initial SOC level. Moreover, the C sequestration efficiencies of wheat straw, pig manure and cattle manure were 17%, 11% and 17%, respectively. The results indicate that the Vertisol has a large potential to sequester SOC with a high efficiency, and applying cattle manure or wheat straw is a recommendable SOC sequestration practice in Vertisols.     

The Role of Dissolved Organic Carbon, Dissolved Organic Nitrogen, and Dissolved Inorganic Nitrogen in a Tropical Wet Forest Ecosystem

Although tropical wet forests play an important role in the global carbon (C) and nitrogen (N) cycles, little is known about the origin, composition, and fate of dissolved organic C (DOC) and N (DON) in these ecosystems. We quantified and characterized fluxes of DOC, DON, and dissolved inorganic N (DIN) in throughfall, litter leachate, and soil solution of an old-growth tropical wet forest to assess their contribution to C stabilization (DOC) and to N export (DON and DIN) from this ecosystem. We found that the forest canopy was a major source of DOC (232 kg C ha^–1 y^–1). Dissolved organic C fluxes decreased with soil depth from 277 kg C ha^–1 y^–1 below the litter layer to around 50 kg C kg C ha^–1 y^–1 between 0.75 and 3.5m depth. Laboratory experiments to quantify biodegradable DOC and DON and to estimate the DOC sorption capacity of the soil, combined with chemical analyses of DOC, revealed that sorption was the dominant process controlling the observed DOC profiles in the soil. This sorption of DOC by the soil matrix has probably led to large soil organic C stores, especially below the rooting zone. Dissolved N fluxes in all strata were dominated by mineral N (mainly NO₃⁻). The dominance of NO₃^– relative to the total amount nitrate of N leaching from the soil shows that NO₃^– is dominant not only in forest ecosystems receiving large anthropogenic nitrogen inputs but also in this old-growth forest ecosystem, which is not N-limited.     

Species and Genotype Effects of Bioenergy Crops on Root Production,Carbon and Nitrogen in Temperate Agricultural Soil

Bioenergy crops have a secondary benefit if they increase soil organic C (SOC) stocks through capture and allocation below-ground. The effects of four genotypes of short-rotation coppice willow (Salix spp., ‘Terra Nova’ and ‘Tora’) and Miscanthus (M.?×?giganteus (‘Giganteus’) and M. sinensis (‘Sinensis’)) on roots, SOC and total nitrogen (TN) were quantified to test whether below-ground biomass controls SOC and TN dynamics. Soil cores were collected under (‘plant’) and between plants (‘gap’) in a field experiment on a temperate agricultural silty clay loam after 4 and 6 years’ management. Root density was greater under Miscanthus for plant (up to 15.5 kg m^?3) compared with gap (up to 2.7 kg m^?3), whereas willow had lower densities (up to 3.7 kg m^?3). Over 2 years, SOC increased below 0.2 m depth from 7.1 to 8.5 kg m^?3 and was greatest under Sinensis at 0–0.1 m depth (24.8 kg m^?3). Miscanthus-derived SOC, based on stable isotope analysis, was greater under plant (11.6 kg m^?3) than gap (3.1 kg m^?3) for Sinensis. Estimated SOC stock change rates over the 2-year period to 1-m depth were 6.4 for Terra Nova, 7.4 for Tora, 3.1 for Giganteus and 8.8 Mg ha^?1 year^?1 for Sinensis. Rates of change of TN were much less. That SOC matched root mass down the profile, particularly under Miscanthus, indicated that perennial root systems are an important contributor. Willow and Miscanthus offer both biomass production and C sequestration when planted in arable soil.     

The Ratios of Carbon,Nitrogen, and Phosphorus in a Wetland Coastal Ecosystem of Southern India

The fertility of the coastal and estuarine waters is of great concern because of its influence on the productivity of these waters. Seasonal variations in the distribution of organic carbon, total nitrogen and total phosphorus in the sediments of Kuttanad Waters, a part of the tropical Cochin Estuary on the south west coast of India, are examined to identify the contribution of sediments to the fertility of the aquatic systems. The adjoining region has considerable agricultural activity. The fresh water zones had higher quantities of silt and clay whereas the estuarine zone was more sandy. Organic carbon, total phosphorus and total nitrogen were higher in the fresh water zones and lower in the estuarine zones. Total phosphorus and organic carbon showed the lowest values during monsoon periods. No significant trends were observed in the seasonal distributions of total nitrogen. Ratios of C/N, C/P and N/P, and the phosphorus and nitrogen content indicate significant modification in the character of the organic matter. Substantial amounts of the organic matter can contribute to reducing conditions and modify diagenetic processes.     

The Effects of Manure and Nitrogen Fertilizer Applications on Soil Organic Carbon and Nitrogen in a High-Input Cropping System

With the goal of improving N fertilizer management to maximize soil organic carbon (SOC) storage and minimize N losses in high-intensity cropping system, a 6-years greenhouse vegetable experiment was conducted from 2004 to 2010 in Shouguang, northern China. Treatment tested the effects of organic manure and N fertilizer on SOC, total N (TN) pool and annual apparent N losses. The results demonstrated that SOC and TN concentrations in the 0-10cm soil layer decreased significantly without organic manure and mineral N applications, primarily because of the decomposition of stable C. Increasing C inputs through wheat straw and chicken manure incorporation couldn''t increase SOC pools over the 4 year duration of the experiment. In contrast to the organic manure treatment, the SOC and TN pools were not increased with the combination of organic manure and N fertilizer. However, the soil labile carbon fractions increased significantly when both chicken manure and N fertilizer were applied together. Additionally, lower optimized N fertilizer inputs did not decrease SOC and TN accumulation compared with conventional N applications. Despite the annual apparent N losses for the optimized N treatment were significantly lower than that for the conventional N treatment, the unchanged SOC over the past 6 years might limit N storage in the soil and more surplus N were lost to the environment. Consequently, optimized N fertilizer inputs according to root-zone N management did not influence the accumulation of SOC and TN in soil; but beneficial in reducing apparent N losses. N fertilizer management in a greenhouse cropping system should not only identify how to reduce N fertilizer input but should also be more attentive to improving soil fertility with better management of organic manure.     

Impacts of Labile Organic Carbon Concentration on Organic and Inorganic Nitrogen Utilization by a Stream Biofilm Bacterial Community

In aquatic ecosystems, carbon (C) availability strongly influences nitrogen (N) dynamics. One manifestation of this linkage is the importance in the dissolved organic matter (DOM) pool of dissolved organic nitrogen (DON), which can serve as both a C and an N source, yet our knowledge of how specific properties of DOM influence N dynamics are limited. To empirically examine the impact of labile DOM on the responses of bacteria to DON and dissolved inorganic nitrogen (DIN), bacterial abundance and community composition were examined in controlled laboratory microcosms subjected to various combinations of dissolved organic carbon (DOC), DON, and DIN treatments. Bacterial communities that had colonized glass beads incubated in a stream were treated with various glucose concentrations and combinations of inorganic and organic N (derived from algal exudate, bacterial protein, and humic matter). The results revealed a strong influence of C availability on bacterial utilization of DON and DIN, with preferential uptake of DON under low C concentrations. Bacterial DON uptake was affected by the concentration and by its chemical nature (labile versus recalcitrant). Labile organic N sources (algal exudate and bacterial protein) were utilized equally well as DIN as an N source, but this was not the case for the recalcitrant humic matter DON treatment. Clear differences in bacterial community composition among treatments were observed based on terminal restriction fragment length polymorphisms (T-RFLP) of 16S rRNA genes. C, DIN, and DON treatments likely drove changes in bacterial community composition that in turn affected the rates of DON and DIN utilization under various C concentrations.     

人工林和农田对东北地区土壤碳、氮含量及相关指标垂直分布的影响

以兴安落叶松(Larix gmelinii)人工林及其附近农田为研究对象,选取8组配对样地不同土层进行相关指标测定。结果发现:多数样地(8组中的7组)0～20 cm土层有机碳含量林地高于农田37%,但深层(20～80 cm)农田高于林地8%～58%;土壤无机碳中所有样地平均显示林地高于农田(林地:1.33 mg·kg-1;农田:1.17 mg·kg-1);表层(0～20 cm)林地土壤全氮和碱解氮多高于农田,平均高出20%和34%,而深层土壤中(20～80 cm)多表现为相反趋势,这使得0～80 cm土层平均林地(6%)<农田(4%)。0～20 cm土层多为林地pH值>农田,林地电导率、容重<农田,而深层多(4～5组样地)多表现为相反趋势,0～80 cm土壤平均显示pH值差异不大,农田电导率>林地约2.22μs·cm-1,而容重差异仅0.02 g·cm-3(1%)。上述结果说明,土地利用对表层和深层影响差异明显,甚至趋势相反,农田和林地土壤碳及相关理化指标发生了明显垂直分布特征变化。过分强调土壤表层而得出的农田使SOC大量减少、土壤肥力下降的结论,在考虑深层土壤后能够明显降低上述数据的大小。这一发现说明需要同时考虑表层和深层土壤碳和氮等指标变化,以得出更科学的结论。     

Impacts of Canopy and Understory Nitrogen Additions on Stomatal Conductance and Carbon Assimilation of Dominant Tree Species in a Temperate Broadleaved Deciduous Forest

Ecosystems - Excess N deposition has aroused concerns about its negative impacts on forest ecosystems. A two-year study was conducted to assess the responses of stomatal conductance (Gc) and carbon...     

Spatial Variability of Organic Carbon,CaCO3 and Nutrient Burial Rates Spanning a Mangrove Productivity Gradient in the Coastal Everglades

Ecosystems - Mangrove wetlands are some of the most important locations of organic carbon (OC) sequestration and storage in the world on a per area basis. The high stocks of soil OC are driven by...     

Hydrologic Drivers and Seasonality of Dissolved Organic Carbon Concentration, Nitrogen Content, Bioavailability, and Export in a Forested New England Stream

We present the results of a full year of high-resolution monitoring of hydrologic event-driven export of stream dissolved organic matter (DOM) from the forested Bigelow Brook watershed in Harvard Forest, Massachusetts, USA. A combination of in situ fluorescent dissolved organic matter (FDOM) measurement, grab samples, and bioassays was utilized. FDOM was identified as a strong indicator of concentration for dissolved organic carbon (DOC, r ² = 0.96), dissolved organic nitrogen (DON, r ² = 0.81), and bioavailable DOC (BDOC, r ² = 0.81). Relationships between FDOM and concentration were utilized to improve characterization of patterns of hydrological event-driven export and the quantification of annual export. This characterization was possible because DOM composition remained relatively consistent seasonally; however, a subtle shift to increased fluorescence per unit absorbance was observed for summer and fall seasons and percent BDOC did increase slightly with increasing concentrations. The majority of export occurred during pulsed hydrological events, so the greatest impact of bioavailable exports may be on downstream aquatic ecosystems. Export from individual events was highly seasonal in nature with the highest flow weighted mean concentrations (DOC_FW) being observed in late summer and fall months, but the highest total export being observed for larger winter storms. Seasonal trends in DOC export coincide with weather driven changes in surface and subsurface flow paths, potential for depletion and rebuilding of a flushable soil organic matter pool, and the availability of terrestrial carbon sources such as leaf litter. Our approach and findings demonstrate the utility of high frequency FDOM measurement to improve estimates of intra-annual temporal trends of DOM export.     

Organic Carbon Composition and Oxygen Metabolism across a Gradient of Seasonally Inundated Limesink and Riparian Wetlands in the Southeast Coastal Plain,USA

A set of three relatively pristine seasonally inundated limesink wetlands and one riparian wetland was studied over a 4–6 month long inundation period in 2001. Patterns in organic matter properties and oxygen consumption in the water column followed a previously documented ecological gradient based on soil composition, vegetation type, and canopy cover. The full canopy, cypress-gum swamp had the highest mean concentrations of dissolved organic carbon (DOC; 26.2 mg/l) and dissolved lignin (sum 6; 299 μg/l) with lower concentrations observed in the partial canopy, cypress savanna (22.0 mg/l DOC; 252 μg/l sum 6) and the open marsh savanna (20.6 mg/l DOC; 135 μg/l sum 6), respectively. During the inundation period, DOC increased in concentration, dissolved lignin decreased, and δ¹³C shifted to more positive values which collectively indicate a large reduction in the percentage of aromatic carbon during the inundation period. All wetlands had very high concentrations of organic matter, yet microbial oxygen consumption was almost always stimulated by the addition of glucose rather than inorganic nutrients. Stimulation by glucose suggests that there were very small pools of highly bioavailable forms of DOC in the wetlands. A larger pool of moderately bioavailable organic matter had the capacity to sustain microbial oxygen consumption rates under dark conditions for at least 15 d. During the inundation period, the cypress-gum swamp had the lowest average rates of whole water oxygen consumption (1.0 μM/h) with increasing rates observed in the cypress savanna (1.3 μM/h), marsh savanna (1.6 μM/h), and riparian wetland (1.9 μM/h), respectively. The lignin compositional fingerprint varied across the gradient of limesink wetlands, and was useful for identifying different sources of vascular plant-derived DOM. Vascular plant production, algal production, microbial respiration, and UV degradation are all important drivers of DOM cycling, and the consistencies observed in this initial assessment of seasonally inundated limesink wetlands suggest they vary in predictable ways across the ecological gradient.     

Some Factors Affecting on Dry Weight,Organic Weight and Concentrations of Carbon and Nitrogen in Freshly Prepared and in Preserved Zooplankton

Effects of rinsing and preservation on dry weight, organic weight, and chemical composition were determined on the chaetognath Sagitta nagae and the copepods Calanus sinicus, Pleuromamma xiphias and Acartia tonsa. Samples rinsed with distilled water were always lighter than comparable samples rinsed with seawater, showing apparent decrease in the organic matter and chemical contents (as total carbon and nitrogen) by weight; these losses increased with increasing volume of the rinse water. The volume of distilled water used for rinsing should be less than 0.3 ml per 1 mg dry weight of sample in order to avoid the significant loss of body substances. Isotonic ammonium formate rinse often resulted in the widest range of variation and on an average the organic weight and chemical contents were slightly lower than those of fresh samples rinsed with filtered seawater. Freeze-drying samples resulted in dry and organic weights as well as chemical compositions that were somewhat greater than oven-dried samples. The organic matter and chemical contents by weight decreased significantly within 2.5 days of being placed in Borax-buffered formaldehyde-sea water solution. Loss of organic weight from fresh samples exceeded 20% after one week of the preservation. Hexamine-buffered formaldehyde solution showed a result which was different from the Borax-buffered solution.     

Effects of the Dry–Wet Hydrological Shift on Dissolved Organic Carbon Dynamics and Fate Across Stream–Riparian Interface in a Mediterranean Catchment

Abstract The stream–riparian interface, characterized by a dynamic and complex hydrology, is an important control point for nutrient fluxes and processing between terrestrial and aquatic systems. Predicted alterations in the discharge regime in Mediterranean climate regions make it necessary to understand the effects of abrupt hydrological transition between dry and wet conditions on the transport and fate of dissolved organic carbon (DOC) across the stream–riparian interface. In this study, the concentrations and fate of total DOC (TDOC) and a subset of four molecular weight fractions (<1 kDa, 1–10 kDa, 10–100 kDa, >100 kDa) were investigated in stream water and riparian groundwater during autumn of 2003 and 2004. The two study periods were characterized by contrasting antecedent hydrological conditions: the streamflow was interrupted in summer 2003 but was permanent in summer 2004. Comparison of the two study periods indicates that an abrupt dry–wet hydrological transition amplifies the water exchange across the stream–riparian interface and favors retention of up to 57% of the TDOC that flows across the interface. Furthermore, the efficiency of DOC retention across the stream-riparian interface also varies greatly depending on DOC molecular size. More than 70% of DOC fractions higher than 10 kDa were retained, whereas the smaller fraction (less than 1 kDa) was nearly conserved. Consequently, our study helps to clarify the effects of extreme hydrological events on DOC transport in running waters in Mediterranean regions.     

Spatial Distribution of Soil Organic Carbon and Total Nitrogen Based on GIS and Geostatistics in a Small Watershed in a Hilly Area of Northern China

The spatial variability of soil organic carbon (SOC) and total nitrogen (STN) levels is important in both global carbon-nitrogen cycle and climate change research. There has been little research on the spatial distribution of SOC and STN at the watershed scale based on geographic information systems (GIS) and geostatistics. Ninety-seven soil samples taken at depths of 0–20 cm were collected during October 2010 and 2011 from the Matiyu small watershed (4.2 km²) of a hilly area in Shandong Province, northern China. The impacts of different land use types, elevation, vegetation coverage and other factors on SOC and STN spatial distributions were examined using GIS and a geostatistical method, regression-kriging. The results show that the concentration variations of SOC and STN in the Matiyu small watershed were moderate variation based on the mean, median, minimum and maximum, and the coefficients of variation (CV). Residual values of SOC and STN had moderate spatial autocorrelations, and the Nugget/Sill were 0.2% and 0.1%, respectively. Distribution maps of regression-kriging revealed that both SOC and STN concentrations in the Matiyu watershed decreased from southeast to northwest. This result was similar to the watershed DEM trend and significantly correlated with land use type, elevation and aspect. SOC and STN predictions with the regression-kriging method were more accurate than those obtained using ordinary kriging. This research indicates that geostatistical characteristics of SOC and STN concentrations in the watershed were closely related to both land-use type and spatial topographic structure and that regression-kriging is suitable for investigating the spatial distributions of SOC and STN in the complex topography of the watershed.     

Growth of Bacteria on 3-Nitropropionic Acid as a Sole Source of Carbon,Nitrogen, and Energy

3-Nitropropionic acid (3NPA) is a widespread nitroaliphatic toxin found in a variety of legumes and fungi. Several enzymes have been reported that can transform the compound, but none led to the mineralization of 3NPA. We report here the isolation of bacteria that grow on 3NPA and its anion, propionate-3-nitronate (P3N), as the sole source of carbon, nitrogen, and energy. Experiments with resting cells, cell extracts, and purified enzymes indicate that the pathway involves conversion of 3NPA to P3N, which upon denitration yields malonic semialdehyde, nitrate, nitrite, and traces of H₂O₂. Malonic semialdehyde is decarboxylated to acetyl coenzyme A. The gene that encodes the enzyme responsible for the denitration of P3N was cloned and expressed, and the enzyme was purified. Stoichiometry of the reaction indicates that the enzyme is a monooxygenase. The gene sequence is related to a large group of genes annotated as 2-nitropropane dioxygenases, but the P3N monooxygenase and closely related enzymes form a cluster within COG2070 that differs from previously characterized 2-nitropropane dioxygenases by their substrate specificities and reaction products. The results suggest that the P3N monooxygenases enable bacteria to exploit 3NPA in natural habitats as a growth substrate.Large-scale release of synthetic nitroaromatic compounds to the biosphere followed the invention of nitrobenzene around 1830. In less than 200 years, microorganisms adapted to the presence of nitroaromatic compounds in the environment by developing catalytic pathways to exploit them as growth substrates. Such rapid development suggests that the pathways did not develop de novo but evolved from preexisting degradation pathways such as might be found in microorganisms that degrade naturally occurring compounds.3-Nitropropionic acid (3NPA) is a widespread naturally occurring nitroaliphatic compound. It is a principal toxic component of Astragalus locoweeds and has been found in hundreds of species of legumes (20, 39) and a variety of fungi (6). The compound causes irreversible inhibition of succinate dehydrogenase, which makes it deadly to eukaryotes (1). Plants that make 3NPA also contain an enzyme, 3NPA oxidase (NPAO) (19, 20), which converts the compound to malonic semialdehyde (MSA) to protect the plant against the toxic effects of the compound (20). Given its widespread occurrence, we hypothesized that there must be bacteria in soil that degrade 3NPA and play a major role in determining the flux of the compound.Although bacteria that degrade 3NPA have previously been sought, the focus has been on organisms that ingest 3NPA-containing plant matter. Rumen microorganisms reduce 3NPA to β-alanine (4), and in the grasshopper gut, 3NPA is bound to glycine to form inert conjugates which are then eliminated (24). The plant enzyme NPAO converts 3NPA and O₂ to MSA, nitrate, nitrite, and hydrogen peroxide (19). It is similar to propionate-3-nitronate (P3N) oxidase (P3NO; EC 1.7.3.5) from Penicillium atrovenetum that converts the P3N form of the compound to MSA (36). The enzymes mentioned above are “orphan enzymes” (28), which means that the gene(s) has not been identified. None of the previously studied microorganisms can use 3NPA as a growth substrate, and the physiological roles of the enzymes have not been established.MSA appears to be a central intermediate in the eukaryotic transformation of 3NPA and its analogs. However, the transformations involve distinctly different types of reactions and metabolites released. P3N and 3NPA release nitrate and nitrite in a 2:1 ratio when attacked by the fungal or plant oxidases. When 3NPA is reduced to β-alanine by rumen microorganisms, β-alanine is further metabolized (4), possibly by deamination to MSA (18).We report here the isolation from soil of aerobic bacteria that grow on 3NPA as the sole source of carbon, nitrogen, and energy. The genes that encode the initial enzymes of the degradation pathway were cloned, and recombinant proteins were purified and partially characterized to allow determination of the initial steps in the catabolic pathway.(Preliminary reports of this work have been presented previously at the 106th General Meeting of the American Society for Microbiology [32a] and the 108th General Meeting of the American Society for Microbiology [32b].)