Nitrogen fixation by biological soil crusts and heterotrophic bacteria in an intact Mojave Desert ecosystem with elevated CO2 and added soil carbon

Fixation of N by biological soil crusts and free-living heterotrophic soil microbes provides a significant proportion of ecosystem N in arid lands. To gain a better understanding of how elevated CO₂ may affect N₂-fixation in aridland ecosystems, we measured C₂H₂ reduction as a proxy for nitrogenase activity in biological soil crusts for 2 yr, and in soils either with or without dextrose-C additions for 1 yr, in an intact Mojave Desert ecosystem exposed to elevated CO₂. We also measured crust and soil δ¹⁵N and total N to assess changes in N sources, and δ¹³C of crusts to determine a functional shift in crust species, with elevated CO₂. The mean rate of C₂H₂ reduction by biological soil crusts was 76.9±5.6 μmol C₂H₄ m⁻² h⁻¹. There was no significant CO₂ effect, but crusts from plant interspaces showed high variability in nitrogenase activity with elevated CO₂. Additions of dextrose-C had a positive effect on rates of C₂H₂ reduction in soil. There was no elevated CO₂ effect on soil nitrogenase activity. Plant cover affected soil response to C addition, with the largest response in plant interspaces. The mean rate of C₂H₂ reduction in soils either with or without C additions were 8.5±3.6 μmol C₂H₄ m⁻² h⁻¹ and 4.8±2.1 μmol m⁻² h⁻¹, respectively. Crust and soil δ¹⁵N and δ¹³C values were not affected by CO₂ treatment, but did show an effect of cover type. Crust and soil samples in plant interspaces had the lowest values for both measurements. Analysis of soil and crust [N] and δ¹⁵N data with the Rayleigh distillation model suggests that any plant community changes with elevated CO₂ and concomitant changes in litter composition likely will overwhelm any physiological changes in N₂-fixation.     

A bradyrhizobial-<Emphasis Type="Italic">Penicillium</Emphasis> spp. biofilm with nitrogenase activity improves N<Subscript>2</Subscript> fixing symbiosis of soybean

A bradyrhizobial-fungal biofilm (i.e. Bradyrhizobium elkanii SEMIA 5019-Penicillium spp.) developed in vitro was assayed for its nitrogenase activity and was evaluated for N₂-fixing symbiosis with soybean under greenhouse conditions. The biofilm showed nitrogenase activity, but the bradyrhizobial strain alone did not. Shoot and root growth, nodulation and N accumulation of soybean increased significantly with an inoculum developed from the biofilm. This study concludes that such biofilmed inoculants can improve N₂-fixing symbiosis in legumes, and can also directly contribute to soil N fertility in the long term. Further studies should be conducted to investigate the performance of these inoculants under field conditions.     

Influence of selenium on oxidoreductive enzymes activity in soil and in plants

The aim of this greenhouse experiment was the assessment of the influence of H₂SeO₃ at soil concentrations of 0.05, 0.15 and 0.45 mmol kg⁻¹, on the activity of selected oxidoreductive enzymes in wheat (Triticum aestivum). The wheat plants were grown in 2 dm³ pots filled with dust-silt black soil of pH 7.7. Applied H₂SeO₃ caused activation of plant nitrate reductase at all concentrations, but activation of plant polyphenol oxidase at only two lower concentrations. The highest concentration caused inhibition of polyphenol oxidase and peroxidase. Plant catalase activity decreased under the influence of 0.15 and 0.45 mmol kg⁻¹ concentration. After the final analysis Se was quantified in plants and soil. The amounts in plants were: control (unamended soil) 1.95 mg kg⁻¹; I dose (0.05 mmol kg⁻¹) 18.27 mg kg⁻¹; II dose (0.15 mmol kg⁻¹) 33.20 mg kg⁻¹ and III dose (0.45 mmol kg⁻¹) 38.37 mg kg⁻¹, in soil: 0.265 mg kg⁻¹; 3.61 mg kg⁻¹; 10.53 mg kg⁻¹; 30.53 mg kg⁻¹; respectively. Simultaneously, a laboratory experiment was performed, where the activity of soil catalase and peroxidase were tested after 1, 3, 7, 14, 28, 56, and 112 days after Se treatment. Peroxidase activity in soil decreased with increasing Se content, over the whole experiment. The lowest dose of Se caused activation a significant 10% increase in catalase activity, but the influence of others doses was unclear.     

Differences in isotopic fractionation of nitrogen in water-saturated and unsaturated soils

Temporal variations in δ¹⁵N of NH₄⁺ and NO₃⁻ in water-saturated and unsaturated soils were examined in a laboratory incubation study. Ammonium sulfate (δ¹⁵N=−2.6‰) was added to 25 g samples of soil at concentrations of 160 mg N kg⁻¹. Soils were then incubated under unsaturated (50% of water holding capacity at saturation, WHC) or saturated (100% of WHC) water conditions for 7 and 36 d, respectively. During 7 d incubation of unsaturated soil, the NH₄⁺-N concentration decreased from 164.8 to 34.4 mg kg⁻¹, and the δ¹⁵N of NH₄⁺ increased from −0.4 to +57.2‰ through nitrification, as evidenced by corresponding increase in NO₃⁻-N concentration and lower δ¹⁵N of NO₃⁻ (product) than that of NH₄⁺ (substrate) at each sampling time. In saturated soil, the concentration of NH₄⁺-N decreased gradually from 162.4 to 24.2 mg kg⁻¹, and the δ¹⁵N values increased from +0.8 to +21.0‰ during 36 d incubation. However, increase in NO₃⁻ concentration was not observed due to loss of NO₃⁻ through concurrent denitrification in anaerobic sites. The apparent isotopic fractionation factors (α_s/p) associated with decrease in NH₄⁺ concentration were 1.04 and 1.01 in unsaturated and saturated soils, respectively. Since nitrification is likely to introduce greater isotope fractionation than microbial immobilization, the higher value for unsaturated soil probably reflected faster nitrification under aerobic conditions. The lower value for saturated soil suggests that immobilization and subsequent remineralization of NH₄⁺ were relatively more dominant than nitrification under the anaerobic conditions.     

Changes in soil biological and biochemical characteristics in a long-term field trial on a sub-tropical inceptisol

Soil organic carbon (SOC), microbial biomass carbon (MBC), their ratio (MBC/SOC) which is also known as microbial quotient, soil respiration, dehydrogenase and phosphatase activities were evaluated in a long-term (31 years) field experiment involving fertility treatments (manure and inorganic fertilizers) and a maize (Zea mays L.)-wheat (Triticum aestivum L.)-cowpea (Vigna unguiculata L.) rotation at the Indian Agricultural Research Institute near New Delhi, India. Applying farmyard manure (FYM) plus NPK fertilizer significantly increased SOC (4.5-7.5 g kg⁻¹), microbial biomass (124-291 mg kg⁻¹) and microbial quotient from 2.88 to 3.87. Soil respiration, dehydrogenase and phosphatase activities were also increased by FYM applications. The MBC response to FYM+100% NPK compared to 100% NPK (193 vs. 291 mg kg⁻¹) was much greater than that for soil respiration (6.24 vs. 6.93 μl O₂ g⁻¹ h⁻¹) indicating a considerable portion of MBC in FYM plots was inactive. Dehydrogenase activity increased slightly as NPK rates were increased from 50% to 100%, but excessive fertilization (150% NPK) decreased it. Acid phosphatase activity (31.1 vs. 51.8 μg PNP g⁻¹ h⁻¹) was much lower than alkali phosphatase activity (289 vs. 366 μg PNP g⁻¹ h⁻¹) in all treatments. Phosphatase activity was influenced more by season or crop (e.g. tilling wheat residue) than fertilizer treatment, although both MBC and phosphatase activity were increased with optimum or balanced fertilization. SOC, MBC, soil respiration and acid phosphatase activity in control (no NPK, no manure) treatment was lower than uncultivated reference soil, and soil respiration was limiting at N alone or NP alone treatments.     

Mycorrhiza‐soil fertility effects on regrowth,nodulation and Nitrogenase activity of siratro (Macroptilium atropurpureum (DC) Urb.)

Siratro (Macroptilium atropurpureum (DC) Urb.) is a vigorous perennial forage legume with good potential for improving pastures in the extensive neotropical regions of the world. It is well adapted to a wide range of soil and climatic conditions. The objective of these studies was to determine effects of Glomus fasciculatum colonization, rigorous defoliation, and soil fertility treatments to a Psammentic Paleustalf (Eufaula) soil on growth, regrowth, nodulation, and nitrogenase activity (C₂H₂ red.) of Siratro inoculated with Rhizobium leguminosarum Frank. Top growth increased significantly with soil K and P amendment and with mycorrhiza colonization. Nodulation and nitrogenase activity were correlated with highly significant increases from G. fasciculatum, P treatments and K additions to 300 mg K kg^‐1 soil. Growth and peduncles of nonclipped plants increased about 4 fold from 90 to 225 day age with mature seed yield increasing about 10 fold; nodule mass and nitrogenase activity levels approximately doubled. Regrowth response of plants defoliated at 45 day intervals, following their initial 90 day age, was somewhat constant between clippings for magnitude of regrowth 12.3–13.8g, development in number of peduncles 4.0–6.8, seed yield 1.4–2.6g, nodulation 2.9–3.7g, and nitrogenase activity 73.9–95.8μ mol C₂H₄g^‐1 nodule. Multiple regression for nitrogenase = 0.55 g top wt. + 0.63 g nodule wt. + 1.91 day age ‐ 0.07 peduncle no., R² = 0.85 and C.V. = 14.3%. Favorable tripartite symbiosis with both effective Rhizobium and endophyte mycorrhiza were essential for high levels of symbiotic nitrogen fixation.     

Controls on nitrification in a water-limited ecosystem: experimental inhibition of ammonia-oxidising bacteria in the Patagonian steppe

We studied controls on nitrification in an undisturbed water-limited ecosystem by inhibiting autotrophic nitrifying bacteria in soils with varying levels of vegetative cover. The activity of nitrifying bacteria was disrupted using nitrapyrin, 2-chloro-6-(trichloromethyl)-pyridine, under field conditions in three microenvironments (underneath shrubs, next to grasses and in bare soil). Ammonia-oxidising bacteria were detected by PCR analysis of DNA in soils. The inhibition of nitrification changed the concentrations of NO₃⁻ and NH₄⁺ in the soil, while the microenvironment was most important in determining the response of bacteria to the inhibitor. Nitrapyrin application resulted in a significant (p<0.05) reduction in soil NO₃⁻ concentration (39%) and a significant increase (p<0.001) in soil NH₄⁺ concentration (41%). Untreated bare-soil microenvironments had the lowest concentrations of NH₄⁺ (1.57 μg/g of dry soil) and NO₃⁻ (0.49 μg/g of dry soil) when compared to the other microenvironments, and showed the highest impacts of nitrification inhibition. For example, NH₄⁺ concentrations increased 288% and NO₃⁻ concentrations decreased 60% in inhibited bare-soil microenvironments. In contrast, untreated microenvironments underneath shrubs had the highest levels of NH₄⁺ (10.01 μg/g of dry soil) and NO₃⁻ (0.69 μg/g of dry soil), but showed no significant effects of inhibition of nitrification on soil nitrogen concentrations.     

Microbial growth efficiencies across a soil moisture gradient assessed using C-acetic acid vapor and N-ammonia gas

One integrative measurement of microbial activity in soils is the efficiency by which microbes convert assimilated carbon (C) into biomass C. This efficiency, called the microbial growth efficiency (Y), is a key physiological characteristic that regulates soil carbon sequestration, nutrient immobilization, and greenhouse gas emissions. Changes in rainfall patterns and soil water content as the result of global climate change have the potential to influence microbial activity and lead to changes in Y and thus, nutrient cycling at the ecosystem level. Unfortunately, little information is available on how environmental variables such as soil moisture influence Y. We have developed a new method for injecting ¹³C-labeled carbon as acetic acid vapor into soil that will allow measurement of microbial growth efficiency (as Y_C) without increasing soil moisture content. We compare Y determined with this new approach with an alternate method where injected ¹⁵N-labeled ammonia gas is used to quantify microbial N immobilization, and microbial growth efficiency is calculated based on microbial C:N and respiration rate (as Y_N). We also include injections of a solution containing labeled ammonium and acetate in our experiment to compare the results of our vapor methods with more commonly employed liquid-based methods. The ¹³C-acetic acid vapor, which was supplied to soils with soil moisture content ranging from 0.05 to 0.21 g H₂O g⁻¹ soil, was readily assimilated and respired by microbes. Between 0.10 and 0.21 g H₂O g⁻¹ soil (−0.60 to −0.04 MPa), values of Y_C averaged 0.46, and were significantly lower than values of Y_N, with average values of 0.58. Over this range, soil moisture content had no significant effect on either Y_C or Y_N. However, at the lowest soil moisture content (0.05 g H₂O g⁻¹ soil; <−6.0 MPa), Y_C and Y_N diverged substantially, suggesting that in very dry soils, constraints on microbial growth cause differential uptake of C and N resources.     

Co-localization of atmospheric H2 oxidation activity and high affinity H2-oxidizing bacteria in non-axenic soil and sterile soil amended with Streptomyces sp. PCB7

Two complementary experimental approaches were utilized to examine the extent to which free soil hydrogenases and H₂-oxidizing bacteria contribute to the soil uptake of atmospheric H₂. First, high affinity hydrogenase activity and H₂-oxidizing bacteria were fractionated in non-axenic soil and axenic soil colonized with the high affinity H₂-oxidizing bacterium Streptomyces sp. PCB7. Non-axenic soil was fractionated by buoyant density centrifugation. High affinity H₂ oxidation activity measured in individual fractions was proportional to the copy number of hhyL gene, specifying the large subunit of putative high affinity [NiFe]-hydrogenases. 2.5% of the hydrogenase activity was recovered in bacteria-free soil extract. Similarly, sequential centrifugation and wet filtrations of strain PCB7-colonized soil dispersed in solubilization buffer caused a loss of the activity, at a ratio proportional to the number of living cells removed. No abiontic hydrogenase activity was detected in bacteria-free fractions. The second experimental approach was designed to verify whether or not the [NiFe]-hydrogenase of strain PCB7 retains high affinity H₂ oxidation activity in soil, under the abiontic state. H₂ oxidation rates of crude enzyme extract of strain PCB7 measured under aerobic and anaerobic conditions were indistinguishable, indicating that the high affinity hydrogenase of strain PCB7 is oxygen-tolerant. The hydrogenase activity of sterile soil spiked with as much as 0.14 mg_(protein) g_(soil-dw)⁻¹ was equivalent to the H₂-oxidation activity of only 10⁶-10⁷ CFU of strain PCB7 g_(soil-dw)⁻¹. Taken together, our results indicate that high affinity hydrogenase activity is proportional to the abundance of H₂-oxidizing bacteria in soil and, that abiontic hydrogenases contribute only a few percent of the total high affinity H₂ oxidation activity detected in soil.     

Short-term effects of clearfelling on soil CO2, CH4, and N2O fluxes in a Sitka spruce plantation

We examined the effects of forest clearfelling on the fluxes of soil CO₂, CH₄, and N₂O in a Sitka spruce (Picea sitchensis (Bong.) Carr.) plantation on an organic-rich peaty gley soil, in Northern England. Soil CO₂, CH₄, N₂O as well as environmental factors such as soil temperature, soil water content, and depth to the water table were recorded in two mature stands for one growing season, at the end of which one of the two stands was felled and one was left as control. Monitoring of the same parameters continued thereafter for a second growing season. For the first 10 months after clearfelling, there was a significant decrease in soil CO₂ efflux, with an average efflux rate of 4.0 g m⁻² d⁻¹ in the mature stand (40-year) and 2.7 g m⁻² d⁻¹ in clearfelled site (CF). Clearfelling turned the soil from a sink (−0.37 mg m⁻² d⁻¹) for CH₄ to a net source (2.01 mg m⁻² d⁻¹). For the same period, soil N₂O fluxes averaged 0.57 mg m⁻² d⁻¹ in the CF and 0.23 mg m⁻² d⁻¹ in the 40-year stand. Clearfelling affected environmental factors and lead to higher daily soil temperatures during the summer period, while it caused an increase in the soil water content and a rise in the water table depth. Despite clearfelling, CO₂ remained the dominant greenhouse gas in terms of its greenhouse warming potential.     

Porous tubing for use in monitoring soil CO2 concentrations

The composition of the soil atmosphere is an indicator of biological processes, and soil CO₂ gradients have been used to estimate CO₂ efflux from the surface. Soil atmosphere samplers, constructed with gas-permeable materials, have been used to quantify soil CO₂ concentrations. The type of material used can influence the perceived real-time concentrations of CO₂ in the soil. Previous works have not directly compared different types of materials under the same conditions. The objective of this study was to determine the diffusion coefficient (D) and time of 95% equilibrium (t_eq) of CO₂ through several materials, and to evaluate the effect of long-term soil burial (183 days) on diffusion characteristics. Materials tested included silicone, expanded Teflon (ePTFE), and ultra high molecular weight polyethylene (PE) tubing. The D of each material was determined using a closed-loop system consisting of a CO₂-enriched (7800 ppm) chamber, a CO₂ analyzer and an inner tube (experimental tubing) placed inside the chamber. Air was re-circulated through the inner tube, and as CO₂ diffused from the chamber into the tubing, the analyzer recorded the increase in concentration. The silicone tubes had values of D ranging from 8.64 to 5.80×10⁻⁶ cm² s⁻¹ with corresponding t_eq between 3.9 and 9.7 h. Diffusion coefficients of the ePTFE (1.25×10⁻⁴ cm² s⁻¹) and PE (7.70×10⁻⁴ cm² s⁻¹) materials were 2 orders of magnitude greater, with t_eq<6 min. Exposure to the soil environment for 183 days did not visibly deteriorate the materials or significantly affect the D or t_eq values. Use of the ePTFE or PE materials, over the silicone materials, may allow for better characterization of dynamic CO₂ concentrations in the soil based on the greater D and lesser t_eq values of these materials.     

Short-term competition between crop plants and soil microbes for inorganic N fertilizer

Agricultural systems that receive high amounts of inorganic nitrogen (N) fertilizer in the form of either ammonium (NH₄⁺), nitrate (NO₃⁻) or a combination thereof are expected to differ in soil N transformation rates and fates of NH₄⁺ and NO₃⁻. Using ¹⁵N tracer techniques this study examines how crop plants and soil microbes vary in their ability to take up and compete for fertilizer N on a short time scale (hours to days). Single plants of barley (Hordeum vulgare L. cv. Morex) were grown on two agricultural soils in microcosms which received either NH₄⁺, NO₃⁻ or NH₄NO₃. Within each fertilizer treatment traces of ¹⁵NH₄⁺ and ¹⁵NO₃⁻ were added separately. During 8 days of fertilization the fate of fertilizer ¹⁵N into plants, microbial biomass and inorganic soil N pools as well as changes in gross N transformation rates were investigated. One week after fertilization 45-80% of initially applied ¹⁵N was recovered in crop plants compared to only 1-10% in soil microbes, proving that plants were the strongest competitors for fertilizer N. In terms of N uptake soil microbes out-competed plants only during the first 4 h of N application independent of soil and fertilizer N form. Within one day microbial N uptake declined substantially, probably due to carbon limitation. In both soils, plants and soil microbes took up more NO₃⁻ than NH₄⁺ independent of initially applied N form. Surprisingly, no inhibitory effect of NH₄⁺ on the uptake and assimilation of nitrate in both, plants and microbes, was observed, probably because fast nitrification rates led to a swift depletion of the ammonium pool. Compared to plant and microbial NH₄⁺ uptake rates, gross nitrification rates were 3-75-fold higher, indicating that nitrifiers were the strongest competitors for NH₄⁺ in both soils. The rapid conversion of NH₄⁺ to NO₃⁻ and preferential use of NO₃⁻ by soil microbes suggest that in agricultural systems with high inorganic N fertilizer inputs the soil microbial community could adapt to high concentrations of NO₃⁻ and shift towards enhanced reliance on NO₃⁻ for their N supply.     

Michaelis constants of soil enzymes

Studies to determine the Michaelis constants (k_m values) for the arylsulfatase and phosphatase activity in Iowa surface soils showed that the value obtained for either activity was different for different soils. When the incubation technique used to determine k_m did not involve shaking of the soil-substrate mixture, the k_m value for arylsulfatase activity in nine soils studied ranged from 1·37 × 10⁻³m to 5·69 × 10⁻³m, and the k_m value for phosphatase activity ranged from 1·26 × 10⁻³m to 4·58 × 10⁻³m. Shaking the soil-substrate mixture during incubation decreased the k_m value obtained for arylsulfatase or phosphatase activity and reduced the variation in k_m among soils. The maximum enzyme reaction velocity (V_max value) for soil arylsulfatase or soil phosphatase activity was markedly different for different soils and usually increased when the soil-substrate mixture was shaken during incubation. The k_m value for soil arylsulfatase or soil phosphatase activity was not significantly correlated with other soil properties studied (pH, cation-exchange capacity, percentage organic carbon, percentage clay, percentage sand).     

Differentiating microbial and stabilized β-glucosidase activity relative to soil quality

Previous research has shown that β-glucosidase activity can detect soil management effects and has potential as a soil quality indicator, but mechanisms for this response are not well understood. A significant amount of hydrolytic enzyme activity comes from extracellular (abiontic) activity that is bound and protected by soil colloids. This study was conducted to determine how management affects the kinetics of this enzyme (K_m, substrate affinity, and V_max, maximum reaction velocity) and its degree of stabilization on soil colloids. Soils were sampled from three sites in Oregon, with a paired comparison within each site of a native, unmanaged soil, and a matching soil under agricultural production (>50 years). Microwave radiation (MW) stress was used to denature the β-glucosidase fraction associated with viable microorganisms in these soils as an estimate of abiontic activity. Total activity and V_max were decreased by both management and MW. The results showed that β-glucosidase activity is sensitive to soil management on a variety of soils and environments (135 vs. 190, 80 vs. 111 and 80 vs. 134 μg PNP g⁻¹ h⁻¹ for managed and unmanaged treatments, respectively, at the three study sites in Oregon). The evidence suggests that this sensitivity to management is not (or minimally) due to differences in isoenzymes (K_m generally was unaffected) but rather due to an overall reduction in the amount of enzyme present (V_max decreased) and that this reduction in activity is reflected more from the activity of enzymes in the stabilized fraction than that associated with viable microbial population. Although β-glucosidase activity after MW irradiation appears to be limited as a soil quality indicator, it maybe useful as research tool to separate abiontic from microbial activity ‘biomass’ β-glucosidase activity correlated with microbial biomass C (r=0.42, P<0.05) but MW irradiated, abiontic, activity did not (r=−0.20^NS).     

Nodulation and nitrogen fixation by arrowleaf clover: Effects of phosphorus and potassium

“Yuchi” arrowleaf clover (Trifolium vesiculosum Sav.) has a potential for high forage productivity with desirable symbiotic nitrogen fixation within most temperate regions. Our objective was to determine the effects of soil fertility on growth, nodulation, nitrogenase and associated enzyme activities of arrowleaf clover. In greenhouse experiments top growth increased with additions of 300 mg K kg^?1 soil with and without 100 mg P kg^?1 soil to a Cumulic Haplustoll (Port silt loam, pH 6.1). Nodule mass without P fertilizer additions increased linearly up to 400 mg K kg^?1 soil. When both P and K fertilizer additions were combined nodule mass increased significantly only up to the 300 mg K addition. However, nodule weight, increased 4-fold with the PK combination treatments. Nitrogenase activity, as measured by C₂H₂ reduction, more than doubled with P additions and increased linearly up to 400 mg K kg^? soil, with and without the P additions. Aspartate amino-transferase (AST) activity of nodule cytosol more than doubled with P additions but increased only with up to 300 mg K without P. Highest AST activities were recorded with the 400 mg K addition when combined with P. Glutamate synthetase (GS) activity increased with up to 300 mg K without P addition, but when combined with P was approximately 3 times higher, increasing linearly to 400 mg K. Differences in glutamate synthetase (GOGAT) activity were not significant with K additions without P, but when combined with P treatments were almost doubled up to the 400 mg K concentrations. Multiple regression for nitrogenase (C₂H₂ red.) as the dependent variable = 5.89 (AST) + 12.79 (GS) + 21.52 (GOGAT) + 13.53 (GDH); R² = 0.92 and C.V. = 15.6%. Nodule cytosol P and K compositions reflected soil treatment levels and combinations. Reciprocal effects of monovalent cations were highly significant, with increased K concentrations reducing Na content; nitrogenase = 0.12 (P) + 0.01(K) + 0.14(Ca) ?0.34 (Na); R² = 0.86 and C.V. = 21.9%.     

Soil fertility effects on growth,yield, nodulation and nitrogenase activity of Austrian winter pea

Austrian winter pea (Pisum sativum subspecies arvense (L.) Poir) is grown as a cool season annual to produce high protein seed and forage as well as for soil fertility improvement. This legume is grown on a wide range of soil types with many different cropping systems. The objective of these studies was to determine the influence of K levels, with and without P and Ca fertilization, for increased growth, yield, nodulation and nitrogenase activity. Results were from 3 years’ field and greenhouse experiments with a Psammentic Paleustalf (Eufaula series) utilizing Rhizobium leguminosarum (Frank), ATCC 10314 as inoculum. Soil fertility effects on composition and histology of field‐grown nodules are presented.

Available soil P was a limiting plant nutrient in field studies with significant response to K resulting with PK combinations for top growth, tillers, pods, seed yield, nodule mass, and nitrogenase activity levels (C₂H₂, red.). Multiple regression for nitrogenase (umol C₂H₄ h^‐1) = 1.09 tiller number + 3.37 nodule weight + 2.29 pod number, R² = 0.837, C.V. = 29.9%. Results from the greenhouse experiments indicated significant responses with increased K application levels when combined with P and Ca fertilization for top growth, nodule weight, number of nodules and nitro‐genase activity. Highly significant correlations resulted with nitrogenase x nodule weight (r=0.538) and nitrogenase x top growth (r=0.359) with multiple regression of treatment effects for nitrogenase (μmol C₂H₄ h^‐1) = 2.73 P + 1.04 K + 4.92 Ca, R² = 0.797 and C.V. = 48.8%. Soil addition of plant nutrients resulted in significantly increased concentrations of those elements within nodules. Magnesium content was not consistently influenced by P, Ca, and K amendments. Sodium decreased with increased K fertilization. Multiple regression of elemental composition (mg g^‐1 nodule) for nitrogenase (pmol C₂H₄ h^‐1) = 0.21 P + 0.86 K + 2.35 Ca ‐ 2.01 Na, R² = 0.772, C.V. = 55.6%. The proportion of plant nutrients in nodules contained within the nodule cytosol was highest for K (56.2%) and lowest for Ca (21.4%) with intermediate levels of Mg (50.2%), P (45.4%), and Na (37.2%).

Practical application from these data include the requirement of adequate available soil K for increased yield and nitrogen fixation with favorable P and Ca soil levels in Austrian winter pea production.     

The wood-decaying fungus Hygrophoropsis aurantiaca increases P availability in acid forest humus soil, while N addition hampers this effect

We evaluated the influence of the brown rot fungus Hygrophoropsis aurantiaca on P solubility in the humus layer of a podzolic forest soil. This fungus is known to exude large amounts of oxalic acid that may stimulate weathering of minerals and increase dissolution of humus, which in turn may increase P availability in the soil surrounding the fungus. Humus was inoculated using small wooden pieces colonised by the fungus. The presence of the fungus resulted in elevated concentration of PO₄⁻ in the humus solution. In a second experiment birch seedlings grown in the same humus were able to utilise the PO₄⁻ mobilised by the fungus to increase their internal P content. The factor determining this increased P uptake and the increased available P might be oxalate produced by fungus. The acid may directly dissolve P or change organic forms of P making it more susceptible to reaction with phosphatases. This fungal effect on P solubility diminished when N was added to the soil in the form of a slow release N fertilizer (methyl urea), or when a soil with a higher soil N concentration was used. We found a strong correlation between NH₄⁺ concentration and total organic carbon in the soil solution at high NH₄⁺ concentrations, suggesting the dissolution of humus as a result of the high NH₄⁺ content in the solution. This study demonstrates that the wood-decaying fungus H. aurantiaca influences nutrient turnover in forest soil, and thereby nutrient uptake by forest trees. An intensified harvest of forest products such as whole tree harvesting may decrease the active biomass of the wood decomposers and may thereby change the availability of P and the leaching of N.     

Changes in soil organic carbon dynamics in an Eastern Chinese coastal wetland following invasion by a C4 plant Spartina alterniflora

The exotic C₄ grass Spartina alterniflora was intentionally introduced to tidal coastal wetlands in Jiangsu province of China in 1982. Since then it has rapidly replaced the native C₃ plant Suaeda salsa, becoming one of the dominant vegetation types in the coastal wetlands of China. Although plant invasion can change soil organic carbon (SOC) storage, little is known about how plant invasion influences C storage within soil fractions. We investigated how S. alterniflora invasion across an 8, 12 and 14-year chronosequence affected SOC and soil nitrogen (N), using soil fractionation and stable δ¹³C isotope analyses. SOC and N concentrations at 0-10 cm depth in S. alterniflora soil increased during the S. alterniflora invasion chronosequence, ranging from 3.67 to 4.90 g C kg⁻¹ soil, and from 0.307 to 0.391 g N kg⁻¹ soil. These were significantly higher than the values in the Suaeda salsa community, by 27.0-69.6% for SOC, and 21.8-55.2% for total N. The S. alterniflora-derived SOC varied from 0.40 to 0.92 g C kg⁻¹ according to mixing calculations, assuming the two possible SOC sources of S. alterniflora and S. salsa, and accounted for 10.8-18.7% of total SOC in the colonized soils. The estimated accumulative rate of SOC from C₄ (S. alterniflora) was 64.1 C kg⁻¹ soil year⁻¹ and from C₃ sources was 78.1 mg C kg⁻¹. The concentration of S. alterniflora-derived SOC significantly decreased from coarse fraction to fine fraction, and linearly increased as the period of S. alterniflora invasion increased. The highest accumulative rate of SOC from a C₄ source occurred in macroaggregates, while the highest rate from C₃ was in microaggregates. The storage of SOC derived from S. alterniflora in the macroaggregates was 0.27-0.44 g C kg⁻¹ soil, accounting for 43.1-49.1% of the total C₄derived SOC in the soil. Our results suggest that S. alterniflora invasion in coastal wetlands could facilitate SOC storage, because of the high potential for accumulation of the C which has been newly derived from S. alterniflora litter and roots.     

Partitioning root and microbial contributions to soil respiration in Leymus chinensis populations

Based on the enclosed chamber method, soil respiration measurements of Leymus chinensis populations with four planting densities (30, 60, 90 and 120 plants/0.25 m²) and blank control were made from July 31 to November 24, 2003. In terms of soil respiration rates of L. chinensis populations with four planting densities and their corresponding root biomass, linear regressive equations between soil respiration rates and dry root weights were obtained at different observation times. Thus, soil respiration rates attributed to soil microbial activity could be estimated by extrapolating the regressive equations to zero root biomass. The soil microbial respiration rates of L. chinensis populations during the growing season ranged from 52.08 to 256.35 mg CO₂ m⁻² h⁻¹. Soil microbial respiration rates in blank control plots were also observed directly, ranging from 65.00 to 267.40 mg CO₂ m⁻² h⁻¹. The difference of soil microbial respiration rates between the inferred and the observed methods ranged from −26.09 to 9.35 mg CO₂ m⁻² h⁻¹. Some assumptions associated with these two approaches were not completely valid, which might result in this discrepancy. However, these two methods' application could provide new insights into separating root respiration from soil microbial respiration. The root respiration rates of L. chinensis populations with four planting densities could be estimated based on measured soil respiration rates, soil microbial respiration rates and corresponding mean dry root weight, and the highest values appeared at the early stage, then dropped off rapidly and tended to be constant after September 10. The mean proportions of soil respiration rates of L. chinensis populations attributable to the inferred and the observed root respiration rates were 36.8% (ranging from 9.7 to 52.9%) and 30.0% (ranging from 5.8 to 41.2%), respectively. Although root respiration rates of L. chinensis populations declined rapidly, the proportion of root respiration to soil respiration still increased gradually with the increase of root biomass.     

Microbial community response to nitrogen deposition in northern forest ecosystems

The productivity of temperate forests is often limited by soil N availability, suggesting that elevated atmospheric N deposition could increase ecosystem C storage. However, the magnitude of this increase is dependent on rates of soil organic matter formation as well as rates of plant production. Nonetheless, we have a limited understanding of the potential for atmospheric N deposition to alter microbial activity in soil, and hence rates of soil organic matter formation. Because high levels of inorganic N suppress lignin oxidation by white rot basidiomycetes and generally enhance cellulose hydrolysis, we hypothesized that atmospheric N deposition would alter microbial decomposition in a manner that was consistent with changes in enzyme activity and shift decomposition from fungi to less efficient bacteria. To test our idea, we experimentally manipulated atmospheric N deposition (0, 30 and 80 kg NO₃⁻-N) in three northern temperate forests (black oak/white oak (BOWO), sugar maple/red oak (SMRO), and sugar maple/basswood (SMBW)). After one year, we measured the activity of ligninolytic and cellulolytic soil enzymes, and traced the fate of lignin and cellulose breakdown products (¹³C-vanillin, catechol and cellobiose).In the BOWO ecosystem, the highest level of N deposition tended to reduce phenol oxidase activity (131±13 versus 104±5 μmol h⁻¹ g⁻¹) and peroxidase activity (210±26 versus 190±21 μmol h⁻¹ g⁻¹) and it reduced ¹³C-vanillin and ¹³C-catechol degradation and the incorporation of ¹³C into fungal phospholipids (p<0.05). Conversely, in the SMRO and SMBW ecosystems, N deposition tended to increase phenol oxidase and peroxidase activities and increased vanillin and catechol degradation and the incorporation of isotope into fungal phospholipids (p<0.05). We observed no effect of experimental N deposition on the degradation of ¹³C-cellulose, although cellulase activity showed a small and marginally significant increase (p<0.10). The ecosystem-specific response of microbial activity and soil C cycling to experimental N addition indicates that accurate prediction of soil C storage requires a better understanding of the physiological response of microbial communities to atmospheric N deposition.