Effect of inoculation with wild type Azospirillum brasilense and A. irakense strains on development and nitrogen uptake of spring wheat and grain maize

Under the controlled conditions of the greenhouse and by varying some biotic and abiotic factors, we tried to identify some of the factors critical to obtain successful Azospirillum inoculation. Spring wheat and grain maize were inoculated with different concentrations of the wild type strains A. brasilense Sp245 and A. irakense KBC1, and grown in a substrate with varying concentrations of organic matter (OM) and N fertiliser. The inoculum concentration was one of the factors that influenced most the outcome of an inoculation experiment on wheat, with lower inoculum concentrations (10⁵-10⁶ cfu plant^-1) stimulating root development and plant dry weight and higher inoculum concentrations (10⁷-10⁸ cfu plant^-1) having no effect or sometimes even inhibiting root development. The effect of inoculation was most pronounced at low to intermediate N fertilisation levels, while the OM content of the substrate had no effect. Inoculation was found to affect early plant and root development, plant and root dry weight, grain yield and the N-uptake efficiency of plants. However, inoculation did not change the N concentration in plants or grains. In addition, a difference in the ability of both strains to stimulate plant growth and N uptake of wheat and maize was observed, with A. brasilense Sp245 having most effect on spring wheat and A. irakense KBC1 being more effective on grain maize. The significance of the obtained results for agriculture is discussed.     

Cultivable heterotrophic N<Subscript>2</Subscript>-fixing bacterial diversity in rice fields in the Yangtze River Plain

Heterotrophic N₂-fixing bacteria are a potentially important source of N₂ fixation in rice fields due to the moist soil conditions. This study was conducted at eight sites along a geographic gradient of the Yangtze River Plain in central China. A nitrogen-free solid malate-sucrose medium was used to isolate heterotrophic N₂-fixing bacteria. Numbers of the culturable N₂-fixing bacteria expressed as CFU (colony forming units) ranged between 1.41ǂ.42᎒⁶ and 1.24ǂ.23᎒⁸ in the sampled paddy field sites along the plain. Thirty strains with high ARA (acetylene reduction activity) were isolated and purified; ARA of the strains varied from 0.9 to 537.8 nmol C₂H₄ culture^-1 h^-1, and amounts of ¹⁵N fixed ranged between 0.008 and 0.4866 mg·culture^-1·day^-1. According to morphological and biochemical characteristics, 14 strains were identified as the genus Bacillus, 2 as Burkholderia, 1 as Agrobacterium, 4 as Pseudomonas, 2 as Derxia, 1 as Alcaligenes, 1 as Aeromonas, 2 as Citrobacter, and 3 strains belonged to the corynebacter-form group.     

Nitrous oxide emissions from the wheat-growing season in eighteen Chinese paddy soils: an outdoor pot experiment

To identify the key soil parameters influencing N₂O emission from the wheat-growing season, an outdoor pot experiment with a total of 18 fertilized Chinese soils planted with wheat was conducted in Nanjing, China during the 2000/2001 wheat-growing season. Average seasonal N₂O-N emission for all 18 soils was 610 mg m^-2, ranging from 193 to 1,204 mg m^-2, approximately a 6.2-fold difference between the maximum and the minimum. Correlation analysis indicated that the seasonal N₂O emission was negatively correlated with soil organic C (r²=0.5567, P<0.001), soil total N (r²=0.4684, P<0.01) and the C:N ratio (r²=0.4530, P<0.01), respectively. A positive dependence of N₂O emission on the soil pH (r²=0.3525, P<0.01) was also observed. No clear relationships existed between N₂O emission and soil texture, soil trace elements of Fe, Cu and Mg, and above-ground biomass of the wheat crop at harvest. A further investigation suggested that the seasonal N₂O-N emission (E, mg m^-2) can be quantitatively explained by E=1005-34.2SOC+4.1S_a (R²=0.7703, n=18, P=0.0000). SOC and S_a represent the soil organic C (g kg^-1) and available S (mg kg^-1), respectively.     

Naidid worms (Oligochaeta, Naididae) in paddy soils as affected by the application of legume mulch and/or tillage practice

Reproduction, intrinsic rate of natural increase and population density of naidid worms were investigated in submerged paddy fields and the laboratory. No tillage plus legume-mulching increased the population density of naidid worms. Soil treatments with neither tillage nor legume mulch, and tillage practice alone, did not increase the number of worms. Dero dorsalis Ferronnière was dominant in soil of the no-tillage treatment. In laboratory experiments, legume-mulching with the proper amount of dissolved O₂ accelerated asexual reproduction of D. dorsalis through zooid budding. For the legume and aeration treatment, (N_i+1-N_i) N_i^-1 values (where N_i and N_i+1 are the populations at times t=i and t=i+1) were plotted against N_i+1. Utilizing this linear relation, this data fitted the logistic curve (r²=0.885, P<0.05). Based on the linear relation, the intrinsic rate of natural increase (r), carrying capacity (K), and doubling time (T) were calculated as 0.2125 day^-1, 12,666 m^-2, and 3.26 days, respectively. The amounts of legumes applied were highly correlated with the population of D. dorsalis, indicating that the weight of legume is a limiting factor with respect to carrying capacity. A literature review indicated a significant correlation (P<0.01) between intrinsic rate of natural increase and maximum body length of naidids with temperature conversion of the growth rate. Sexually mature worms were rarely found in submerged paddy fields. Sexual reproduction seems to be adopted in response to soil desiccation after paddy field drainage.     

Ecotoxicity of mercury to Eisenia fetida, Enchytraeus albidus and Folsomia candida

Despite growing concerns about the potential adverse effects of elevated mercury concentrations in the environment, only a few toxicity data are available for soil invertebrates. The chronic toxicity of mercury (II) was therefore assessed for Eisenia fetida, Enchytraeus albidus and Folsomia candida using standard test protocols. The 21-day EC₅₀ (the concentration causing 50% effect) for the cocoon production of E. fetida was 9.16 mg Hg kg^-1 dry wt. Based on the reproduction, a 42-day EC₅₀ of 22.0 mg Hg kg^-1 dry wt was observed for E. albidus while for F. candida the 28-day EC₅₀ was 3.26 (2.45-4.05) mg Hg kg^-1 dry wt. Although these data can be considered as a step forward in the assessment of the potential risks of mercury in terrestrial environments, further research is needed to evaluate the influence of soil parameters on the toxicity of mercury and to quantify the effect of ageing on the bioavailability.     

Carbon partitioning in plant and soil,carbon dioxide fluxes and enzyme activities as affected by cutting ryegrass

The effect of a single cut (simulated grazing) and regrowth of Lolium perenne on CO₂ efflux from soil (loamy Haplic Luvisol), on below-ground C translocation and on the distribution of plant C among different soil particle size fractions was investigated under controlled conditions with and without N fertilization by pulse labelling with ¹⁴C 7 times (four before and three after the cutting). The amount of ¹⁴C respired from the rhizosphere of Lolium decreased by a factor of about 3 during 1 month of growth. At the same time the amount of ¹⁴C stored in soil increased. Cut and non-fertilized plants respired less C in the rhizosphere compared to the uncut plants and cut fertilized plants. About 80% of the root-derived CO₂ efflux originated from the C assimilated after defoliation, and 20% originated from the C assimilated before cutting. N fertilization decreased the below-ground C losses (root respiration and exudation) during regrowth. The shoot is the main sink of assimilated C before and after the defoliation. N fertilization led to higher C incorporation into the shoot parts growing after defoliation compared to unfertilized plants. A lower incorporation of ¹⁴C was observed in the roots of N fertilized plants. The relative growth rates (expressed as ¹⁴C specific activity) of roots and stubble were minimal and that of shoot parts growing after defoliation was maximal. Twelve percent of ¹⁴C was found in the newly grown leaves after regrowth; nevertheless, 4.7% and 2.4% of ¹⁴C in the new shoot parts were translocated from the root and shoot reserves of unfertilized and fertilized plants, respectively. Most of the C retranslocated into the new Lolium leaves originates from the stubble and not from the roots. Between 0.5% and 1.7% of ¹⁴C recovered in shoots and below-ground C pools was found in the soil microbial biomass. Cutting and fertilization did not change ¹⁴C incorporation into the microbial biomass and did not affect xylanase, invertase, and protease activities. Tracing the assimilated C in particle size fractions revealed maximal incorporation for the sand and clay fraction.     

Dehydrogenase activity, redox potential, and emissions of carbon dioxide and nitrous oxide from Cambisols under flooding conditions

Samples from topsoils (0-10 cm) of 16 Polish arable Cambisols developed from different parent materials (sand, silt, sandy gravel, loess, loam and clay), were incubated under flooded conditions with NO₃^-. Dehydrogenase activity, redox potential (Eh), and emissions of CO₂ and N₂O were measured. According to dehydrogenase activity, the soils were divided into two groups: those of low activity (I), where the final dehydrogenase activity was <0.03 nmol triphenylformazan (TPF) g^-1 min^-1, and those with high final dehydrogenase activity (II), >0.03 nmol TPF g^-1 min^-1. Generation of CO₂ and of N₂O under flooded conditions was shown to be significantly related to dehydrogenase activity. Soil dehydrogenase activity increased curvilinearly with organic matter content, showed a maximum at pH 7.1, and decreased curvilinearly with Eh. The final cumulative CO₂ production increased linearly with soil organic matter content and curvilinearly with dehydrogenase activity and decreased linearly with Eh. The most significant relationship was found with dehydrogenase activity (R²=0.74, P<0.001). The final cumulative N₂O production decreased linearly with Eh and increased curvilinearly with pH and dehydrogenase activity but linearly with organic matter content; the most significant relation being found with dehydrogenase activity (R²=0.69, P<0.001). The CO₂:N₂O ratio in the gases evolved increased curvilinearly with Eh and decreased with dehydrogenase activity and N₂O and CO₂ production.     

Cultivar-specific dinitrogen fixation in Vicia faba studied with the nitrogen-15 natural abundance method

N fixation by different faba bean (Vicia faba) cultivars was studied using the natural abundance method. The delta ¹⁵N ('¹⁵N) values of the faba beans and the reference plants differed by 4.6-7.0‰. The non-nodulating V. faba cv. F48 seems to be the best reference plant for nodulated and N₂-fixing V. faba. Significant differences occurred in the quantity of N₂ fixation of six V. faba cultivars. The average fraction of N derived from air (FNdfa) estimated from leaf material ranged between 69 and 80%. Shoot-based estimates of N fixation varied between 200 and 360 kg N ha^-1. N fixation was affected more by differences in FNdfa than by differences in total N accumulation. Fixation data calculated with the non-nodulated reference plant V. faba cv. F48 were lower than those calculated with cabbage (Brassica oleracea) and ryegrass (Lolium perenne) as reference plants. Of all reference plants, non-N₂-fixing V. faba cv. F48 has a root system and temporal pattern of N assimilation that is the one most similar to that of N₂-fixing V. faba plants. Cv. F48 showed senescence as did the other V. faba cultivars after pod-fill was complete, whereas cabbage, ryegrass and camomile had a later senescence period. N fixation during pod-filling appears more important for a good yield than N₂ fixation abilities in the earlier growth period. The best V. faba cultivars left about 100 kg N ha^-1 in residual material on the field as fertilization for the following crops.     

Influence of soil parameters on the effect of 3,4-dimethylpyrazole-phosphate as a nitrification inhibitor

Nitrification inhibitors specifically retard the oxidation of NH₄⁺ to NO₂^- during the nitrification process in soil. In this study, the influence of soil properties on the nitrification-inhibiting effect of 3,4-dimethylpyrazole-phosphate (DMPP), a newly developed nitrification inhibitor, has been investigated. Based on short-term incubation experiments, where the degradation of DMPP could be largely disregarded, the oxidation of the applied NH₄⁺ was more inhibited in sandy soils compared with loamy soils. The influence of soil parameters on the relative NO₂^- formation could be described by a multiple regression model including the sand fraction, soil H⁺ concentration and soil catalase activity (R²=0.62). Adsorption studies showed that the binding behaviour of DMPP was influenced markedly by soil textural properties, viz. the clay fraction (r²=0.61). The adsorption of DMPP was found to be an important factor for the inhibitory effect on NH₄⁺ oxidation in a short-term incubation (r²=0.57). It is concluded that the evaluated soil properties can be used to predict the short-term inhibitory effect of DMPP in different soils. The significance of these results for long-term experiments under laboratory and field conditions needs further investigation.     

Different behavioral patterns of the earthworms Octolasion tyrtaeum and Diplocardia spp. in tallgrass prairie soils: potential influences on plant growth

This study addressed differences between Diplocardia spp. (a native North American earthworm) and Octolasion tyrtaeum (an introduced European species), with respect to behavior, influence on soil microbial biomass, and plant uptake of N in tallgrass prairie soils. We manipulated earthworms in PVC-encased soil cores (20 cm diameter) over a 45-day period under field conditions. Treatments included: (1) control with no earthworms, (2) Diplocardia spp. only, and (3) O. tyrtaeum only. Prior to addition of earthworms, seedlings of Andropogon gerardii (a dominant tallgrass) were established in each core, and a dilute solution of ¹³C-labeled glucose and ¹⁵N-labeled (NH₄)₂SO₄ was added to the soil to facilitate examination of earthworm/microbe/plant interactions. We found that Diplocardia spp. were significantly more active than O. tyrtaeum, and quickly assimilated ¹³C and ¹⁵N from the tracer. Individuals of Diplocardia spp. were present at shallower soil depths than O. tyrtaeum throughout the study. Contrary to expectation, this greater activity of Diplocardia spp. did not result in increased plant productivity. Rather, the activity of Diplocardia spp. was associated with less plant growth and smaller amounts of N acquired by A. gerardii seedlings compared to controls or O. tyrtaeum treatments. We observed few significant influences of earthworm treatments on microbial biomass C or N pool sizes, but the microbial C/N ratio was consistently greater in the presence of Diplocardia spp. relative to O. tyrtaeum. Results of this study indicate that activity of earthworms may enhance competition for N between microbes and plants during the growing season in tallgrass prairie.     

Bioavailability of triazine herbicides in a sandy soil profile

The bioavailability of atrazine was evaluated in a Danish soil profile (Drengsted) using a combination of soil sorption, transport and mineralisation methods as well as inoculation using Pseudomonas ADP. Sorption of atrazine decreased markedly with depth as indicated by K_d values of 5.2 l kg^-1 for the upper soil and 0.1 l kg^-1 for the subsoils. The transport of atrazine was evaluated using soil TLC plates and the resulting R_f values were 0.1 for the upper soil and 0.9 for the subsoil. Only a relatively small amount of atrazine leached through undisturbed soil columns taken from the upper 60 cm. Inoculating with Pseudomonas strain ADP (1᎒⁶ CFU g^-1 dry weight soil) revealed that the degradation of 0.01 ppm atrazine was fully completed (80% mineralisation) within 10 days in the subsoil, while it reached less than 15% in the upper soil. Over a period of 500 days, a total mineralisation of 37% of added atrazine in the upper soil was found (2 mg kg^-1 incubated at 20° C). However, in the subsurface soil where 0.02 mg kg^-1 of atrazine was incubated at 10°C, the degradation was slower, only reaching about 12%. Terbuthylazine mineralisation was found to be temperature-dependent and low (less than 5%) in the upper soil and very much lower in the subsoil. Desethylterbuthylazine was the most frequently found metabolite. Finally, Pseudomonas strain ADP inoculated into soils from different depths increased the mineralisation of terbuthylazine dramatically. Modelling using a "two-compartment model" indicated that desorption of terbuthylazine is the limiting step for its mineralisation.     

Denitrification, N2O and CO2 fluxes in rice-wheat cropping system as affected by crop residues, fertilizer N and legume green manure

Use of renewable N and C sources such as green manure (GM) and crop residues in rice-wheat cropping systems of South Asia may lead to higher crop productivity and C sequestration. However, information on measurements of gaseous N losses (N₂O+N₂) via denitrification and environmental problems such as N₂O and CO₂ production in rice-wheat cropping systems is not available. An acetylene inhibition-intact soil core technique was employed for direct measurement of denitrification losses, N₂O and CO₂ production, in an irrigated field planted to rice (Oryza sativa L.) and wheat (Triticum aestivum L.) in an annual rotation. The soil was a coarse-textured Tolewal sandy loam soil (Typic Ustochrept) and the site a semi-arid subtropical Punjab region of India. Wheat residue (WR, C:N=94) was incorporated at 6 t ha^-1 and sesbania (Sesbania aculeata L.) was grown as GM crop for 60 days during the pre-rice fallow period. Fresh biomass of GM (C:N.=18) at 20 or 40 t ha^-1 was incorporated into the soil 2 days before transplanting rice. Results of this study reveal that (1) denitrification is a significant N loss process under wetland rice amounting to 33% of the prescribed dose of 120 kg N ha^-1 applied as fertilizer urea-N (FN); (2) integrated management of 6 t WR ha^-1 and 20 t GM ha^-1 supplying 88 kg N ha^-1 and 32 kg FN ha^-1 significantly reduced cumulative gaseous N losses to 51.6 kg N ha^-1 as compared with 58.2 kg N ha^-1 for 120 kg FN ha^-1 alone; (3) application of excessive N and C through applying 40 t GM ha^-1 (176 kg N ha^-1) resulted in the highest gaseous losses of 70 kg N ha^-1; (4) the gaseous N losses under wheat were 0.6% to 2% of the applied 120 kg FN ha^-1 and were eight- to tenfold lower (5-8 kg N ha^-1) than those preceding rice; (5) an interplay between the availability of NO₃^- and organic C largely controlled denitrification and N₂O flux during summer-grown flooded rice whereas temperature and soil aeration status were the primary regulators of the nitrification-denitrification processes and gaseous N losses during winter-grown upland wheat; (6) the irrigated rice-wheat system is a significant source of N₂O as it emits around 15 kg N₂O-N ha^-1 year^-1; (7) incorporation of WR in rice and rice residue (C:N=63) in wheat increased soil respiration, and increased CO₂ production in WR- and GM-amended soils under anaerobic wetland rice coincided with enhanced rates of denitrification; and (8) with adequate soil moisture, most of the decomposable C fraction of added residues was mineralized within one crop-growing season and application of FN and GM further accelerated this process.     

Nitrous oxide emissions from kharif and rabi legumes grown on an alluvial soil

Nitrous oxide (N₂O) emissions were monitored for a period of 60 days in a pot culture study, from two kharif (June-September) and two rabi (October-March) season legumes, which were grown on a Typic Ustochrept, alluvial sandy loam soil. Black gram (Vigna mungo L. Hepper), var. T-9, and soybean (Glycine max L. Merril), var. Punjab 1, were taken up in kharif season whereas lentil (Lens esculenta Moench), var. JLS-1, and Bengal gram (Cicer arietinum L.), var. BGD-86, were grown in rabi season. All the crops were grown with and without urea and one pot (containing soil but with no fertilizer or crop) was used as a control. Nitrous oxide emissions were significantly higher in unfertilized cropped soil than in the control, while the addition of urea to the crops further increased the emissions. Significant emissions occurred during third and seventh week after sowing for all the treatments in both kharif and rabi seasons. In kharif, soil cropped with soybean had higher total N₂O-N emission than soil sown with black gram both under fertilized and unfertilized conditions; while in rabi, lentil had a higher total N₂O-N emission than Bengal gram under both fertilized and unfertilized conditions. In kharif, total N₂O-N emissions ranged from 0.53 (control) to 3.84 kg ha^-1 (soybean+urea), while in rabi it ranged from 0.45 (control) to 3.06 kg ha^-1 (lentil+urea). Higher N₂O-N emissions in kharif than in rabi was probably due to the favorable effect of temperature on nitrification and denitrification in the former season. The results of the study indicated that legume crops may lead to an increase in N₂O formation and emission from soils, the extent of which varies from crop to crop.     

Effects of nitrogen fertilization on soil nitrogen pools and microbial properties in a hoop pine (Araucaria cunninghamii) plantation in southeast Queensland, Australia

A field study was conducted to investigate the effects of N fertilization on soil N pools and associated microbial properties in a 13-year-old hoop pine (Araucaria cunninghamii) plantation of southeast Queensland, Australia. The treatments included: (1) control (without N application); (2) 300 kg N ha^-1 applied as NH₄NO₃; and (3) 600 kg N ha^-1 as NH₄NO₃. The experiment employed a randomized complete block design with four replicates. Soil samples were taken approximately 5 years after the N application. The results showed that application of 600 kg N ha^-1 significantly increased concentrations of NH₄⁺-N in 0-10 cm soil compared with the control and application of 300 kg N ha^-1. Concentrations of NO₃^--N in soil (both 0-10 cm and 10-20 cm) with an application rate of 600 kg N ha^-1 were significantly higher compared with the control. Application of 600 kg N ha^-1 significantly increased gross N mineralization and immobilization rates (0-10 cm soil) determined by ¹⁵N isotope dilution techniques under anaerobic incubation, compared with the control. However, N application did not significantly affect the concentrations of soil total C and total N. N application appeared to decrease microbial biomass C and N and respiration, and to increase the metabolic quotient (qCO₂) in 0-10 cm soil, but these effects were not statistically significant. The lack of statistical significance in these microbial properties between the treatments might have been associated with large spatial variability between the replicate plots at this experimental site. Spatial variability in soil microbial biomass C and N was found to relate to soil moisture, total C and total N.     

Nitrogen mineralization potential of dairy manures and its relationship to composition

The objectives of this study were to determine the variability in mineralization of dairy manure N, to determine if N mineralization can be predicted by compositional factors or by near- or mid-infrared reflectance spectroscopy. Dairy manures (n =107) were collected from farms in Maryland, Virginia, Pennsylvania, New York, and Connecticut. The composition of these manures ranged from 14 to 386 g dry matter kg^-1, 0.9 to 9.5 kg total N/m³, and 0.3 to 4.7 kg NH₄⁺-N/m³. Manure-amended soil was aerobically incubated at 25°C and concentrations of NH₄⁺-N and NO₃^--N were determined at day 2 and day 56. The manures were highly variable in their N mineralization characteristics, ranging from a net mineralization of 54.9% to a net immobilization of 29.2% of the organic N. When compositional parameters were individually regressed against percentage mineralized organic N, the highest correlation coefficient (r) was 0.164. A stepwise regression of all 11 variables yielded a maximal r of 0.486. These results suggest that the availability of dairy manure organic N is highly variable and that the availability cannot be predicted from simple compositional parameters. No relationship was found between near-infrared spectral characteristics and N mineralization suggesting that no simple relationship exists between N mineralization and compositional characteristics. There appears to be some potential for the use of mid-infrared for determining the mineralization potential of manures.     

Effects of dicyandiamide, farmyard manure and irrigation on crop yields and ammonia volatilization from an alluvial soil under a rice (Oryza sativa L.)-wheat (Triticum aestivum L.) cropping system

Volatilization of NH₃ from soil is a major N-loss mechanism that reduces the efficiency of applied N fertilizers, and causes environmental pollution. Strategies are needed to reduce the loss. The influences of dicyandiamide (DCD), farmyard manure (FYM) and irrigation on NH₃ volatilization from an alluvial soil in rice (Oryza sativa L.)-wheat (Triticum aestivum L.) cropping system was studied using the acid trap method. The loss of NH₃ in the rice-wheat system ranged from 38.6 kg N ha^-1 from the unfertilized soil to 69.0 kg N ha^-1 in the treatment with urea+DCD. Substitution of 50% N provided through urea by FYM reduced NH₃-N volatilization by 10% in rice and wheat as compared to the urea treatment. Application of DCD increased NH₃ volatilization in wheat by 7% but in rice it had no effect. The irrigation level had no effect on NH₃ volatilization in rice but fewer irrigations with fewer splits of N in wheat resulted in higher NH₃ volatilization. Application of DCD and FYM with urea had similar effects on grain yield and N uptake by rice and wheat as that of the urea treatment. The study showed that integrated use of organic manure and chemical fertilizer has the potential to reduce the loss of N due to volatilization and thereby minimize environmental pollution. Nitrification inhibitors, which are reported to be useful in increasing the N-use efficiency by reducing the leaching and denitrification losses of N, however, may increase N loss due to volatilization.     

Use of principal component analysis to assess factors controlling net N mineralization in deciduous and coniferous forest soils

Net N mineralization was studied in three different forest sites (Belgium): a mixed deciduous forest with oak (Quercus robur L. and Quercus rubra L.) and birch (Betula pendula Roth) as dominant species, a deciduous stand of silver birch (Betula pendula) and a coniferous stand of Corsican pine (Pinus nigra ssp. Laricio). The organic (F + H) layer and mineral soil at different depths (0-10, 10-20 and 20-30 cm) were sampled at three locations in the mixed deciduous forest (GE, GF1, GF2), at one location in the silver birch stand (SB) and one in the Corsican pine stand (CP). All samples were incubated over 10 weeks under controlled temperature and moisture conditions. The net N mineralization rates in the organic and upper mineral layer (0-10 cm) were found to be significantly different from the other layers and accounted for 66-95% of the total mineralization over the first 30 cm. Net N mineralization rates in the organic layer ranged from 4.2 to 27.3 mg N m^-2 day^-1. Net N mineralization and nitrification rates were positively correlated. For the mineral soil, net N mineralization rates decreased with depth and the upper 10 cm showed significantly higher rates, ranging from 8.9 to 33.5 mg N m^-2 day^-1. The rates of the 10-20 cm and 20-30 cm sublayers were similar, ranging from 1.2 to 7.4 mg N m^-2 day^-1. The net N mineralization rates for the total mineral layer (0-30 cm) ranged from 17.4 mg N m^-2 day^-1 (SB) to 36.1 mg N m^-2 day^-1 (CP). Both from PCA and multiple regression analysis, we could conclude that net N mineralization rates were closely related to the initial mineral N content (N_initial). Furthermore, significant correlations were observed between the net N mineralization rate, the total carbon (TC) and NH₄⁺-N content for the mineral layers and between net N mineralization rate, total nitrogen (TN), hemicellulose content and C/N for the organic layers.     

Impact of soil nitrogen concentration on Glomus spp.-Sinorhizobium interactions as affecting growth, nitrate reductase activity and protein content of Medicago sativa

Our objective was to evaluate how increasing levels of N in the medium (0, 4, 8 and 16 mmol N added kg^-1 soil) affect the interaction between Sinorhizobium and arbuscular mycorrhiza (AM) fungi in the tripartite symbiosis with Medicago sativa. Growth response, nutrient acquisition, protein content, and nitrate reductase (NR) activity were measured both in plant shoots and roots. Results showed that N levels in soil did not affect mycorrhizal colonization but they strongly influenced nodulation, particularly of mycorrhizal plants. Mycorrhizal colonization was required for a proper nodulation when no N was applied to soil. In contrast, the addition of 4 mmol N kg^-1 soil reduced nodulation only in mycorrhizal plants and 8 mmol N added kg^-1 soil allowed nodule formation only in non-mycorrhizal plants. Nodulation was totally inhibited in all treatments with the addition of 16 mmol N added kg^-1 soil. N addition enhanced NR activity in all the treatments, while AM colonization increased the proportion of NR allocated to roots. This effect was more pronounced under the lowest N levels in the medium. The two AM fungal species showed different distribution pattern of enzymatic activities in plant tissues indicating specific physiological traits. Protein content as well as the relative proportion of protein in roots were greatly increased after mycorrhizal colonization. Glomus intraradices-colonized plants had the highest protein content in shoot and root. Mycorrhizal effects on growth, N acquisition and biochemical variables cannot be interpreted as an indirect P-mediated effect since P content was lower in mycorrhizal plants than in those which were P fertilized. Mycorrhizal colonization increased the N content in plants irrespective of the N level, but the effectiveness of AM fungi on plant N acquisition depended on the AM fungus involved, G. intraradices being the most effective, particularly at the highest N rate. N₂ fixation, enhanced by AM colonization, contributed to N acquisition when a moderate N quantity was available in the soil. Nevertheless, under a high N amount the nodulating process and/or fixing capacity by Sinorhizobium was reduced in AM plants. In contrast, the AM fungal mycelium from a particular mycorrhizal fungus may continue to contribute efficiently to the N uptake from the soil even at high N levels. These results demonstrate the particular sensitivity of AM fungal species in terms of their growth and/or function to increasing N amounts in the medium. A selection of AM fungi used to address specific environmental conditions, such as N fertilization regimes comparable to those used in agronomic practices, is required for a better use of N applied to soil.     

Earthworm excreta (mucus and urine) affect the distribution of springtails in forest soils

The collembolan species Heteromurus nitidus, exclusively located in soils at pH>5, can be cultured in acidic humus. As this species is attracted to the excreta of earthworms from calcic mull, its distribution is supposed to be determined only indirectly by soil pH through the distribution of earthworms. Higher densities and biomasses of Lumbricidae were observed in a calcic mull (pH 7.8) than in an acidic mull (pH 4.8) and a moder humus (pH 4.2). Choice experiments were performed to compare the attraction of H. nitidus to the mucus-urine mix of five earthworm species from the calcic mull and the acidic mull. H. nitidus was attracted to the excreta of the five species, whatever their ecological category and the humus form from which they originated. The collembolan Heteromurus major, which was indifferent to soil pH, was not attracted to earthworm excreta, which emphasizes the significance of this phenomenon for the distribution of H. nitidus over a pH range. The attraction of H. nitidus to earthworm excreta tended to be weaker and more variable when earthworms originated from acidic mull compared to calcic mull, particularly in the case of Lumbricus terrestris. Increased earthworm density reinforced by better mucus quality and quantity could determine the distribution of H. nitidus according to soil acidity. The only urine compound capable of attracting H. nitidus was NH₃ at a low concentration (0.03 g l^-1). The NH₃ content of the mucus-urine mix which attracted H. nitidus was 0.037 g l^-1, and was therefore responsible, at least partly, for the attraction.     

Plant growth enhancement and suppression of Macrophomina phaseolina causing charcoal rot of peanut by fluorescent Pseudomonas

A plant growth-promoting bacterial strain, GRC₂, was isolated from potato rhizosphere and characterized as fluorescent Pseudomonas. It produced a hydroxamate-type siderophore in iron-deficient tryptic soy medium. The production of hydrocyanic acid and indole acetic acid was also recorded under normal growth conditions. The strain showed a strong antagonistic effect against Macrophomina phaseolina, a charcoal rot pathogen of peanut, when co-cultured on tryptic soy agar medium. The growth inhibition of M. phaseolina was 74% after 5 days of incubation. Bacterization of peanut seeds with fluorescent Pseudomonas GRC₂ resulted in increased seed germination, early seedling growth, fresh nodule weight, grain yield and reduced charcoal rot disease of peanut in M. phaseolina-infested soil as compared with control. A streptomycin-resistant marker of the bacterium GRC₂^strep+ was used to monitor root colonization, which positively confirmed the efficient colonization of peanut root. Seed bacterization reduced charcoal rot disease in M. phaseolina-infested soil as compared with the control by 99%, making the organism a potential biocontrol agent against charcoal rot of peanut.