Distribution of monoterpenes between organic resources in upper soil horizons under monocultures of Picea abies, Picea sitchensis and Pinus sylvestris

The aim of this study was to compare the monoterpene content and distribution in litters and roots of three conifer species: Picea abies (L.) Karst, Picea sitchensis (Bong.) Carr. and Pinus sylvestris (L.). We analysed the monoterpene content of green needles, needle litter, F (fermentation) layer material and roots collected from monoculture plots. The rate of loss of monoterpenes from freshly fallen litter in the field was also studied at two monthly intervals over 10 months, to assess the length of time that monoterpenes entering the litter layer remain. Monoterpene analysis was carried out by extracting homogenised samples in hexane and identifying and quantifying the resulting monoterpenes using gas chromatography with flame ionisation detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Mean total monoterpene concentrations varied significantly between the three species examined (e.g. in freshly fallen litter 1531 ± 96, 100 ± 5 and 1175 ± 122 μg g⁻¹ d. wt for P. abies, P. sitchensis and P. sylvestris); each species had distinctive and consistent monoterpene profiles associated with each type of tissue, and total monoterpene concentrations in green needles varied between individual trees of the same species, particularly for P. sitchensis. A substantial proportion of the monoterpene content of green needles remained in the needles after litter fall for P. abies (42%), P. sitchensis (11%) and P. sylvestris (30%). Although rates of monoterpene loss from needle litters varied initially (P. sylvestris > P. abies > P. sitchensis), the majority of the monoterpene content was lost after 4-6 months. Maximum monoterpene emission rates from decaying litter were calculated of 39 (P. abies), 1.7 (P. sitchensis) and 39 μg m⁻² h⁻¹ (P. sylvestris). Monoterpene concentrations in F layer material were very low (<10 μg g⁻¹ d. wt). Roots, particularly in P. sylvestris, represented a significant pool of monoterpenes (185 ± 16, P. abies; 258 ± 54, P. sitchensis; 2133 ± 200 μg g⁻¹ d. wt, P. sylvestris). The monoterpene profile was similar between roots and litter of P. sylvestris (α-pinene most abundant), and for P. sitchensis, (limonene and α-pinene most abundant), although a different pattern was observed between needle litter (most abundant β-pinene) and roots (most abundant myrcene) of P. abies. The relatively high concentrations and different profiles of monoterpenes characterised in upper organic soil horizons here emphasise the need for their influence on soil ecological processes to be assessed.     

Microbial colonisation of roots as a function of plant species

Fifteen plants species were grown in the greenhouse on the same soil and sampled at flowering to obtain rhizosphere soil and root material. In both fractions, the data on fungal and bacterial tissue obtained by amino sugar analysis were compared with the total microbial biomass based on fumigation-extraction and ergosterol data. The available literature on glucosamine concentrations in fungi and on muramic acid concentrations in bacteria was reviewed to prove the possibility of generating conversion values for general use in root material. All microbial properties analysed revealed strong species-specific differences in microbial colonisation of plant roots. The root material contained considerable amounts of microbial biomass C and biomass N, reaching mean levels of 10.9 and 1.4 mg g⁻¹ dry weight, respectively. However, the majority of CHCl₃ labile C and N, i.e. 89 and 55% was root derived. The average amount of ergosterol was 13 μg g⁻¹ dry weight and varied between 0.0 for Phacelia roots and 45.5 μg g⁻¹ dry weight for Vicia roots. The ergosterol content in root material of mycorrhizal and non-mycorrhizal plant species did not differ significantly. Fungal glucosamine was converted to fungal C by multiplication by 9 giving a range of 7.1-25.9 mg g⁻¹ dry weight in the root material. Fungal C and ergosterol were significantly correlated. Bacterial C was calculated by multiplying muramic acid by 45 giving a range from 1.7 to 21.6 mg g⁻¹ dry weight in the root material. In the root material of the 15 plant species, the ratio of fungal C-to-bacterial C ranged from 1.0 in mycorrhizal Trifolium roots to 9.5 in non-mycorrhizal Lupinus roots and it was on average 3.1. These figures mean that the microbial tissue in the root material consists on average of 76% fungal C and 24% bacterial C. The differences in microbial colonisation of the roots were reflected by differences in microbial indices found in the rhizosphere soil, most strongly for microbial biomass C and ergosterol, but to some extent also for glucosamine and muramic acid.     

Boreal pine forest floor biogenic volatile organic compound emissions peak in early summer and autumn

In boreal forests, canopy-scale emissions of biogenic volatile organic compounds (BVOCs) are rather well characterised, but knowledge of ecosystem-scale BVOC emissions is still inadequate. We used adsorbent tubes to measure BVOCs from a boreal Scots pine (Pinus sylvestris L.) forest floor in southern Finland and analysed the compounds with a gas chromatograph-mass spectrometer. The most abundant compound group was the monoterpenes (averaging 5.04 μg m⁻² h⁻¹), in which α-pinene, Δ³-carene and camphene contributed over 90% of the emissions. Emissions of other terpenoids (isoprene and sesquiterpenes) were low (averaging 0.05 and 0.04 μg m⁻² h⁻¹, respectively). BVOC emissions from the forest floor varied seasonally, peaking in early summer and autumn, with most of the compounds following similar patterns. The emission pattern was sustained throughout the measurement period, suggesting that the main sources of the emissions remained more or less stable. We compared the BVOC fluxes with environmental parameters such as temperature, precipitation and PAR, and with fluxes of other trace gases (CO₂, CH₄, N₂O), as well as with ground vegetation photosynthesis and with litter input. Several of these parameters were correlated with the presence of BVOCs. The sources of soil BVOC emissions are very poorly understood, but our results suggest, that changes in litter quantity and quality, soil microbial activity and the physiological stages of plants are linked with changes in BVOC fluxes.     

Subtropical plantations are large carbon sinks: Evidence from two monoculture plantations in South China

Quantifying the net carbon (C) storage of forest plantations is required to assess their potential to offset fossil fuel emissions. In this study, a biometric approach was used to estimate net ecosystem productivity (NEP) for two monoculture plantations in South China: Acacia crassicarpa and Eucalyptus urophylla. This approach was based on stand-level net primary productivity (NPP, based on direct biometric inventory) and heterotrophic respiration (R_h). In comparisons of R_h determination based on trenching vs. tree girdling, both trenching and tree girdling changed soil temperature and soil moisture relative to undisturbed control plots, and we assess the effects of corrections for disturbances of soil moisture and soil moisture on the estimation of soil CO₂ efflux partitioning. Soil microbial biomass and dissolved organic carbon were significantly lower in trenched plots than in tree girdled plots for both plantations. Annual soil CO₂ flux in trenched plots (R_h-t) was significantly lower than in tree-girdled plots (R_h-g) in both plantations. The estimates of R_h-t and R_h-g, expressed as a percentage of total soil respiration, were 58 ± 4% and 74 ± 6%, respectively, for A. crassicarpa, and 64 ± 3% and 78 ± 5%, respectively, for E. urophylla. By the end of experiment, the difference in soil CO₂ efflux between the trenched plots and tree-girdled plots had become small for both plantations. Annual R_h (mean of the annual R_h-t and R_h-g) and net primary production (NPP) were 470 ± 25 and 800 ± 118 g C m⁻² yr⁻¹, respectively, for A. crassicarpa, and 420 ± 35 and 2380 ± 187 g C m⁻² yr⁻², respectively, for E. urophylla. The two plantations in the developmental stage were large carbon sinks: NEP was 330 ± 76 C m⁻² yr⁻¹ for A. crassicarpa and 1960 ± 178 g C m⁻² yr⁻¹ for E. urophylla.     

Profiling of PLFA: Implications for nonlinear spatial gradient of PCP degradation in the vicinity of Lolium perenne L. roots

An investigation was conducted using phospholipid fatty acids (PLFAs) profiles to follow the spatial response of the microbial community at the millimeter scale with the purpose of illustrating the mechanism of nonlinear spatial dependence of PCP degradation on the distance from the root surface in the rhizosphere of Lolium perenne L. A laminar rhizobox was designed to allow the harvest of intact layers of root compartment, near-(1, 2, 3, 4, 5 mm) and far-(>5 mm) rhizosphere soil from root surfaces without the removal of the root material itself. Lolium perenne L. was grown in environmental chambers for 53 days with soil spiked with 8.7 and 18 mg kg⁻¹ PCP. PLFA profiles were found to be affected by the distance from the rhizosphere, indicating a distance-dependent selective enrichment of competent species that may be responsible for efficient PCP degradation. In particular, the five fatty acids 16:1ω5, 16:0, i17:0, a17:0 and 10Me18:0 emerged as microbiological biomarkers that may be used for assessing phytoremediation processes of PCP in soil. Their synergistic effects were shown to be most responsive to the nonlinear spatial patterns of PCP degradation in the vicinity of Lolium perenne L. roots. The results suggest that root exudates induced modifications of microbial communities in the PCP contaminated rhizosphere and spatially modified the dominant species within these communities, resulting in the nonlinear PCP degradation pattern.     

Assessing the uncertainty of estimated annual totals of net ecosystem productivity: A practical approach applied to a mid latitude temperate pine forest

Values for annual NEP of micrometeorological tower sites are usually published without an estimate of associated uncertainties. Few authors quantify total uncertainty of annual NEP. Moreover, different methods to assess total uncertainty are applied, usually addressing only one aspect of the uncertainty. This paper presents a robust and easy to apply method to quantify uncertainty of annual totals of Net Ecosystem Productivity (NEP), related to multiple factors involved therein. The method was applied to NEP observations for a Scots pine forest (Loobos) in the Netherlands. Total uncertainty of annual NEP for the Loobos site was on average ±32 g C m⁻² a⁻¹ (±8% of NEP), which is a quarter of the standard deviation of annual NEP (127 g C m⁻² a⁻¹).     

Rapid quantification of Bacillus subtilis antibiotics in the rhizosphere

Biological control agents like Bacillus subtilis offer an alternative and supplement to synthetic pesticides. Antibiotic production by biocontrol strains of B. subtilis can play a major role in plant disease suppression. Our current understanding of B. subtilis antibiosis comes from culture media measurements of antibiotic production and in vitro suppression of pathogens. Quantifying the antibiotic metabolite chemistry of B. subtilis biofilms growing on root surfaces provides a more accurate understanding of in vivo antibiotic production. An analytical method based on solid-phase extraction (SPE) with high-performance liquid chromatography (HPLC) and mass spectroscopy (MS) has been developed to quantify antibiotics produced by B. subtilis growing on plant roots. Cucumber (Cucumis sativus) was grown in composted soil and potting media inoculated with B. subtilis strain QST 713 (AgraQuest, USA). Two important B. subtilis antibiotics, surfactin and iturin A, were extracted from root and rhizosphere soil using acidified organic solvents followed by cleaning and concentration using SPE. HPLC and HPLC-MS were used to measure surfactin and iturin A. Rhizosphere concentrations of both antibiotics increased with plant age. For plants grown in peat-based potting media, surfactin concentrations increased from 9 μg g⁻¹ root fresh weight (RFW) at 15 d to 30 μg g⁻¹ RFW at 43 d. Iturin concentrations were 7 μg g⁻¹ RFW at 15 d and 180 μg g⁻¹ RFW at 43 d. In an initial field trial in a composted fine sandy loam, we demonstrated rhizosphere production of surfactin and iturin under competition and predation by the myriad macro- and microfauna existing in a fertile high-organic soil, with mature B. subtilis-inoculated cucumber roots yielding 33 μg g⁻¹ RFW surfactin and 630 μg g⁻¹ RFW iturin at 78 d.     

Specific flavonoids as interconnecting signals in the tripartite symbiosis formed by arbuscular mycorrhizal fungi, Bradyrhizobium japonicum (Kirchner) Jordan and soybean (Glycine max (L.) Merr.)

Many legume plants benefit from the tripartite symbiosis of arbuscular mycorrhizal fungi (AMF) and rhizobia. Beneficial effects for the plant have been assumed to rely on increased P supply through the mycorrhizas. Recently, we demonstrated that P does not regulate the establishment of the tripartite symbiosis. Flavonoids appear to play a role as early signals for both rhizobia and AMF. Four soybean lines known to express different concentrations of the isoflavones genistein, daidzein and glycitein in the seed were used to test three hypotheses: (i) The establishment of the tripartite symbiosis is not dependent of a nutrient mediated effect; (ii) There is a positive correlation between seed isoflavone concentrations of different soybean lines and the progress of the tripartite symbiosis; (iii) Specific flavonoids control the establishment of the tripartite symbiosis in that a change in flavonoid root accumulation resulting from the development of one microbial partner can stimulate colonization of soybean roots by the other. Disturbed versus undisturbed soil treatments were produced to vary the potential for indigenous AMF colonization of soybean. In contrast, the potential for Bradyrhizobium was kept identical in both soil disturbance treatments. The uptake of P and Zn and the concentration of flavonoids in mycorrhizal soybean roots at 10 d after emergence were analysed either separately of Bradyrhizobium or in context of the tripartite symbiosis. Zinc nutrition did not differ between AMF treatments which supports the first hypothesis. The concentration of daidzein was at least four times greater in the root than in the seed reaching 3958±249 μg g⁻¹ dry across soybean lines. Coumestrol, which was absent in the seed, was synthesized to reach 2154±64 μg g⁻¹ dry. Conversely, the concentration of genistein was approximately three times smaller in the root that in the seed (301±15 μg g⁻¹ dry), while glycitein and formononetin were never detected. The establishment of the tripartite symbiosis was identical across soybean lines which does not support the second hypothesis. Concentrations of flavonoids were significantly greater in roots under disturbed soil, for which both symbioses were not as developed as in plants from undisturbed soil. This clearly supports the third hypothesis. This research provides the first data linking the function of different flavonoids to the establishment of the tripartite symbiosis, and suggests that these compounds are produced and released into the rhizosphere as a function of the colonization process.     

Community composition and activities of denitrifying bacteria from adjacent agricultural soil, riparian soil, and creek sediment in Oregon, USA

We examined denitrifying bacteria from wet soils and creek sediment in an agroecosystem in Oregon, USA that received inputs of nitrogen (N) fertilizer. Our objective was to determine the variation in denitrifying community composition and activities across three adjacent habitats: a fertilized agricultural field planted to perennial ryegrass, a naturally vegetated riparian area, and creek sediment. Using C₂H₂ inhibition, denitrifying enzyme and N₂O-reductase activities were determined in short-term incubations of anaerobic slurries. A key gene in the denitrification pathway, N₂O reductase (nosZ), served as a marker for denitrifiers. Mean denitrifying enzyme activity (DEA) was similar among habitats, ranging from 0.5 to 1.8 μg N g⁻¹ dry soil h⁻¹. However, the ratio of N₂O production, without C₂H₂, to DEA was substantially higher in riparian soil (0.64±0.02; mean±standard error, n=12) than in agricultural soil (0.19±0.02) or creek sediment (0.32±0.03). Mean N₂O-reductase activity ranged from 0.5 to 3.2 μg N g⁻¹ dry soil h⁻¹, with greater activity in agricultural soil than in riparian soil. Denitrifying community composition differed significantly among habitats based on nosZ terminal-restriction fragment length polymorphisms. The creek sediment community was unique. Communities in the agricultural and riparian soil were more closely related but distinct. A number of unique nosZ genotypes were detected in creek sediment. Sequences of nosZ obtained from riparian soil were closely related to nosZ from Bradyrhizobium japonicum. Although nosZ distribution and N₂O-reductase activity differed among habitats, relationships between activity and community composition appeared uncoupled across the agroecosystem.     

Brassica genotypes differ in growth, phosphorus uptake and rhizosphere properties under P-limiting conditions

Compared to other crops, Brassicas are generally considered to grow well in soils with low P availability, however, little is known about genotypic differences within Brassicas in this respect. To assess the role of rhizosphere properties in growth and P uptake by Brassicas, three Brassica genotypes (mustard, Brassica juncea cv Chinese greens and canola, Brassica napus cvs Drum and Outback) were grown in an acidic soil with low P availability at two treatments of added P: 25 and 100 mg P kg⁻¹ as FePO₄ (P25 and P100). The plants were harvested at the 6-leaf stage, at flowering and at maturity. Shoot and root dry weight (dry weight) and root length increased with time and were lower in P25 than in P100. In P25, shoot dry weight was lowest in Outback and highest in Chinese greens. In the P100 treatment, Chinese greens had a higher shoot dry weight than the two canola cultivars. Chinese greens had a lower root dry weight and root length at flowering and maturity than the canola genotypes in both P treatments. Irrespective of P treatment, shoot P concentration was lower in Chinese greens than in the two canola genotypes. Specific P uptake (μg P m⁻¹ root length) decreased with time. In P25, Chinese greens had the lowest specific P uptake at the 6-leaf stage but it was higher than in the two canola genotypes at flowering and maturity. In P100, Outback had the lowest specific P uptake. Available P in the rhizosphere (resin P) decreased over time with the greatest decrease from the 6-leaf stage to flowering. In P25, resin P in the rhizosphere was greatest in Chinese greens at the 6-leaf stage and flowering and smallest in Outback at flowering. Microbial P and acid phosphatase activity changed little over time, were not affected by P treatment and there were only small differences between the genotypes. The rhizosphere microbial community composition [assessed by fatty acid methyl ester (FAME) analysis] of Outback and Chinese greens differed from that of the other two genotypes at the 6-leaf stage and flowering, respectively. At maturity, all three genotypes had distinct microbial communities. Plant traits such as production of high biomass at low shoot P concentrations as well as the capacity to maintain high P availability in the rhizosphere by P mobilisation can explain the observed differences in plant growth and P uptake among the Brassica genotypes.     

Additions of maize root mucilage to soil changed the structure of the bacterial community

The organic compounds released from roots (rhizodeposits) stimulate the growth of the rhizosphere microbial community. They may be responsible for the differences in the structure of the microbial communities commonly observed between the rhizosphere and the bulk soil. Rhizodeposits consists of a broad range of compounds including root mucilage. The aim of this study was to investigate if additions of maize root mucilage, at a rate of 70 μg C g⁻¹ day⁻¹ for 15 days, to an agricultural soil could affect the structure of the bacterial community. Mucilage additions moderately increased microbial C (+23% increase relative to control), which suggests that the turnover rate of microorganisms consuming this substrate was high. Consistent with this, the number of cultivable bacteria was enhanced by +450%. Catabolic (Biolog^® GN2) and 16S-23S intergenic spacer fingerprints exhibited significant differences between control and mucilage treatments. These data indicate that mucilage can affect both the metabolic and genetic structure of the bacterial community as shown by a greater catabolic potential for carbohydrates. We concluded that mucilage is likely to significantly contribute to differences in the structure of the bacterial communities present in the rhizosphere compared to the bulk soil.     

Toxicological effects of TiO2 and ZnO nanoparticles in soil on earthworm Eisenia fetida

Nanoparticles (NPs) of TiO₂ and ZnO are receiving increasing attention due to their widespread applications. To evaluate their toxicities to the earthworm Eisenia fetida (Savigny, 1826) in soil, artificial soil systems containing distilled water, 0.1, 0.5, 1.0 or 5.0 g kg⁻¹ of NPs were prepared and earthworms were exposed for 7 days. Contents of Zn and Ti in earthworm, activities of antioxidant enzymes, DNA damage to earthworm, activity of cellulase and damage to mitochondria of gut cells were investigated after acute toxicity test. The results from response of the antioxidant system combined with DNA damage endpoint (comet assay) indicated that TiO₂ and ZnO NPs could induce significant damage to earthworms when doses were greater than 1.0 g kg⁻¹. We found that Ti and Zn, especially Zn, were bioaccumulated, and that mitochondria were damaged at the highest dose in soil (5.0 g kg⁻¹). The activity of cellulase was significantly inhibited when organisms were exposed to 5.0 g kg⁻¹ of ZnO NPs. Our study demonstrates that both TiO₂ and ZnO NPs exert harmful effects to E. fetida when their levels are higher than 1.0 g kg⁻¹ in soil and that toxicity of ZnO NPs was higher than TiO₂.     

NO, N2O, CH4 and CO2 fluxes in winter barley field of Japanese Andisol as affected by N fertilizer management

The study was carried out at the experimental station of the Japan International Research Center for Agricultural Sciences to investigate gas fluxes from a Japanese Andisol under different N fertilizer managements: CD, a deep application (8 cm) of the controlled release urea; UD, a deep application (8 cm) of the conventional urea; US, a surface application of the conventional urea; and a control, without any N application. NO, N₂O, CH₄ and CO₂ fluxes were measured simultaneously in a winter barley field under the maize/barley rotation. The fluxes of NO and N₂O from the control were very low, and N fertilization increased the emissions of NO and N₂O. NO and N₂O from N fertilization treatments showed different emission patterns: significant NO emissions but low N₂O emissions in the winter season, and low NO emissions but significant N₂O emissions during the short period of barley growth in the spring season. The controlled release of the N fertilizer decreased the total NO emissions, while a deep application increased the total N₂O emissions. Fertilizer-derived NO-N and N₂O-N from the treatments CD, UD and US accounted for 0.20±0.07%, 0.71±0.15%, 0.62±0.04%, and 0.52±0.04%, 0.50±0.09%, 0.35±0.03%, of the applied N, respectively, during the barley season. CH₄ fluxes from the control were negative on most sampling dates, and its net soil uptake was 33±7.1 mg m⁻² during the barley season. The application of the N fertilizer decreased the uptake of atmospheric CH₄ and resulted in positive emissions from the soil. CO₂ fluxes were very low in the early period of crop growth while higher emissions were observed in the spring season. The N fertilization generally increased the direct CO₂ emissions from the soil. N₂O, CH₄ and CO₂ fluxes were positively correlated (P<0.01) with each other, whereas NO and CO₂ fluxes were negatively correlated (P<0.05). The N fertilization increased soil-derived global warming potential (GWP) significantly in the barley season. The net GWP was calculated by subtracting the plant-fixed atmospheric CO₂ stored in its aboveground parts from the soil-derived GWP in CO₂ equivalent. The net GWP from the CD, UD, US and the control were all negative at −243±30.7, −257±28.4, −227±6.6 and −143±9.7 g C m⁻² in CO₂ equivalent, respectively, in the barley season.     

Interannual and interseasonal soil CO2 efflux and VOC exchange rates in a Mediterranean holm oak forest in response to experimental drought

Climate models predict drier conditions in the next decades in the Mediterranean basin. Given the importance of soil CO₂ efflux in the global carbon balance and the important role of soil monoterpene and volatile organic compounds (VOCs) in soil ecology, we aimed to study the effects of the predicted drought on soil CO₂, monoterpenes and other VOC exchange rates and their seasonal and interannual variations. We decreased soil water availability in a Mediterranean holm oak forest soil by means of an experimental drought system performed since 1999 to the present. Measurements of soil gas exchange were carried out with IRGA, GC and PTR-MS techniques during two annual campaigns of contrasting precipitation. Soil respiration was twice higher the wet year than the dry year (2.27±0.26 and 1.05±0.15, respectively), and varied seasonally from 3.76±0.85 μmol m⁻² s⁻¹ in spring, to 0.13±0.01 μmol m⁻² s⁻¹ in summer. These results highlight the strong interannual and interseasonal variation in CO₂ efflux in Mediterranean ecosystems. The drought treatment produced a significant soil respiration reduction in drought plots in the wet sampling period. This reduction was even higher in wet springs (43% average reduction). These results show (1) that soil moisture is the main factor driving seasonal and interannual variations in soil respiration and (2) that the response of soil respiration to increased temperature is constrained by soil moisture. The results also show an additional control of soil CO₂ efflux by physiology and phenology of trees and animals. Soil monoterpene exchange rates ranged from −0.01 to 0.004 nmol m⁻² s⁻¹, thus the contribution of this Mediterranean holm oak forest soil to the total monoterpenes atmospheric budget seems to be very low. Responses of individual monoterpenes and VOCs to the drought treatment were different depending on the compound. This suggests that the effect of soil moisture reduction in the monoterpenes and VOC exchange rates seems to be dependent on monoterpene and VOC type. In general, soil monoterpene and other VOC exchange rates were not correlated with soil CO₂ efflux. In all cases, only a low proportion of variance was explained by the soil moisture changes, since almost all VOCs increased their emission rates in summer 2005, probably due to the effect of high soil temperature. Results indicate thus that physical and biological processes in soil are controlling soil VOC exchange but further research is needed on how these factors interact to produce the observed VOCs exchange responses.     

Elevation of atmospheric CO2 and N-nutritional status modify nodulation, nodule-carbon supply, and root exudation of Phaseolus vulgaris L.

Increased root exudation and a related stimulation of rhizosphere-microbial growth have been hypothesised as possible explanations for a lower nitrogen- (N-) nutritional status of plants grown under elevated atmospheric CO₂ concentrations, due to enhanced plant-microbial N competition in the rhizosphere. Leguminous plants may be able to counterbalance the enhanced N requirement by increased symbiotic N₂ fixation. Only limited information is available about the factors determining the stimulation of symbiotic N₂ fixation in response to elevated CO₂.In this study, short-term effects of elevated CO₂ on quality and quantity of root exudation, and on carbon supply to the nodules were assessed in Phaseolus vulgaris, grown in soil culture with limited (30 mg N kg⁻¹ soil) and sufficient N supply (200 mg N kg⁻¹ soil), at ambient (400 μmol mol⁻¹) and elevated (800 μmol mol⁻¹) atmospheric CO₂ concentrations.Elevated CO₂ reduced N tissue concentrations in both N treatments, accelerated the expression of N deficiency symptoms in the N-limited variant, but did not affect plant biomass production. ¹⁴CO₂ pulse-chase labelling revealed no indication for a general increase in root exudation with subsequent stimulation of rhizosphere microbial growth, resulting in increased N-competition in the rhizosphere at elevated CO₂. However, a CO₂-induced stimulation in root exudation of sugars and malate as a chemo-attractant for rhizobia was detected in 0.5-1.5 cm apical root zones as potential infection sites. Particularly in nodules, elevated CO₂ increased the accumulation of malate as a major carbon source for the microsymbiont and of malonate with essential functions for nodule development. Nodule number, biomass and the proportion of leghaemoglobin-producing nodules were also enhanced. The release of nod-gene-inducing flavonoids (genistein, daidzein and coumestrol) was stimulated under elevated CO₂, independent of the N supply, and was already detectable at early stages of seedling development at 6 days after sowing.     

Controls over pathways of carbon efflux from soils along climate and black spruce productivity gradients in interior Alaska

Small changes in C cycling in boreal forests can change the sign of their C balance, so it is important to gain an understanding of the factors controlling small exports like water-soluble organic carbon (WSOC) fluxes from the soils in these systems. To examine this, we estimated WSOC fluxes based on measured concentrations along four replicate gradients in upland black spruce (Picea mariana [Mill.] BSP) productivity and soil temperature in interior Alaska and compared them to concurrent rates of soil CO₂ efflux. Concentrations of WSOC in organic and mineral horizons ranged from 4.9 to 22.7 g C m⁻² and from 1.4 to 8.4 g C m⁻², respectively. Annual WSOC fluxes (4.5-12.0 g C m⁻² y⁻¹) increased with annual soil CO₂ effluxes (365-739 g C m⁻² y⁻¹) across all sites (R²=0.55, p=0.02), with higher fluxes occurring in warmer, more productive stands. Although annual WSOC flux was relatively small compared to total soil CO₂ efflux across all sites (<3%), its relative contribution was highest in warmer, more productive stands which harbored less soil organic carbon. The proportions of relatively bioavailable organic fractions (hydrophilic organic matter and low molecular weight acids) were highest in WSOC in colder, low-productivity stands whereas the more degraded products of microbial activity (fulvic acids) were highest in warmer, more productive stands. These data suggest that WSOC mineralization may be a mechanism for increased soil C loss if the climate warms and therefore should be accounted for in order to accurately determine the sensitivity of boreal soil organic C balance to climate change.     

Earthworm activity as a determinant for N2O emission from crop residue

Earthworm activity may have an effect on nitrous oxide (N₂O) emissions from crop residue. However, the importance of this effect and its main controlling variables are largely unknown. The main objective of this study was to determine under which conditions and to what extent earthworm activity impacts N₂O emissions from grass residue. For this purpose we initiated a 90-day (experiment I) and a 50-day (experiment II) laboratory mesocosm experiment using a Typic Fluvaquent pasture soil with silt loam texture. In all treatments, residue was applied, and emissions of N₂O and carbon dioxide (CO₂) were measured. In experiment I the residue was applied on top of the soil surface and we tested (a) the effects of the anecic earthworm species Aporrectodea longa (Ude) vs. the epigeic species Lumbricus rubellus (Hoffmeister) and (b) interactions between earthworm activity and bulk density (1.06 vs. 1.61 g cm⁻³). In experiment II we tested the effect of L. rubellus after residue was artificially incorporated in the soil. In experiment I, N₂O emissions in the presence of earthworms significantly increased from 55.7 to 789.1 μg N₂O-N kg⁻¹ soil (L. rubellus; p<0.001) or to 227.2 μg N₂O-N kg⁻¹ soil (A. longa; p<0.05). This effect was not dependent on bulk density. However, if the residue was incorporated into the soil (experiment II) the earthworm effect disappeared and emissions were higher (1064.2 μg N₂O-N kg⁻¹ soil). At the end of the experiment and after removal of earthworms, a drying/wetting and freezing/thawing cycle resulted in significantly higher emissions of N₂O and CO₂ from soil with prior presence of L. rubellus. Soil with prior presence of L. rubellus also had higher potential denitrification. We conclude that the main effect of earthworm activity on N₂O emissions is through mixing residue into the soil, switching residue decomposition from an aerobic and low denitrification pathway to one with significant denitrification and N₂O production. Furthermore, A. longa activity resulted in more stable soil organic matter than L. rubellus.     

Influence of earthworm activity on aggregate-associated carbon and nitrogen dynamics differs with agroecosystem management

Earthworms are known to be important regulators of soil structure and soil organic matter (SOM) dynamics, however, quantifying their influence on carbon (C) and nitrogen (N) stabilization in agroecosystems remains a pertinent task. We manipulated population densities of the earthworm Aporrectodea rosea in three maize-tomato cropping systems [conventional (i.e., mineral fertilizer), organic (i.e., composted manure and legume cover crop), and an intermediate low-input system (i.e., alternating years of legume cover crop and mineral fertilizer)] to examine their influence on C and N incorporation into soil aggregates. Two treatments, no-earthworm versus the addition of five A. rosea adults, were established in paired microcosms using electro-shocking. A ¹³C and ¹⁵N labeled cover crop was incorporated into the soil of the organic and low-input systems, while ¹⁵N mineral fertilizer was applied in the conventional system. Soil samples were collected during the growing season and wet-sieved to obtain three aggregate size classes: macroaggregates (>250 μm), microaggregates (53-250 μm) and silt and clay fraction (<53 μm). Macroaggregates were further separated into coarse particulate organic matter (cPOM), microaggregates and the silt and clay fraction. Total C, ¹³C, total N and ¹⁵N were measured for all fractions and the bulk soil. Significant earthworm influences were restricted to the low-input and conventional systems on the final sampling date. In the low-input system, earthworms increased the incorporation of new C into microaggregates within macroaggregates by 35% (2.8 g m⁻² increase; P=0.03), compared to the no-earthworm treatment. Within this same cropping system, earthworms increased new N in the cPOM and the silt and clay fractions within macroaggregates, by 49% (0.21 g m⁻²; P<0.01) and 38% (0.19 g m⁻²; P=0.02), respectively. In the conventional system, earthworms appeared to decrease the incorporation of new N into free microaggregates and macroaggregates by 49% (1.38 g m⁻²; P=0.04) and 41% (0.51 g m⁻²; P=0.057), respectively. These results indicate that earthworms can play an important role in C and N dynamics and that agroecosystem management greatly influences the magnitude and direction of their effect.     

Facilitation of pentachlorophenol degradation in the rhizosphere of ryegrass (Lolium perenne L.)

The phytoremediation of xenobiotics depends upon plant-microbe interactions in the rhizosphere, but the extent and intensity of these effects are currently unknown. To investigate rhizosphere effects on the biodegradation of xenobiotics, a glasshouse experiment was conducted using a specially designed rhizobox where ryegrass seedlings were grown for 53 days in a soil spiked with pentachlorophenol (PCP) at concentrations of 8.7±0.5 and 18±0.5 mg kg⁻¹ soil. The soil in the rhizobox was divided into six separate compartments at various distances from the root surface. Changes in PCP concentrations with increasing distance from the root compartment of the rhizobox were then assessed. The largest and most rapid loss of PCP in planted soil was at 3 mm from the root zone where total PCP decreased to 0.20 and 0.65 mg kg⁻¹, respectively with the two PCP treatments. The degradation gradient followed the order: near-rhizosphere>root compartment>far-rhizosphere soil zones for both concentrations where ryegrass was grown. In contrast, there was no difference in PCP concentration with distance in the unplanted soil. The increases in both soil microbial biomass carbon and the activities of soil urease and phosphatase were accompanied by the enhanced degradation of PCP, which was higher in the near-rhizosphere than far-rhizosphere soil. The results suggest that the effect of root proximity is important in the degradation of xenobiotics such as PCP in soil.     

Performance of para-nitrophenyl phosphate and 4-methylumbelliferyl phosphate as substrate analogues for phosphomonoesterase in soils with different organic matter content

Phosphomonoesterase (PMEase) activity plays a key role in nutrient cycling and is a potential indicator of soil condition and ecosystem stress. We compared para-nitrophenyl phosphate (pNPP) and 4-methylumbelliferyl phosphate (MUP) as substrate analogues for PMEase in 7 natural ecosystem soils and 8 agricultural top soils with contrasting C contents (8.0-414 g kg⁻¹ C) and pH (3.0-7.5). PMEase activities obtained with pNPP (0.05-5 μmol g⁻¹ h⁻¹) were significantly less than activities obtained with MUP (0.9-13 μmol g⁻¹ h⁻¹), especially in soils with a high organic matter content (>130 g kg⁻¹). Only PMEase activities assayed with MUP correlated significantly with total C and total N (r=0.7, P<0.01 all), and pH (r=−0.71, P<0.01). PMEase activities obtained with the two substrate analogues were correlated when expressed on a C-content basis (r=0.8, P<0.001), but not when expressed on an oven-dry soil weight basis. This indicated that interference by organic matter is related to the quantity rather than to the quality of organic matter. Overall, assaying with MUP was more sensitive compared to assaying with pNPP, particularly in the case of high organic and acid soils.