Comparison of soil fungal/bacterial ratios in a pH gradient using physiological and PLFA-based techniques

We have compared the total microbial biomass and the fungal/bacterial ratio estimated using substrate-induced respiration (SIR) in combination with the selective inhibition technique and using the phospholipid fatty acid (PLFA) technique in a pH gradient (3.0-7.2) consisting of 53 mature broad-leaved forest soils. A fungal/bacterial biomass index using the PLFA technique was calculated using the PLFA 18:2ω6,9 as an indicator of fungal biomass and the sum of 13 bacterial specific PLFAs as indicator of the bacterial biomass. Good linear correlation (p<0.001) was found between the total microbial biomass estimated with SIR and total PLFAs (totPLFA), indicating that 1 mg biomass-C was equivalent to 130 nmol totPLFA. Both biomass estimates were positively correlated to soil pH. The fungal/bacterial ratio measured using the selective inhibition technique decreased significantly with increasing pH from about 9 at pH 3 to approximately 2 at pH 7, while the fungal/bacterial biomass index using PLFA measurements tended to increase slightly with increasing soil pH. Good correlation between the soil content of ergosterol and of the PLFA 18:2ω6,9 indicated that the lack of congruency between the two methods in estimating fungal/bacterial ratios was not due to PLFA 18:2ω6,9-related non-fungal structures to any significant degree. Several PLFAs were strongly correlated to soil pH (R² values >0.8); for example the PLFAs 16:1ω5 and 16:1ω7c increased with increasing soil pH, while i16:0 and cy19:0 decreased. A principal component analysis of the total PLFA pattern gave a first component that was strongly correlated to soil pH (R²=0.85, p<0.001) indicating that the microbial community composition in these beech/beech-oak forest soils was to a large extent determined by soil pH.     

The microbial PLFA composition as affected by pH in an arable soil

The influence of soil pH on the phospholipid fatty acid (PLFA) composition of the microbial community was investigated along the Hoosfield acid strip, Rothamsted Research, UK - a uniform pH gradient between pH 8.3 and 4.5. The influence of soil pH on the total concentration of PLFAs was not significant, while biomass estimated using substrate induced respiration decreased by about 25%. However, the PLFA composition clearly changed along the soil pH gradient. About 40% of the variation in PLFA composition along the gradient was explained by a first principal component, and the sample scores were highly correlated to pH (R² = 0.97). Many PLFAs responded to pH similarly in the Hoosfield arable soil compared with previous assessments in forest soils, including, e.g. monounsaturated PLFAs 16:1ω5, 16:1ω7c and 18:1ω7, which increased in relative concentrations with pH, and i16:0 and cy19:0, both of which decreased with pH. Some PLFAs responded differently to pH between the soil types, e.g. br18:0. We conclude that soil pH has a profound influence on the microbial PLFA composition, which must be considered in all applications of this method to detect changes in the microbial community.     

Substrate inputs and pH as factors controlling microbial biomass, activity and community structure in an arable soil

Our aim was to determine whether the smaller biomasses generally found in low pH compared to high pH arable soils under similar management are due principally to the decreased inputs of substrate or whether some factor(s) associated with pH are also important. This was tested in a soil incubation experiment using wheat straw as substrate and soils of different pHs (8.09, 6.61, 4.65 and 4.17). Microbial biomass ninhydrin-N, and microbial community structure evaluated by phospholipid fatty acids (PLFAs), were measured at 0 (control soil only), 5, 25 and 50 days and CO₂ evolution up to 100 days. Straw addition increased biomass ninhydrin-N, CO₂ evolution and total PLFA concentrations at all soil pH values. The positive effect of straw addition on biomass ninhydrin-N was less in soils of pH 4.17 and 4.65. Similarly total PLFA concentrations were smallest at the lowest pH. This indicated that there is a direct pH effect as well as effects related to different substrate availabilities on microbial biomass and community structure. In the control soils, the fatty acids 16:1ω5, 16:1ω7c, 18:1ω7c&9t and i17:0 had significant and positive linear relationships with soil pH. In contrast, the fatty acids i15:0, a15:0, i16:0 and br17:0, 16:02OH, 18:2ω6,9, 17:0, 19:0, 17:0c9,10 and 19:0c9,10 were greatest in control soils at the lowest pHs. In soils given straw, the fatty acids 16:1ω5, 16:1ω7c, 15:0 and 18:0 had significant and positive linear relationships with pH, but the concentration of the monounsaturated 18:1ω9 PLFA decreased at the highest pHs. The PLFA profiles indicative of Gram-positive bacteria were more abundant than Gram-negative ones at the lowest pH in control soils, but in soils given straw these trends were reversed. In contrast, straw addition changed the microbial community structures least at pH 6.61. The ratio: [fungal PLFA 18:2w6,9]/[total PLFAs indicative of bacteria] indicated that fungal PLFAs were more dominant in the microbial communities of the lowest pH soil. In summary, this work shows that soil pH has marked effects on microbial biomass, community structure, and response to substrate addition.     

Soil microbial community dynamics and phenanthrene degradation as affected by rape oil application

The effects of rape oil application on soil microbial communities and phenanthrene degradation were characterized by examining phenanthrene concentrations, changes in microbial composition and incorporation of [¹³C] phenanthrene-derived carbon into phospholipid fatty acids (PLFAs). A Haplic Chernozem was incubated with and without rape oil in combination with and without phenanthrene over 60 days. High-performance liquid chromatography (HPLC) analysis showed a net reduction in extractable phenanthrene in the soils treated with rape oil but no net reduction in the soils without rape oil. Rape oil application increased the total PLFA content and changed microbial community composition predominantly due to growth of fungal groups and Gram-positive bacterial groups. Under rape oil and phenanthrene amendment all detected microbial groups grew until day 24 of incubation. The ¹³C PLFA profiles showed ¹³C enrichment for the PLFAs i14:0, 15:0, 18:0, 18:1ω5 and the fungal biomarker 18:2ω6,9 under rape oil application. Fungal PLFA growth was highest among detected all PLFAs, but its ¹³C incorporation was lower compared to the Gram-positive and Gram-negative bacteria PLFAs. Our results demonstrate the effect of rape oil application on the abundance of microbial groups in soil treated with phenanthrene and its impact on phenanthrene degradation.     

Incorporation of C-labeled rice-straw-derived carbon into microbial communities in submerged rice field soil and percolating water

Rice straw is a major organic material applied to rice fields. The microorganisms growing on rice-straw-derived carbon have not been well studied. Here, we applied ¹³C-labeled rice straw to submerged rice soil microcosms and analyzed phospholipid fatty acids (PLFAs) in the soil and percolating water to trace the assimilation of rice-straw-derived carbon into microorganisms. PLFAs in the soil and water were markedly enriched with ¹³C during the first 3 days of incubation, which indicated immediate incorporation of rice-straw-derived carbon into microbial biomass. The enrichment of PLFAs in the percolating water with ¹³C suggested that microorganisms other than the population colonizing rice straw also assimilated rice-straw-derived carbon or that some bacterial groups were selectively released from the straw. The microbial populations could be categorized into two communities based on the carbon isotope data of the PLFAs: those derived from rice straw and those derived from soil organic matter (SOM). The composition of the PLFAs from the two communities differed, which indicated the assimilation of rice-straw-derived carbon by a subset of microbial populations. The composition of rice-straw-derived PLFAs in the percolating water was also distinct from that in the soil.     

Incorporation of urea-derived 13C into microbial communities in four different agriculture soils

The objective of this study was to investigate changes in the composition of the soil microbial community brought about by urea application and differences in the incorporation of urea-derived C into the soil phospholipid fatty acid (PLFA) pool at differing soil pH. We selected four soils which ranged in pH from 3.9 to 7.8. ¹³C-labeled urea was applied at two concentrations 100 and 200 mg N kg^?1 which represents commonly used and high levels of application. Significant hydrolysis of applied urea occurred within 2 h; less than 2 % of urea-C was retained in the soil with one exception, the fluvo-aquic soil at pH 7.8 amended with 200 mg kg^?1 urea-N 3 days after urea application. According to principal component analysis (PCA), the effect of urea and incubation time on microbial community composition was far weaker than differences between the four soils due to their large differences in basic properties; the scores of PC2 were significantly correlated with pH values. The incorporation of ¹³C-urea to PLFAs increased with soil pH; this may be related to increases in the speciation of inorganic C into bicarbonate.¹³C label was primarily incorporated into 16:1ω5c, 16:0, and cy19:0 in red soil, pH 3.9; and into 16:1ω7c, 16:0, and 16:1ω5c in fluvo-aquic soil, pH 7.8. A wider range of PLFAs became labeled in the two paddy soils at pH 5.2 and 6.7. This suggests that the profile of PLFAs labeled from the application of ¹³C-urea may be affected by redox potential.     

Linking dynamics of soil microbial phospholipid fatty acids to carbon mineralization in a C natural abundance experiment: Impact of heavy metals and acid rain

A ¹³C natural abundance experiment including GC-c-IRMS analysis of phospholipid fatty acids (PLFAs) was conducted to assess the temporal dynamics of the soil microbial community and carbon incorporation during the mineralization of plant residues under the impact of heavy metals and acid rain. Maize straw was incorporated into (i) control soil, (ii) soil irrigated with acid rain, (iii) soil amended with heavy metal-polluted filter dust and (iv) soil with both, heavy metal and acid rain treatment, over a period of 74 weeks. The mineralization of maize straw carbon was significantly reduced by heavy metal impact. Reduced mineralization rate of the added carbon likely resulted from a reduction of the microbial biomass due to heavy metal stress, while the efficiency of ¹³C incorporation into microbial PLFAs was hardly affected. Since acid rain did not significantly change soil pH, little impact on soil microorganisms and mineralization rate was found. Temporal dynamics of labelling of microbial PLFAs were different between bacterial and fungal PLFA biomarkers. Utilization of maize straw by bacterial PLFAs peaked immediately after the application (2 weeks), while labelling of the fungal biomarker 18:2ω6,9 was most pronounced 5 weeks after the application. In general, ¹³C labelling of microbial PLFAs was closely linked to the amounts of maize carbon present in the soil. The distinct higher labelling of microbial PLFAs in the heavy metal-polluted soils 74 weeks after application indicated a large fraction of available maize straw carbon still present in the soil.     

Soil microbial communities and activities in sand dunes of subtropical coastal forests

To understand the soil microbial activities and community structures in different forests in a sand-dune ecosystem, we conducted a study of 2 topographic conditions, upland and lowland, under a Casuarina forest. As well, in the lowland site, we compared forest soil microbial properties under 3 coastal forests (Casuarina, Hibiscus and mixed stand). The soil microbial biomass did not significantly differ between the upland and lowland Casuarina forest sites. At the lowland site, the soil microbial biomass was higher in the Hibiscus than Casuarina forest soil. Cellulase, xylanase, phosphatase and urease activities did not show a consistent trend by topography or vegetation. Analysis of phospholipid fatty acids (PLFAs) of bacteria and actinomycetes revealed a significant difference in microbial community structure by both topography and vegetation. PLFA content was higher at upland than lowland sites in the Casuarina forest. At the lowland site, the level of PLFAs was higher in Hibiscus than Casuarina forest soil. In addition, we examined the ratios 16:1ω7t/16:1ω7c and, cy17:0/16:1ω7c as indicators of physiological stress; the soil in the Casuarina forest had the highest values, which suggests that the microbial community in the Casuarina forest soil is under physiological stress or starvation conditions. Comparison of soil microbial properties suggest that planting Hibiscus may help to enrich soil fertility and increase microbial activities in coastal sand-dune Casuarina forest.     

Soil microbial communities of no-till dryland agroecosystems across an evapotranspiration gradient

In degraded agricultural soils, organic C levels can be increased and conserved by adopting alternative management strategies such as no-tillage and increased cropping intensity. However, soil microbial community responses to increased soil organic C (SOC) may be constrained due to water limitations in semi-arid dryland agroecosystems. The purpose of this study was to assess SOC, microbial biomass C (MBC) and community ester-linked fatty acid methyl ester (EL-FAME) composition under winter wheat (Triticum aestivum L.) in no-till systems of wheat–corn (Zea mays L.)–fallow (WCF), wheat–wheat–corn–millet (Panicum miliaceum L.) (WWCM), wheat–corn–millet (WCM), opportunity cropping (OPP), and perennial grass across a potential evapotranspiration gradient in eastern Colorado. Rotations of WWCM and OPP, in which crops are chosen based on available soil water at the time of planting rather than according to a predetermined rotation schedule, increased levels of SOC to those measured under perennial grass. However, MBC under OPP cropping accounted for the smallest fraction (2.0–3.6%) of SOC compared to other systems, in which MBC ranged from 2.4 to 6.3% of SOC. Microbial community structure was most divergent between OPP-cropped and perennial grass soils, whereas few differences were observed among microbial communities of the WCF, WCM, and WWCM rotations. Compared to perennial grass and other cropping systems, microbial biomass in OPP-cropped soil was low and contained less of the arbuscular mycorrhizal fungal biomarker 16:1ω5c. Microbial stress, as indicated by the ratio of 17:0 cy to 16:1ω7c, was greatest under OPP and WCF cropping. In contrast, soils under perennial grass contained lower ratios of bacterial:fungal EL-FAMEs and higher levels of MBC, ratios of MBC:SOC, and relative abundances of 16:1ω5c. Across locations, SOC and moisture content increased as soil texture became finer, whereas trends in MBC and community structure followed the potential evapotranspiration gradient. Soil from the high potential evapotranspiration site contained the lowest level of MBC but greater relative amounts of 16:1ω5c and lower ratios of stress indicator and bacterial:fungal EL-FAMEs compared to soil located at the moderate and low potential evapotranspiration sites. Indistinct microbial communities under WCF, WCM, and WWCM could be explained by EL-FAME limitations to detecting slight differences in microbial community structure or to the overwhelming response of microbial communities to environmental rather than management conditions. Further research is needed to assess potential legacy effects of long-term agricultural management that may mask microbial responses to recent management change, as well as to identify conditions that lead to high microbial community resiliency in response to management so that communities are similar under a given crop despite different preceding crops.     

Distinct microbial and faunal communities and translocated carbon in Lumbricus terrestris drilospheres

Lumbricus terrestris is a deep-burrowing anecic earthworm that builds permanent, vertical burrows with linings (e.g., drilosphere) that are stable and long-lived microhabitats for bacteria, fungi, micro- and mesofauna. We conducted the first non-culture based field study to assess simultaneously the drilosphere (here sampled as 0–2 mm burrow lining) composition of microbial and micro/mesofaunal communities relative to bulk soil. Our study also included a treatment of surface-applied ¹³C- and ¹⁵N-labeled plant residue to trace the short-term (40 d) translocation of residue C and N into the drilosphere, and potentially the assimilation of residue C into drilosphere microbial phospholipid fatty acids (PLFAs). Total C concentration was 23%, microbial PLFA biomass was 58%, and PLFAs associated with protozoa, nematodes, Collembola and other fauna were 200-to-300% greater in the drilosphere than in nearby bulk soil. Principal components analysis of community PLFAs revealed that distributions of Gram-negative bacteria and actinomycetes and other Gram-positive bacteria were highly variable among drilosphere samples, and that drilosphere communities were distinct from bulk soil communities due to the atypical distribution of PLFA biomarkers for micro- and mesofauna. The degree of microbial PLFA ¹³C enrichment in drilosphere soils receiving ¹³C-labeled residue was highly variable, and only one PLFA, 18:1ω9c, was significantly enriched. In contrast, 11 PLFAs from diverse microbial groups where enriched in response to residue amendment in bulk soil 0–5 cm deep. Among control soils, however, a significant δ¹³C shift between drilosphere and bulk soil at the same depth (5–15 cm) revealed the importance of L. terrestris for translocating perennial ryegrass-derived C into the soil at depth, where we estimated the contribution of the recent grass C (8 years) to be at least 26% of the drilosphere soil C. We conclude that L. terrestris facilitates the translocation of plant C into soil at depth and promotes the maintenance of distinct soil microbial and faunal communities that are unlike those found in the bulk soil.     

High clay content accelerates the decomposition of fresh organic matter in artificial soils

Clay is generally considered an important stabiliser that reduces the rate of decomposition of organic matter (OM) in soils. However, several recent studies have shown trends contradicting this widely held view, emphasising our poor understanding of the mechanisms underlying the clay effects on OM decomposition. Here, an incubation experiment was conducted using artificial soils differing in clay content (0, 5, and 50%) at different temperatures (5, 15, and 25 °C) to determine the effects of clay content, temperature and their interaction on fresh OM decomposition. CO₂ efflux was measured throughout the experiment. Phospholipid fatty acids (PLFAs), enzyme activities, microbial biomass carbon (MBC), and dissolved organic carbon (DOC) were also measured at the end of the pre-incubation and incubation periods in order to follow changes in microbial community structure, functioning, and substrate availability. The results showed that higher clay contents promoted OM decomposition probably by increasing substrate availability and by sustaining a greater microbial biomass, albeit with a different community structure and with higher activities of most of the extracellular enzymes assayed. Higher clay content induced increases in the PLFA contents of all bacterial functional groups relative to fungal PLFA content. However, clay content did not change the temperature sensitivity (Q₁₀) of OM decomposition. The higher substrate availability in the high clay artificial soils sustained more soil microbial biomass, resulting in a different community structure and different functioning. The higher microbial biomass, as well as the changed community structure and functions, accelerated OM decomposition. From these observations, an alternative pathway to understanding the effects of clay on OM decomposition is proposed, in which clay may not only accelerate the decomposition of organic materials in soils but also facilitate the SOM accumulation as microbial products in the long term. Our results highlight the importance of clay content as a control over OM decomposition and greater attention is required to elucidate the underlying mechanisms.     

Microbial community composition and rhizodeposit-carbon assimilation in differently managed temperate grassland soils

Rhizodeposit-carbon provides a major energy source for microbial growth in the rhizosphere of grassland soils. However, little is known about the microbial communities that mediate the rhizosphere carbon dynamics, especially how their activity is influenced by changes in soil management. We combined a ¹³CO₂ pulse-labeling experiment with phospholipid fatty acid (PLFA) analysis in differently managed Belgian grasslands to identify the active rhizodeposit-C assimilating microbial communities in these grasslands and to evaluate their response to management practices. Experimental treatments consisted of three mineral N fertilization levels (0, 225 and 450 kg N ha⁻¹ y⁻¹) and two mowing frequencies (3 and 5 times y⁻¹). Phospholipid fatty acids were extracted from surface (0-5 cm) bulk (BU) and root-adhering (RA) soil samples prior to and 24 h after pulse-labeling and were analyzed by gas chromatography-combustion-isotope ratio mass spectrometry (GC-c-IRMS). Soil habitats significantly differed in microbial community structure (as revealed by multivariate analysis of mol% biomarker PLFAs) as well as in gram-positive bacterial rhizodeposit-C uptake (as revealed by greater ¹³C-PLFA enrichment following pulse-labeling in RA compared to BU soil in the 450N/5M treatment). Mowing frequency did not significantly alter the relative abundance (mol%) or activity (¹³C enrichment) of microbial communities. In the non-fertilized treatment, the greatest ¹³C enrichment was seen in all fungal biomarker PLFAs (C16:1ω5, C18:1ω9, C18:2ω6,9 and C18:3ω3,6,9), which demonstrates a prominent contribution of fungi in the processing of new photosynthate-C in non-fertilized grassland soils. In all treatments, the lowest ¹³C enrichment was found in gram-positive bacterial and actinomycetes biomarker PLFAs. Fungal biomarker PLFAs had significantly lower ¹³C enrichment in the fertilized compared to non-fertilized treatments in BU soil (C16:1ω5, C18:1ω9) as well as RA soil (all fungal biomarkers). While these observations clearly indicated a negative effect of N fertilization on fungal assimilation of plant-derived C, the effect of N fertilization on fungal abundance could only be detected for the arbuscular mycorrhizal fungal (AMF) PLFA (C16:1ω5). On the other hand, increases in the relative abundance of gram-positive bacterial PLFAs with N fertilization were found without concomitant increases in ¹³C enrichment following pulse-labeling. We conclude that in situ¹³C pulse-labeling of PLFAs is an effective tool to detect functional changes of those microbial communities that are dominantly involved in the immediate processing of new rhizosphere-C.     

温度和秸秆质量调节与秸秆分解相关的微生物磷脂脂肪酸（PLFA）组成

Variations in temperature and moisture play an important role in soil organic matter (SOM) decomposition. However, relationships between changes in microbial community composition induced by increasing temperature and SOM decomposition are still unclear. The present study was conducted to investigate the effects of temperature and moisture levels on soil respiration and microbial communities involved in straw decomposition and elucidate the impact of microbial communities on straw mass loss. A 120-d litterbag experiment was conducted using wheat and maize straw at three levels of soil moisture (40%, 70%, and 90% of water-holding capacity) and temperature (15, 25, and 35°C). The microbial communities were then assessed by phospholipid fatty acid (PLFA) analysis. With the exception of fungal PLFAs in maize straw at day 120, the PLFAs indicative of Gram-negative bacteria and fungi decreased with increasing temperatures. Temperature and straw C/N ratio significantly affected the microbial PLFA composition at the early stage, while soil microbial biomass carbon (C) had a stronger effect than straw C/N ratio at the later stage. Soil moisture levels exhibited no significant effect on microbial PLFA composition. Total PLFAs significantly influenced straw mass loss at the early stage of decomposition, but not at the later stage. In addition, the ratio of Gram-negative and Gram-positive bacterial PLFAs was negatively correlated with the straw mass loss. These results indicated that shifts in microbial PLFA composition induced by temperature, straw quality, and microbial C sources could lead to changes in straw decomposition.     

Rice rhizodeposition and its utilization by microbial groups depends on N fertilization

Rhizodeposits have received considerable attention, as they play an important role in the regulation of soil carbon (C) sequestration and global C cycling and represent an important C and energy source for soil microorganisms. However, the utilization of rhizodeposits by microbial groups, their role in the turnover of soil organic matter (SOM) pools in rice paddies, and the effects of nitrogen (N) fertilization on rhizodeposition are nearly unknown. Rice (Oryza sativa L.) plants were grown in soil at five N fertilization rates (0, 10, 20, 40, or 60 mg N kg^?1 soil) and continuously labeled in a ¹³CO₂ atmosphere for 18 days during tillering. The utilization of root-derived C by microbial groups was assessed by ¹³C incorporation into phospholipid fatty acids. Rice shoot and root biomass strongly increased with N fertilization. Rhizodeposition increased with N fertilization, whereas the total ¹³C incorporation into microorganisms, as indicated by the percentage of ¹³C recovered in microbial biomass, decreased. The contribution of root-derived ¹³C to SOM formation increased with root biomass. The ratio of ¹³C in soil pools (SOM and microbial biomass) to ¹³C in roots decreased with N fertilization showing less incorporation and faster turnover with N. The ¹³C incorporation into fungi (18:2ω6,9c and 18:1ω9c), arbuscular mycorrhizal fungi (16:1ω5c), and actinomycetes (10Me 16:0 and 10Me 18:0) increased with N fertilization, whereas the ¹³C incorporation into gram-positive (i14:0, i15:0, a15:0, i16:0, i17:0, and a17:0) and gram-negative (16:1ω7c, 18:1ω7c, cy17:0, and cy19:0) bacteria decreased with N fertilization. Thus, the uptake and microbial processing of root-derived C was affected by N availability in soil. Compared with the unfertilized soil, the contribution of rhizodeposits to SOM and microorganisms increased at low to intermediate N fertilization rates but decreased at the maximum N input. We conclude that belowground C allocation and rhizodeposition by rice, microbial utilization of rhizodeposited C, and its stabilization within SOM pools are strongly affected by N availability: N fertilization adequate to the plant demand increases C incorporation in all these polls, but excessive N fertilization has negative effects not only on environmental pollution but also on C sequestration in soil.     

Changes in soil microbial community structure by Allonychiurus kimi (Collembola) and their interactive effects on glyphosate degradation

Collembolans have been known to be involved in various soil ecosystem functions. However, the role of Collembola in organic contaminant degradation has not been sufficiently elucidated to assess its contribution. In this study, varying densities of Allonychiurus kimi (Lee, 1973) (0, 10, and 30 individuals per 30 g of soil) were introduced into glyphosate-contaminated soils (74.1 mg glyphosate kg⁻¹ soil). This study investigated changes in the microbial community and the residual glyphosate concentration in soils over incubation time to elucidate the effects of A. kimi on the glyphosate degradation through its influence on the microbial community. Furthermore, the investigation was conducted in soils collected in May and September 2018 to assess the contribution of A. kimi to glyphosate degradation in soils with varying microbial compositions and biomass. Autoclaved soil was used as a control to minimize the influence of indigenous soil microorganisms on glyphosate degradation. We hypothesize that as the initial density of A. kimi increases, the effects of A. kimi on the soil microbial community become pronounced, altering the degradation kinetics of glyphosate in the soil. The composition and biomass of the soil microorganisms were quantified using the phospholipid fatty acid (PLFA) method. Our study determined that the presence of A. kimi altered the microbial community structure by increasing the bacterial and total microbial, but not fungal, biomass. After seven days of treatment, the bacterial and total microbial biomass in the treatment with A. kimi were >2.0-fold and 1.5-fold greater, respectively, compared to those in the treatments without A. kimi. Specifically, the concentration of PLFA 18:1ω7c, i15:0, and 16:1ω7c was positively correlated with A. kimi density. The residual glyphosate concentration decreased exponentially over time as A. kimi density increased. At the end of the experiment, the remaining portions (%) of glyphosate in the May soil samples were 26.3, 20.1, and 6.2, with A. kimi densities of 0, 10, and 30 per vessel, respectively, and the portions in the September soil samples were 13.4, 12.7, and 2.2, respectively. The DT₅₀s (time required for 50 % degradation) decreased significantly with increasing A. kimi density, ranging from 6.8 to 10.1 days at an A. kimi density of 30 to 12.9–19.4 days without A. kimi. However, in the autoclaved soil, a similar effect was not apparent (i.e., DT₅₀s ranged from 23.3 to 27.4 days). Our study demonstrated that Collembola can enhance organic contaminant degradation in soils by altering the microbial community structure.     

Stability and composition of soil organic matter control respiration and soil enzyme activities

Relationships between soil organic matter (SOM) molecular composition, thermal stability and decomposability by soil enzymes and microbes are largely unknown. We incubated soils from unfertilized and NPK-fertilized neighboring field plots of a long-term rye (Secale cereale) monoculture experiment and investigated relationships between changes in the molecular-chemical composition of SOM, the CO₂ flux and the activities of enzymes. Pyrolysis-field ionization mass spectrometry (Py-FIMS) showed larger ion intensities in the NPK-fertilized than in the unfertilized soil at start of the incubation, only small changes in composition and thermal stability in the unfertilized soil, and a preferential reduction in thermally stable components as well as general shifts towards lower pyrolysis temperature after three weeks of incubation in the NPK-treatment. We found evidence that thermally labile and stable proportions of various compound classes were differently susceptible to decomposition, depending on the fertilization history of the soil. Irrespective of fertilization treatment, peaks in xylanase activity after 7 days of incubation followed by decreasing values were reflected by the ratio of xylan (m/z 114) to xylose (m/z 132) marker signals in the Py-FI mass spectra. Thus, the study proved that (1) SOM composition was changed due to long-term rye cropping without and with NPK-fertilization, (2) the modified SOM composition affected the decomposability and microbial parameters under optimized conditions and (3) the thermal properties of individual compound classes derived from Py-FI mass spectra can be sensitive predictors of microbial decomposition.     

Distribution of microbial biomass and phospholipid fatty acids in Podzol profiles under coniferous forest

Microbial‐derived phospholipid fatty acids (PLFAs) can be used to characterize the microbial communities in soil without the need to isolate individual fungi and bacteria. They have been used to assess microbial communities of humus layers under coniferous forest, but nothing is known of their distribution in the deeper soil. To investigate the vertical distribution we sampled nine Podzol profiles on a 100‐m‐long transect in a coniferous forest and analysed for their microbial biomass and PLFA pattern to a depth of 0.4 m. The transect covered a fertility gradient from Vaccinium vitis‐idaea forest site type to Vaccinium myrtillus forest site type. The cores were divided into humus (O) and eluvial (E) layers and below that into 10‐cm sections and designated as either illuvial (B) or parent material (C), or as a combination (BC). Two measures of microbial biomass analyses were applied: substrate‐induced respiration (SIR) to determine microbial biomass C (C_mic), and the sum of the extracted microbial‐derived phospholipid fatty acids (totPLFA). The soil fertility had no effect on the results. The C_mic correlated well with totPLFA (r= 0.86). The microbial biomass decreased with increasing depth. In addition the PLFA pattern changed with increased depth as assessed with principal component analysis, indicating a change in the microbial community structure. The composition of the PLFAs in the O layer differed from that in the E layer and both differed from the upper part of the B layer and from the rest of the BC layers. The deeper parts of the B layer (BC₁, BC₂ and BC₃) were similar to one other. The O layer had more 18:2ω6, a PLFA indicator of fungi, whereas the E layer contained relatively more of the PLFAs 16:1ω9, 18:1ω7 and cy19:0 common in gram‐negative bacteria. With increased depth the relative amount of 10Me18:0, the PLFA indicator for actinomycetes, increased. We conclude that the PLFA method is a promising discriminator between the microbial community structures of the horizons in Podzols.     

Soil respiration is not limited by reductions in microbial biomass during long-term soil incubations

Declining rates of soil respiration are reliably observed during long-term laboratory incubations. However, the cause of this decline is uncertain. We explored different controls on soil respiration to elucidate the drivers of respiration rate declines during long-term soil incubations. Following a long-term (707 day) incubation (30 °C) of soils from two sites (a cultivated and a forested plot at Kellogg Biological Station, Hickory Corners, MI, USA), soils were significantly depleted of both soil carbon and microbial biomass. To test the ability of these carbon- and biomass-depleted (“incubation-depleted”) soils to respire labile organic matter, we exposed soils to a second, 42 day incubation (30 °C) with and without an addition of plant residues. We controlled for soil carbon and microbial biomass depletion by incubating field fresh (“fresh”) soils with and without an amendment of wheat and corn residues. Although respiration was consistently higher in the fresh versus incubation-depleted soil (2 and 1.2 times higher in the fresh cultivated and fresh forested soil, respectively), the ability to respire substrate did not differ between the fresh and incubation-depleted soils. Further, at the completion of the 42 day incubation, levels of microbial biomass in the incubation-depleted soils remained unchanged, while levels of microbial biomass in the field-fresh soil declined to levels similar to that of the incubation-depleted soils. Extra-cellular enzyme pools in the incubation-depleted soils were sometimes slightly reduced and did not respond to addition of labile substrate and did not limit soil respiration. Our results support the idea that available soil organic matter, rather than a lack microbial biomass and extracellular enzymes, limits soil respiration over the course of long-term incubations. That decomposition of both wheat and corn straw residues did not change after major changes in the soil biomass during extended incubation supports the omission of biomass values from biogeochemical models.     

Incorporation of 13C-labelled rice rhizodeposition carbon into soil microbial communities under different water status

The overall processes by which carbon is fixed by plants in photosynthesis then released into the soil by rhizodeposition and subsequently utilized by soil micro-organisms, links the atmospheric and soil carbon pools. The objective of this study was to determine the plant derived ¹³C incorporated into the phospholipid fatty acid (PLFA) pattern in paddy soil, to test whether utilization of rice rhizodeposition carbon by soil micro-organisms is affected by soil water status. This is essential to understand the importance of flooded conditions in regulating soil microbial community structure and activity in wetland rice systems. Rice plants were grown in soil derived from a paddy system under controlled irrigation (CI), or with continuous waterlogging (CW). Most of the ¹³C-labelled rice rhizodeposition carbon was distributed into the PLFAs 16:0, 18:1ω7 and 18:1ω9 in both the CW and CI treatments. The bacterial PLFAs i15:0 and a15:0, both indicative of gram positive bacteria, were relatively more abundant in the treatments without rice plants. When rice plants were present rates of ¹³C-incorporation into i15:0 and a15:0 was slow; the microbes containing these PLFAs may derive most of their carbon from more recalcitrant C (soil organic matter). PLFAs, 18:1ω7 and 16:1ω7c, indicative of gram negative bacteria showed a greater amount incorporation of labelled plant derived carbon in the CW treatment. In contrast, 18:2ω6,9 indicative of fungi and 18:1ω9 indicative of aerobes but also potentially fungi and plant roots had greater incorporation in the CI treatment. The greater root mass concomitant with lower incorporation of ¹³C into the total PLFA pool in the CW treatment suggests that the microbial communities in wetland rice soil are limited by factors other than substrate availability in flooded conditions. In this study differing soil microbial communities were established through manipulating the water status of paddy soils. Steady state ¹³C labelling enabled us to determine that the microbial community utilizing plant derived carbon was also affected by water status.     

Changes in soil microbial community composition in response to fertilization of paddy soils in subtropical China

Repeated fertilizer applications to cultivated soils may alter the composition and activities of microbial communities in terrestrial agro-ecosystems. In this study, we investigated the effects of different long term fertilization practices (control (CK), three levels of mineral fertilizer (N₁P₁K₁, N₂P₂K₂, and N₃P₃K₃), and organic manure (OM)) on soil environmental variables and microbial communities by using phospholipid fatty acid (PLFA) biomarkers analysis in subtropical China. Study showed that OM treatment led to increases in soil organic carbon (SOC), total nitrogen (TN) and total phosphorus (TP) contents, while the mineral fertilizer treatment led to increases in dissolved organic carbon (DOC) content. Changes in soil microbial communities (eg. bacteria, actinomycetes) were more noticeable in soils subjected to organic manure applications than in the control soils or those treated with mineral fertilizer applications. Fungal PLFA biomarkers responded differently from the other PLFA groups, the numerical values of fungal PLFA biomarkers were similar for all the OM and mineral fertilizer treatments. PCA analysis showed that the relative abundance of most PLFA biomarkers increased in response to OM treatment, and that increased application rates of the mineral fertilizer changed the composition of one small fungal PLFA biomarker group (namely 18:3ω6c and 16:1ω5c). Further, from the range of soil environmental factors that we examined, SOC, TN and TP were the key determinants affecting soil microbial community. Our results suggest that organic manure should be recommended to improve soil microbial activity in subtropical agricultural ecosystems, while increasing mineral fertilizer applications alone will not increase microbial growth in paddy soils.