Response of microbial diversity to C:N:P stoichiometry in fine root and microbial biomass following afforestation

Soil samples were collected in June and October from areas with three land-use types, i.e., Robinia pseudoacacia L. (RP), Caragana korshinskii Kom. (CK), and abandoned land (AL), of which the former two were afforested areas, whereas the latter was not. These areas were converted from similar farmlands 40 years prior. Illumina sequencing of 16S rRNA gene and fungal ITS gene was used to analyze soil bacterial and fungal diversity. Additionally, plant communities, soil properties, fine root biomass, and C, N, and P levels in fine root and microbial biomass were estimated. Compared to AL, the C:N:P stoichiometry in fine root and microbial biomass in the afforested lands was synchronously changed, especially the N:P ratio. Soil microbial diversities were affected by afforestation and were more related to N:P ratio than C:P and C:N ratios. Moreover, Alpha-proteobacteria, Gamma-proteobacteria, and Bacteroidetes were significantly more abundant in afforested soils than in the AL soil, and the abundances of Actinobacteria, Chloroflexi, Cyanobacteria, and Nitrospirae ranked as AL > RP or CK. For fungal taxa, Ascomycota abundance responded positively to afforestation, whereas Basidiomycota abundance responded negatively. Changes of soil microbial taxa were significantly correlated with the N:P ratio in fine root and microbial biomass, which explained 54.1 and 55% of the total variation in bacterial and fungal taxa, respectively. Thus, our results provide evidence that compositions of soil microbial communities are linked to the N:P ratio in the plant-soil system.     

Taxon-specific responses of soil microbial communities to different soil priming effects induced by addition of plant residues and their biochars

Purpose

This work investigated changes in priming effects and the taxonomy of soil microbial communities after being amended with plant feedstock and its corresponding biochar.

Materials and methods

A soil incubation was conducted for 180 days to monitor the mineralization and evolution of soil-primed C after addition of maize and its biochar pyrolysed at 450 °C. Responses of individual microbial taxa were identified and compared using the next-generation sequencing method.

Results and discussion

Cumulative CO₂ showed similar trends but different magnitudes in soil supplied with feedstock and its biochar. Feedstock addition resulted in a positive priming effect of 1999 mg C kg^?1 soil (+253.7 %) while biochar gave negative primed C of ?872.1 mg C kg^?1 soil (?254.3 %). Linear relationships between mineralized material and mineralized soil C were detected. Most priming occurred in the first 15 days, indicating co-metabolism. Differences in priming may be explained by differences in properties of plant material, especially the water-extractable organic C. Predominant phyla were affiliated to Acidobacteria, Actinobacteria, Chloroflexi, Gemmatimonadetes, Firmicutes, Planctomycetes, Proteobacteria, Verrucomicrobia, Ascomycota, Basidiomycota, Blastocladiomycota, Chytridiomycota, Zygomycota, Euryarchaeota, and Thaumarchaeota during decomposition. Cluster analysis resulted in separate phylogenetic grouping of feedstock and biochar. Bacteria (Acidobacteria, Firmicutes, Gemmatimonadetes, Planctomycetes), fungi (Ascomycota), and archaea (Euryarchaeota) were closely correlated to primed soil C (R ²?=??0.98, ?0.99, 0.84, 0.81, 0.91, and 0.91, respectively).

Conclusions

Quality of plant materials (especially labile C) shifted microbial community (specific microbial taxa) responses, resulting in a distinctive priming intensity, giving a better understanding of the functional role of soil microbial community as an important driver of priming effect.

     

Changes in soil microbial substrate utilization in response to altered litter diversity and precipitation in a Mediterranean shrubland

This study aimed at quantifying the consequences of reduced precipitation and plant diversity on soil microbial community functioning in a Mediterranean shrubland of southern France. Across a natural gradient of shrub species diversity, we established a total of 92 plots (4 × 4 m) with and without a moderate rain exclusion treatment of about 12 % of total precipitation. Shrub diversity included all possible combinations of the four dominant species (Cistus albidus, Quercus coccifera, Rosmarinus officinalis, and Ulex parviflorus). Respective leaf litter mixtures of these species combinations were exposed in all plots over 2 years. We quantified how litter species richness and the reduction in precipitation affected the soil microbial substrate utilization (measured by CO₂ evolution using the MicroResp method) on soil samples collected underneath each individual litter mixture after 1 and 2 years of decomposition. Moderate precipitation reduction had a minor impact, but litter species richness and the dissimilarity in phenolic concentrations (estimated using Rao’s quadratic entropy) showed a positive effect on the diversity of substrates metabolized by the microbial communities. Moreover, litter species richness increased soil microbial activity by increasing the catabolic diversity of the soil microbial community. These effects were mostly driven by the presence of Quercus and Ulex leaf litter, which at the same time reduced microbial metabolic dominance, while the presence of Rosmarinus had opposite effects. Our data suggest that plant species loss can have stronger effects on the functioning of soil microbial communities than moderate drought, with potentially important feedbacks on biogeochemical cycling in Mediterranean shrubland ecosystems.     

Effects of vermicompost amendment as a basal fertilizer on soil properties and cucumber yield and quality under continuous cropping conditions in a greenhouse

Purpose

We examined the effects of vermicompost application as a basal fertilizer on the properties of a sandy loam soil used for growing cucumbers under continuous cropping conditions when compared to inorganic or organic fertilizers.

Materials and methods

A commercial cucumber (Cucumis sativus L.) variety was grown on sandy loam soil under four soil amendment conditions: inorganic compound fertilizer (750 kg/ha,), replacement of 150 kg/ha of inorganic compound fertilizer with 3000 kg/ha of organic fertilizer or vermicompost, and untreated control. Experiments were conducted in a greenhouse for 4 years, and continuous planting resulted in seven cucumber crops. The yield and quality of cucumber fruits, basic physical and chemical properties of soil, soil nutrient characteristics, and the soil fungal community structure were measured and evaluated.

Results and discussion

Continuous cucumber cropping decreased soil pH and increased electrical conductivity. However, application of vermicompost significantly improved several soil characteristics and induced a significant change in the rhizosphere soil fungal community compared to the other treatments. Notably, the vermicompost amendments resulted in an increase in the relative abundance of Ascomycota, Chytridiomycota, Sordariomycetes, Eurotiomycetes, and Saccharomycetes, and a decrease in Glomeromycota, Zygomycota, Dothideomycetes, Agaricomycetes, and Incertae sedis. Compared to the organic fertilizer treatment, vermicompost amendment increased the relative abundance of beneficial fungi and decreased those of pathogenic fungi. Cucumber fruit yield decreased yearly under continuous cropping conditions, but both inorganic and organic fertilizer amendments increased yields. Vermicompost amendment maintained higher fruit yield and quality under continuous cropping conditions.

Conclusions

Continuous cropping decreased cucumber yield in a greenhouse, but basic fertilizer amendment reduced this decline. Moreover, basal fertilizer amendment decreased beneficial and pathogenic fungi, and the use of vermicompost amendment in the basic fertilizer had a positive effect on the health of the soil fungal community.

     

Responses of soil microbial community to nitrogen fertilizer and precipitation regimes in a semi-arid steppe

Purpose

Changes of nitrogen (N) cycle caused by N fertilization and precipitation regimes have affected the key ecosystem structure and functions in temperate steppe, which may modify the structure of soil microbial communities involved in N transformation. This paper was designated to examine the response of soil ammonia oxidizers and denitrifiers to the N fertilization and precipitation regimes in a semi-arid steppe where N and water contents are major limiting factors of the grassland productivity.

Materials and methods

This study was based on a long-term N fertilization and precipitation regimes experiment in Inner Mongolia (116° 17′ 20″ E, 42° 2′ 29″ N). The treatments including CK (control), R (reduced precipitation), W (30% increase in precipitation), N (10 g N m^?2 y^?1), RN (reduced precipitation and 10 g N m^?2 y^?1), and WN (30% increase in precipitation and 10 g N m^?2 y^?1). Soil basic chemical properties and microbial activities were analyzed. Molecular methods were applied to determine the abundance, structure and diversity of ammonia oxidizers and denitrifiers. Statistical analysis detected the main and interactive effect of treatments on soil microbial communities and revealed the relationship between soil microbial community structures and environmental factors.

Results and discussion

N fertilization significantly increased ammonia-oxidizing bacteria (AOB) abundance. Ammonia-oxidizing archaea (AOA) community structure was markedly changed in N fertilizer treatment and strongly affected by soil pH, while soil nitrate and water content correlated with AOB community structure. Soil nitrate was the key factor influencing nirK gene community structure, while soil pH and water content explained much of the variations of nosZ gene community. AOB-amoA and nosZ gene community diversities were influenced by precipitation regimes and interaction of N fertilization and precipitation regimes, respectively.

Conclusions

N fertilization and precipitation regimes had significant influences on the changes of soil properties and microbial functional communities. Soil nitrification was mainly driven by AOB in the semi-arid grassland. Changes of substrate content and soil pH were the key factors in shifting functional microbial communities. The non-synergistic effects of N fertilization and precipitation regimes on the microbial functional groups indicated that the negative effect of lower pH induced by N fertilization would be alleviated by precipitation regimes, which should be well considered in grassland restoration.

     

Long-term and legacy effects of manure application on soil microbial community composition

We analyzed soil prokaryotic and fungal community composition in soils with varying histories of cattle manure application. The manure treatments were (i) annual application for 43 years (MF), (ii) annual application for 14 years followed by 29 years without application (MF14), and (iii) annual application for 30 years followed by 13 years without application (MF30). An annual application of chemical nitrogen (N) fertilizer (CNF) and a non-amended control (Con) were also included. Soil prokaryotic evenness and diversity significantly decreased in MF relative to other treatments in fall, but were similar to the other fertilizer treatments in spring and summer. Distinct prokaryotic and fungal community composition was observed in MF compared to other treatments across fall, spring, and summer seasons. The MF treatment significantly increased the relative abundance of Firmicutes, Gammaproteobacteria, and Gemmatimonadetes, but significantly decreased the relative abundance of Acidobacteria. In fall, the soil prokaryotic and fungal community composition with MF30 was significantly different than the other fertilization treatments. Overall, the study showed that annual manure application (MF) led to a different microbial community composition than the other fertilizer treatments. Soil without manure application for 13 years (MF30) had a significantly different microbial community composition from other fertilizer treatments in fall, while the soil without manure application for 29 years (MF14) resembled a microbial community that had never received manure.     

<Emphasis Type="Italic">Trichoderma</Emphasis> improves the growth of <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Bio-fertilizer application has been proposed as a strategy for enhancing soil fertility, regulating soil microflora composition, and improving crop yields, and it has been widely applied in the agricultural yields. However, the application of bio-fertilizer in grassland has been poorly studied. We conducted in situ and pot experiments to investigate the practical effects of different fertilization regimes on Leymus chinensis growth, with a focus on the potential microecological mechanisms underlying the responses of soil microbial composition. L. chinensis biomass was significantly (P?<?0.05) increased by treatment with 6000 kg ha^?1 of Trichoderma bio-fertilizer compared with other treatments. We found a positive (R² =?0.6274, P <?0.001) correlation between bacterial alpha diversity and L. chinensis biomass. Hierarchical cluster analysis and nonmetric multidimensional scaling (NMDS) revealed that soil bacterial and fungal community compositions were all separated according to the fertilization regime used. The relative abundance of the most beneficial genera in bio-fertilizer (BOF) (6000 kg ha^?1Trichoderma bio-fertilizer) was significantly higher than in organic fertilizer (OF) (6000 kg ha^?1 organic fertilizer) or in CK (non-amend fertilizer), there the potential pathogenic genera were reduced. There were significant negative (P?<?0.05) correlations between L. chinensis biomass and the relative abundance of several potential pathogenic genera. However, the relative abundance of most beneficial genera were significantly (P?<?0.05) positively correlated with L. chinensis biomass. Soil properties had different effects on these beneficial and on these pathogenic genera, further influencing L. chinensis biomass.     

Straw amendment to paddy soil stimulates denitrification but biochar amendment promotes anaerobic ammonia oxidation

Purpose

Organic matter amendment is usually used to improve soil physicochemical properties and to sequester carbon for counteracting climate change. There is no doubt that such amendment will change microbial activity and soil nitrogen transformation processes. However, the effects of straw and biochar amendment on anammox and denitrification activity and on community structure in paddy soil are unclear.

Materials and methods

We conducted a 30-day pot experiment using rice straw and rice straw biochar to deepen our understanding about the activity, microbial abundance, and community structure associated with soil nitrogen cycling during rice growth.

Results and discussion

Regarding activity, anammox contributed 3.1–8.1% of N₂ production and denitrification contributed 91.9–96.9% of N₂ production; straw amendment resulted in the highest denitrification rate (38.9 nmol N g^?1 h^?1), while biochar amendment resulted in the highest anammox rate (1.60 nmol N g^?1 h^?1). Both straw and biochar amendments significantly increased the hzsB and nosZ gene abundance (p < 0.05). Straw amendment showed the highest nosZ gene abundance, while biochar amendment showed the highest hzsB gene abundance. Phylogenetic analysis of the anammox bacteria 16S rRNA genes indicated that Candidatus Brocadia and Kuenenia were the dominant genera detected in all treatments.

Conclusions

Straw and biochar amendments have different influences on anaerobic ammonia oxidation and denitrification within paddy soil. Our results suggested that the changes in denitrification and anammox rates in the biochar and straw treatments were mainly linked to functional gene abundance rather than microbial community structure and that denitrification played the more major role in N₂ production in paddy soil.

     

Effects of cover crop in an apple orchard on microbial community composition,networks, and potential genes involved with degradation of crop residues in soil

Changes in the soil microbial communities and networks were monitored after planting the cover crop for 9 years. The field experiment included plots with a cover crop and without a cover crop but with weed control, and two subplots with or without chemical fertilizer (192 kg N ha^?1, 108 kg P₂O₅ ha^?1, and 168 kg K₂O ha^?1 each year). After applying the cover crop and chemical fertilizer for 9 years, the composition and activity of bacterial and fungal communities changed significantly (p?<?0.05), with the cover crop had greater effects than the chemical fertilizer on the composition of the soil microbial community. The relative abundances of 22 selected genera (in Firmicutes and Bacteroidetes) and two selected classes (Ascomycota) related to cover crop residue degradation increased significantly in the presence of the cover crop (p?<?0.05). Network analysis showed that the cover crop decreased the number of positive links between bacterial and fungal taxa by 25.33%, and increased the negative links by 22.89%. The positive links among bacterial taxa increased by 16.63% with the cover crop, mainly among Proteobacteria (increase of 39), Firmicutes (16), Actinobacteria (five), and Bacteroidetes (10). The links among fungal taxa were less than among bacterial taxa and were not significantly affected by cover crop. Taxa such as Thaumarchaeota, unidentified_Nitrospiraceae, unidentified_Nitrosomonadaceae, Faecalibacterium, Coprococcus_3, and Ruminococcaceae_NK4A214_group dominated the network without the cover crop but they were not dominant with the cover crop. The relative abundances of potential genes involved with the degradation of cellulose, hemicellulose, and cello-oligosaccharides increased significantly with the cover crop. Therefore, the SOC and TN contents were enhanced by the cover crop with the increase of the soil enzyme activities. Thus, the apple yield was improved by the cover crop.     

Long-term fertilisation regimes affect the composition of the alkaline phosphomonoesterase encoding microbial community of a vertisol and its derivative soil fractions

Alkaline phosphomonoesterase (ALP) mainly originates from soil microbial secretion and plays a crucial role in the turnover of soil phosphorus (P). To examine the response of ALP-encoding microbial communities (analysed for the biomarker of the ALP gene, phoD) of soils and derivative soil fractions to different fertilisation regimes, soil samples were collected from a long-term experimental field (over 35 years). The different organic P (Po) pools of soil fractions and the ALP activity of soil were also determined. Compared with chemical-only fertilised soils, the ALP activity was 232–815% higher in organic-amended soils, and the highest enzyme activity was observed in the organic-only fertilised treatment. The abundance of the phoD gene harbouring in soil fractions, determined by quantitative PCR (qPCR), was affected by different fertilisations. The highest abundance of the phoD gene was generally detected in the 2–63-μm-sized fraction (silt), but most phoD-encoding microbial species were associated to the 0.1–2-μm-sized fraction (clay) in the chemical-only fertilised soil. The contents of labile Po (LPo), moderately labile Po (MLPo) and fulvic acid-associated Po (FAPo) were significantly correlated with the phoD gene abundance, whereas only LPo content was significantly correlated with the ALP activity. The dominant phoD-encoding phylas were Actinobacteria and Proteobacteria, according to a high-throughput sequencing. Bradyrhizobium, a N₂-fixer identified as a phoD-encoding genus, showed the highest abundance in fertilised soils. The abundance of Bradyrhizobium, Streptomyces, Modestobacter, Lysobacter, Frankia and Burkholderia increased with the organic-only amendment and was significantly correlated with the ALP activity. According to structure equation models (SEM), pH and LPo content significantly and directly affected the ALP activity; the soil organic C (C_org) content was related to composition and abundances of phoD-harbouring microbial communities; since both microbial properties were correlated to the ALP activity, the C_org content was indirectly related to the ALP activity. In conclusion, soil management practices can be used to optimise the contents of soil available P and the organic P with regulation of soil ALP activity and the community composition of corresponding microbes.     

Recovery of Amoebae Community in the Root Soil of <Emphasis Type="Italic">M. sativa</Emphasis> after a Strong Contamination Pulse with n-Hexane

Microbial food webs tolerate toxic compounds depending on individualistic species resistance and their ability of using alternate food sources. Soil polluted with low-molecular weight volatile organics, such as hexane, diminishes bacterial and fungal communities despite its short residence time. Survival of microbial species depends on perturbation intensity, which in turn restricts resources for amoebae survival in polluted soil. Soil functional recovery from anthropogenic perturbations depends on microbial organic matter (OM) metabolization of pollutants. However, reconfiguration of amoebae community after soil exposure remains largely unknown. A microcosms study was carried out to determine the effects of hexane on the community structure of soil amoebae as well as the importance of Medicago sativa on amoebae community recovering. Hexane had a negative impact on species richness and structure of the amoebae community 24 h after pollution. There was a significant increase in species richness and number of amoebae 30 days after contamination. These two parameters further increased after 60 days from contamination. After 30 days of the initial trophozoites extinction caused by Hexane, M. sativa’s. Root zone showed a significant increase of both species richness and number of individuals. This recovery trend was kept after 60 days when the highest values in species richness and abundance of individuals were shown in both polluted and non-polluted microcosms. In conclusion, M. sativa’s root zone speeds up recovery of the amoebae community structure after pollution exposure.     

Geographic distance and amorphous iron affect the abundance and distribution of <Emphasis Type="Italic">Geobacteraceae</Emphasis> in paddy soils in China

Purpose

Geobacteraceae are important dissimilatory Fe (III)-reducing microorganisms, influencing the cycling of metals, nutrients as well as the degradation of organic contaminants. However, little is known about their distribution, diversity, and abundance of Geobacteraceae and the effects of environment factors and geographic distance on the distribution and diversity of Geobacteraceae in paddy soils remain unclear. Therefore, the objectives of this study were to investigate the distribution, diversity, and abundance of Geobacteraceae in paddy soils and to determine key factors in shaping the Geobacteraceae distribution, environmental factors, geographic distance, or both and to quantify their contribution to Geobacteraceae variation.

Materials and methods

Illumina sequencing and quantitative real-time PCR using a primer set targeting 16S rRNA genes of bacteria affiliated with the family Geobacteraceae were employed to measure the community composition, diversity, and abundance patterns of 16S rRNA genes of Geobacteraceae in 16 samples collected from north to south of China. MRT, Mantel test, and VPA were used to analyze the relationship between communities of Geobacteraceae and environmental factors and geographic distance.

Results and discussion

Quantitative PCR showed that the abundance of 16S rRNA genes of Geobacteraceae ranged from (1.20?±?0.18)?×?10⁸ to 1.13?×?10⁹?±?2.25?×?10⁸ copies per gram of soil (dry weight) across different types of soils. Illumina sequencing results showed Geobacter was the dominant genus within the family of Geobacteraceae. Multivariate regression tree (MRT) analysis showed that soil amorphous iron contributed more (22.46 %) to the variation of dominant species of Geobacteraceae than other examined soil chemical factors such as pH (14.52 %), ammonium (5.12 %), and dissolved organic carbon (4.74 %). Additionally, more geographically distant sites harbored less similar communities. Variance partitioning analysis (VPA) showed that geographic distance contributed more to the variation of Geobacteraceae than any other factor, although the environmental factors explained more variation when combined. So, we detected the uneven distribution of Geobacteraceae in paddy soils of China and demonstrated that Geobacteraceae community composition was strongly associated with geographic distance and soil chemical factors including aFe, pH, Fe, DOC, C:N, and NO₃ ^?-N. These results greatly expand the knowledge of the distribution of Geobacteraceae in environments, particularly in terrestrial ecosystems.

Conclusions

Our results showed that geographic distance and amorphous iron played important roles in shaping Geobacteraceae community composition and revealed that both geographic distance and soil properties governed Geobacteraceae biogeography in paddy soils. Our findings will be critical in facilitating the prediction of element cycling by incorporating information on functional microbial communities into current biogeochemical models.

     

Soil microbial abundance and community structure vary with altitude and season in the coniferous forests,China

Purpose

Soil microbes control the bioelement cycles and energy transformation in forest ecosystems, and are sensitive to environmental change. As yet, the effects of altitude and season on soil microbes remain unknown. A 560 m vertical transitional zone was selected along an altitude gradient from 3023, 3298 and 3582 m, to determine the potential effects of seasonal freeze-thaw on soil microbial community.

Materials and methods

Soil samples were collected from the three elevations in the growing season (GS), onset of freezing period (FP), deeply frozen period (FPD), thawing period (TP), and later thawing period (TPL), respectively. Real-time qPCR and polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) were used to measure the abundance and structure of soil microbial community.

Results and discussion

The bacterial, archaeal, and fungal ribosomal DNA (rDNA) copy numbers decreased from GS to freezing stage (FP and FPD) and then increased in thawing stage (TP and TPL). Similarly, the diversity of microbial community varied with seasonal freeze-thaw processes. The diversity index (H) of the bacterial and archaeal communities decreased from GS to FP and then increased to TPL. The fungal community H index increased in the freezing process.

Conclusions

Our results suggested that abundance and structure of soil microbial community in the Tibetan coniferous forests varied by season and bacterial and archaeal communities respond more promptly to seasonal freeze-thaw processes relative to fungal community. This may have important implications for carbon and nutrient cycles in alpine forest ecosystems. Accordingly, future warming-induced changes in seasonal freeze-thaw patterns would affect soil nutrient cycles via altering soil microbial properties.

     

The effects of silicon fertilizer on denitrification potential and associated genes abundance in paddy soil

This study evaluated the effect of silicate fertilizer on denitrification and associated gene abundance in a paddy soil. A consecutive trial from 2013 to 2015 was conducted including the following treatments: control (CK), mineral fertilizer (NPK), NPK plus sodium metasilicate (NPK + MSF), and NPK plus slag-based silicate fertilizer (NPK + SSF). Real-time quantitative PCR (qPCR) was used to analyze the abundances of nirS, nirK, and nosZ genes. Potential N₂O emissions and ammonium and nitrate concentrations were related to the nirS and nirK gene abundance. Compared with the NPK treatments, the addition of a Si fertilizer decreased N₂O emission rates and denitrification potential by 32.4–66.6 and 22.0–59.2%, respectively, which were probably related to increased rice productivity, soil Fe availability, and soil N depletion. The abundances of nirS and nirK genes were decreased by 17.7–35.8% and 21.1–43.5% with addition of silicate fertilizers, respectively. Rates of total N₂O and N₂O from denitrification (DeN₂O) emission were positively correlated with the nirS and nirK gene abundance. Nitrate, exchangeable NH₄ ⁺, and Fe concentrations were the main factors regulating the nirS and nirK gene abundance. Silicate fertilization during rice growth may serve as an effective approach to decreasing N₂O emissions.     

Enrichment of functional microbes and genes during pyrene degradation in two different soils

Purpose

Stimulating microbial degradation is a promising strategy for the remediation of soils contaminated with polycyclic aromatic hydrocarbons (PAHs). To better understand the functional microbial populations and processes involved in pyrene biodegradation in situ, the dynamics of pyrene degradation and functional microbial abundance were monitored during pyrene incubation in soils. We hope our findings will provide new insights into in situ pyrene biodegradation in soils and help to identify functional microbes from soils.

Materials and methods

Pyrene (60 mg kg^?1) was incubated with two different soils, one is lower PAH-containing agricultural soil (LS), and the other is higher PAH-containing industrial soil (HS). During incubation, triplicate samples were collected on days 0, 3, 7, 14, and 35. Pyrene in soil samples was analyzed using an Agilent gas chromatograph (7890A) equipped with a mass-selective detector (model 5897). DNA in soils was extracted with a FastDNA Spin kit for soil (Bio101, USA). The abundance of functional microbes and genes was monitored by a Taqman or SYBR Green based real-time PCR quantification using an iCycler iQ5 themocycler (Bio-Rad, USA). The diversity of PAH-RHDα GP genes was evaluated by constructing clone libraries and sequencing.

Results and discussion

In both soils, more than 80 % of the added pyrene was degraded within 35 days. After 35-day incubation, there was a significant enrichment of Gram-positive bacteria harboring PAH-ring hydroxylation dioxygenase (PAH-RHD_α GP) genes, and the abundance of Mycobacterium increased significantly. In PAH-RHD_α GP clone libraries from two soils, Mycobacterium was detected, while most sequences were closely related to uncultured Gram-positive bacteria. In addition, two pyrene catabolic pathways might be involved in pyrene degradation, as pyrene dioxygenase genes, nidA and nidA3, were dramatically enriched during incubation. Moreover, the abundance and diversity of potential degraders in two soils showed significantly difference in responding to pyrene stress. This result indicates that soil condition can significantly affect functional microbial populations and biological process for pyrene biodegradation.

Conclusions

These results revealed that Mycobacterium as well as uncultured Gram-positive PAH-RHD_α genotypes may be the important group of pyrene degraders in soils, and two pyrene catabolic pathways, targeted by nidA and nidA3, might potentially contribute to in situ biodegradation of pyrene. This study characterized the response pattern of potential pyrene degraders to pyrene stress in two different soils, which would increase our understanding of the indigenous processes of pyrene biodegradation in soil environment.

     

Effects of biochar on copper immobilization and soil microbial communities in a metal-contaminated soil

Purpose

Copper (Cu) contamination has been increasing in land ecosystems. Biochars (BCs) and arbuscular mycorrhizal fungi (AMF) are known to bind metals, and metallophyte can remove metals from soils. Will BC in combination with AMF contain the Cu uptake by a metallophyte growing in a metal-contaminated soil? The objective of this study was to investigate the effects of BCs on the Cu immobilization and over soil microbial communities in a metal-contaminated soil in the presence of AMF and metallophyte.

Materials and methods

Two BCs were produced from chicken manure (CMB) and oat hull (OHB). A Cu-contaminated sandy soil (338 mg kg^?1) was incubated with CMB and OHB (0, 1, and 5 % w/w) for 2 weeks. Metallophyte Oenothera picensis was grown in pots (500 mL) containing the incubated soils in a controlled greenhouse for 6 months. A number of analyses were conducted after the harvest. These include plant biomass weight, microbial basal respiration, and dehydrogenase activity (DHA), AMF root colonization, spore number, and glomalin production; changes in fungal and bacterial communities, Cu fractions in soil phases, and Cu uptake in plant tissues.

Results and discussion

The BCs increased the soil pH, decreased easily exchangeable fraction of Cu, and increased organic matter and residual fraction of Cu. The BCs provided favorable habitat for microorganisms, thereby increasing basal respiration. The CMB increased DHA by ～62 and ～574 %, respectively, for the low and high doses. Similarly, the OHB increased soil microbial activity by ～68 and ～72 %, respectively, for the low and high doses. AMF root colonization, spore number, and total glomalin-related soil protein (GRSP) production increased by ～3, ～2, and ～3 times, respectively, in soils treated with 1 % OHB. Despite being a metalophyte, O. picensis could not uptake Cu efficiently. Root and shoot Cu concentrations decreased or changed insignificantly in most BC treatments.

Conclusions

The results show that the BCs decreased bioavailable Cu, decreased Cu uptake by O. picensis, improved habitat for microorganisms, and enhanced plant growth in Cu-contaminated soil. This suggests that biochars may be utilized to remediate Cu-contaminated soils.

     

Soil pre-fumigation could effectively improve the disease suppressiveness of biofertilizer to banana <Emphasis Type="Italic">Fusarium</Emphasis> wilt disease by reshaping the soil microbiome

Two seasonal pot experiments were conducted to investigate the effect of biofertilizer application after mixture of lime and ammonium bicarbonate (LA) fumigation, on banana Fusarium wilt disease suppression and soil microbial community composition. Biofertilizer application after LA fumigation decreased 80% of disease incidence compared to control of biofertilizer application to non-fumigated soil. Biofertilizer application after fumigation clearly manipulated soil microbial community composition as revealed by non-metric multidimensional scaling and Venn diagram. LA fumigation significantly reduced the abundance of F. oxysporum while biofertilizer application after fumigation could further decrease it. Furthermore, indigenous microbes, e.g., Bacillus, Pseudomonas, and Mortierella, were associated with disease suppression. Biofertilizer application after fumigation significantly (p?<?0.05) increased the soil pH and content of soil total C and available P and K, and this probably reshaped soil microbial community as revealed by redundancy analysis and variance partitioning analysis. The observed disease suppression due to biofertilizer application after soil fumigation can be attributed to the reduced abundance of F. oxysporum by general suppression resulting from manipulated soil properties and recovered soil microbiome.     

Long-term P fertilisation of pasture soil did not increase soil organic matter stocks but increased microbial biomass and activity

The soil organic matter (OM) content of soils in a long-term fertiliser field trial (Winchmore, New Zealand) are similar (P > 0.05) despite >60 years application of different phosphorus (P) rates. As the net primary productivity increased with P addition, greater losses of carbon (C) occur concomitantly with increased P fertility. Several hypotheses have been proposed to explain the mechanisms, including C leaching, increased earthworm activity or elevated rates of microbial activity. In this study, we found support for both direct and secondary effects of soil P on soil C through impacts on the soil microbial community. Microbial biomass, inferred through quantification of hot water extractable C, increased with soil P status and decreased with C/P ratio (P < 0.001). However, the microbial biomass had no relationship with soil organic C content (P = 0.485). Mineralisation of C substrates added to soil also increased with soil P status (total P, R ² = 0.84; P < 0.001). These results indicated potential conditioning of the microbial community for rapid C cycling. Utilisation of different C compounds was clustered by cophenetic similarity; a distinct group of ten carbon compounds was identified for which rates of mineralisation were strongly associated with soil P status and microbial biomass. However, this alteration of microbial community size and activity was not reflected in abundances of selected oligotrophic and copiotrophic taxa. As such, the alteration may be due to changes in the abundances of all taxa, i.e. a general community response.     

Enhanced Degradation of Atrazine by Soil Microbial Fuel Cells and Analysis of Bacterial Community Structure

Atrazine degradation in soil microbial fuel cells (MFCs) under different anode depths and initial concentrations is investigated for different redox soil conditions, and the microbial communities in the anode and different layers are evaluated. Atrazine degradation is fastest in the upper layer (aerobiotic), followed by the lower layer (anaerobic). A removal efficiency and a half-life of 91.69% and 40 days, respectively, are reported for an anode depth of 4 cm. The degradation rate is found to be dependent on current generation in the soil MFCs rather than on electrode spacing. Furthermore, the degradation rate is inhibited when the initial atrazine concentration is increased from 100 to 750 mg/kg. Meanwhile, the exoelectrogenic bacteria, Deltaproteobacteria and Geobacter, are enriched on the anode and the lower layer in the soil MFCs, while atrazine-degrading Pseudomonas is only observed in very low proportions. In particular, the relative abundances of Deltaproteobacteria and Geobacter are higher for lower initial atrazine concentrations. These results demonstrate that the mechanism of atrazine degradation in soil MFCs is dependent on bioelectrochemistry rather than on microbial degradation.