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.     

Microbial inoculation of seeds characteristically shapes the rhizosphere microbiome in <Emphasis Type="Italic">desi</Emphasis> and <Emphasis Type="Italic">kabuli</Emphasis> chickpea types

Purpose

Chickpea is generally cultivated after seed treatment with host-specific Mesorhizobium ciceri, the nitrogen-fixing bacterium forming root nodules. Some species of free-living cyanobacteria are capable of nitrogen fixation. We examined the rhizosphere microbiota changes and the potential for plant growth promotion by applying a free-living, nitrogen-fixing cyanobacterium and the biofilm formulation of cyanobacterium with M. ciceri, relative to M. ciceri applied singly, to two each of desi and kabuli varieties of chickpea.

Materials and methods

Denaturing gradient gel electrophoresis (DGGE) profiles of archaeal, bacterial and cyanobacterial communities and those of phospholipid fatty acids (PLFAs) were obtained to evaluate the changes of the microbial communities in the chickpea rhizosphere. Plant growth attributes, including the pod yields and the availabilities of soil macronutrients and micronutrients, were monitored.

Results and discussion

The DGGE profiles showed distinct and characteristic changes due to the microbial inoculation; varietal differences exerted a marked influence on the archaeal and cyanobacterial communities. However, bacterial communities were modulated more by the type of microbial inoculants. Abundance of Gram-negative bacteria (in terms of notional PLFAs) differed between the desi and the kabuli varieties inoculated with M. ciceri alone, and the principal component analysis of PLFA profiles confirmed the characteristic effect of microbial inoculants tested. Microbial inoculation led to increases in the 100-seed weight and differential effects on the concentrations of available nitrogen and phosphorus, and those of iron, zinc and copper, suggesting their increased cycling in the rhizosphere.

Conclusions

Microbial inoculation of chickpea brought out the characteristic changes in rhizosphere microbiota. Consequently, the growth promotion of chickpea and nutrient cycling in its rhizosphere distinctively differed. Further studies are needed to analyse the association and dynamic changes in the microbial communities to define the subset of microorganisms selected by chickpea in its rhizosphere and the influence of microbial inoculation.

     

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.     

Microbial and Physico-chemical Characteristics Associated with the Incidence of <Emphasis Type="Italic">Legionella</Emphasis> spp. and <Emphasis Type="Italic">Acanthamoeba</Emphasis> spp. in Rainwater Harvested from Different Roofing Materials

The incidence of Legionella and Acanthamoeba spp. was correlated to microbial indicator analysis and physico-chemical characteristics of rainwater harvested from catchment areas constructed from galvanized zinc, Chromadek®, and asbestos, respectively. Quantitative PCR (qPCR) analysis indicated that no significant difference (p?>?0.05) in copy numbers of Legionella spp. and Acanthamoeba spp. was recorded in tank water samples collected from the respective roofing materials. However, significant positive Spearman (ρ) correlations were recorded between the occurrences of Legionella spp. gene copies vs. nitrites and nitrates (p?=?0.05) in all tank water samples. Significant positive correlations were also established between Acanthamoeba spp. vs. barium (p?=?0.03), magnesium (p?=?0.02), sodium (p?=?0.02), silicon (p?=?0.05), arsenic (p?=?0.03), and phosphate (p?=?0.01), respectively. Additionally, while no significant correlations were observed between Legionella spp. vs. the indicator bacteria (p?>?0.05), positive correlations were observed between Acanthamoeba spp. vs. total coliforms (p?=?0.01) and Acanthamoeba spp. vs. Escherichia coli (p?=?0.02), respectively. Results obtained in the current study thus indicate that the incidence of Acanthamoeba and Legionella spp. in harvested rainwater was not influenced by the roofing material utilized. Moreover, it is essential that the microbial quality of rainwater be assessed before this water source is implemented for potable and domestic uses as untreated harvested rainwater may lead to legionellosis and amoebae infections.     

Effect of <Emphasis Type="Italic">Silene vulgaris</Emphasis> and Heavy Metal Pollution on Soil Microbial Diversity in Long-Term Contaminated Soil

In this study, we analysed the impact of heavy metals and plant rhizodeposition on the structure of indigenous microbial communities in rhizosphere and bulk soil that had been exposed to heavy metals for more than 150 years. Samples of the rhizosphere of Silene vulgaris and non-rhizosphere soils 250 and 450 m from the source of emission that had different metal concentrations were collected for analyses. The results showed that soils were collected 250 m from the smelter had a higher number of Cd-resistant CFU compared with the samples that were collected from 450 m, but no significant differences were observed in the number of total and oligotrophic CFU or the equivalent cell numbers between rhizosphere and non-rhizosphere soils that were taken 250 and 450 m from the emitter. Unweighted pair group method with arithmetic mean (UPGMA) cluster analysis of the denaturing gradient gel electrophoresis (DGGE) profiles, as well as a cluster analysis that was generated on the phospholipid fatty acid (PLFA) profiles, showed that the bacterial community structure of rhizosphere soils depended more on the plant than on the distance and metal concentrations. The sequencing of the 16S rDNA fragments that were excised from the DGGE gel revealed representatives of the phyla Bacteroidetes, Acidobacteria, Gemmatimonadetes, Actinobacteria and Betaproteobacteria in the analysed soil with a predominance of the first three groups. The obtained results demonstrated that the presence of S. vulgaris did not affect the number of CFUs, except for those of Cd-resistant bacteria. However, the presence of S. vulgaris altered the soil bacterial community structure, regardless of the sampling site, which supported the thesis that plants have a higher impact on soil microbial community than metal contamination.     

Bacterial diversity in the rhizosphere of two phylogenetically closely related plant species across environmental gradients

Purpose

Rhizosphere soil bacterial communities are crucial to plant growth, health, and stress resistance. In order to detect how bacterial communities associated with the rhizosphere of phylogenetically related plant species vary in terms of composition, function, and diversity, we investigated the rhizosphere bacterial community structure of two perennial shrub species, Caragana jubata and Caragana roborovskyi, under natural field conditions in northwest China and analyzed the influence of soil properties and environmental factors.

Materials and methods

Eighteen root samples, eight for C. jubata, and ten for C. roborovskyi, along with any adherent soil particles, were collected from multiple sites in northwest China. The rhizosphere soil was washed from the roots, and bacterial communities were analyzed using Illumina MiSeq sequencing of 16S rRNA gene amplicons. Then, α-diversity and β-diversity were calculated using QIIME.

Results and discussion

Across species, Proteobacteria (29 %), Actinobacteria (15 %), Chloroflexi (10 %), Acidobacteria (10 %), Bacteroidetes (8 %), Firmicutes (8 %), Planctomycetes (7 %), Gemmatimonadetes (4 %), and Verrucomicrobia (3 %) were the most abundant phyla in the rhizosphere of C. jubata and C. roborovskyi. However, principal co-ordinates analysis indicated strong interspecific patterns of bacterial rhizosphere communities. Further, the richness of Proteobacteria, Acidobacteria, Bacteroidetes, Verrucomicrobia, Firmicutes, and Nitrospirae was significantly higher in the rhizosphere of C. jubata compared with C. roborovskyi, while the opposite was found for Actinobacteria and Cyanobacteria. However, the Shannon index showed no significant difference in α-diversity between C. jubata and C. roborovskyi. Distance-based redundancy analysis indicated that soil properties and environmental factors exerted strong influences on the structure of the rhizosphere bacterial community and explained 47 and 46 % of community variances between samples, respectively.

Conclusions

Our results showed strong interspecific clustering of the bacterial rhizosphere communities of C. roborovskyi and C. jubata. Altitude explained most of the variation in the composition of bacterial rhizosphere communities of C. roborovskyi and C. jubata, followed by soil pH, water content, organic matter content, total nitrogen content, and mean annual rainfall.

     

Altered precipitation seasonality impacts the dominant fungal but rare bacterial taxa in subtropical forest soils

How soil microbial communities respond to precipitation seasonality change remains poorly understood, particularly for warm-humid forest ecosystems experiencing clear dry-wet cycles. We conducted a field precipitation manipulation experiment in a subtropical forest to explore the impacts of reducing dry-season rainfall but increasing wet-season rainfall on soil microbial community composition and enzyme activities. A 67% reduction of throughfall during the dry season decreased soil water content (SWC) by 17–24% (P < 0.05), while the addition of water during the wet season had limited impacts on SWC. The seasonal precipitation redistribution had no significant effect on the microbial biomass and enzyme activities, as well as on the community composition measured with phospholipid fatty acids (PLFAs). However, the amplicon sequencing revealed differentiated impacts on bacterial and fungal communities. The dry-season throughfall reduction increased the relative abundance of rare bacterial phyla (Gemmatimonadetes, Armatimonadetes, and Baoacteriodetes) that together accounted for only 1.5% of the total bacterial abundance by 15.8, 40, and 24% (P < 0.05), respectively. This treatment also altered the relative abundance of the two dominant fungal phyla (Basidiomycota and Ascomycota) that together accounted for 72.4% of the total fungal abundance. It increased the relative abundance of Basidiomycota by 27.4% while reduced that of Ascomycota by 32.6% (P < 0.05). Our results indicate that changes in precipitation seasonality can affect soil microbial community composition at lower taxon levels. The lack of community-level responses may be ascribed to the compositional adjustment among taxonomic groups and the confounding effects of other soil physicochemical variables such as temperature and substrate availability.     

The foliar spray of <Emphasis Type="Italic">Rhodopseudomonas palustris</Emphasis> grown under <Emphasis Type="Italic">Stevia</Emphasis> residue extract promotes plant growth via changing soil microbial community

Purpose

The extract of Stevia residue is an ideal substitute for cultivation of the purple nonsulfur bacterium, like Rhodopseudomonas palustris (R. palustris). But the influence of R. palustris grown under residue extract on its downstream application is still not well-characterized. The objective of this study was to assess the effect of foliar spray of R. palustris grown under Stevia residue extract on the plant growth and soil microbial properties.

Materials and methods

A pot experiment was carried out under the greenhouse condition, consisting of four treatments varying in the sprayed substances: sterilized water (control), R. palustris grown under the chemical medium supplemented with L-tryptophan (SyT), R. palustris grown under Stevia residue extract supplemented with L-tryptophan (ExT), and R. palustris grown under Stevia residue extract supplemented with NH₄Cl (ExT). The net photosynthesis rate of the uppermost leaves was measured with a portable photosynthesis system. Soil microbial activity was analyzed by microcalorimetry. Soil bacterial community components were determined by real-time quantitative PCR (qPCR) and high-throughput sequencing techniques.

Results and discussion

Compared with SyT, the R. palustris grown under Stevia residue extract not only improved the plant biomass and the net photosynthetic rate to a large extent, but also increased soil microbial metabolic activity and altered community compositions as well. The treatments receiving R. palustris, especially ExT and ExN, increased the relative abundances of some functional guilds involved in C turnover and nutrient cycling in soil, including Acidobacteria, Actinobacteria, Proteobacteria, Gemmatimonadaetes, Nitrospirae, and Planctomycetes.

Conclusions

R. palustris grown under the Stevia residue extract showed advantages over that under the chemical medium on both plant growth and soil microbial properties. One of the possible reasons could result from the increases in microbial activity and several bacterial keystone guilds involved into C and nutrient cycling, both of which potentially contribute to the improved plant growth. The results would be conducive to the downstream application of R. palustris in an economical way.

     

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.

     

Coupled Effects of Electrical Stimulation and Antibiotics on Microbial Community in Three-Dimensional Biofilm-Electrode Reactors

Antibiotics are often misused or overused, resulting in large residue inputs in the environment. Electricity and antibiotics were regarded as potentially important factors, which impact on the microbial community during treatment of antibiotics in three-dimensional biofilm-electrode reactors (3D-BERs). Unfortunately, only a few studies have been reported yet. Four 3D-BERs and one 3D-BR (reactor with biological sludge and no voltage) were designed to assess the effect of low current, sulfamethoxazole (SMX), and tetracycline (TC) on microbial populations. The 3D-BERs achieved excellent removal efficiencies of 72.20–93.52 and 82.61–95.80% for SMX and TC, respectively. Microorganisms were classified into 58 phyla, 125 classes, 166 orders, 187 families, and 220 genera. Proteobacteria held the overwhelming predominance, followed by Bacteroidetes, Chloroflexi, Actinobacteria, Verrucomicrobia, Firmicutes, and Acidobacteria. The 3D-BERs achieved higher richness of microbial composition compared with the 3D-BR. Microbial communities and relative abundance at the phyla level were affected by low current. The microbial communities in the 3D-BERs were similar at the genus level, even with different antibiotic concentrations. However, the relative abundances and compositions of the microbial communities decreased during the treatment of antibiotics. To increase the performance of 3D-BERs, the function of microorganisms should be investigated in future studies.     

Bacterial Community Composition and Genes for Herbicide Degradation in a Stormwater Wetland Collecting Herbicide Runoff

Stormwater wetlands collect and attenuate runoff-related herbicides, limiting their transport into aquatic ecosystems. Knowledge on wetland bacterial communities with respect to herbicide dissipation is scarce. Previous studies showed that hydrological and hydrochemical conditions, including pesticide removal capacity, may change from spring to summer in stormwater wetlands. We hypothesized that these changes alter bacterial communities, which, in turn, influence pesticide degradation capacities in stormwater wetland. Here, we report on bacterial community changes in a stormwater wetland exposed to pesticide runoff, and the occurrence of trz, atz, puh, and phn genes potentially involved in the biodegradation of simazine, diuron, and glyphosate. Based on T-RFLP analysis of amplified 16S rRNA genes, a response of bacterial communities to pesticide exposure was not detected. Changes in stormwater wetland bacterial community mainly followed seasonal variations in the wetland. Hydrological and hydrochemical fluctuations and vegetation development in the wetland presumably contributed to prevent detection of effects of pesticide exposure on overall bacterial community. End point PCR assays for trz, atz, phn, and puh genes associated with herbicide degradation were positive for several environmental samples, which suggest that microbial degradation contributes to pesticide dissipation. However, a correlation of corresponding genes with herbicide concentrations could not be detected. Overall, this study represents a first step to identify changes in bacterial community associated with the presence of pesticides and their degradation in stormwater wetland.     

Spatial patterns of microbial denitrification genes change in response to poultry litter placement and cover crop species in an agricultural soil

Subsurface-banding manure and winter cover cropping are farming techniques designed to reduce N loss. Little is known, however, about the effects of these management tools on denitrifying microbial communities and the greenhouse gases they produce. Abundances of bacterial (16S), fungal (ITS), and denitrification genes (nirK, nirS, nosZ-I, and nosZ-II) were measured in soil samples collected from a field experiment testing the combination of cereal rye and hairy vetch cover cropping with either surface-broadcasted or subsurface-banded poultry litter. The spatial distribution of genes was mapped to identify potential denitrifier hotspots. Spatial distribution maps showed increased 16S rRNA genes around the manure band, but no denitrifier hotspots. Soil depth and nitrate concentration were the strongest drivers of gene abundance, but bacterial gene abundance also differed by gene, soil characteristics, and management methods. Gene copy number of nirK was higher under cereal rye than hairy vetch and positively associated with soil moisture, while nirS gene copies did not differ between cover crop species. The nirS gene copies increased when manure was surface broadcasted compared to subsurface banded and was positively associated with pH. Soil moisture and pH were positively correlated to nosZ-II but not to nosZ-I gene copy numbers. We observed stronger correlations between nosZ-I and nirS, and nosZ-II and nirK gene copies compared to the reverse pairings. Agricultural management practices differentially affect spatial distributions of genes coding for denitrification enzymes, leading to changes in the composition of the denitrifying community.     

Biomineralization of atrazine and analysis of 16S rRNA and catabolic genes of atrazine degraders in a former pesticide mixing site and a machinery washing area

Purpose

The purpose of this study was to determine the first-order rate constants and half-lives of aerobic and anaerobic biomineralization of atrazine in soil samples from an agricultural farm site that had been previously used for mixing pesticide formulations and washing application equipment. Atrazine catabolic genes and atrazine-degrading bacteria in the soil samples were analyzed by molecular methods.

Materials and methods

Biomineralization of atrazine was measured in soil samples with a [U-ring-¹⁴C]-atrazine biometer technique in soil samples. Enrichment cultures growing with atrazine were derived from soil samples and they were analyzed for bacterial diversity by constructing 16S rDNA clone libraries and sequencing. Bacterial isolates were also obtained and they were screened for atrazine catabolic genes.

Results and discussion

The soils contained active atrazine-metabolizing microbial communities and both aerobic and anaerobic biomineralization of [U-ring-¹⁴C]-atrazine to ¹⁴CO₂ was demonstrated. In contrast to aerobic incubations, anaerobic biometers displayed considerable differences in the kinetics of atrazine mineralization between duplicates. Sequence analysis of 16S rDNA clone libraries constructed from the enrichment cultures revealed a preponderance of Variovorax spp. (51 %) and Schlesneria (16 %). Analysis of 16S rRNA gene sequences from pure cultures (n?=?12) isolated from enrichment cultures yielded almost exclusively Arthrobacter spp. (83 %; 10/12 isolates). PCR screening of pure culture isolates for atrazine catabolic genes detected atzB, atzC, trzD, trzN, and possibly atzA. The presence of a complete metabolic pathway was not demonstrated by the amplification of catabolic genes among these isolates.

Conclusions

The soils contained active atrazine-metabolizing microbial communities. The anaerobic biometer data showed variable response of atrazine biomineralization to external electron acceptor conditions. Partial pathways are inevitable in soil microbial communities, with metabolites linking into other catabolic and assimilative pathways of carbon and nitrogen. There was no evidence for the complete set of functional genes of the known pathways of atrazine biomineralization among the isolates.

     

Tillage system affects fertilizer-induced nitrous oxide emissions

Since the development of effective N₂O mitigation options is a key challenge for future agricultural practice, we studied the interactive effect of tillage systems on fertilizer-derived N₂O emissions and the abundance of microbial communities involved in N₂O production and reduction. Soil samples from 0–10 cm and 10–20 cm depth of reduced tillage and ploughed plots were incubated with dairy slurry (SL) and manure compost (MC) in comparison with calcium ammonium nitrate (CAN) and an unfertilized control (ZERO) for 42 days. N₂O and CO₂ fluxes, ammonium, nitrate, dissolved organic C, and functional gene abundances (16S rRNA gene, nirK, nirS, nosZ, bacterial and archaeal amoA) were regularly monitored. Averaged across all soil samples, N₂O emissions decreased in the order CAN and SL (CAN?=?748.8?±?206.3, SL?=?489.4?±?107.2 μg kg^?1) followed by MC (284.2?±?67.3 μg kg^?1) and ZERO (29.1?±?5.9 μg kg^?1). Highest cumulative N₂O emissions were found in 10–20 cm of the reduced tilled soil in CAN and SL. N₂O fluxes were assigned to ammonium as source in CAN and SL and correlated positively to bacterial amoA abundances. Additionally, nosZ abundances correlated negatively to N₂O fluxes in the organic fertilizer treatments. Soils showed a gradient in soil organic C, 16S rRNA, nirK, and nosZ with greater amounts in the 0–10 than 10–20 cm layer. Abundances of bacterial and archaeal amoA were higher in reduced tilled soil compared to ploughed soils. The study highlights that tillage system induced biophysicochemical stratification impacts net N₂O emissions within the soil profile according to N and C species added during fertilization.     

Changes in methane emission and methanogenic and methanotrophic communities in restored wetland with introduction of <Emphasis Type="Italic">Alnus trabeculosa</Emphasis>

Purpose

The dynamics and uncertainties in wetland methane budgets affected by the introduction of Alnus trabeculosa H. necessitate research on production of methane by methanogenic archaea and consumption by methane-oxidizing microorganisms simultaneously.

Materials and methods

This study investigated methane emission in situ by the closed chamber method, and methanogenic and methanotrophic communities using denatured gradient gel electrophoresis (DGGE) and quantitative PCR based on mcrA (methyl coenzyme M reductase), pmoA (particulate methane monooxygenase) genes in the rhizosphere and non-rhizosphere soils in the indigenous pure Phragmites australis T., and A. trabeculosa–P. australis mixed communities in Chongxi wetland.

Results and discussion

Methane flux rate from the pure P. australis community was 2.4 times larger than that of A. trabeculosa–P. australis mixed community in the rhizosphere and 1.7 times larger in the non-rhizosphere, respectively. The abundance of methanogens was lower in the mixed community soils (3.56?×?10³–6.90?×?10³ copies g^?1 dry soil) compared with the P. australis community (1.47?×?10⁴–1.89?×?10⁴ copies g^?1 dry soil), whereas the methanotrophs showed an opposite trend (2.08?×?10⁶–1.39?×?10⁶ copies g^?1 dry soil for P. australis and 6.20?×?10⁶–1.99?×?10⁶ copies g^?1 dry soil for mixed community soil). A liner relationship between methane emission rates against pmoA/mcrA ratios (R ²?=?0.5818, p?<?0.05, n?=?15) was observed. The community structures of the methane-cycling microorganism based on mcrA and pmoA suggested that acetoclastic methanogens belonging to Methanosarcinaceae and a particular type II methanotroph, Methylocystis, were dominant in these two plant communities.

Conclusions

The introduction of A. trabeculosa would promote the proliferation of methanotrophs, especially the dominant Methylocystis, but not methanogens, ultimately diminishing methane emission in the wetland.

     

Effects of saline water irrigation and fertilization regimes on soil microbial metabolic activity

Purpose

Irrigation and fertilization can change soil environment, which thereby influence soil microbial metabolic activity (MMA). How to alleviate the adverse effects by taking judicious saline water irrigation and fertilization regimes is mainly concerned in this research.

Materials and methods

Here, we conducted a field orthogonal designed test under different saline water irrigation amount, water salinity, and nitrogen fertilizer application. The metabolic profiles of soil microbial communities were analyzed by using the Biolog method.

Results and discussion

The results demonstrated that irrigation amount and fertilizer application could significantly change MMA while irrigation water salinity had no significant effect on it. Medium irrigation amount (30 mm), least (50 kg ha^?1) or medium (350 kg ha^?1) N fertilizer application, and whatever irrigation water salinity could obtain the optimal MMA. Different utilization rates of carbohydrates, amino acids, carboxylic acids, and polymers by soil microbial communities caused the differences of the effects, and D-galactonic acid γ-lactone, L-arginine, L-asparagine, D-glucosaminic acid, Tween 80, L-threonine, and D-galacturonic acid were the indicator for distinguishing the effects.

Conclusions

The results presented here demonstrated that by regulating irrigation water amount and fertilizer application, the effects of irrigation salinity on MMA could be alleviated, which offered an efficient approach for guiding saline water irrigation.

     

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.     

Effect of P stoichiometry on the abundance of nitrogen-cycle genes in phosphorus-limited paddy soil

Previous studies have shown that phosphorus addition to P-limited soils increases gaseous N loss. A possible explanation for this phenomenon is element stoichiometry (specifically of C:N:P) modifying linked nutrient cycling, leading to enhanced nitrification and denitrification. In this study, we investigated how P stoichiometry influenced the dynamics of soil N-cycle functional genes. Rice seedlings were planted in P-poor soils and incubated with or without P application. Quantitative PCR was then applied to analyze the abundance of ammonia-oxidizing (amoA) and denitrifying (narG nirK, nirS, nosZ) genes in soil. P addition reduced bacterial amoA abundance but increased denitrifying gene abundance. We suggest this outcome is due to P-induced shifts in soil C:P and N:P ratios that limited ammonia oxidization while enhancing P availability for denitrification. Under P application, the rhizosphere effect raised ammonia-oxidizing bacterial abundance (amoA gene) and reduced nirK, nirS, and nosZ in rhizosphere soils. The change likely occurred through greater C input and O₂ release from roots, thus altering C availability and redox conditions for microbes. Our results show that P application enhances gaseous N loss potential in paddy fields mainly through stimulating denitrifier growth. We conclude that nutrient availability and elemental stoichiometry are important in regulating microbial gene responses, thereby influencing key ecosystem processes such as denitrification.

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Graphical abstract ?

     

Effects of ectomycorrhizal fungus <Emphasis Type="Italic">Boletus edulis</Emphasis> and mycorrhiza helper <Emphasis Type="Italic">Bacillus cereus</Emphasis> on the growth and nutrient uptake by <Emphasis Type="Italic">Pinus thunbergii</Emphasis>

The present greenhouse study was undertaken to evaluate the effects of co-inoculating the ectomycorrhizal (ECM) fungus Boletus edulis with the mycorrhiza helper bacterium Bacillus cereus HB12 or HB59 on the growth and nutrient uptake of Pinus thunbergii. The inoculation with mycorrhiza helper bacterium significantly (P?≤?0.05) increased the ectomycorrhizal colonization. Treatments with dual inoculum (the mycorrhiza helper bacterium plus mycorrhiza) significantly (P?≤?0.05) increased the P. thunbergii growth. Bacteria–mycorrhizae interactions resulted in a great utilization of phosphate and potassium. The single inoculation resulted in a higher root activity than the control while the co-inoculation led to the highest root activity. The 6-CFDA staining assay showed that B. cereus enhanced fungal activity in ectomycorrhizal symbiosis. The results conclusively suggest that B. cereus isolated from the rhizosphere of P. thunbergii can potentially be used as individual inoculant or co-inoculated with ECM fungi to increase the production in sustainable ecological systems. These results support the potential use of B. cereus (HB12 or HB59) and B. edulis as mixed inoculants stimulating growth of P. thunbergii.     

Chemical nature of soil organic carbon under different long-term fertilization regimes is coupled with changes in the bacterial community composition in a Calcaric Fluvisol

Fertilization is an important factor influencing the chemical structure of soil organic carbon (SOC) and soil microbial communities; however, whether any connection exists between the two under different fertilization regimes remains unclear. Soils from a 27-year field experiment were used to explore potential associations between SOC functional groups and specific bacterial taxa, using quantitative multiple cross-polarization magic-angle spinning ¹³C nuclear magnetic resonance and 16S rRNA gene sequencing. Treatments included balanced fertilization with organic materials (OM) and with nitrogen (N), phosphorus (P), and potassium (K) mineral fertilizers (NPK); unbalanced fertilization without one of the major elements (NP, PK, or NK); and an unamended control. These treatments were divided into four distinct groups, namely OM, NPK, NP plus PK, and NK plus control, according to their bacterial community composition and SOC chemical structure. Soil total P, available P, and SOC contents were the major determinants of bacterial community composition after long-term fertilization. Compared to NPK, the OM treatment generated a higher aromatic C–O and OCH₃ and lower alkyl C and OCH abundance, which were associated with the enhanced abundance of members of the Acidobacteria subgroups 6 and 5, Cytophagaceae, Chitinophagaceae, and Bacillus sp.; NP plus PK treatments resulted in a higher OCH and lower aromatic C–C abundance, which showed a close association with the enrichment of unclassified Chloracidobacteria, Syntrophobacteraceae, and Anaerolineae and depletion of Bacillales; and NK plus control treatments resulted in a higher abundance of aromatic C–C, which was associated with the enhanced abundance of Bacillales. Our results indicate that different fertilization regimes changed the SOC chemical structure and bacterial community composition in different patterns. The results also suggest that fertilization-induced variations in SOC chemical structure were strongly associated with shifts in specific microbial taxa which, in turn, may be affected by changes in soil properties.