Biodiversity of network modules drives ecosystem functioning in biochar-amended paddy soil.

Introduction: Soil microbes are central in governing soil multifunctionality and driving ecological processes. Despite biochar application has been reported to enhance soil biodiversity, its impacts on soil multifunctionality and the relationships between soil taxonomic biodiversity and ecosystem functioning remain controversial in paddy soil. Methods: Herein, we characterized the biodiversity information on soil communities, including bacteria, fungi, protists, and nematodes, and tested their effects on twelve ecosystem metrics (including functions related to enzyme activities, nutrient provisioning, and element cycling) in biochar-amended paddy soil. Results: The biochar amendment augmented soil multifunctionality by 20.1 and 35.7% in the early stage, while the effects were diminished in the late stage. Moreover, the soil microbial diversity and core modules were significantly correlated with soil multifunctionality. Discussion: Our analysis revealed that not just soil microbial diversity, but specifically the biodiversity within the identified microbial modules, had a more pronounced impact on ecosystem functions. These modules, comprising diverse microbial taxa, especially protists, played key roles in driving ecosystem functioning in biochar-amended paddy soils. This highlights the importance of understanding the structure and interactions within microbial communities to fully comprehend the impact of biochar on soil ecosystem functioning in the agricultural ecosystem.

The addition of discrimination inhibitors stimulations discrimination potential and N2O emissions were linked to predation among microorganisms in long term nitrogen application and straw returning systems.

Introduction: Ammonia oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) have been proven to be key microorganisms driving the ammonia oxidation process. However, under different fertilization practices, there is a lack of research on the impact of interaction between predators and AOA or AOB on nitrogen cycling at the multi-trophic level. Methods: In this study, a network-oriented microscopic culture experiment was established based on four different long-term fertilization practices soils. We used the nitrification inhibitors 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxide-3-oxyl (PTIO) and 3, 4-Dimethylpyrazole phosphate (DMPP) inhibited AOA and AOB, respectively, to explore the impact of interaction between protists and AOA or AOB on nitrogen transformation. Results: The results showed that long-term nitrogen application promoted the potential nitrification rate (PNR) and nitrous oxide (N2O) emission, and significantly increased the gene abundance of AOB, but had no obvious effect on AOA gene abundance. DMPP significantly reduced N2O emission and PNR, while PTIO had no obvious effect on them. Accordingly, in the multi-trophic microbial network, Cercozoa and Proteobacteria were identified as keystone taxa of protists and AOB, respectively, and were significantly positively correlated with N2O, PNR and nitrate nitrogen. However, Nitrososphaerota archaeon as the keystone species of AOA, had an obvious negative linkage to these indicators. The structural equation model (SEM) showed that AOA and AOB may be competitors to each other. Protists may promote AOB diversity through direct trophic interaction with AOA. Conclusion: The interaction pattern between protists and ammonia-oxidizing microorganisms significantly affects potential nitrification rate and N2O emission, which has important implications for soil nitrogen cycle.

Effects of mineral additives on antibiotic resistance genes and related mechanisms during chicken manure composting.

In this study, two typical minerals (diatomite and bentonite) were applied during composting, and their influences on antibiotics, antibiotic resistance genes (ARGs), intI1 and the bacterial communities were investigated. The relative abundance of total ARGs decreased by 53.72% and 59.54% in diatomite and bentonite addition compared with control on day 42. The minerals addition also reduced the relative abundance of intI1, as much as 41.41% and 59.81% in diatomite and bentonite treatments. Proteobacteria and Firmicutes were the dominant candidate hosts of the major ARGs. There was a significant correlation between total ARGs and intI1 during the composting. Structural equation models further demonstrated that intI1 and antibiotics were the predominant direct factors responsible for ARG variations, and composting properties and bacterial community composition also shifted the variation of ARG profiles by influencing intI1. Overall, these findings suggest that diatomite and bentonite could decrease the potential proliferation of ARGs in chicken manure.

Nanoscale zero-valent iron inhibits the horizontal gene transfer of antibiotic resistance genes in chicken manure compost.

Livestock manure has been identified as a significant hotspot for antibiotic resistance genes (ARGs). However, the impact of nanoscale zero-valent iron (nZVI) on the fate of ARGs during livestock manure composting remains poorly understood. Here, we investigated the evolution of ARGs in chicken manure compost exposed to 100 and 600 mg kg-1 nZVI. The results showed that nZVI addition reduced the concentration of some antibiotics such as doxycycline and sulfamethoxazole. Furthermore, nZVI addition decreased the abundances of most ARGs at the end of composting, but nZVI dosage did not have any significant effect. The abundances of the dominant ARGs (sul1 and sul2) were significantly correlated to the class 1 integron-integrase gene (intI1). A network analysis revealed that the genera Bacteroides, Bacillus, Corynebacterium, Thiopseudomonas and Pseudomonas were the main potential hosts for multiple ARGs, and the decreased abundance of these bacteria contributed to the removal of ARGs. Structural equation models demonstrated that the reduction in intI1 played a predominant role in ARG removal. The nZVI also had direct effects on the intI1 abundance. These findings suggest that the addition of nZVI is a promising strategy to minimize ARG release in chicken manure compost.

Additive quality influences the reservoir of antibiotic resistance genes during chicken manure composting.

Aerobic composting is commonly used to dispose livestock manure and is an efficient way to reduce antibiotic resistance genes (ARGs). Here, the effects of different quality substrates on the fate of ARGs were assessed during manure composting. Results showed that the total relative abundances of ARGs and intI1 in additive treatments were lower than that in control, and high quality treatment with low C/N ratio and lignin significantly decreased the relative abundance of tetW, ermB, ermC, sul1 and sul2 at the end of composting. Additionally, higher quality treatment reduced the relative abundances of some pathogens such as Actinomadura and Pusillimonas, and some thermotolerant degrading-related bacteria comprising Pseudogracilibacillus and Sinibacillus on day 42, probably owing to the change of composting properties in piles. Structural equation models (SEMs) further verified that the physiochemical properties of composting were the dominant contributor to the variations in ARGs and they could also indirectly impact ARGs by influencing bacterial community and the abundance of intI1. Overall, these findings indicated that additives with high quality reduced the reservoir of antibiotic resistance genes of livestock manure compost.

The behavior of antibiotic-resistance genes and their relationships with the bacterial community and heavy metals during sewage sludge composting.

The main contributors to antibiotic resistance genes (ARGs) profiles during the composting process under the addition of biochar (BC) and peat (PT) were not fully explored. This study investigated the influence of BC and PT amendment on ARGs fate, the bacterial community and heavy metals in sewage sludge compost. Compared to control, BC and PT declined the total abundances of ARGs by 17.6% and 43.0% after composting. Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria were the dominant phyla across the composting process, among which, members of Firmicutes (mainly Bacillaceae) were the potential hosts for multiple ARGs. BC and PT addition declined the abundance of bacterial pathogens such as Bacteroides and Pseudomonas. Besides, the concentrations of copper (Cu), zinc (Zn) and lead (Pb) were less in BC and PT treatments than control on day 40, and these metals displayed significant positive correlations to sul1 and intI1. Furthermore, variation partitioning analysis (VPA) revealed that the bacterial community exhibited the most contribution to the ARG patterns, as much as 34.0%, followed by heavy metals (10.8%) and intI1 gene (1.5%). These results suggested that biochar and peat can reduce the risks of ARGs in sewage sludge mainly by sharping the bacterial composition.

Horizontal gene transfer is a key determinant of antibiotic resistance genes profiles during chicken manure composting with the addition of biochar and zeolite.

Livestock manure is an important reservoir of antibiotic resistance genes (ARGs). Biochar and zeolite are commonly used to improve the quality of compost, however, little is known about the impacts of these additives on the fate of ARGs during composting and the underlying mechanisms involved. In this study, zeolite (ZL), biochar (BC), or zeolite and biochar (ZB) simultaneously were added to chicken manure compost to evaluate their effects on the ARGs patterns. After composting, the abundance of ARGs reduced by 92.6% in control, while the reductions were 95.9%, 98.7% and 98.2% for ZL, BC, ZB, respectively. Co-occurrence network analysis indicated that the potential hosts for most ARGs were predominantly affiliated to Firmicutes such as Lactobacillus and Fastidiosipila. Furthermore, shifts in ARGs were significantly correlated with class 1 integrase gene (intI1), and structural equation models further revealed that intI1 gene contributed most (standardized total effect 0.92) to the ARGs-removal, which was trigged by horizontal gene transfer. Together these results suggest that the addition of zeolite and biochar mitigate the accumulation and spread of ARGs during composting, and the crucial role of horizontal gene transfer (HGT) on the behaviors of ARGs should pay more attention to in the future.

Silencing of cyp-33C9 Gene Affects the Reproduction and Pathogenicity of the Pine Wood Nematode, Bursaphelenchus xylophilus.

Cytochrome P450 genes are very important for plant-parasitic nematodes to reproduce and to metabolize xenobiotic compounds generated by their host plants. The pine wood nematode (PWN), Bursaphelenchus xylophilus, causes very high annual economic losses by killing large numbers of pine trees across Asia and into Europe. In this study, we used RNA interference (RNAi) to analyze the function of the cyp-33C9 gene of PWN. Our results showed that expression of the cyp-33C9 gene was suppressed successfully after soaking nematodes for 24 h in cyp-33C9 double-stranded RNA (dsRNA). The silencing of the cyp-33C9 gene significantly decreased the feeding, reproduction, oviposition and egg hatch of B. xylophilus. Meanwhile, the migration speed of B. xylophilus in Pinus thunbergii was reduced in the early stages when the cyp-33C9 gene was silenced in the nematodes. Moreover, knockdown of the cyp-33C9 gene in B. xylophilus caused a decrease in pathogenicity to pine trees. These results suggest that the cyp-33C9 gene plays an important role in the reproduction and pathogenicity of B. xylophilus. This discovery identified several functions of the cyp-33C9 gene in B. xylophilus and provided useful information for understanding the molecular mechanism behind pine wilt disease caused by PWN.

Effects of seaweed fertilizer on enzyme activities, metabolic characteristics, and bacterial communities during maize straw composting.

The objective of this study was to investigate microbial responses when seaweed fertilizer was added to maize straw compost. The metabolic characteristics and bacterial communities were assessed using 16S rRNA sequencing and Biolog analysis. Results showed that the urease, dehydrogenase, and metabolic activities improved on day 21 after the addition of seaweed fertilizer. Firmicutes, Proteobacteria, and Actinobacteria were the dominant phyla in the compost. The relative abundances of Proteobacteria and Actinobacteria were increased by addition of seaweed fertilizer during the early composting stage. Temperature, NH4+-N, NO3--N concentrations, and carbon source metabolism had close relationships with bacterial community variation during composting. A network analysis revealed that NO3--N content had a positive association with Aeromicrobium, and seaweed fertilizer application improved the growth of keystone species related to N cycling. These results suggested that seaweed fertilizer influenced bacterial community succession through its effects on N concentrations during the composting process.

Succession of organics metabolic function of bacterial community in response to addition of earthworm casts and zeolite in maize straw composting.

Succession and metabolism functions of bacterial communities were determined in maize straw composting with earthworm casts and zeolite addition by using high-throughput sequencing, Biolog and PICRUSt. Results showed that earthworm casts and zeolite addition increased the temperature, decreased NH4+ contents and affected bacterial community structure. The relative abundances of Firmicutes and Betaproteobacteria increased with earthworm casts and zeolite addition in the late stage. The abundances of genes related to carbohydrate metabolism, amino acid metabolism, and energy metabolism were increased by these two additives in the early stage, but decreased in the late stage. Network analysis demonstrated that members of Bacillaceae were identified as the keystone taxa. Temperature showed negative relationship with Georgenia, while NH4+ exhibited positive associations with Georgenia, Devosia, Ruania and Mycobacterium. These results indicated that earthworm casts and zeolite addition benefitted the keystone species and enhanced the metabolism capacity of bacterial community, thereby improved the quality of compost.

Biochar influences the succession of microbial communities and the metabolic functions during rice straw composting with pig manure.

The present study determined the dynamic changes of bacterial community structures and its metabolism functions in rice straw composting process with different types of biochar addition. Results showed that both wheat and maize straw biochar addition greatly increased the temperatures and germination index. Biochar addition influenced microbial community structure and metabolic characteristics of the compost. Firmicutes, Actinobacteria and Proteobacteria were the dominant phyla throughout the process, and both biochar addition significantly increased the relative abundance of Actinobacteria phylum, especially genus Saccharomonospora. The abundance of bacterial genes related to amino acid metabolism, carbohydrate metabolism and energy metabolism were also increased with biochar addition. Moreover, total nitrogen was strongly affected by biochar addition and had the greatest influence on the bacterial community structure. These results indicated that biochar addition improved the maturity and fertility of the compost product as well as significantly regulated the microbial community structure and functions during composting process.

Microbial community responses to biochar addition when a green waste and manure mix are composted: A molecular ecological network analysis.

High-throughput sequencing and network analysis were used to investigate the dynamic changes and the effects on bacterial community structures in green waste compost after biochar addition. Biochar addition led to higher thermophilic temperatures and total nitrogen (TN) concentrations and a longer thermophilic period compared with no biochar addition. Biochar also greatly influenced the composition of the bacterial community. Nitriliruptoraceae and Bacillaceae abundances increased in the poultry manure after biochar treatment, and Alcaligenaceae, Rhodispirillaceae and Xanthomonadaceae were more abundant in the cattle manure. Dissolved organic carbon (DOC) and TN emerged as the key determinants of bacterial community composition. Network analysis revealed that DOC had strong positive associations with some taxa (e.g. Comamonas, Leucobacter and Acidimicrobiales), whereas TN had negative associations with other taxa (e.g. Microbacteriaceae and Aeromicrobium). This study has revealed the key taxa related to the carbon and nitrogen cycle during composting with biochar.

Influence of enhanced ultraviolet-B radiation during rice plant growth on rice straw decomposition with nitrogen deposition.

Although straw decomposition is important for ecosystem fertility and carbon balance, influence of ultraviolet-B (UV-B) radiation and nitrogen (N) deposition on this process is unclear. In this study, UV-B-exposed rice straw was decomposed under different N addition treatments for 15 months to investigate the indirect effects of UV-B radiation on straw chemistry and direct effects of N deposition on decomposition. UV-B exposure during rice plant growth changed the rice straw chemical composition, increasing the concentrations of acid-insoluble fraction (AIF), acid-soluble fraction, and UV-B-absorbing compounds. High N content had a negative effect on decomposition of rice straw exposed to enhanced and ambient UV-B radiation. Both AIF concentration and FTIR peak intensities indicated that lignin in rice straw was selectively preserved following N addition and UV-B radiation, reducing straw decomposition rate, which corresponded to lower activities of lignin-degrading enzymes in the later stage of decomposition. Thus, enhanced UV-B radiation during rice plant growth produced more recalcitrant substrates (lignin) and N reacted with lignin to produce more resistant compounds, further decreasing straw decomposition rate. UV-B radiation during plant growth and N deposition inhibit litter decomposition in agroecosystem, and their effects should be considered when establishing biogeochemical models in response to global changes.

Decomposing litter and associated microbial activity responses to nitrogen deposition in two subtropical forests containing nitrogen-fixing or non-nitrogen-fixing tree species.

Atmospheric nitrogen (N) deposition has caused concern due to its effects on litter decomposition in subtropical regions where N-fixing tree species are widespread. However, the effect of N deposition on litter decomposition in N-fixing plantations remains unclear. We investigated the effects of a 2-year N deposition treatment on litter decomposition, microbial activity, and nutrient release in two subtropical forests containing Alnus cremastogyne (AC, N-fixing) and Liquidambar formosana (LF, non-N-fixing). The decomposition rate in AC was faster than in LF when there was no experimental N deposition. In AC, the initial decomposition rate was faster when additional N was applied and was strongly linked to higher cellulose-degrading enzyme activities during the early decomposition stage. However, N deposition reduced litter decomposition and inhibited lignin-degrading enzyme activities during the later decomposition stage. Nitrogen deposition enhanced carbohydrate and alcohol utilization, but suppressed amino acid and carboxylic acid uptake in the AC plantation. However, it did not significantly affect litter decomposition and microbial activity in the LF plantation. In conclusion, N deposition could inhibit litter decomposition by changing microbial enzyme and metabolic activities during the decomposition process and would increase carbon accumulation and nitrogen retention in subtropical forests with N-fixing tree species.

Co-metabolic degradation of iomeprol by a Pseudomonas sp. and its application in biological aerated filter systems.

The non-ionic water-soluble X-ray contrast agent iomeprol (IOM) enters the water supply through sewage treatment plants, which can cause considerable environmental harm. In this study, Pseudomonas sp. I-24 (I-24) was tested for its ability to remove IOM from water via co-metabolic pathways. The optimum removal rate of IOM by I-24 was 38.43% ± 3.70% when starch served as the source of external carbon, and its co-metabolism of IOM conformed to the first-order kinetics. The highest activity of intracellular enzyme (degrading enzyme) extracted from I-24 was 0.143 ± 0.005 mU in starch condition. The Michaelis constant of the degrading enzyme was found to be 91.08 µmol L-1. However, glucose and maltose showed the best promotive effects on the growth and electron transport activity of I-24, indicating that overgrowth may result in competitive inhibition and a reduced degradation rate of IOM. Adding I-24 and degrading enzymes to biological aerated filters increased IOM removal rates without affecting CODMn removal.

Effects of Endobacterium (Stenotrophomonas maltophilia) on Pathogenesis-Related Gene Expression of Pine Wood Nematode (Bursaphelenchus xylophilus) and Pine Wilt Disease.

Pine wilt disease (PWD) caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, is responsible for devastating epidemics in pine trees in Asia and Europe. Recent studies showed that bacteria carried by the PWN might be involved in PWD. However, the molecular mechanism of the interaction between bacteria and the PWN remained unclear. Now that the whole genome of B. xylophilus (Bursaphelenchus xylophilus) is published, transcriptome analysis is a unique method to study the role played by bacteria in PWN. In this study, the transcriptome of aseptic B. xylophilus, B. xylophilus treated with endobacterium (Stenotrophomonas maltophilia NSPmBx03) and fungus B. xylophilus were sequenced. We found that 61 genes were up-regulated and 830 were down-regulated in B. xylophilus after treatment with the endobacterium; 178 genes were up-regulated and 1122 were down-regulated in fungus B. xylophilus compared with aseptic B. xylophilus. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were used to study the significantly changed biological functions and pathways for these differentially expressed genes. Many pathogenesis-related genes, including glutathinone S-transferase, pectate lyase, ATP-binding cassette transporter and cytochrome P450, were up-regulated after B. xylophilus were treated with the endobacterium. In addition, we found that bacteria enhanced the virulence of PWN. These findings indicate that endobacteria might play an important role in the development and virulence of PWN and will improve our understanding of the regulatory mechanisms involved in the interaction between bacteria and the PWN.

Influence of Bxpel1 Gene Silencing by dsRNA Interference on the Development and Pathogenicity of the Pine Wood Nematode, Bursaphelenchus xylophilus.

As the causal agent of pine wilt disease (PWD), the pine wood nematode (PWN), Bursaphelenchus xylophilus, causes huge economic losses by devastating pine forests worldwide. The pectate lyase gene is essential for successful invasion of their host plants by plant-parasitic nematodes. To demonstrate the role of pectate lyase gene in the PWD process, RNA interference (RNAi) is used to analyze the function of the pectate lyase 1 gene in B. xylophilus (Bxpel1). The efficiency of RNAi was detected by real-time PCR. The result demonstrated that the quantity of B. xylophilus propagated with control solution treatment was 62 times greater than that soaking in double-stranded RNA (dsRNA) after B. xylophilus inoculation in Botrytis cinerea for the first generation (F1). The number of B. xylophilus soaking in control solution was doubled compared to that soaking in Bxpel1 dsRNA four days after inoculation in Pinus thunbergii. The quantity of B. xylophilus was reduced significantly (p < 0.001) after treatment with dsRNAi compared with that using a control solution treatment. Bxpel1 dsRNAi reduced the migration speed and reproduction of B. xylophilus in pine trees. The pathogenicity to P. thunbergii seedling of B. xylophilus was weaker after soaking in dsRNA solution compared with that after soaking in the control solution. Our results suggest that Bxpel1 gene is a significant pathogenic factor in the PWD process and this basic information may facilitate a better understanding of the molecular mechanism of PWD.

Specifically expressed genes of the nematode Bursaphelenchus xylophilus involved with early interactions with pine trees.

As the causal agent of pine wilt disease (PWD), the pine wood nematode (PWN), Bursaphelenchus xylophilus, causes huge economic losses by devastating pine forests worldwide. However, the pathogenesis-related genes of B. xylophilus are not well characterized. Thus, DNA microarrays were used to investigate differential gene expression in PWN where Pinus thunbergii was inoculated with nematodes, compared with those cultured on Botrytis cinerea. The microarrays comprised 31121 probes, 1310 (4.2%) of which were differentially regulated (changes of >2-fold, P < 0.01) in the two growth conditions. Of these 1310 genes, 633 genes were upregulated, whereas 677 genes were downregulated. Gene Ontology (GO) categories were assigned to the classes Cellular Component, Molecular Function, and Biological Process. The comparative gene expression analysis showed that a large number of the pathogenesis-related genes of B. xylophilus, such as pectate lyase genes, cytochrome P450s, UGTs, and ABC transporter genes, were highly expressed when B. xylophilus infected P. thunbergii. Annotation analysis indicated that these genes contributed to cell wall degradation, detoxification, and the reproduction process. The microarray results were validated using quantitative RT-PCR (qRT-PCR). The microarray data confirmed the specific expression of B. xylophilus genes during infection of P. thunbergii, which provides basic information that facilitates a better understanding of the molecular mechanism of PWD.