Spatial and temporal dynamics of the bacterial community under experimental warming in field-grown wheat.

Climate change may lead to adverse effects on agricultural crops, plant microbiomes have the potential to help hosts counteract these effects. While plant-microbe interactions are known to be sensitive to temperature, how warming affects the community composition and functioning of plant microbiomes in most agricultural crops is still unclear. Here, we utilized a 10-year field experiment to investigate the effects of warming on root zone carbon availability, microbial activity and community composition at spatial (root, rhizosphere and bulk soil) and temporal (tillering, jointing and ripening stages of plants) scales in field-grown wheat (Triticum aestivum L.). The dissolved organic carbon and microbial activity in the rhizosphere were increased by soil warming and varied considerably across wheat growth stages. Warming exerted stronger effects on the microbial community composition in the root and rhizosphere samples than in the bulk soil. Microbial community composition, particularly the phyla Actinobacteria and Firmicutes, shifted considerably in response to warming. Interestingly, the abundance of a number of known copiotrophic taxa, such as Pseudomonas and Bacillus, and genera in Actinomycetales increased in the roots and rhizosphere under warming and the increase in these taxa implies that they may play a role in increasing the resilience of plants to warming. Taken together, we demonstrated that soil warming along with root proximity and plant growth status drives changes in the microbial community composition and function in the wheat root zone.

Crop rotation increases root biomass and promotes the correlation of soil dissolved carbon with the microbial community in the rhizosphere.

As essential approaches for conservation agricultural practices, straw residue retention and crop rotation have been widely used in the Mollisols of Northeast China. Soil organic carbon, root development and microbial community are important indicators representing soil, crop and microbiota, respectively, and these factors work together to influence soil fertility and crop productivity. Studying their changes and interactions under different conservation practices is crucial to provide a theoretical basis for developing rational agricultural practices. The experiment in this study was conducted using the conventional practice (continuous maize without straw retention, C) and three conservation practices, namely, continuous maize with straw mulching (CS), maize-peanut rotation (R), and maize-peanut rotation with straw mulching (RS). Straw mulching (CS) significantly increased soil total organic carbon (TOC), active organic carbon (AOC), and microbial biomass carbon (MBC), but did not promote maize yield. Maize-peanut rotation (R and RS) significantly increased dissolved organic carbon (DOC) in the rhizosphere by promoting root growth, and maize yield (increased by 10.2%). For the microbial community structure, PERMANOVA and PCoA indicated that the bacterial community differed significantly between rhizosphere soil and bulk soil, but the fungal community shifted more under different agricultural practices. The correlation analysis indicated that the rotation system promoted the association between the soil DOC and the microbial community (especially the bacterial community), and straw mulching enhanced the connection between the soil TOC and the fungal community. Some plant growth-promoting rhizobacteria (including Bacillus, Streptomyces, Rhizobium, and Pseudomonas) were enriched in the rhizosphere soil and were increased in the rotation system (R and RS), which might be due to an increase in the soil rhizosphere DOC level. These beneficial microbes had significantly negative correlations with several fungal groups (such as Mycosphaerella, Penicillium, Paraphoma and Torula) that were classified as plant pathotrophs by FUNGuild. These results indicated that ensuring plant root development and improving root-bacteria interactions are of great importance to guarantee crop yield when implementing conservation tillage practices.

Lytic Bacteriophage PZL-Ah152 as Biocontrol Measures Against Lethal Aeromonas hydrophila Without Distorting Gut Microbiota.

Phage therapy is an alternative approach to overcome the problem of multidrug resistance in bacteria. In this study, a bacteriophage named PZL-Ah152, which infects Aeromonas hydrophila, was isolated from sewage, and its biological characteristics and genome were studied. The genome contained 54 putative coding sequences and lacked known putative virulence factors, so it could be applied to phage therapy. Therefore, we performed a study to (i) investigate the efficacy of PZL-Ah152 in reducing the abundance of pathogenic A. hydrophila strain 152 in experimentally infected crucian carps, (ii) evaluate the safety of 12 consecutive days of intraperitoneal phage injection in crucian carps, and (iii) determine how bacteriophages impact the normal gut microbiota. The in vivo and in vitro results indicated that the phage could effectively eliminate A. hydrophila. Administering PZL-Ah152 (2 × 109 PFU) could effectively protect the fish (2 × 108 CFU/carp). Furthermore, a 12-day consecutive injection of PZL-Ah152 did not cause significant adverse effects in the main organs of the treated animals. We also found that members of the genus Aeromonas could enter and colonize the gut. The phage PZL-Ah152 reduced the number of colonies of the genus Aeromonas. However, no significant changes were observed in α-diversity and ß-diversity parameters, which suggested that the consumed phage had little effect on the gut microbiota. All the results illustrated that PZL-Ah152 could be a new therapeutic method for infections caused by A. hydrophila.

Proteiniclasticum sediminis sp. nov., an obligate anaerobic bacterium isolated from anaerobic sludge.

An obligate anaerobic bacterial BAD-10 T was isolated from anaerobic acetochlor-degrading sludge. The strain was Gram-stain negative, curved rod-shaped, non-motile and non-spore-forming. Growth was observed in PYT medium at pH 6.0-9.0 (optimum, pH 7.5), at 25-47 °C (37 °C) and with 0-1.0% NaCl (w/v, 0%). Strain BAD-10 T could degrade acetochlor. The major fermentation products from peptone-yeast (PY) medium were acetate and butyrate. The predominant cellular fatty acids were iso-C15:0 FAME, anteiso-C15:0 FAME and C16:0 FAME. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain BAD-10 T showed closest affiliation to Proteiniclasticum ruminis D3RC-2 T, with a sequence similarity of 97.6%. Genome sequencing revealed a genome size of 2,983,986 bp, a G + C content of 51.4 mol% and protein-coding genes of 3,102. The average nucleotide identity and in silico DNA-DNA hybridization values between strain BAD-10 T and Proteiniclasticum ruminis D3RC-2 T were 71.0% and 20.4%, respectively, which were below the standard thresholds for species differentiation. On the basis of phenotypic, physiological and phylogenetic evidence, strain BAD-10 T represents a novel species in the genus Proteiniclasticum, for which the name Proteiniclasticum sediminis sp. nov. is proposed. Strain BAD-10 T (= CCTCC AB 2021091 T = KCTC 25288 T) is the type strain of the proposed novel species.

Soil bacterial community dynamics following bioaugmentation with Paenarthrobacter sp. W11 in atrazine-contaminated soil.

Atrazine is one of the most widely used herbicides, however it and its metabolites cause widespread contamination in soil and ground water. Bioaugmentation is an effective method for remediation of environmental organic pollutants. High-throughput sequencing provides an important tool for understanding the changes of microbial community and function in response to pollutants degradation based on bioaugmentation. In this study, the effect of biodegradation with Paenarthrobacter sp. W11 and the change of microbial community during atrazine degradation were investigated. The results showed that bioaugmentation significantly accelerated the degradation rate of atrazine in soil and reduced the toxic effect of atrazine residues on wheat growth. The extra available NH4+ through atrazine mineralization could serve as a nitrogen source to increase microbial numbers. High-throughput sequencing further revealed that the microbial community restored a new balance. The function of microbial community predicted by PICRUSt2 suggested that the biodegradation process of atrazine affected not only the atrazine degradation pathway, but also the nitrogen metabolism pathway. Methylobacillus and Pseudomonas were considered as the most important indigenous atrazine-degrading microorganisms, because their relative abundances were positively correlated with the relative abundance of Paenarthrobacter and atrazine degradation pathway. This study provides insight into the cooperation between indigenous microorganisms and external inoculums on atrazine degradation process.

Fifteen-Year Application of Manure and Chemical Fertilizers Differently Impacts Soil ARGs and Microbial Community Structure.

Manure, which contains large amounts of antibiotics and antibiotic resistance genes (ARGs), is widely used in agricultural soils and may lead to the evolution and dispersal of ARGs in the soil environment. In the present study, soils that received manure or chemical fertilizers for 15 years were sampled on the North China Plain (NCP), which is one of the primary areas of intensive agriculture in China. High-throughput quantitative PCR and sequencing technologies were employed to assess the effects of long-term manure or chemical fertilizer application on the distribution of ARGs and microbial communities. A total of 114 unique ARGs were successfully amplified from all soil samples. Manure application markedly increased the relative abundance and detectable numbers of ARGs, with up to 0.23 copies/16S rRNA gene and 81 unique ARGs. The increased abundance of ARGs in manure-fertilized soil was mainly due to the manure increasing the abundance of indigenous soil ARGs. In contrast, chemical fertilizers only moderately affected the diversity of ARGs and had no significant effect on the relative abundance of the total ARGs. In addition, manure application increased the abundance of mobile genetic elements (MGEs), which were significantly and positively correlated with most types of ARGs, indicating that horizontal gene transfer via MGEs may play an important role in the spread of ARGs. Furthermore, the application of manure and chemical fertilizers significantly affected microbial community structure, and variation partitioning analysis showed that microbial community shifts represented the major driver shaping the antibiotic resistome. Taken together, our results provide insight into the long-term effects of manure and chemical fertilization on the dissemination of ARGs in intensive agricultural ecosystems.

Root-associated microbiomes of wheat under the combined effect of plant development and nitrogen fertilization.

BACKGROUND: Plant roots assemble microbial communities both inside the roots and in the rhizosphere, and these root-associated microbiomes play pivotal roles in plant nutrition and productivity. Although it is known that increased synthetic fertilizer input in Chinese farmlands over the past 50 years has resulted in not only increased yields but also environmental problems, we lack a comprehensive understanding of how crops under elevated nutrient input shape root-associated microbial communities, especially through adjusting the quantities and compositions of root metabolites and exudates. METHODS: The compositions of bacterial and fungal communities from the roots and rhizosphere of wheat (Triticum aestivum L.) under four levels of long-term inorganic nitrogen (N) fertilization were characterized at the tillering, jointing and ripening stages. The root-released organic carbon (ROC), organic acids in the root exudates and soil organic carbon (SOC) and soil active carbon (SAC) in the rhizosphere were quantified. RESULTS: ROC levels varied dramatically across wheat growth stages and correlated more with the bacterial community than with the fungal community. Rhizosphere SOC and SAC levels were elevated by long-term N fertilization but varied only slightly across growth stages. Variation in the microbial community structure across plant growth stages showed a decreasing trend with N fertilization level in the rhizosphere. In addition, more bacterial and fungal genera were significantly correlated in the jointing and ripening stages than in the tillering stage in the root samples. A number of bacterial genera that shifted in response to N fertilization, including Arthrobacter, Bacillus and Devosia, correlated significantly with acetic acid, oxalic acid, succinic acid and tartaric acid levels. CONCLUSIONS: Our results indicate that both plant growth status and N input drive changes in the microbial community structure in the root zone of wheat. Plant growth stage demostrated a stronger influence on bacterial than on fungal community composition. A number of bacterial genera that have been described as plant growth-promoting rhizobacteria (PGPR) responded positively to N fertilization, and their abundance correlated significantly with the organic acid level, suggesting that the secretion of organic acids may be a strategy developed by plants to recruit beneficial microbes in the root zone to cope with high N input. These results provide novel insight into the associations among increased N input, altered carbon availability, and shifts in microbial communities in the plant roots and rhizosphere of intensive agricultural ecosystems.

High-Quality Draft Genome Sequence of Pseudomonas songnenensis L103, a Denitrifier Isolated from a 100-Meter-Deep Aquifer in a Heavily Nitrogen-Fertilized Agricultural Area.

Pseudomonas songnenensis strain L103 was isolated from a 100-m-deep aquifer in the North China Plain, a heavily nitrogen-fertilized agricultural area. The genome is 4.8 Mb and contains 4,409 protein-coding genes, including a full set of genes (nar, nir, nor, and nos) for complete denitrification.

Electrodes Donate Electrons for Nitrate Reduction in a Soil Matrix via DNRA and Denitrification.

Microbial strains and indigenous microbiota in soil slurries have been reported to use electrons from electrodes for nitrate (NO3-) reduction. However, few studies have confirmed this in a soil matrix hitherto. This study investigated if, and how, an electric potential affected NO3- reduction in a soil matrix. The results showed that, compared to a control treatment, applying an electric potential of -0.5 V versus the standard hydrogen electrode (SHE) significantly increased the relative abundance of NO3--reducing microbes (e.g., Alcaligenaceae and Pseudomonadaceae) and the abundances of the nrfA, nirK, nirS, and nosZ genes in soil matrices. Meanwhile, the electric potential treatment doubled the NO3- reduction rate and significantly increased the rates of production of ammonium (NH4+), dinitrogen (N2), and nitrous oxide (N2O). The amount of NO3--N reduced under the electric potential treatment was comparable to the sum of the amounts of N observed in the increased N2O, N2, NH4+, and nitrite (NO2-) pools. An open-air experiment showed that the electric potential treatment promoted soil NO3- reduction with a spatial scale of at least 38 cm. These results demonstrated that an electric potential treatment could enhance NO3- reduction via both denitrification and dissimilatory NO3- reduction to ammonium (DNRA) in the soil matrix. The mechanisms revealed in this study have implications for the future development of potential techniques for enhancing NO3- reduction in the vadose zone and consequently reducing the risk of NO3- leaching.

Long-Term Nitrogen Fertilization Elevates the Activity and Abundance of Nitrifying and Denitrifying Microbial Communities in an Upland Soil: Implications for Nitrogen Loss From Intensive Agricultural Systems.

The continuous use of nitrogen (N) fertilizers to increase soil fertility and crop productivity often results in unexpected environmental effects and N losses through biological processes, such as nitrification and denitrification. In this study, multidisciplinary approaches were employed to assess the effects of N fertilization in a long-term (~20 years) field experiment in which a fertilizer gradient (0, 200, 400, and 600 kg N ha-1 yr-1) was applied in a winter wheat-summer maize rotation cropping system in the North China Plain, one of the most intensive agricultural regions in China. The potential nitrification/denitrification rates, bacterial community structure, and abundances of functional microbial communities involved in key processes of the N cycle were assessed during both the summer maize (SM) and winter wheat (WW) seasons. Long-term N fertilization resulted in a decrease in soil pH and an increase in soil organic matter (OM), total N and total carbon concentrations. Potential nitrification/denitrification and the abundances of corresponding functional N cycling genes were positively correlated with the fertilization intensity. High-throughput sequencing of the 16S rRNA gene revealed that the increased fertilization intensity caused a significant decrease of bacterial diversity in SM season, while changed the microbial community composition such as increasing the Bacteroidetes abundance and decreasing Acidobacteria abundance in both SM and WW seasons. The alteration of soil properties markedly correlated with the variation in microbial structure, as soil pH and OM were the most predominant factors affecting the microbial structure in the SM and WW seasons, respectively. Furthermore, consistently with the results of functional gene quantification, functional prediction of microbial communities based on 16S rRNA sequence data also revealed that the abundances of the key nitrificaiton/denitrification groups were elevated by long-term N inputs. Taken together, our results suggested that soil microbial community shifted consistently in both SM and WW seasons toward a higher proportion of N-cycle microbes and exhibited higher N turnover activities in response to long-term elevated N fertilizer. These findings provided new insights into the molecular mechanisms responsible for N loss in intensively N fertilized agricultural ecosystems.

Response of Nitrifier and Denitrifier Abundance and Microbial Community Structure to Experimental Warming in an Agricultural Ecosystem.

Soil microbial community plays an important role in terrestrial carbon and nitrogen cycling. However, the response of the soil nitrifier and denitrifier communities to climate warming is poorly understood. A long-term field warming experiment has been conducted for 8 years at Luancheng Experimental Farm Station on the North China Plain; we used this field to examine how soil microbial community structure, nitrifier, and denitrifier abundance respond to warming under regular irrigation (RI) and high irrigation (HI) at different soil depths (0-5, 5-10, and 10-20 cm). Nitrifier, denitrifier, and the total bacterial abundance were assessed by quantitative polymerase chain reaction of the functional genes and 16S rRNA gene, respectively. Bacterial community structure was studied through high throughput sequencing of the 16S rRNA gene. Under RI, warming significantly (P < 0.05) increased the potential nitrification rate and nitrate concentration and decreased the soil moisture. In most of the samples, warming increased the ammonia-oxidizing bacteria abundance but decreased the ammonia-oxidizing archaea (AOA) and denitrifier (nirK, nirS, and nosZ genes) abundance. Under HI, there was a highly increased AOA and 16S rRNA gene abundance and a slightly higher denitrifier abundance compared with RI. Warming decreased the bacterial diversity and species richness, and the microbial community structure differed greatly between the warmed and control plots. The decrease in bacterial diversity was higher in RI than HI and at the 0-5 cm depths than at the 5-10 and 10-20 cm soil depths. Warming led to an increase in the relative abundance of Actinobacteria, Bacteroidetes, and TM7 but a decrease in Acidobacteria, Alphaproteobacteria, Deltaproteobacteria, Nitrospira, and Planctomycetes. The greater shift in microbial community structure was observed only in RI at the 0-5 cm soil depth. This study provides new insight into our understanding of the nitrifier and denitrifier activity and microbial community response to climate warming in agricultural ecosystems.

Organic carbon availability limiting microbial denitrification in the deep vadose zone.

Microbes in the deep vadose zone play an essential role in the mitigation of nitrate leaching; however, limited information is available on the mechanisms of microbial denitrification due to sampling difficulties. We experimentally studied the factors that affect denitrification in soils collected down to 10.5 meters deep along the soil profile. After an anoxic pre-incubation, denitrification rates moderately increased and the N2 O/(N2 O + N2 ) ratios declined while the microbial abundance and diversity did not change significantly in most of the layers. Denitrification rate was significantly enhanced and the abundance of the denitrification genes was simultaneously elevated by the increased availability of organic carbon in all studied layers, to a greater extent in the subsurface layers than in the surface layers, suggesting the severe scarcity of carbon in the deep vadose zone. The genera Pseudomonas and Bacillus, which are made up of a number of species that have been previously identified as denitrifiers in soil, were the major taxa that respond to carbon addition. Overall, our results suggested that the limited denitrification in the deep vadose zone is not because of the lack of denitrifiers, but due to the low abundance of denitrifiers which is caused by low carbon availability.

A secure RFID authentication protocol adopting error correction code.

RFID technology has become popular in many applications; however, most of the RFID products lack security related functionality due to the hardware limitation of the low-cost RFID tags. In this paper, we propose a lightweight mutual authentication protocol adopting error correction code for RFID. Besides, we also propose an advanced version of our protocol to provide key updating. Based on the secrecy of shared keys, the reader and the tag can establish a mutual authenticity relationship. Further analysis of the protocol showed that it also satisfies integrity, forward secrecy, anonymity, and untraceability. Compared with other lightweight protocols, the proposed protocol provides stronger resistance to tracing attacks, compromising attacks and replay attacks. We also compare our protocol with previous works in terms of performance.