[Effects of water-nitrogen interactions on NH3 and N2O emissions and yield in winter wheat cropland]. / 水氮互作对冬小麦农田NH3和N2O排放及产量的影响.

To investigate the effects of different irrigation and nitrogen application modes on nitrogen gaseous loss in winter wheat farmland, we conducted a field experiment at Changqing Irrigation Experiment Station in Shandong Province, with two irrigation levels (80%-90% θf(I1) and 70%-80% θf(I2)) and three nitrogen application levels (conventional nitrogen application of 240 kg·hm-2(N1), nitrogen reduction of 12.5% (N2), and nitrogen reduction of 25% (N3)). The results showed that ammonia volatilization and nitrous oxide emission rate peak appeared within 2-4 days after fertilization or irrigation. The ammonia volatilization rate during the chasing fertilizer period was significantly higher than that during the basal fertilizer period. Compared with other treatments, the ave-rage ammonia volatilization rate of I2N2 treatment during the chasing fertilizer period was reduced by 10.1%-51.6%, and the average nitrous oxide emission rate over the whole growth period was reduced by 15.4%-52.2%. The ammonia volatilization rate was significantly positively associated with surface soil pH value and ammonium nitrogen content, while the nitrous oxide emission rate was significantly positively associated with nitrate content in topsoil. The accumulation amount of soil ammonia volatilization and nitrous oxide emission ranged from 0.83-1.42 and 0.11-0.33 kg·hm-2, respectively. Moderate reduction of irrigation water and nitrogen input could effectively reduce cumulative amounts of ammonia volatilization and nitrous oxide emission from winter wheat farmland. The cumulative amounts of ammonia volatilization and nitrous oxide emission under I1N3 and I2N2 treatments were signi-ficantly lower than those under other treatments. The highest winter wheat yield (5615.6 kg·hm-2) appeared in I2N2 treatment. The irrigation water utilization efficiency of I2 was significantly higher than that of I1, with the maximum increase rate of 45.2%. Compared with N1 and N3 treatments, the maximum increase rate of nitrogen fertilizer productivity and agricultural utilization efficiency in N2 reached 15.2% and 31.8%, respectively. In conclusion, the treatment with 70%-80% θf irrigation level and 210 kg·hm-2 nitrogen input could effectively improve the utilization efficiency of irrigation water and nitrogen fertilization and reduce gaseous loss from winter wheat farmland.

Determinants of photochemical characteristics of the photosynthetic electron transport chain of maize.

Introduction: The photosynthetic electron transport chain (ETC) is the bridge that links energy harvesting during the photophysical reactions at one end and energy consumption during the biochemical reactions at the other. Its functioning is thus fundamental for the proper balance between energy supply and demand in photosynthesis. Currently, there is a lack of understanding regarding how the structural properties of the ETC are affected by nutrient availability and plant developmental stages, which is a major roadblock to comprehensive modeling of photosynthesis. Methods: Redox parameters reflect the structural controls of ETC on the photochemical reactions and electron transport. We conducted joint measurements of chlorophyll fluorescence (ChlF) and gas exchange under systematically varying environmental conditions and growth stages of maize and sampled foliar nutrient contents. We utilized the recently developed steady-state photochemical model to infer redox parameters of electron transport from these measurements. Results and discussion: We found that the inferred values of these photochemical redox parameters varied with leaf macronutrient content. These variations may be caused either directly by these nutrients being components of protein complexes on the ETC or indirectly by their impacts on the structural integrity of the thylakoid and feedback from the biochemical reactions. Also, the redox parameters varied with plant morphology and developmental stage, reflecting seasonal changes in the structural properties of the ETC. Our findings will facilitate the parameterization and simulation of complete models of photosynthesis.

Secondary succession of shrub-herb communities in the hilly area of Taihang Mountain.

Introduction: To document the successional processes of shrub-herb communities after large-scale human disturbance, and understand how changing environmental conditions affect species replacement in semi-humid hilly areas. Methods: Utilizing the established permanent plots in the hilly area of Taihang Mountain, we evaluated temporal patterns of vegetation and soil following grass-to-shrub succession. Results and Discussion: Along secondary succession, Vitex negundo var. heterophylla gradually dominated in dry sunny slope and shared the dominance with Leptodermis oblonga in shaded slope. Herbaceous dominant species in shrub-herb communities switched from Themeda japonica, Bothriochloa ischaemum, Artemisia sacrorum, and Cleistogenes chinensis in 1986 census to B. ischaemum and A. sacrorum in 2008 census, but herb was no longer dominant in 2020 census. As succession progresses, species dominance increased while richness decreased generally, and herb cover and aboveground biomass decreased, whereas shrub height, cover, and aboveground biomass increased significantly. Soil organic matter (SOM), total nitrogen (TN), total phosphorus (TP), and total potassium (TK) in topsoil increased significantly while pH declined by 1.04 units over the past three decades. Plant communities transitioned from perennial herbs to shrub-herb and then shrub communities, and V. negundo var. heterophylla dominated in the succession of shrub-herb communities. Climate and soil properties, combined with plant attributes, together drive post-disturbance secondary succession. From a management perspective, the tight coupling between vegetation and soil under local climatic conditions should be considered to improve the fragile ecosystem in the hilly area of Taihang Mountain.

Advanced maternal age: copy number variations and pregnancy outcomes.

Objective: Adverse pregnancy outcomes are closely related to advanced maternal age (AMA; age at pregnancy ≥35 years). Little research has been reported on aneuploid abnormalities and pathogenic copy number variations (CNVs) affecting pregnancy outcomes in women with AMA. The purpose of this study was to assess CNVs associated with AMA in prenatal diagnosis to determine the characteristics of pathogenic CNVs and assist with genetic counseling of women with AMA. Methods: Among 277 fetuses of women with AMA, 218 (78.7%) were isolated AMA fetuses and 59 (21.3%) were non-isolated AMA fetuses and showed ultrasound anomalies from January 2021 to October 2022. Isolated AMA was defined as AMA cases without sonographic abnormalities. Non-isolated AMA was defined as AMA cases with sonographic abnormalities such as sonographic soft markers, widening of the lateral ventricles, or extracardiac structural anomalies. The amniotic fluid cells underwent routine karyotyping followed by single nucleotide polymorphism array (SNP-array) analysis. Results: Of the 277 AMA cases, karyotype analysis identified 20 chromosomal abnormalities. As well as 12 cases of chromosomal abnormalities corresponded to routine karyotyping, the SNP array identified an additional 14 cases of CNVs with normal karyotyping results. There were five pathogenetic CNVs, seven variations of uncertain clinical significance (VOUS), and two benign CNVs. The detection rate of abnormal CNVs in non-isolated AMA cases was increasing (13/59; 22%) than in isolated AMA cases (13/218; 5.96%) (p < 0.001). We also determined that pathogenic CNVs affected the rate of pregnancy termination in women with AMA. Conclusion: Aneuploid abnormalities and pathogenic CNVs affect pregnancy outcomes in women with AMA. SNP array had a higher detection rate of genetic variation than did karyotyping and is an important supplement to karyotype analysis, which enables better informed clinical consultation and clinical decision-making.

Microbial gradual shifts during the process of species replacement in Taihang Mountain.

Introduction: Understanding microbial gradual shifts along species replacement can help elucidate the mechanisms driving secondary succession, and predict microbial responses to changing environments. However, how climate-induced species replacement alters microbial processes, and whether microbial shifts follow predictable assembly trajectories remain unclear. Methods: Using space-for-time substitution approach, we studied shifts in bacterial and fungal communities in the succession from Leptodermis oblonga to Vitex negundo var. heterophylla shrubland in Taihang Mountain. Results and Discussion: Species replacement, induced by climate related environmental change, significantly increased the above-ground biomass of shrublands, and TP and TK contents in topsoil. The succession from L. oblonga to V. negundo var. heterophylla communities resulted in the gradually replacement of cold-tolerant microbes with warm-affinity ones, and alterations of microbial communities involved in soil biogeochemical processes. Soil and plant variables, such as above-ground biomass, soil pH, total phosphorus, and total potassium, well explained the variations in microbial communities, indicating that the coordinated changes in plant communities and soil properties during secondary succession caused accompanied shifts in microbial diversity and composition.

Responses of Nitrous Oxide Emissions and Bacterial Communities to Experimental Freeze-Thaw Cycles in Contrasting Soil Types.

Nitrous oxide (N2O) pulse emissions are detected in soils subjected to freeze-thaw cycles in both laboratory and field experiments. However, the mechanisms underlying this phenomenon are poorly understood. In this study, a laboratory incubation experiment that included freeze-thaw cycles (FTC), freezing (F) and control (CK) treatments was performed on three typical Chinese upland soils, namely, fluvo-aquic soil (FS), black soil (BS) and loess soil (LS). A higher similarity in soil properties and bacterial community structure was discovered between FS and LS than between FS and BS or LS and BS, and the bacterial diversity of FS and LS was higher than that of BS. FTC significantly increased the denitrification potential and the proportion of N2O in the denitrification gas products in FS and LS but decreased the denitrification potential in BS. Accordingly, with the increasing number of freeze-thaw cycles, the bacterial community composition in the FTC treatments in FS and LS diverged from that in CK but changed little in BS. Taxa that responded to FTC or correlated with denitrification potential were identified. Taken together, our results demonstrated that the effects of FTC on N2O emissions are soil-type-dependent and that the shift in the microbial community structure may contribute to the elevated N2O emissions.

Anthropogenic N input increases global warming potential by awakening the "sleeping" ancient C in deep critical zones.

Even a small net increase in soil organic carbon (SOC) mineralization will cause a substantial increase in the atmospheric CO2 concentration. It is widely recognized that the SOC mineralization within deep critical zones (2 to 12 m depth) is slower and much less influenced by anthropogenic disturbance when compared to that of surface soil. Here, we showed that 20 years of nitrogen (N) fertilization enriched a deep critical zone with nitrate, almost doubling the SOC mineralization rate. This result was supported by corresponding increases in the expressions of functional genes typical of recalcitrant SOC degradation and enzyme activities. The CO2 released and the SOC had a similar 14C age (6000 to 10,000 years before the present). Our results indicate that N fertilization of crops may enhance CO2 emissions from deep critical zones to the atmosphere through a previously disregarded mechanism. This provides another reason for markedly improving N management in fertilized agricultural soils.

Nitrate-Nitrite-Nitric Oxide Pathway: A Mechanism of Hypoxia and Anoxia Tolerance in Plants.

Oxygen (O2) is the most crucial substrate for numerous biochemical processes in plants. Its deprivation is a critical factor that affects plant growth and may lead to death if it lasts for a long time. However, various biotic and abiotic factors cause O2 deprivation, leading to hypoxia and anoxia in plant tissues. To survive under hypoxia and/or anoxia, plants deploy various mechanisms such as fermentation paths, reactive oxygen species (ROS), reactive nitrogen species (RNS), antioxidant enzymes, aerenchyma, and adventitious root formation, while nitrate (NO3-), nitrite (NO2-), and nitric oxide (NO) have shown numerous beneficial roles through modulating these mechanisms. Therefore, in this review, we highlight the role of reductive pathways of NO formation which lessen the deleterious effects of oxidative damages and increase the adaptation capacity of plants during hypoxia and anoxia. Meanwhile, the overproduction of NO through reductive pathways during hypoxia and anoxia leads to cellular dysfunction and cell death. Thus, its scavenging or inhibition is equally important for plant survival. As plants are also reported to produce a potent greenhouse gas nitrous oxide (N2O) when supplied with NO3- and NO2-, resembling bacterial denitrification, its role during hypoxia and anoxia tolerance is discussed here. We point out that NO reduction to N2O along with the phytoglobin-NO cycle could be the most important NO-scavenging mechanism that would reduce nitro-oxidative stress, thus enhancing plants' survival during O2-limited conditions. Hence, understanding the molecular mechanisms involved in reducing NO toxicity would not only provide insight into its role in plant physiology, but also address the uncertainties seen in the global N2O budget.

The ecological clusters of soil organisms drive the ecosystem multifunctionality under long-term fertilization.

Long-term fertilization is known to impact the biodiversity and community structures of soil organisms, which are responsible for multiple soil ecosystem functions (multifunctionality). However the relationship between the alterations of soil organisms and ecosystem multifunctionality remains unclear, especially in the case of long-term fertilization. To explore the contribution of soil organismal biodiversity and community structures to ecosystem multifunctionality, we took soil samples from a nearly 25-year field fertilization experiment. Organic matter significantly improved the soil ecosystem multifunctionality. Ecosystem multifunctionality was found to be closely linked to the biodiversity and communities of soil organisms within the major ecological clustering of soil organisms (Module 1) according to the trophic co-occurrence network, rather than the entire community of soil organisms. This indicated that ecological clusters of soil organisms within the network were critical in maintaining soil ecosystem multifunctionality. The application of organic fertilization could enrich specialized soil organisms and increase interactions of soil organisms in the ecological cluster. As a result, our findings emphasize the role of ecological clusters in the soil organismal co-occurrence network in controlling soil multifunctionality after long-term fertilization, presenting a novel perspective on the link between soil biodiversity and ecosystem multifunctionality.

[Clinical and genetic analysis of three children patients with Kleefstra syndrome].

OBJECTIVE: To explore the genetic basis of three children with unexplained developmental delay/intellectual disability (DD/ID). METHODS: Peripheral blood samples were collected from the patients and subjected to chromosomal microarray analysis (CMA). RESULTS: Patient 1 was found to harbor a 190 kb deletion at 9q34.3, which encompassed most of EHMT1 (OMIM 607001), the key gene for Kleefstra syndrome (OMIM 610253). Patients 2 and 3 were siblings. CMA showed that they have shared four chromosomal copy number variations (CNVs) including a deletion at 9q34.3 which spanned 154 kb and 149 kb, respectively, and encompassed the EHMT1 and CACNA1B (OMIM 601012) genes. The remaining 3 CNVs were predicted to be with no clinical significance. CONCLUSION: Microdeletions at 9q33.4 probably underlay the pathogenesis of DD/ID in the three children, for which EHMT1 may be the key gene.

Plants are a natural source of nitrous oxide even in field conditions as explained by 15N site preference.

Plants are either recognized to produce nitrous oxide (N2O) or considered as a medium to transport soil-produced N2O. To date, it is not clear whether in their habitat plants conduit N2O produced in soil or are a natural source. We aimed to understand role of plants in N2O emissions in field conditions. Therefore, rubber plants (Ficus elastica) were planted in the field; then plant and soil chambers were deployed simultaneously to collect gas samples, and 15N site preference (SP) of N2O was evaluated. The mean SP values of plant and soil emitted N2O were -20.85 ± 2.8‰ and -8.85 ± 1.08‰, respectively, and were significantly different (p < 0.0001); while bulk 15N of plant and soil emitted N2O were -10.83 ± 3.33‰ and -22.56 ± 3.37‰, respectively and were similar (p = 0.06). In the current study, soil always acted as a source of N2O, while plants were both source and sink. Plant and soil N2O fluxes had significant positive exponential relationship with both soil and air temperature. Soil water-filled pore space (WFPS) had significant negative linear relationship with only soil N2O fluxes. Plant N2O fluxes had significant positive linear relationship with plant respiration rates and negative linear relationship with plant surface areas. Based on the relationship between plant respiration rates and N2O fluxes, we suggest that mitochondria are the possible sites of N2O formation in plant cells while the relationship between plant surface areas and N2O fluxes suggests that roots are the parts of its formation in natural and field conditions. Our results suggest that plants are a natural source of N2O even at field conditions and challenge a view that plants are a medium to transport soil-produced N2O into the atmosphere.

Long-term warming and nitrogen fertilization affect C-, N- and P-acquiring hydrolase and oxidase activities in winter wheat monocropping soil.

The enzymatic activities and ratios are critical indicators for organic matter decomposition and provide potentially positive feedback to carbon (C) loss under global warming. For agricultural soils under climate change, the effect of long-term warming on the activities of oxidases and hydrolases targeting C, nitrogen (N) and phosphorus (P) and their ratios is unclear, as well as whether and to what extend the response is modulated by long-term fertilization. A 9-year field experiment in the North China Plain, including an untreated control, warming, N fertilization, and combined (WN) treatment plots, compared the factorial effect of warming and fertilization. Long-term warming interacted with fertilization to stimulate the highest activities of C, N, and P hydrolases. Activities of C and P hydrolase increased from 8 to 69% by N fertilization, 9 to 53% by warming, and 28 to 130% by WN treatment compared to control, whereas the activities of oxidase increased from 4 to 16% in the WN soils. Both the warming and the WN treatments significantly increased the enzymatic C:N ratio from 0.06 to 0.16 and the vector length from 0.04 to 0.12 compared to the control soil, indicating higher energy and resource limitation for the soil microorganisms. Compared to WN, the warming induced similar ratio of oxidase to C hydrolase, showing a comparable ability of different microbial communities to utilize lignin substrates. The relationship analyses showed mineralization of organic N to mediate the decomposition of lignin and enzyme ratio in the long-term warming soil, while N and P hydrolases cooperatively benefited to induce more oxidase productions in the soil subject to both warming and N fertilization. We conclude that coupled resource limitations induced microbial acclimation to long-term warming in the agricultural soils experiencing high N fertilizer inputs.

Predominant frequency of HLA-B*27 in patients with ankylosing spondylitis in southeastern China.

OBJECTIVES: This study was to investigate the polymorphism and distribution of alleles of HLA-B*27 in patients with ankylosing spondylitis (AS) in Han population of southeastern China. METHODS: A total of 89 peripheral blood samples from southeastern Chinese Han patients with AS that diagnosed according to Modified New York criteria were subtyped using the high-resolution PCR-SSP.Exon 2-3 of HLA-B*27 gene was amplified and sequenced to further confirm the HLA-B*27 subtype. RESULTS: The frequency of HLA-B*27 was 99.87% in AS patients. Three subtypes, HLA-B*2704, HLA-B*2705, and HLA-B*2706 were identified. The frequencies for these three alleles were HLA-B*2704 in 84/88 (95.46%), HLA-B*2705 in 3/88(3.41%), and HLA-B*2706 in 1/88 (1.13%) of the HLA-B*27 positive patients, respectively. CONCLUSIONS: Our study shows that HLA-B*2704 has an overwhelming frequency in southeastern Chinese Han AS patients. A combined analysis including previous studies of HLA-B*27-subtype distributions in Chinese Han populations showed that HLA-B*2704 may originate from the southern Han and then migrate and spread to the northern areas, and HLA-B*2705 show the opposite result.

Characterization of tetracycline-resistant microbiome in soil-plant systems by combination of H218O-based DNA-Stable isotope probing and metagenomics.

The emergence and spread of antibiotic resistance have been considered as a global health threat. However, effective methods to identify antibiotic-resistant bacteria (ARB) in complex microbial community are lacking, and the potential transmission pathways of ARB and antibiotic resistance genes (ARGs) in the soil-plant system remain scarce. Here in this study, tetracycline was chosen as the target antibiotic due to its globally wide usage and clinical significance. DNA-based stable isotope probing with H218O was applied to identify the tetracycline-resistant bacteria from soil-plant systems. Eighteen-year organic fertilization significantly changed the composition of the tetracycline-resistant microbiome in the soil-wheat system and resulted in a higher relative abundance of ARGs in the wheat endophyte. Rhizosphere harboring the most diverse ARGs and mobile genetic elements was identified as a hot spot for horizontal gene transfer and an important bridge between bulk soil and wheat endophyte. Micrococcaceae and Sphingomonadaceae carrying ARGs associated with abundant mobile genetic elements, were identified as the core bacterial taxa in long-term manure-amended and untreated soil-wheat systems, respectively. This method contributes to a more precise track of ARB in the environment, and our work depicts the high potential of ARG transfer in the rhizosphere mediated by the core species.

An improved microelectrode method reveals significant emission of nitrous oxide from the rhizosphere of a long-term fertilized soil in the North China Plain.

Microsensors are able to accurately quantify nitrous oxide (N2O) emissions in microenvironments at high spatio-temporal resolution; yet, limited studies have been conducted on agricultural soils due to the inability to obtain electrical signal under conditions of low soil moisture. This study improved the calibration of a microelectrode for measuring N2O emissions from agricultural soil. The microelectrode was applied to evaluate the effect of long-term fertilization with mineral fertilizer (NPK), complemented with pig manure (MNPK), straw (SNPK), or without fertilizer (CK), all with and without urea addition, on N2O emissions from the soil, with explicit separation of the rhizosphere and the bulk soil compartments. The use of soil solution instead of pure water for calibration of the microelectrode doubled the signal and significantly improved the sensor sensitivity. The optimal electrolytic concentration of the soil solution, expressed as water: soil ratio, was found at the maximum vertex of the quadratic equation fitted on the slope values of the calibration equations for different soil solutions. The application of the calibrated microelectrode revealed significantly higher N2O emission from the rhizosphere compared to the bulk soil, accounting for 60% of the total emission. For the bulk soil, MNPK significantly increased N2O emissions compared to SNPK and NPK, whereas the differences between these treatments for the rhizosphere soil were insignificant. The statistical modeling revealed significant relation of the N2O emission with soil inorganic nitrogen contents and an additive effect of treatment (MNPK and SNPK), urea addition and rhizosphere soil. This study provides novel insights into the use of microelectrodes for measuring N2O emissions from the soil microenvironment and also points on the rhizosphere compartment and the management practices of agroecosystems able to reduce the N2O emission from agriculture.

Changes of δ15N values during the volatilization process after applying urea on soil.

Ammonia (NH3) volatilized from soils plays an important role in N cycle and air pollution, thus it is important to trace the emission source and predict source contributions to development strategies mitigating the environmental harmful of soil NH3 volatilization. The measurements of 15N natural abundance (δ15N) could be used as a complementary tool for apportioning emissions sources to resolve the contribution of multiple NH3 emission sources to air NH3 pollution. However, information of the changes of δ15N-NH3 values during the whole volatilization process under different N application rates are currently lacking. Hence, to fill this gap, we conducted a 15-day incubation experiment included different urea-N application rates to determine δ15N values of NH3 during volatilization process. Results showed that volatilization process depleted 15N in NH3. The average δ15N value of NH3 volatilized from the 0, 20, 180, and 360 kg N ha-1 treatment was -16.2 ± 7.3‰, -26.0 ± 5.4‰, -34.8 ± 4.8‰, and -40.6 ± 5.7‰. Overall, δ15N-NH3 values ranged from -46.0‰ to -4.7‰ during the whole volatilization process, with lower in higher urea-N application treatments than those in control. δ15N-NH3 values during the NH3 volatilization process were much lower than those of the primary sources, soil (-3.4 ± 0.1‰) and urea (-3.6 ± 0.1‰). Therefore, large isotopic fractionation may occur during soil volatilization process. Moreover, negative relationships between soil NH4+-N and NH3 volatilization rate and δ15N-NH3 values were observed in this study. Our results could be used as evidences of NH3 source apportionments and N cycle.

Nitrogen isotopic signatures and fluxes of N2O in response to land-use change on naturally occurring saline-alkaline soil.

The conversion of natural grassland to semi-natural or artificial ecosystems is a large-scale land-use change (LUC) commonly occurring to saline-alkaline land. Conversion of natural to artificial ecosystems, with addition of anthropogenic nitrogen (N) fertilizer, influences N availability in the soil that may result in higher N2O emission along with depletion of 15N, while converting from natural to semi-natural the influence may be small. So, this study assesses the impact of LUC on N2O emission and 15N in N2O emitted from naturally occurring saline-alkaline soil when changing from natural grassland (Phragmites australis) to semi-natural [Tamarix chinensis (Tamarix)] and to cropland (Gossypium spp.). The grassland and Tamarix ecosystems were not subject to any management practice, while the cropland received fertilizer and irrigation. Overall, median N2O flux was significantly different among the ecosystems with the highest from the cropland (25.3 N2O-N µg m-2 h-1), intermediate (8.2 N2O-N µg m-2 h-1) from the Tamarix and the lowest (4.0 N2O-N µg m-2 h-1) from the grassland ecosystem. The 15N isotopic signatures in N2O emitted from the soil were also significantly affected by the LUC with more depleted from cropland (- 25.3 ‰) and less depleted from grassland (- 0.18 ‰). Our results suggested that the conversion of native saline-alkaline grassland with low N to Tamarix or cropland is likely to result in increased soil N2O emission and also contributes significantly to the depletion of the 15N in atmospheric N2O, and the contribution of anthropogenic N addition was found more significant than any other processes.

Potential Pathway of Nitrous Oxide Formation in Plants.

Plants can produce and emit nitrous oxide (N2O), a potent greenhouse gas, into the atmosphere, and several field-based studies have concluded that this gas is emitted at substantial amounts. However, the exact mechanisms of N2O production in plant cells are unknown. Several studies have hypothesised that plants might act as a medium to transport N2O produced by soil-inhabiting microorganisms. Contrarily, aseptically grown plants and axenic algal cells supplied with nitrate (NO3) are reported to emit N2O, indicating that it is produced inside plant cells by some unknown physiological phenomena. In this study, the possible sites, mechanisms, and enzymes involved in N2O production in plant cells are discussed. Based on the experimental evidence from various studies, we determined that N2O can be produced from nitric oxide (NO) in the mitochondria of plants. NO, a signaling molecule, is produced through oxidative and reductive pathways in eukaryotic cells. During hypoxia and anoxia, NO3 in the cytosol is metabolised to produce nitrite (NO2), which is reduced to form NO via the reductive pathway in the mitochondria. Under low oxygen condition, NO formed in the mitochondria is further reduced to N2O by the reduced form of cytochrome c oxidase (CcO). This pathway is active only when cells experience hypoxia or anoxia, and it may be involved in N2O formation in plants and soil-dwelling animals, as reported previously by several studies. NO can be toxic at a high concentration. Therefore, the reduction of NO to N2O in the mitochondria might protect the integrity of the mitochondria, and thus, protect the cell from the toxicity of NO accumulation under hypoxia and anoxia. As NO3 is a major source of nitrogen for plants and all plants may experience hypoxic and anoxic conditions owing to soil environmental factors, a significant global biogenic source of N2O may be its formation in plants via the proposed pathway.

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.