The effects of warming and elevated ozone (O₃) concentrations on nitrous oxide (N₂O) emission from cropland has received increasing attention; however, the small number of studies on this topic impedes understanding. A field experiment was performed to explore the role of warming and elevated O₃ concentrations on N₂O emission from wheat-soybean rotation cropland from 2012 to 2013 using open-top chambers (OTCs). Experimental treatments included ambient temperature (control), elevated temperature (+2 °C), elevated O₃ (100 ppb), and combined elevated temperature (+2 °C) and O₃ (100 ppb). Results demonstrate that warming significantly increased the accumulative amount of N₂O (AAN) emitted from the soil-winter wheat system due to enhanced nitrification rates in the wheat farmland and nitrate reductase activity in wheat leaves. However, elevated O₃ concentrations significantly decreased AAN emission from the soil-soybean system owing to reduced nitrification rates in the soybean farmland. The combined treatment of warming and elevated O₃ inhibited the emission of N₂O from the soybean farmland. Additionally, both the warming and combined treatments significantly increased soil nitrification rates in winter wheat and soybean croplands and decreased denitrification rates in the winter wheat cropping system. Our results suggest that global warming and elevated O₃ concentrations will strongly affect N₂O emission from wheat-soybean rotation croplands.

The performance, nitrogen removal rate, microbial enzymatic activity and extracellular polymeric substances (EPS) of activated sludge were assessed under nickel (Ni(II)) stress. The organic matter and NH₄⁺-N removal efficiencies were stable at less than 10 mg/L Ni(II) and subsequently decreased with the increment of Ni(II) concentration from 10 to 30 mg/L. The specific oxygen uptake rate and dehydrogenase activity kept stable at less than 5 mg/L Ni(II) and then declined at 5–30 mg/L Ni(II). Both specific ammonia-oxidizing rate (SAOR) and specific nitrite-oxidizing rate (SNOR) decreased with the increment of Ni(II) concentration. The changing trends of ammonia monooxygenase and nitrite oxidoreductase activities were matched those of SAOR and SNOR, respectively. The nitrite-reducing rate and nitrate-reducing rate illustrated a similar variation tendency to the nitrite reductase activity and nitrate reductase activity, respectively. Ni(II) impacted on the production, chemical composition and functional group of EPS. The relation between the sludge volume index and the EPS production exhibited a better linear function with a negative slope, demonstrating that Ni(II) improved the sludge settleability despite of the increase of EPS production.

Hydrogen gas (H₂) has been shown as an important factor in plant tolerance to abiotic stresses, but the underlying mechanisms remain unclear. In the present study, the effects of H₂ and its interaction with nitric oxide (NO) on alleviating cadmium (Cd) stress in Brassica campestris seedlings were investigated. NO donor (SNP) or hydrogen-rich water (HRW) treatment showed a significant improvement in growth of Cd-stressed seedlings. Cd treatment upregulated both endogenous NO and H₂ (36% and 66%, respectively), and the increase of H₂ was prior to NO increase. When treated with NO scavenger (PTIO) or NO biosynthesis enzyme inhibitors (L-NAME and Gln), HRW-induced alleviation under Cd stress was prevented. Under Cd stress, HRW pretreatment significantly enhanced the NO accumulation, and together up-regulated the activity of NR (nitrate reductase) and expression of NR. HRW induced lower reactive oxygen species (ROS), higher AsA content, enhanced activity of POD (peroxidase) and SOD (superoxide dismutase) in seedling roots were inhibited by PTIO, L-NAME and Gln. Through proteomic analysis, the level of 29 proteins were changed in response to H₂ and NO-induced amelioration of Cd stress. Nearly half of them were involved in oxidation-reduction processes (about 20%) or antioxidant enzymes (approximately 20%). These results strongly indicate that in Cd-stressed seedlings, pretreatment with HRW induces the accumulation of H₂ (biosynthesized or permeated), which further stimulates the biosynthesis of NO through the NR pathway. Finally, H₂ and NO together enhance the antioxidant capabilities of seedlings in response to Cd toxicity.

This study investigated the effects of arsenic on the in vitro activities of the enzymes (nitrate reductase and nitrite reductase) involved in nitrate metabolism in the roots, rhizomes, and fronds of four-month old Pteris vittata (arsenic - hyperaccumulator) and Pteris ensiformis (non-arsenic--hyperaccumulator) plants. The arsenic treatments (0, 150, and 300 μM as sodium arsenate) in hydroponics had adverse effects on the root and frond dry weights, and this effect was more evident in P. ensiformis than in P. vittata. Nitrate reductase and nitrite reductase activities of arsenate-treated plants were reduced more in P. ensiformis than in P. vittata. This effect was accompanied by similar decreases in tissue NO₃⁻ concentrations. Therefore, this decrease is interpreted as being indirect, i.e., the consequence of the reduced NO₃⁻ uptake and translocation in the plants. The study shows the difference in the tolerance level of the two Pteris species with varying sensitivity to arsenic. Arsenic reduced the activity of nitrate and nitrite reductase more in Pteris ensiformis than Pteris vittata.

Soil antibiotic resistome and the nitrogen cycle are affected by florfenicol addition to manured soils but their interactions have not been fully described. In the present study, antibiotic resistance genes (ARGs) and nitrogen cycle genes possessed by soil bacteria were characterized using real-time fluorescence quantification PCR (qPCR) and metagenomic sequencing in a short-term (30 d) soil model experiment. Florfenicol significantly changed in the abundance of genes conferring resistance to aminoglycosides, β-lactams, tetracyclines and macrolides. And the abundance of Sphingomonadaceae, the protein metabolic and nitrogen metabolic functions, as well as NO reductase, nitrate reductase, nitrite reductase and N₂O reductase can also be affected by florfenicol. In this way, ARG types of genes conferring resistance to aminoglycosides, β-lactamases, tetracyclines, colistin, fosfomycin, phenicols and trimethoprim were closely associated with multiple nitrogen cycle genes. Actinobacteria, Chlorobi, Firmicutes, Gemmatimonadetes, Nitrospirae, Proteobacteria and Verrucomicrobia played an important role in spreading of ARGs. Moreover, soil physicochemical properties were important factors affecting the distribution of soil flora. This study provides a theoretical basis for further exploration of the transmission regularity and interference mechanism of ARGs in soil bacteria responsible for nitrogen cycle.

The impact of antibiotics on denitrification has attracted widespread attention recently. Norfloxacin, as a representative of fluoroquinolone antibiotics, is extensively detected in groundwater. However, whether the release of norfloxacin into the groundwater poses potential risks to denitrification remains unclear. In this study, effect of norfloxacin on denitrification was investigated. The results showed that increasing norfloxacin from 0 to 100 μg/L decreased nitrate removal rate from 0.68 to 0.44 mg/L/h, but enhanced N₂O emission by 177 folds. Additionally, 100 μg/L of norfloxacin decreased nitrite accumulation by 50.6%. Corresponding inhibition of norfloxacin on bacterial growth, carbon source utilization, electron transport system activity and genes expression was revealed. Furthermore, denitrifying enzyme dynamic monitoring results showed that norfloxacin inhibited nitrate reductase activity, and enhanced nitrite reductase activity to some extent in denitrification process, which was consistent with the variations of nitrate and nitrite. Meanwhile, sensitivity analysis demonstrated that nitrate reductase was more easily affected by norfloxacin than nitrite reductase. Overall, this study suggests that multiple regulation of denitrifying enzyme activity contributes to evaluating the comprehensive effects of antibiotics on groundwater denitrification.

The open top chamber (OTC) method was used in a farmland to study the influence of different levels of O3 concentrations (40 ppb, 80 ppb and 120 ppb) on the enzymatic activity and metabolite contents of the antioxidation system of the winter wheat leaves during the jointing, heading and milk stage. The protective effect of exogenous spermidine (Spd) against the antioxidation of winter wheat under the O3 stress was investigated. With the increasing O3 concentrations and fumigation time, the injuries of the winter wheat leaves were observed to be more serious. For instance, when the O3 concentration reached 120 ppb, the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and nitrate reductase (NR) in the jointing stage decreased by 50.3%, 64.9%, 75.5% and 92.9%, respectively; peroxidase (POD) and glutathione reductase (GR) increased by 45.1% and 80.5%, respectively; the contents of malondialdehyde (MDA), ascorbic acid (AsA) and reduced glutathione (GSH) increased by 314.3%, 8.4% and 31.7%, respectively; and the soluble protein (SP) content decreased by 47.5%. The O3 stress also had significant impact on the contents of proline (Pro), NO3––N and NH4+–N of the winter wheat leaves. During the heading stage, when the O3 concentration was 40 ppb and 80 ppb, the content of Pro was 163.9% and 173.2% higher than that in the control group, respectively. But under 120 ppb, it was decreased by 42.4%. Exogenous application of Spd increased the activities of SOD, POD, CAT, APX and GR, as well as the contents of GSH and SP, but decreased the contents of MDA and AsA. This indicates that Spd is an effective antioxidant to relieve the O3 stress on winter wheat leaves, thereby might be applicable to protect winter wheat from the harm of O3.

The present study investigated nitrogen process in rhizosphere of Kandelia obovata under nitrogen input. Results showed that nitrogen additions significantly increased 4 kinds of enzyme activities (Urease, Nitrate reductase, Nitrite reductase and hydroxylamine reductase). The pH value increased to 7.1 under ammonium addition, but decreased to 6.9 under nitrate addition. Potential Nitrification Intensity (PNI) increased 200–1500% compared with control under ammonium addition, and Potential Denitrification Intensity (PDI) increased more than 200% under nitrate addition. Ten types of organic acids were detected from root exudates, which mainly included oxalic acid, tartaric acid, formic acid, acetic acid, and citric acid. The abundance of 5 kinds of microbial functional groups (nifH, AOA, AOB, nirS, nirK) responded differently. Total nitrogen in organs of K. obovata increased more than 200%. This indicated that nitrogen additions accelerated the transformation of nitrogen directly and stimulated the exudation of root exudates and 5 kinds of genes indirectly.

The aim of this study was to investigate the effect of nitrate on nitrite toxicity to Microcystis aeruginosa. Short-term uptake experiment revealed nitrate could depress the nitrite active uptake. The long-term experiment of nitrate effects on nitrite toxicity showed a negative relationship between the growth and intracellular nitrite levels of M. aeruginosa. The net nitrite uptake displayed a decline in the range of 0–150mg NO₃ ⁻-NL⁻¹ and an elevation at 200mg NO₃ ⁻-NL⁻¹, whereas the activities of nitrate reductase (NR) were more sensitive to nitrate than nitrite reductase (NiR). As a result, the total intracellular nitrite levels tended to decrease up to 100mg NO₃ ⁻-NL⁻¹ then increased at 150–200mg NO₃ ⁻-NL⁻¹. These results suggested the existence of external nitrate could affect the toxicity of nitrite to alga through changing intracellular nitrite content.

Solar ultraviolet radiation (UVR, 280–400nm) is known to inhibit the photosynthesis of macroalgae, whereas nitrogen availability may alter the sensitivity of the algae to UVR. Here, we show that UV-B (280–315nm) significantly reduced the net photosynthetic rate of Gracilaria lemaneiformis. This inhibition was alleviated by enrichment with ammonia, which also caused a decrease in dark respiration. The presence of both UV-A (315–400nm) and UV-B stimulated the accumulation of UV-absorbing compounds. However, this stimulation was not affected by enrichment with ammonia. The content of phycoerythrin (PE) was increased by the enrichment of ammonia only in the absence of UVR. Ammonia uptake and the activity of nitrate reductase were repressed by UVR. However, exposure to UVR had an insignificant effect on the rate of nitrate uptake. In conclusion, increased PE content associated with ammonia enrichment played a protective role against UVR in this alga, and UVR differentially affected the uptake of nitrate and ammonia.