nirK Gene Copy Research Articles

Dual mechanism of membrane covering on GHG and NH3 mitigation during industrial-scale experiment on dairy manure composting: Inhibiting formation and blocking emissions

An industrial-scale experiment on dairy manure composting with the control group (Ctrl) and the membrane covering group (CM) was conducted to explore the effects of functional membrane covering on gas emissions, the conversion of carbon and nitrogen, and revealing the underlying mechanisms. Results indicated that CM achieved the synergistic effects on gas mitigation and improved compost product quality. CO2, CH4, N2O, and NH3 emissions were reduced by 81.8%, 87.0%, 82.6%, and 82.2%, respectively. The micro-aerobic condition formed in membrane covering compost pile together with the covering inhibiting effect dominated the mitigation effect. CM significantly downregulated the mcrA gene copies and the value of mcrA/pmoA (p < 0.01), which reduced CH4 emission. CM decreased the nirS and nirK gene copies and increased the nosZ gene copies to reduce N2O emission. Functional Annotation of Prokaryotic Taxa showed that membrane covering effectively amended part of carbon and nitrogen cycles, which stimulated the degradation of organic matter, accelerated compost maturity and reduced the gaseous emissions.

Maize lowers the N2O emissions from maize/soybean intercropping

Interspecific interactions between intercropped maize and soybean are expected to cycle soil nitrogen efficiently and avoid N2O emissions. However, the unique interactions of maize and soybean with nitrogen cycling bacteria makes it hard to predict the crop-specific impact on soil N2O production. We conducted a field microcosm experiment with root separation to simulate monoculture and intercropping with maize and soybean. Intercropped maize reduced the N2O emissions by 16–41%, compared to monoculture maize. This was related to reduced nitrification by ammonia-oxidizing bacteria and denitrification reactions (as indicated by the abundance of nirS and nirK gene copies), as well as drier soil conditions and lower soil NO3−-N levels. Soil N2O emissions were the same in monoculture and intercropped soybean, suggesting stable denitrification (based on nirS, nirK and nosZ genes) with constant soil physicochemical conditions. As monoculture maize can stimulate soil N2O emission through denitrification, this work justifies the adoption of maize-soybean intercropping as a low N2O-emitting practice in sustainable agriculture, due to its beneficial effects on soil biology and biochemistry.

Effect of n-hexadecanoic acid on N2O emissions from vegetable soil and its synergism with Pseudomonas stutzeri NRCB010

Recently, it has been recommended to utilize specific plant root metabolites to decrease nitrous oxide (N2O) emissions from agricultural soil. However, only a limited number of plant root metabolites have been shown to decrease soil N2O emissions, and their combined effects with plant growth-promoting rhizobacteria, as well as the mechanisms involved in mitigating N2O emissions, remain to be explored. This study investigated the impact and microbial mechanisms of three tomato root exudate metabolites, namely, methyl hexadecanoate, methyl stearate, and n-hexadecanoic acid, on N2O emissions. In a pure culture experiment, n-hexadecanoic acid significantly enhanced Pseudomonas stutzeri NRCB010 auxin production, nitrogen removal, N2O reduction capacity, and N2O reductase activity. Methyl stearate significantly promoted NRCB010 N removal and the N2O reduction capacity. In soil microcosm and greenhouse pot experiments, n-hexadecanoic acid decreased N2O cumulative emissions by 34.3 % and 46.6 %, respectively, and further decreases were observed when combined with NRCB010. Additionally, n-hexadecanoic acid, both independently and in combination with NRCB010, significantly promoted tomato growth. N-hexadecanoic acid, alone or with NRCB010, also altered soil microbial communities and nitrogen-cycle functional gene abundance. Structural equation models revealed that n-hexadecanoic acid, either alone or with NRCB010, decreased N2O cumulative emissions by modifying the environmental conditions to be more conducive to N2O mitigation (e.g., higher soil pH and organic matter content), inhibiting N2O production (through a decrease in ammonia-oxidizing bacteria and nirK gene copy numbers), and promoting N2O reduction (by increasing nosZII gene copy numbers). Our results support the potential use of n-hexadecanoic acid in enhancing crop productivity and mitigating N2O emissions in agricultural soils.

Seasonal Variability of Cultivable Nitrate-Reducing and Denitrifying Bacteria and Functional Gene Copy Number in Fresh Water Lake.

This study describes the seasonal course of denitrifying and nitrate-reducing bacteria in a dimictic mesotrophic lake (Lake Scharmützelsee, Brandenburg, Germany) within a three-year period from 2011 to 2013. The bacterial cell numbers were quantified by the fluorescence microscopy, most probable number (MPN) and PCR-dependent quantification of the chromosomal 16S rDNA and of the nirS and nirK gene copy number. The highest seasonal differences (up to three orders of magnitudes) have been measured using MPN in the epilimnion. This variation was not reflected by PCR-dependent approaches or direct microscopical enumeration. At adverse conditions (low temperature and/or low nitrate concentrations), the differences between MPN and gene copy numbers increased by up to five orders of magnitudes and decreased to one magnitude at favourable environmental conditions. These results can be explained best by an increasing ratio of viable but not cultivable (VBNC) cells or dead cells at impairing conditions. In the hypolimnion, the courses of MPN and nir gene copy numbers were similar. This can be explained by a higher feeding pressure and therefore smaller amounts of dormant cells. In the pelagial in general, the total cell numbers enumerated by either microscopical or molecular approaches were similar. In the sediment, more than 99% of the DNA was obviously not related to viable bacteria but was rather DNA in dead cells or adsorbed to particle surfaces.

Biochar Amendment in Vermi-Wetland for Enhancing Nitrification during Excess Sludge Recycling

Vermi-wetland is a sustainable technology for recycling excess sludge in small-town areas. Although biochar (BC) amendment into the vermi-wetland could considerably boost the effectiveness of treating sludge, its impact on the nitrogen transformation in vermi-wetland remains unclear. Hence, this study aimed to explore the mechanism and performance of BC amendment into the vermi-wetland for enhancing nitrogen transformation during excess sludge recycling. The semi-aquatic plant Acorus calamus and the earthworm Eisenia fetida were planted in the designed vertical vermi-reactor, with corncob BC added to the upper and lower layers of one vermi-reactor, in comparison with the vermi-reactor without BC. The vermi-reactor with BC significantly lowered (p &lt; 0.05) ammonia nitrogen (NH4+-N) and nitrite nitrogen (NO2−-N) in the effluent by 1.63 and 4.85-fold, respectively, and increased considerably nitrate nitrogen (NO3−-N) in the effluent by 1.5-fold. The numbers of ammonia-oxidizing bacteria (AOB) and archaea (AOA) in the vermi-reactor with BC were greatly enriched by 6 and 1.42-fold, compared with their counterparts (p &lt; 0.05). Moreover, nirS and nirK gene copies in the vermi-reactor with BC were considerably improved (p &lt; 0.05) by 2.03 and 1.82-fold, respectively. BC significantly enhanced the growth of earthworms by 6.92-fold and promoted plant growth by 1.28-fold. In addition, the AOB members like Nitrosomonas and Nitrosospira and the AOA members like Crenarchaeota and Thaumarchaeota cohabited in BC. Overall, these results suggest that a vermi-reactor amended with BC could enhance the nitrification processes of excess sludge, thereby improving the treatment performance of vermi-wetland.

Long-term effects of Ag NPs on denitrification in sediment: Importance of Ag NPs exposure ways in aquatic ecosystems

The widespread use of silver nanoparticles (Ag NPs) inevitably leads to their increasing release into aquatic systems, with studies indicating that the mode of Ag NPs entry into water significantly affects their toxicity and ecological risks. However, there is a lack of research on the impact of different exposure ways of Ag NPs on functional bacteria in sediment. This study investigates the long-term influence of Ag NPs on denitrification process in sediments by comparing denitrifies responses to single (pulse injection of 10 mg/L) and repetitive (1 mg/L × 10 times) Ag NPs treatments over 60-day incubation. Results showed that a single exposure of 10 mg/L Ag NPs caused an obvious toxicity on activity and abundance of denitrifying bacteria on the first 30 days, reflecting by the decreased NADH amount, ETS activity, NIR and NOS activity, and nirK gene copy number, which resulted in a significant decline of denitrification rate in sediments (from 0.59 to 0.64 to 0.41–0.47 μmol15N L−1 h−1). While inhibition was mitigated with time and denitrification process recovered to the normal at the end of the experiment, the accumulated nitrate generated in the system showed that the recovery of microbial function did not mean the restoration of aquatic ecosystem after pollution. Differently, the repetitive exposure of 1 mg/L Ag NPs exhibited the evident inhibition on metabolism, abundance, and function of denitrifiers on Day 60, due to the accumulated amount of Ag NPs with the increased dosing number, indicating that the accumulated toxicity on functional microorganic community of repetitive exposure in less toxic concentration. Our study highlights the importance of Ag NPs entry pathways into aquatic ecosystem on their ecological risks, which affected dynamic responses of microbial function to Ag NPs.

Citric acid modified biochar application at a low dosage can synchronically mitigate the nitrogenous gas pollutants emission from rice paddy soils

Raw biochar with high pH possibly stimulated ammonia (NH3) volatilization in the agricultural soil. We hypothesized that the modified biochar (MBC) with low pH can synchronically decrease the NH3 and nitrous oxide (N2O) losses. We performed a two-year experiment to clarify how citric acid MBC influence the NH3 volatilization and N2O emission as well as the underlying mechanisms. Two typical paddy soils, i.e., Hydragric Anthrosol and Haplic Acrisol, receiving equal urea N with 240 kg ha−1 but varied rates of MBC with 0, 5, 10, and 20 t ha−1 (named Urea, Urea + MBC5, Urea + MBC10, and Urea + MBC20, respectively) were studied. The results showed that MBC-amended treatments effectively mitigated the NH3 volatilization from Hydragric Anthrosol and Haplic Acrisol by 29.6%–57.9% and 30.5%–62.4% in 2017, and by 16.5%–21.0% and 24.5%–35.0% in 2018, respectively, compared to Urea treatment. In addition, significantly lower N2O emissions with averaged 38.3% and 43.1% in 2017, and 51.7% and 26.7% were recorded under Hydragric Anthrosol and Haplic Acrisol, respectively, following the MBC application (P < 0.05). Increased MBC addition performed higher efficacy on mitigating NH3 volatilization, particularly in the first rice season, while this “dosage effect” was not found for N2O reduction. Lowered pH in overlying water, enhanced adsorption of NH4+-N and its nitrification rate likely contributed to the lower NH3 volatilization as result of MBC addition. The nirS and nosZ gene copies were not changed by MBC, while the nirK gene copies were decreased as result of MBC amendment by 8.3%–25.2% under Hydragric Anthrosol and by 21.8%–24.9% under Haplic Acrisol. Consequent lower ratio of nirK/(nirS + nosZ) explained the mitigation effect of MBC on N2O emission. In conclusion, the present two-year study recommends that MBC applied at a low dosage can perform positive effect on controlling the nitrogenous gas pollutants from paddy soil.

Saline-alkali soil applied with vermicompost and humic acid fertilizer improved macroaggregate microstructure to enhance salt leaching and inhibit nitrogen losses

High salinity, poor soil structure and nutrient deficiencies are the main limitations in saline-alkali soils. We investigated the effects on the microstructure, and the aerobic and anaerobic bacterial communities in the macroaggregates following addition of vermicompost and humic acid fertilizer to saline-alkali soil. These effects could enhance salt leaching and inhibit nitrogen losses in the topsoil. Vermicompost and humic acid fertilizer improved soil aggregate stability and the microstructure of macroaggregates (porosity, path tortuosity and effective porosity), which in turn increased soil permeability and salt leaching in saline-alkali topsoil. The improvement in aggregate microstructure affected the soil bacterial community of the macroaggregates by increasing the relative abundances of aerobic heterotrophs, such as Chloroflexi and Bacteroidetes. Additionally, vermicompost and humic acid fertilizer influenced nitrogen cycling by the macroaggregate microbial community, and stimulated nitrification by increasing archaeal and bacterial amoA gene copy numbers in the topsoil, but inhibited nitrite-reducing bacteria by decreasing nirS and nirK gene copy numbers. Furthermore, these amendments increased microbial N2O reduction by increasing nosZ gene copy numbers, suggesting that vermicompost and humic acid fertilizer have the potential to reduce N2O emissions in saline-alkali soil. In summary, vermicompost and humic acid fertilizer can ameliorate saline-alkali soils by replenishing organic matter, improving aggregate microstructure, and regulating the microbial community, which could increase salt leaching and inhibit nitrogen losses.

Effects of repeated applications of urea with DMPP on ammonia oxidizers, denitrifiers, and non-targeted microbial communities of an agricultural soil in Queensland, Australia

Nitrification inhibitors have been reported to reduce nitrous oxide emission and nitrate leaching in agricultural systems. The effects of repeated applications of urea alone or in combination with nitrification inhibitors on nitrogen (N) cycling microbes involved in nitrification and denitrification together with non-targeted microbes are not well understood. Therefore, the objective of this study was to investigate the effects of repeated application of urea and DMPP on soil physio-chemistry, ammonia oxidizers and total bacteria in the soil. We collected soil samples from a 4.5-year field experiment under crop rotation with repeated application of seven treatments, namely control (CK), Urea (U), Urea + DMPP (UE) applied at 40, 80 and 120 kg N ha−1, each treatment with three replicates. Ammonia-oxidizing bacteria (AOB) gene copy numbers increased as the N application rate increased (from 0 to 120 kg N ha−1). The use of DMPP significantly reduced AOB and nirK gene copy numbers compared to urea alone at an application rate of 120 kg N ha−1. There was no treatment effect on the abundance of ammonia-oxidizing archaea (AOA), Comammox clade A and B, nosZ and bacterial 16S rRNA genes. The community composition of AOB and AOA changed with N addition and use of DMPP but increasing N addition rate changed the composition of AOB only. Addition of N increased potential nitrification rates at 80 and 120 kg N ha−1. There was no significant treatment effect on the relative abundance of bacteria at the phylum level. This experiment demonstrated that the application of N (with or without DMPP) at rates lower than 120 kg N ha−1 would not result in significant impacts on soil archaeal and bacterial ecology.

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

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

Nitrous oxide emission and denitrifier communities in drip-irrigated calcareous soil as affected by chemical and organic fertilizers

The effects of consecutive application of chemical fertilizer with or without organic fertilizer on soil N2O emissions and denitrifying community structure in a drip-irrigated field were determined. The four fertilizer treatments were (i) unfertilized, (ii) chemical fertilizer, (iii) 60% chemical fertilizer plus cattle manure, and (iv) 60% chemical fertilizer plus biofertilizer. The treatments with organic amendments (i.e. cattle manure and biofertilizer) reduced cumulative N2O emissions by 4.9–9.9%, reduced the N2O emission factor by 1.3–42%, and increased denitrifying enzyme activities by 14.3–56.2%. The nirK gene copy numbers were greatest in soil which received only chemical fertilizer. In contrast, nirS- and nosZ-copy numbers were greatest in soil amended with chemical fertilizer plus biofertilizer. Chemical fertilizer application with or without organic fertilizer significantly changed the community structure of nirK-type denitrifiers relative to the unfertilized soil. In comparison, the nirS- and nosZ-type denitrifier genotypes varied in treatments receiving organic fertilizer but not chemical fertilizer alone. The changes in the denitrifier communities were closely associated with soil organic carbon (SOC), NO3−, NH4+, water holding capacity, and soil pH. Modeling indicated that N2O emissions in this soil were primarily associated with the abundance of nirS type denitrifying bacteria, SOC, and NO3−. Overall, our findings indicate that (i) the organic fertilizers increased denitrifying enzyme activity, increased denitrifying-bacteria gene copy numbers, but reduced N2O emissions, and (ii) nirS- and nosZ-type denitrifiers were more sensitive than nirK-type denitrifiers to the organic fertilizers.

Optimization of moving bed biofilm reactors for oil sands process-affected water treatment: The effect of HRT and ammonia concentrations

Two moving bed biofilm reactors (MBBRs) were optimized to improve the biodegradation of organic compounds in raw and ozonated OSPW by changing the hydraulic retention time (HRT) and the influent ammonia concentrations. During the five stages, the average COD removal reached 50.8±3.4%, 52.8±6.5%, 54.7±4.3%, 56.3±2.2%, and 58.0±2.3% respectively in raw OSPW MBBR, and 54.6±3.8%, 57.2±7.1%, 55.5±5.8%, 58.3±2.2%, and 60.7±2.3% respectively in ozonated OSPW MBBR. Welch's weighted ANOVA tests show that the increase in ammonia levels significantly improved the COD removal in the two systems, while the HRT was an important parameter for COD decrease in the raw OSPW MBBR. Compared to the HRT, the increase in ammonia concentrations were more beneficial for acid extractable fraction (AEF) degradation and the average AEF removal reached 29.80% (raw OSPW MBBR) and 16.50% (ozonated OSPW MBBR) by the end of the optimization (Stage V; HRT=96h, 60mg/L NH4+-N). >98% of the NH4+-N was removed in the two MBBR systems, showing good nitrification. Microtoxicity tests showed that no significant correlations were found between HRT/ammonia levels and the OSPW toxicity changes toward V. fischeri. Spearman's rank correlation analysis was applied for q-PCR data, showing that positive correlations between the removal efficiencies of AEF and NSR and NirK gene copies were observed in the raw OSPW MBBR system, while positive correlations between AEF removal efficiency and total bacteria gene, NSR, Nitro, and NirK gene copies were observed in the ozonated OSPW MBBR system.

Simulated animal trampling of a free‐draining stony soil stimulated denitrifier growth and increased nitrous oxide emissions

AbstractWinter forage grazing systems in New Zealand cause compaction of soil by grazing animals, especially when the soil is wet. However, there is little information on the effects of animal trampling on denitrifiers in soil, despite their importance for N2O production. Here, we report a field study of the abundance of the denitrifying genes nirS, nirK, and nosZ and N2O emissions following the application of dairy cow urine in a free‐draining stony soil. Importantly, we found that simulated animal trampling altered some of the denitrifying microbial communities, thus leading to increased N2O emissions. Over the 111 day measurement period, the abundance of nitrite (NO2−)‐reducing nirS gene copy numbers increased significantly by 87% in the trampled soil with urine (P &lt; 0.01) and increased by 40% in the trampled soil without urine (P &lt; 0.05), but the nirS gene abundance did not change significantly in the nontrampled soil. The abundance of NO2− reducing nirK gene copy numbers was not affected by trampling, but increased significantly following urine application. The abundance of N2O‐reducing nosZ clade I and nosZ clade II gene copy numbers increased significantly in the trampled soil, but did not change significantly in the nontrampled soil. N2O emissions from the trampled soil were about twice that from the nontrampled soil without urine (1.20 and 0.62 kg N2O‐N per ha, respectively) and about eight times greater (6.24 kg N2O‐N per ha) than from nontrampled soil (0.80 kg N2O‐N per ha) when urine was applied. These results strongly suggest that animal trampling during winter forage grazing can have a major impact on denitrifying communities in soil, which in turn stimulate greater denitrification with increased N2O emissions.

An endophytic bacterium Acinetobacter calcoaceticus Sasm3-enhanced phytoremediation of nitrate–cadmium compound polluted soil by intercropping Sedum alfredii with oilseed rape

Intensive agricultural system with high input of fertilizer results in high agricultural output. However, excessive fertilization in intensive agricultural system has great potential to cause nitrate and heavy metal accumulation in soil, which is adverse to human health. The main objective of the present study was to observe the effects of intercropping and inoculation of endophytic bacterium Acinetobacter calcoaceticus Sasm3 on phytoremediation of combined contaminated soil in oilseed rape (Brassica napus L.). The results showed that with Sasm3 inoculation, the biomass of rape was increased by 10-20% for shoot, 64% for root, and 23-29% for seeds while the nitrate accumulation in rape was decreased by 14% in root and by 12% in shoot. The cadmium concentration in rape increased significantly with mono-inoculating treatment, whereas it decreased significantly after intercropping treatment. By denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR analysis, the diversity of bacterial community and the number of nirS and nirK gene copies increased significantly with inoculation or/and intercropping treatment. In conclusion, the endophytic bacterium Sasm3-inoculated intercropping system not only improved the efficiency of clearing cadmium from soil without obstructing crop production, but also improved the quality of crop.

Influence of soil bulk density and matric potential on microbial dynamics, inorganic N transformations, N2O and N2 fluxes following urea deposition

Transformation of ruminant urine-nitrogen (N) can contribute to negative environmental outcomes such as nitrate leaching which leads to eutrophication of waterways and production of nitrous oxide (N2O), a greenhouse gas. Although abiotic factors influencing urine-N processing have been well studied, detailed studies of the soil microbial community dynamics following urine application are required to improve mitigation strategies for reducing harmful N fluxes from urine deposition. A factorial laboratory experiment using packed silt-loam soil cores with two levels each of urea (±), soil matric potential (ψ −6.0 or −0.2 kPa) and soil bulk density (ρb 1.1 or 1.5 g cm−3) was performed to study the interaction of urea and soil physical conditions on both soil inorganic N transformations and the abundance of ammonia-oxidizers and denitrifiers. Soil ψ and ρb treatments had an immediate impact on soil pH, dissolved organic carbon, inorganic N pools and emissions of N2O and N2 following urea deposition, and microorganisms carrying the nosZ gene were present in lower numbers in the most aerobic soil (−6.0 kPa and 1.1 g cm−3) from day 7. In all treatments, both bacterial amoA and denitrifier nirS, nirK and nosZ gene copy numbers increased within 1 day following urea application. Dynamics in the NH4+ concentrations were significantly correlated with cumulative changes in the abundance of the ammonia-oxidizers, but no relation was found between cumulative changes in the denitrifier nirS, nirK and nosZ gene copy numbers and the dynamics in soil inorganic N, N2O or N2 emissions. Throughout most of the study period the specific soil conditions, induced by the ψ and ρb treatments, determined nitrifier and denitrifier activity rather than the size of the microbial communities involved. However, by day 35 soil ψ and ρb treatments exerted large treatment effects on bacterial amoA, nirS and nirK gene copy numbers. Thus, although nitrate concentrations and N2O emissions following urea deposition were determined by the soil ψ and ρb conditions in the short-term, our results indicate that changes in the population sizes of denitrifiers and AOB in ruminant urine patches may influence environmental N fluxes in the long-term.

Bacterial community structure of nirK-bearing denitrifiers and the development of properties of soils in created mitigation wetlands

We investigated the abundance and genetic heterogeneity of bacterial nitrite reductase genes (nir) and soil structural properties in created and natural freshwater wetlands in the Virginia piedmont. Soil attributes included soil organic matter (SOM), total organic carbon (TOC), total nitrogen (TN), pH, gravimetric soil moisture (GSM), and bulk density (Db). A subset of soil attributes were analyzed across the sites, using euclidean cluster analysis, resulting in three soil condition (SC) groups of increasing wetland soil development (i.e., SC1<SC2<SC3; less to more developed or matured) as measured by accumulation of TOC, TN, the increase of GSM, and the decrease of Db. There were no difference found in the bacterial community diversity between the groups (p=0.4). NirK gene copies detected ranged between 3.6×104 and 3.4×107copiesg−1 soil and were significantly higher in the most developed soil group, SC3, than in the least developed soil group, SC1. However, the gene copies were lowest in SC2 that had a significantly higher soil pH (~6.6) than the other two SC groups (~5.3). The same pattern was found in denitrifying enzyme activity (DEA) on a companion study where DEA was found negatively correlated with soil pH. Gene fragments were amplified and products were screened by terminal restriction fragment length polymorphism (T-RFLP) analysis. Among 146 different T-RFs identified, fourteen were dominant and together made up more than 65% of all detected fragments. While SC groups did not relate to whole nirK communities, most soil properties that identified SC groups did significantly correlate to dominant members of the community.

Indirect Effects and Biological Control of Mosquitoes by Mosquitofish

The community structure and abundance of nirK-type denitrifying bacteria in different soil layers (0-20 cm and 20-40 cm) under various fertilization regimes in Wuwei, Gansu Province were investigated by the combination of terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR. Results showed that the nirK-type denitrifying bacteria community structure was significantly affected by fertilization regimes, especially for 70, 156 and 190 bp T-RFs that represented the dominant populations in greenhouse soil. Fertilization regimes significantly influenced the abundance of nirK gene in the 0-20 cm soil layer with the highest abundance of nirK gene copy number (2.16 x 10(7) copies x g(-1) soil) detected in the manure treatment (M), which was 2.04 and 2.02 times of that in the control (CK) and chemical fertilizer (NPK) treatments, respectively. Both the dominant population and abundance of nirK-type denitrifying bacteria in the greenhouse soil were significantly different between the 0-20 cm and 20-40 cm soil layers, and the nirK-type denitrifying bacteria community structure and abundance in the greenhouse soil were obviously different from that in the field. Soil pH, soil organic matter content and nitrate-N content had the greatest influence on the bacterial community composition. Phylogenetic analysis indicated that there were not only anaerobic nirK-type denitrifying bacteria in greenhouse soil, but also aerobic denitrifying bacteria, such as Rhizobium, Ochrobactrum, Agrobacterium.