Denitrification Genes nirS Research Articles

Enhancing nitrogen removal through macrophyte harvest and installation of woodchips-based floating beds in surface-flow constructed wetlands

Wetland management maintains nitrogen (N) removal capacity in mature and overgrown constructed wetlands (CWs). We evaluated whether CW management by macrophyte harvesting, and subsequent installation of woodchips-based floating beds (WFBs) planted with Glyceria maxima and Filipendula ulmaria improved N removal. In sixteen heavily overgrown experimental CWs, we applied four treatments: i) only macrophyte harvesting, ii) 5% of the harvested-CW surface covered with WFBs, iii) 20% WFBs cover, and iv) a control treatment (heavily overgrown). N removal was determined in all wetlands at nine occasions. Plant biomass accrual, N assimilation, and denitrification genes nirS, nirK, nosZI and nosZII on plant roots and woodchips from WFBs were estimated. Macrophyte harvesting improved N removal of heavily overgrown CWs, whereas subsequent WFB installation only sometimes improved N removal. Mean N removal efficiencies (± standard deviation) overall were 41 ± 15 %, 45 ± 20 %, 46 ± 16 % and 27 ± 8.3 % for treatments i to iv, respectively. Relative biomass production, root length and root surface area for G.maxima (mean ± standard deviation: 234 ± 114 %, 40 ± 6.5 cm, 6308 ± 1059 cm2g-1, respectively) were higher than those for F. ulmaria (63 ± 86 %, 28 ± 12 cm, 3131 ± 535 cm2g-1, respectively) whereas biomass N assimilation was higher for F. ulmaria (1.8 ± 0.9 gNm−2 of WFB) than for G. maxima (1.3 ± 0.5 gNm−2 of WFB). Denitrification gene abundance was higher on plant roots than on woodchips while G. maxima hosted higher root denitrification gene abundance than F. ulmaria. We conclude that macrophyte harvesting improves N removal in heavily overgrown CWs. WFBs installation has the potential to support plant growth and denitrification in surface-flow constructed wetlands. Further studies need to evaluate the long-term effects of macrophyte harvesting and WFB installation on N removal in CWs.

Microbial Responses to Biochar Soil Amendment and Influential Factors: A Three-Level Meta-Analysis.

Biochar is a multifunctional soil conditioner capable of enhancing soil health and crop production while reducing greenhouse gas emissions. Understanding how soil microbes respond to biochar amendment is a vital step toward precision biochar application. Here, we quantitatively synthesized 3899 observations of 24 microbial responses from 61 primary studies worldwide. Biochar significantly boosts microbial abundance [microbial biomass carbon (MBC) > colony-forming unit (CFU)] and C- and N-cycling functions (dehydrogenase > cellulase > urease > invertase > nirS) and increases the potential nitrification rate by 40.8% while reducing cumulative N2O by 12.7%. Biochar derived at lower pyrolysis temperatures can better improve dehydrogenase and acid phosphatase and thus nutrient retention, but it also leads to more cumulative CO2. Biochar derived from lignocellulose or agricultural biomass can better inhibit N2O through modulating denitrification genes nirS and nosZ; repeated biochar amendment may be needed as inhibition is stronger in shorter durations. This study contributes to our understanding of microbial responses to soil biochar amendment and highlights the promise of purpose-driven biochar production and application in sustainable agriculture such that biochar preparation can be tuned to elicit the desired soil microbial responses, and an amendment plan can be optimized to invoke multiple benefits. We also discussed current knowledge gaps and future research needs.

Nitrogen-cycling genes in oyster reefs and surrounding sediments: Relationships with environmental factors and respective nitrogen rates

To investigate nitrogen (N) cycling in oyster reef habitats along the East coast of Australia, we assessed N-cycling gene abundances in oyster shell biofilms and surrounding sediments, and explored their correlation with environmental factors and respective N rates. We found higher abundances of the denitrification gene nosZII in oyster shell biofilms, while there were not significant differences in the denitrification genes nirS and nirK between oyster biofilms and sediments. Additionally, oyster shell biofilms had a lower (nirS + nirK)/nosZII ratio, indicating a greater capacity for N removal and limited nitrous oxide release compared to sediments. Abundance of nirS, nirK, and dissimilatory nitrate reduction to ammonium (nrfA) genes in sediments decreased with increasing content of organic material, suggesting the influence of large-scale environmental conditions. N-cycling gene abundances did not relate to N rates, emphasising the importance of investigating microbial genes to enhance our understanding of the N cycle in oyster reef habitats.

Soil amendments promoting nitrifying bacteria recovery faster than the denitrifying bacteria at post soil fumigation

Chloropicrin (CP) is a soil fumigant that not only reduces disease-causing pathogenic microbes but regrettably also those that benefit soil quality and crop health.However, we have few knowledge on rapidly restoring populations of beneficial microbes suppressed by CP fumigation. Here we used genetic sequencing technology to monitor changes in the recovery of soil bacteria in response to ammonium sulfate added to the soil following CP fumigation. The results showed that regardless of the N fertilizer addition rate, the accumulated NH4+-N in CP fumigated soil was rapidly consumed within 42 d. The rapid reduction in NH4+-N coincided with the observed recovery nitrogen-cycling microorganisms, especially the nitrification bacteria AOA and AOB that contributed to the formation of NH4+-N. Additionally, we further observed that the resilience index of nitrifying bacteria (AOB and AOA) was greater than the resilience index of denitrifying bacteria that contain the denitrification genes nirS, nirK and nosZ (0.12 to 0.55 vs. −0.27 to 0.073). These results revealed that N fertilizer stimulated the recovery of nitrifying bacteria more than denitrifying bacteria. Our research suggests that ammonium sulfate applied to CP fumigated soil could be used commercially to improve soil health as a result of an increase in beneficial microbes.

Ammonia oxidation and nitrate reduction marker genes are key indicators of nitrogen losses in temperate forest catchments.

Chronic nitrogen inputs can alleviate N limitation and potentially impose N losses in forests, indicated by soil enrichment in 15 N over 14 N. However, the complexity of the nitrogen cycle hinders accurate quantification of N fluxes. Simultaneously, soil ecologists are striving to find meaningful indicators to characterise the "openness" of the nitrogen cycle. We integrate soil δ15 N with constrained ecosystem N losses and the functional gene potential of the soil microbiome in 14 temperate forest catchments. We show that N losses are associated with soil δ15 N and that δ15 N scales with the abundance of soil bacteria. The abundance of the archaeal amoA gene, representing the first step in nitrification (ammonia oxidation to nitrite), followed by the abundance of narG and napA genes, associated with the first step in denitrification (nitrate reduction to nitrite), explains most of the variability in soil δ15 N. These genes are more informative than the denitrification genes nirS and nirK, which are directly linked to N2 O production. Nitrite formation thus appears to be the critical step associated with N losses. Furthermore, we show that the genetic potential for ammonia oxidation and nitrate reduction is representative of forest soil 15 N enrichment and thus indicative of ecosystem N losses.

Recovery trajectories and community resilience of biofilms in receiving rivers after wastewater treatment plant upgrade

Wastewater treatment plant (WWTP) upgrades can reduce both nutrient and micropollutant emissions into receiving rivers, thus modifying the composition and function of biological communities. However, how microbial communities vary and whether they can be restored to levels found in less-polluted rivers remains uncertain. Aquatic biofilms are sensitive to environmental change and respond rapidly to bottom-up pressure. Thus, we used 12 flumes configured in three experimental treatments to mimic the dynamic processes of biofilm microbial communities occurring in a wastewater-receiving river following WWTP upgrade, with rivers containing two levels of nutrients and micropollutants used as references. We compared the biofilm microbial biomass, carbon source utilization, and community composition among the three “blocks”. Results showed that the metabolic patterns of the carbon sources and composition of the biofilm bacterial communities in the flumes mimicking a receiving river with WWTP upgrade recovered over time to those mimicking a less-disturbed river. The restoration of potential carboxylic acid-consuming denitrifying bacteria (i.e., Zoogloea, Comamonas, Dechloromonas, and Acinetobacter) likely played a significant role in this process. Combining quantitative analysis of the denitrification genes nirS and nosZ, we confirmed that the denitrification function of the river biofilms recovered after WWTP upgrade, consistent with our previous field investigation.

Relative importance of plant uptake and plant associated denitrification for removal of nitrogen from mine drainage in sub-arctic wetlands

Reactive nitrogen (N) species released from undetonated ammonium-nitrate based explosives used in mining or other blasting operations are an emerging environmental problem. Wetlands are frequently used to treat N-contaminated water in temperate climate, but knowledge on plant–microbial interactions and treatment potential in sub-arctic wetlands is limited. Here, we compare the relative importance of plant uptake and denitrification among five plant species commonly occurring in sub-arctic wetlands for removal of N in nitrate-rich mine drainage in northern Sweden. Nitrogen uptake and plant associated potential denitrification activity and genetic potential for denitrification based on quantitative PCR of the denitrification genes nirS, nirK, nosZI and nosZII were determined in plants growing both in situ and cultivated in a growth chamber. The growth chamber and in situ studies generated similar results, suggesting high relevance and applicability of results from growth chamber experiments. We identified denitrification as the dominating pathway for N-removal and abundances of denitrification genes were strong indicators of plant associated denitrification activity. The magnitude and direction of the effect differed among the plant species, with the aquatic moss Drepanocladus fluitans showing exceptionally high ratios between denitrification and uptake rates, compared to the other species. However, to acquire realistic estimates of N-removal potential of specific wetlands and their associated plant species, the total plant biomass needs to be considered. The species-specific plant N-uptake and abundance of denitrification genes on the root or plant surfaces were affected by the presence of other plant species, which show that both multi- and inter-trophic interactions are occurring. Future studies on N-removal potential of wetland plant species should consider how to best exploit these interactions in sub-arctic wetlands.

Isolation and characterization of a denitrifying Acinetobacter baumannii H1 using NO2--N as nitrogen source from shrimp farming ponds

Recent studies have revealed that the use of denitrifying bacteria is an alternative to eliminate nitrite accumulation in the wastewaters. However, no information is available about Acinetobacter species as a denitrifying agent for reducing nitrite in aquaculture. In the present study, a potential denitrifying strain H1 was isolated from shrimp farming ponds using nitrite as nitrogen source, which displayed almost complete nitrite removal. It was initially identified as Acinetobacter baumannii using API identification kits, and confirmed to be A. baumannii strain (GenBank accession no. JF751054) by phylogenetic analysis. Three key denitrification genes nirS, norB and nirK were detected from strain H1 by PCR amplification. In addition, it was demonstrated to be safe for mammals and giant freshwater prawns, and showed in vitro high nitrite removal activities at a wide range of initial pH 5-9 and 15 to 35°C with a final cell density of 105-107 cfu/ml. To the best of our knowledge, this is the first report on a promising denitrifying A. baumannii strain from shrimp farming ponds. Key words: Characterization, nitrite removal, Acinetobacter baumannii, shrimp farming.

Response of denitrification genes nirS, nirK, and nosZ to irrigation water quality in a Chinese agricultural soil

Denitrification is an important biochemical process in global nitrogen cycle, with a potent greenhouse gas product N(2)O. Wastewater irrigation can result in the changes of soil properties and microbial communities of agricultural soils. The purpose of this study was to examine how the soil denitrification genes responded to different irrigation regimes. Soil samples were collected from three rural districts of Beijing (China) with three different irrigation regimes: clean groundwater (CW), reclaimed water (RW), and wastewater (WW). The abundance and diversity of three denitrification microbial genes (nirS, nirK, and nosZ) were examined by real-time polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) molecular approaches. The abundance of nirS in the WW treatment was higher than that in the CW treatment, and no significant difference was found between the RW and CW or WW treatments. The abundance of nirK gene of the RW and WW treatments was higher than that of the CW treatment. There was no difference for nosZ gene among the three treatments. Correspondence analysis based on the DGGE profiles showed that there was no obvious difference in the nosZ gene composition, but nirS and nirK genes changed with different irrigation regimes. Irrigation with unclean water sources enhanced the soil NO (3) (-) content and changed the abundance and composition of soil denitrifiers, and different functional genes had different responses. Irrigation with unclean water sources increased the abundance of nirK gene and changed the community structures of nirS and nirK genes, while nosZ gene was relatively stable in the soil. These results could be helpful to explore the mechanisms of the variation of denitrification processes under long-term wastewater irrigation and partially explain the reason of more N(2)O output in the field with wastewater irrigation.