Impacts of continuous excessive fertilization on soil potential nitrification activity and nitrifying microbial community dynamics in greenhouse system

Purpose

Sampling and analysis of greenhouse soils were conducted in Shouguang, China, to study continuous excessive fertilization effect on nitrifying microbial community dynamics in greenhouse environment.

Materials and methods

Potential nitrification activity (PNA), abundance, and structure of nitrifying microbial communities as well as the correlations with soil properties were investigated.

Results and discussion

Short-term excessive fertilization increased soil nutrient contents and the diversity of nitrifying microbial communities under greenhouse cultivation. However, the abundance and diversity of nitrifying communities decreased greatly due to the increase of soil acidity and salinity after 14 years of high fertilization in greenhouse. There was a significant positive correlation between soil PNA and the abundance of ammonia-oxidizing bacteria (AOB) but not that of ammonia-oxidizing archaea (AOA) in topsoil (0–20 cm) when pH ≥7. Soil PNA and AOB were strongly influenced by soil pH. The groups of Nitrososphaeraceae, Nitrosomonadaceae, and Nitrospiraceae were predominant in the AOA, AOB, and nitrite-oxidizing bacteria (NOB) communities, respectively. Nitrifying community structure was significantly correlated with soil electrical salinity (EC), organic carbon (OC), and nitrate nitrogen (NO₃ ^?–N) content by redundancy analysis (RDA).

Conclusions

Nitrification was predominated by AOB in greenhouse topsoil with high fertilizer loads. Soil salinity, OC, NO₃ ^?–N content, and pH affected by continuous excessive fertilization were the major edaphic factors in shaping nitrifying community structure in greenhouse soils.

     

Management practices have a major impact on nitrifier and denitrifier communities in a semiarid grassland ecosystem

Purpose

Nitrification and denitrification, two of the key nitrogen (N) transformation processes in the soil, are carried out by a diverse range of microorganisms and catalyzed by a series of enzymes. Different management practices, such as continuous grazing, mowing, and periodic fencing off from grazing, dramatically influenced grassland ecosystems. This study aimed to examine the effects of management practices on the abundance and community structure of nitrifier and denitrifier communities in grassland ecosystems.

Materials and methods

Soil samples were collected from a semiarid grassland ecosystem in Xilingol region, Inner Mongolia, where long-term management practices including free-grazing, different periods of enclosure from grazing, and different frequencies of mowing were conducted. Real-time quantitative polymerase chain reaction (Q-PCR), denaturing gradient gel electrophoresis (DGGE), sequencing, and phylogenetic analysis were applied to estimate the abundance and composition of amoA, nirS, nirK, and nosZ genes.

Results and discussion

The ammonia-oxidizing archaea (AOA) amoA copies were in the range 5.99?×?10⁸ to 8.60?×?10⁸, while those of ammonia-oxidizing bacteria (AOB) varied from 3.02?×?10⁷ to 4.61?×?10⁷. The abundance of AOA was substantially higher in the light grazing treatment (LG) than in the mowing treatments. The quantity and intensity of DGGE bands of AOA varied with pasture management. In stark contrast, AOB population abundance and community structure remained largely unchanged in all the soils irrespective of the management practices. All these results suggested that ammonia oxidizers were dominated by AOA. The higher gene abundance and greater intensity of DGGE bands of nirS and nosZ under the enclosure treatments would suggest greater stimulated denitrification. The ratio of nosZ/(nirS?+?nirK) was higher in mowing treatments than in the free-grazing and enclosure treatments, possibly leading to more complete denitrification. Correlation analysis indicated that soil moisture and inorganic nitrogen content were the two main soil environmental variables that influence the community structure of nitrifiers and denitrifiers.

Conclusions

In this semiarid neutral to alkaline grassland ecosystem under low temperature conditions, AOA mainly affiliated with Nitrososphaera dominated nitrification. These results clearly demonstrate that grassland management practices can have a major impact on nitrifier and denitrifier communities in this semiarid grassland ecosystem, under low temperature conditions.

     

Determinants of the biodiversity patterns of ammonia-oxidizing archaea community in two contrasting forest stands

Purpose

The variation in soil microbial community patterns is primarily influenced by ecological processes associated with spatial distance and environmental heterogeneities. However, the relative importance of these processes in determining the patterns of soil microbial biodiversity in different successional forests remains unclear.

Materials and methods

Based on the species data from denaturing gradient gel electrophoresis (DGGE) analysis, we described the composition and beta diversity of ammonia-oxidizing archaea (AOA) community, an important functional microbial group in regulating nitrogen cycle, in a middle-succeed stand (60 years of secondary succession) and an undisturbed native stand in a subtropical forest in southern China. The composition pattern was examined using a multi-response permutation procedure (MRPP), and the beta diversity was described using the Sørensen index. The relative influence of edaphic, vegetational, spatial, and topographical factors on AOA composition and beta diversity was assessed by variation partitioning and multiple regression on distance matrices (MRM), respectively.

Results and discussion

We did not find any stand-specific patterns in AOA community composition in the two stands; however, the influential variables were different between the two stands; 7.3 and 4.5 % of the total variation in AOA community composition could be explained by edaphic (i.e., available potassium and total phosphorus) and spatial variables, respectively, in the middle-succeed stand, while 3.7 and 2.8 % of the variation were explained by spatial variable and available phosphorus, respectively, in the native stand. Soil total phosphorus influenced the beta diversity of AOA community most in the middle-succeed stand, while genetic distance of tree species was found to be the most important factor in driving the beta diversity pattern in the native stand.

Conclusions

Soil nutrients influenced the beta diversity of AOA community in the middle-succeed stand more than that in the native stand, while vegetation is more important in the native stand. The substantial unexplained variations were possibly due to the effects of other unmeasured variables. Nevertheless, dispersal process is more important in controlling AOA community composition in the native stand, while processes associated with environmental heterogeneities are more important in the middle-succeed stand in this subtropical forest.

     

Comammox—a newly discovered nitrification process in the terrestrial nitrogen cycle

Purpose

Nitrification, the microbial oxidation of ammonia to nitrate via nitrite, is a pivotal component of the biogeochemical nitrogen cycle. Nitrification was conventionally assumed as a two-step process in which ammonia oxidation was thought to be catalyzed by ammonia-oxidizing archaea (AOA) and bacteria (AOB), as well as nitrite oxidation by nitrite-oxidizing bacteria (NOB). This long-held assumption of labour division between the two functional groups, however, was challenged by the recent unexpected discovery of complete ammonia oxidizers within the Nitrospira genus that are capable of converting ammonia to nitrate in a single organism (comammox). This breakthrough raised fundamental questions on the niche specialization and differentiation of comammox organisms with other canonical nitrifying prokaryotes in terrestrial ecosystems.

Materials and methods

This article provides an overview of the recent insights into the genomic analysis, physiological characterization and environmental investigation of the comammox organisms, which have dramatically changed our perspective on the aerobic nitrification process. By using quantitative PCR analysis, we also compared the abundances of comammox Nitrospira clade A and clade B, AOA, AOB and NOB in 300 forest soil samples from China spanning a wide range of soil pH.

Results and discussion

Comammox Nitrospira are environmentally widespread and numerically abundant in natural and engineered habitats. Physiological data, including ammonia oxidation kinetics and metabolic versatility, and comparative genomic analysis revealed that comammox organisms might functionally outcompete other canonical nitrifiers under highly oligotrophic conditions. These findings highlight the necessity in future studies to re-evaluate the niche differentiation between ammonia oxidizers and their relative contribution to nitrification in various terrestrial ecosystems by including comammox Nitrospira in such comparisons.

Conclusions

The discovery of comammox and their broad environmental distribution added a new dimension to our knowledge of the biochemistry and physiology of nitrification and has far-reaching implications for refined strategies to manipulate nitrification in terrestrial ecosystems and to maximize agricultural productivity and sustainability.

     

Responses of soil microbial community to nitrogen fertilizer and precipitation regimes in a semi-arid steppe

Purpose

Changes of nitrogen (N) cycle caused by N fertilization and precipitation regimes have affected the key ecosystem structure and functions in temperate steppe, which may modify the structure of soil microbial communities involved in N transformation. This paper was designated to examine the response of soil ammonia oxidizers and denitrifiers to the N fertilization and precipitation regimes in a semi-arid steppe where N and water contents are major limiting factors of the grassland productivity.

Materials and methods

This study was based on a long-term N fertilization and precipitation regimes experiment in Inner Mongolia (116° 17′ 20″ E, 42° 2′ 29″ N). The treatments including CK (control), R (reduced precipitation), W (30% increase in precipitation), N (10 g N m^?2 y^?1), RN (reduced precipitation and 10 g N m^?2 y^?1), and WN (30% increase in precipitation and 10 g N m^?2 y^?1). Soil basic chemical properties and microbial activities were analyzed. Molecular methods were applied to determine the abundance, structure and diversity of ammonia oxidizers and denitrifiers. Statistical analysis detected the main and interactive effect of treatments on soil microbial communities and revealed the relationship between soil microbial community structures and environmental factors.

Results and discussion

N fertilization significantly increased ammonia-oxidizing bacteria (AOB) abundance. Ammonia-oxidizing archaea (AOA) community structure was markedly changed in N fertilizer treatment and strongly affected by soil pH, while soil nitrate and water content correlated with AOB community structure. Soil nitrate was the key factor influencing nirK gene community structure, while soil pH and water content explained much of the variations of nosZ gene community. AOB-amoA and nosZ gene community diversities were influenced by precipitation regimes and interaction of N fertilization and precipitation regimes, respectively.

Conclusions

N fertilization and precipitation regimes had significant influences on the changes of soil properties and microbial functional communities. Soil nitrification was mainly driven by AOB in the semi-arid grassland. Changes of substrate content and soil pH were the key factors in shifting functional microbial communities. The non-synergistic effects of N fertilization and precipitation regimes on the microbial functional groups indicated that the negative effect of lower pH induced by N fertilization would be alleviated by precipitation regimes, which should be well considered in grassland restoration.

     

Response of archaeal communities to water regimes under simulated warming and drought conditions in Tibetan Plateau wetlands

Purpose

Understanding how archaeal communities are affected by water-table drawdown is essential for predicting soil functional responses to future climate change and the consequences of the responses on the soil carbon cycle.

Material and methods

We investigated the effect of water-table drawdown, warming, drought, and combinations thereof on archaeal communities using terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR.

Results and discussion

Methanosarcinales, Methanosaeta, Methanomicrobiales, Methanobacteriales, uncultured Rice Cluster II (RC-II), and uncultured Crenarchaeota were detected. Water-table drawdown and drought exhibited significant effects on the archaeal communities. When the water table was at or above 10 cm, the archaeal abundance at 10 cm remained high (approximately 10⁹ cells per gram dry soil), whereas the archaeal abundance at 10 cm was reduced to approximately 10⁸ cells per gram dry soil where the water table was lowered to 20 cm or below. When the water table kept constant, warming caused a significant reduction in the archaeal abundance, whereas drought only caused a decrease in archaeal abundance when the water table was higher than ?20 cm.

Conclusions

Results suggest that changes in water table may directly impact archaeal community abundance and assemblage which can in turn influence methane emissions, potentially on a large scale. Our results also indicate that archaeal communities response to water-table drawdowns that are dependent on the initial ecohydrology.

     

A hollow bacterial diversity pattern with elevation in Wolong Nature Reserve,Western Sichuan Plateau

Purpose

The pattern of eukaryotic macroorganisms varies with altitude is well-documented; by contrast, very little is known of how a bacterial pattern in soils varies with the elevation in a montane ecosystem. Mostly, previous studies on soil bacteria have either found a diversity decline, no trend, or a hump-back trend with increasing elevation. The aim of this study was to investigate the bacterial community composition and diversity patterns of Mount Nadu in Wolong Nature Reserve, Western Sichuan Plateau (3000–3945 masl).

Materials and methods

In total, 30 soil samples from the mountain at 10 sampling elevational zones (every 100 m from the baseband to the summit) were collected. High-throughput pyrosequencing approach was performed of soil bacterial 16S rRNA targeting V3?+?V4 region by MiSeq PE300 and taxonomically classified based on Silva database. Bacterial community composition and diversity patterns were detected, and bacterial data were correlated with environmental factors to determine which factors influenced bacterial community composition.

Results and discussion

We obtained an average of 30,172 sequences per soil and found that the relative abundance of Acidobacteria and Proteobacteria count more than 70 % of the whole bacteria. Cooperative network analysis also revealed that Acidobacteria and Proteobacteria were important hubs in the community. Bacterial diversity pattern was found to be a significant hollow trend along altitudinal gradients and diversity of the dominant phyla (e.g., Acidobacteria, Proteobacteria) followed the results of the whole bacterial diversity. Moreover, distance-based linear model identified that soil pH and TN significantly provided 7.40 and 6.01 % of the total variation.

Conclusions

The hollow trend of bacterial diversity has rarely been observed in nature. It indicated that no unifying bacterial diversity pattern can be expected along elevational gradients among the mountain system, and our result suggested the importance of environmental factors in structuring bacterial communities in this montane ecosystem.

     

Land contamination by toxic elements in abandoned mine areas in Italy

Purpose

The present paper concerns the distribution and mobility of heavy metals (Cu, Pb, Zn and Fe) in the soils of some abandoned mine sites in Italy and their transfer to wild flora.

Materials and methods

Soils and plants were sampled from mixed sulphide mine dumps in different parts of Italy, and the concentrations of heavy metals were determined.

Results and discussion

The phytoremediation ability of Salix species (Salix eleagnos, Salix purpurea and Salix caprea), Taraxacum officinale and P?lantago major for heavy metals and, in particular, zinc was estimated. The results showed that soils affected by mining activities presented total Zn, Cu, Pb and Fe concentrations above the internationally recommended permissible limits. A highly significant correlation occurred between metal concentrations in soils.

Conclusions

The obtained results confirmed the environmental effects of mine waste; exploring wild flora ability to absorb metals, besides metal exploitation, proved a useful tool for planning possible remediation projects.

     

Response of ammonia oxidizers and denitrifiers to repeated applications of a nitrification inhibitor and a urease inhibitor in two pasture soils

Purpose

The nitrification inhibitor 3,4-dimethylpyrazol-phosphate (DMPP) and the urease inhibitor N-(n-butyl) thiophosphoric triamide (nBTPT) can mitigate N losses through reducing nitrification and ammonia volatilization, respectively. However, the impact of repeated applications of these inhibitors on nitrogen cycling microorganisms is not well documented. This study aimed to investigate the changes in the abundance and community structure of the functional microorganisms involved in nitrification and denitrification in Australian pasture soils after repeated applications of DMPP and nBTPT.

Materials and methods

Soil was collected in autumn and spring, 2014 from two pasture sites where control, urea, urea ammonium nitrate, and urea-coated inhibitors had been repeatedly applied over 2 year. Soil samples were analyzed to determine the potential nitrification rates (PNRs), the abundances of amoA, narG, nirK and bacterial 16S rRNA genes, and the community structure of ammonia oxidizers.

Results and discussion

Two years of urea application resulted in a significantly lower soil pH at Terang and a significant decrease in total bacterial 16S rRNA gene abundance at Glenormiston and led to significantly higher PNRs and abundances of ammonia oxidizers compared to the control. Amendment with either DMPP or nBTPT significantly decreased PNRs and the abundance of amoA and narG genes. However, there was no fertilizer- or inhibitor-induced change in the community structure of ammonia oxidizers.

Conclusions

These results suggest that there were inhibitory effects of DMPP and nBTPT on the functional groups mediating nitrification and denitrification, while no significant impact on the community structure of ammonia oxidizers was observed. The application of nitrification or urease inhibitor appears to be an effective approach targeting specific microbial groups with minimal effects on soil pH and the total bacterial abundance.

     

Archaeal communities in the sediments of three contrasting mangroves

Purpose

This study evaluates the presence and diversity of 16S rRNA (rrs) and amoA genes from archaea in three mangrove sediments under different stages of preservation (one pristine mangrove, one affected by anthropogenic activity, and another contaminated by an oil spill) in the state of São Paulo (Brazil).

Materials and methods

A combination of DGGE, coupled with ordination analysis, and clone libraries of both targeted genes (rrs and amoA) was used to infer the diversity and phylogeny of archaeal communities in the mangrove analyzed samples.

Results and discussion

The DGGE combined with multivariate analysis revealed, based on the ribosomal gene, differences in archaeal communities according to environmental parameters such as mangrove location, anthropogenic activity, and oil contamination. The profiles based on the amoA gene were more similar than those obtained for the gene rrs, with the only difference statistically observed for the community found in the mangrove under anthropogenic pressure. Furthermore, phylogenetic analysis revealed most archaeal groups affiliated to sequences belonging to the Thaumarchaeota (53.1%, 24 OTUs) and Euryarchaeota (29.6%, 14 OTUs) phyla, in addition to 14 sequences affiliated to an unclassified Archaea (16.1%, 8 OTUs). Concerning the analysis of the amoA gene, mangroves harbored sequences affiliated with those previously described in water column and soil/sediment samples, besides two possible clusters specifically found in mangroves.

Conclusions

The findings are that the mangroves act as a reservoir for archaeal diversity, are possibly involved in nitrogen transformation in this ecosystem, and are affected by distinct pressures caused by anthropogenic activities.

     

Shifts of archaeal community structure in soil along an elevation gradient in a reservoir water level fluctuation zone

Purpose

Although archaea play an important role in nutrients cycling, the archaeal community in a reservoir water-level fluctuation zone (WLFZ) remains unclear. An elucidation of archaeal community responding to the environmental variables is essential to understand the nutrients dynamics in WLFZ. This study focused on the response of the archaeal community structure and abundance to the periodic water flooding along an elevation gradient in the WLFZ of the Three Gorges Reservoir.

Materials and methods

Along the elevation gradient (152–175 m) of the study area, soil samples in the beginning and late stages of water flooding were collected to investigate the influence of water flooding on the archaeal community in soil, using quantitative PCR and Illumina high-throughput sequencing approaches.

Results and discussion

An increase of archaeal abundance from 3.8?×?10⁸ to 3.8?×?10⁹ copies (g d.w.s)^?1 on average was observed after water flooding. The archaeal abundance was positively correlated with the contents of ammonium, organic matter, and moisture in soil and with the accumulated flooding time. Higher diversity was observed in dry samples (non-flooded soil samples) rather than wet samples (flooded soil samples). The Thaumarchaeota were predominant in most of the dry samples. Interestingly, high proportions of Candidatus Nitrososphaera were observed in the transition zone, while euryarchaeotal methanogens dominated the wet samples. The proportion of methanogens decreased dramatically in the dry samples at higher elevations, which was associated with the decrease of the moisture content and the probably increase of available oxygen in soil.

Conclusions

Archaeal abundance, diversity, and community composition shifted along an elevation gradient and were influenced by water flooding. The increased archaea abundance after water flooding and elevation related community composition and diversity indicated that water flooding was a key dynamic environmental variable in the WLFZ.

     

Soil microbial abundance and community structure vary with altitude and season in the coniferous forests,China

Purpose

Soil microbes control the bioelement cycles and energy transformation in forest ecosystems, and are sensitive to environmental change. As yet, the effects of altitude and season on soil microbes remain unknown. A 560 m vertical transitional zone was selected along an altitude gradient from 3023, 3298 and 3582 m, to determine the potential effects of seasonal freeze-thaw on soil microbial community.

Materials and methods

Soil samples were collected from the three elevations in the growing season (GS), onset of freezing period (FP), deeply frozen period (FPD), thawing period (TP), and later thawing period (TPL), respectively. Real-time qPCR and polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) were used to measure the abundance and structure of soil microbial community.

Results and discussion

The bacterial, archaeal, and fungal ribosomal DNA (rDNA) copy numbers decreased from GS to freezing stage (FP and FPD) and then increased in thawing stage (TP and TPL). Similarly, the diversity of microbial community varied with seasonal freeze-thaw processes. The diversity index (H) of the bacterial and archaeal communities decreased from GS to FP and then increased to TPL. The fungal community H index increased in the freezing process.

Conclusions

Our results suggested that abundance and structure of soil microbial community in the Tibetan coniferous forests varied by season and bacterial and archaeal communities respond more promptly to seasonal freeze-thaw processes relative to fungal community. This may have important implications for carbon and nutrient cycles in alpine forest ecosystems. Accordingly, future warming-induced changes in seasonal freeze-thaw patterns would affect soil nutrient cycles via altering soil microbial properties.

     

Archaeal communities in the sediments of different mangrove stands at Dongzhaigang,China

Purpose

Knowledge of archaeal communities is essential for understanding of the mechanism of carbon and nitrogen cycle in the mangrove sediment ecosystem. Presently, little is known about archaeal communities in the Dongzhaigang mangrove sediments. This study aimed to characterize the archaeal communities in sediments of different mangrove stands and to find out the correlations between archaeal communities and the environmental factors of sediments.

Materials and methods

Sediment samples were collected from the Dongzhaigang mangrove forest for analysis of soil properties and archaeal communities, by national standard methods and Illumina Miseq archaeal 16S ribosomal RNA (rRNA) gene sequencing, respectively.

Results and discussion

The archaeal community in the Dongzhaigang mangrove forest was constituted by some phyla from “TACK” and “DPANN” supergroups, and dominated by Euryarchaeota. Among sediments of the four mangroves in Dongzhaigang, principal coordinates analysis (PCoA) scatter plot showed a trend of difference in the archaeal community structure in the Bruguiera gymnoihiza and Kandelia candel stands from that in the Laguncularia racemosa and Sonneratia apetala stands. The abundance of the order Methanosarcinales was the highest in the sediments of K. candel mangroves, whereas the order of Methanobacteriales dominated in B. gymnoihiza sediments. The highest richness and diversity values of Archaea occurred in K. candel sediments, while the lowest in B. gymnoihiza. Pearson correlation showed the significant relationships between sediment properties and some dominant genera, with a positive and significant correlation between sediment properties and genus Methanobacterium, coinciding with the maximum values of sediment properties and abundance of Methanobacterium in the sediment of B. gymnoihiza. Such results indicated that the difference of archaeal community structure among mangrove sediments may be caused by the different sediment characteristics. Methanogenic communities in the Dongzhaigang mangrove forest sediments were, at the order level, constituted by Methanobacteriales, Methanomicrobiales, Methanosarcinales, and Methanomassiliicoccales.

Conclusions

The investigation indicated that the Dongzhaigang mangrove sediment ecosystems support diverse archaeal communities and methanogenic communities, and that there was a general trend of difference in the archaeal community structure in the B. gymnoihiza and K. candel mangrove sediments from that in the L. racemosa and S. apetala sediments. Such difference may be caused by the difference in sediment characteristics.

     

Effects of pH and polycyclic aromatic hydrocarbon pollution on thaumarchaeotal community in agricultural soils

Purpose

Thaumarchaeota is an ecologically relevant archaeal phylum which may significantly contribute to global nitrogen cycling. Thaumarchaeotal abundance, composition, and activity can be changed by soil pH and pollutants such as toxic metals. This study aims to examine the responses of thaumarchaeotal community to soil pH variation and polycyclic aromatic hydrocarbon (PAH) pollution which may co-occur in agricultural soils.

Materials and methods

Field soil samples were collected from agricultural land impacted by both acidification and PAH contamination. Thaumarchaeotal abundance and composition were assessed using molecular approaches targeting 16S rRNA or amoA genes and were linked to environmental factors by correlation and canonical correspondence analysis (CCA). To evaluate the short-term responses of Thaumarchaeota to PAHs, additional soil microcosms amended with either three selected PAHs were established. Changes in thaumarchaeotal communities during the incubation were monitored.

Results and discussion

A significant correlation between thaumarchaeotal gene abundance and soil pH was observed within field samples, with the I.1a-associated group enriched when pH <5.0. CCA suggests that the community variation was primarily related to soil pH. In contrast, the effects of PAHs were minimal. In soil microcosms, high concentrations of PAHs persisted after the 4-week incubation. Independent of the PAHs added, thaumarchaeotal amoA abundance slightly increased and the compositions were stable at the end of the incubation. This might be associated with the pollutants bioavailability and potential microbe-PAH interactions in the soil.

Conclusions

Soil pH variation strongly shapes the agricultural soil thaumarchaeotal community, whereas PAH effects appear to be marginal even in the presence of high concentrations of pollutants. The complicated interaction between soil matrix, pollutants, and Thaumarchaeota requires further study.

     

Straw amendment to paddy soil stimulates denitrification but biochar amendment promotes anaerobic ammonia oxidation

Purpose

Organic matter amendment is usually used to improve soil physicochemical properties and to sequester carbon for counteracting climate change. There is no doubt that such amendment will change microbial activity and soil nitrogen transformation processes. However, the effects of straw and biochar amendment on anammox and denitrification activity and on community structure in paddy soil are unclear.

Materials and methods

We conducted a 30-day pot experiment using rice straw and rice straw biochar to deepen our understanding about the activity, microbial abundance, and community structure associated with soil nitrogen cycling during rice growth.

Results and discussion

Regarding activity, anammox contributed 3.1–8.1% of N₂ production and denitrification contributed 91.9–96.9% of N₂ production; straw amendment resulted in the highest denitrification rate (38.9 nmol N g^?1 h^?1), while biochar amendment resulted in the highest anammox rate (1.60 nmol N g^?1 h^?1). Both straw and biochar amendments significantly increased the hzsB and nosZ gene abundance (p < 0.05). Straw amendment showed the highest nosZ gene abundance, while biochar amendment showed the highest hzsB gene abundance. Phylogenetic analysis of the anammox bacteria 16S rRNA genes indicated that Candidatus Brocadia and Kuenenia were the dominant genera detected in all treatments.

Conclusions

Straw and biochar amendments have different influences on anaerobic ammonia oxidation and denitrification within paddy soil. Our results suggested that the changes in denitrification and anammox rates in the biochar and straw treatments were mainly linked to functional gene abundance rather than microbial community structure and that denitrification played the more major role in N₂ production in paddy soil.

     

Vinasse irrigation: effects on soil fertility and arbuscular mycorrhizal fungi population

Purpose

The aim of this research was to determine the vinasse irrigation effects on the arbuscular mycorrhizal fungi (AMF) population (total spore abundance (TSA), richness, relative abundance, and diversity indices) and soil parameters and nutrients at high doses. The irrigation of soil with vinasses derived from sugarcane, beet, or alcohol production is a common practice around the world. Little is known about how this affects the AMF community and soil nutrients.

Materials and methods

The spider plant (Chlorophytum comosum, (Thunb.) Jacques), a mycorrhizable plant, was used to investigate the effect of 4 months of frequent vinasse irrigation (0, 25, 50, 75, and 100% vinasse concentration) on AMF and soil characteristics, e.g., electrical conductivity (EC), pH, mineral N, available P, Na⁺, K⁺, Ca²⁺, and Mg²⁺.

Results and discussion

The vinasse irrigation decreased the TSA, AMF richness and diversity after 4 months, regardless of vinasse concentration. The vinasse irrigation did not acidify the soil, but the EC, mineral N and available P increased. The biomass of C. comosum decreased (77–81%) after vinasse irrigation for 4 months.

Conclusions

Frequent irrigation with vinasse at concentrations ≥50% increases EC, K⁺, Na⁺, Mg²⁺, Ca²⁺ and available P in the soil, and decreases the amount of AMF spores, richness and diversity, which is not desirable in agricultural soils.

     

Contrasting response of two grassland soils to N addition and moisture levels: N<Subscript>2</Subscript>O emission and functional gene abundance

Purpose

Nitrification and denitrification processes dominate nitrous oxide (N₂O) emission in grassland ecosystems, but their relative contribution as well as the abiotic factors are still not well understood.

Materials and methods

Two grassland soils from Duolun in Inner Mongolia, China, and Canterbury in New Zealand were used to quantitatively compare N₂O production and the abundance of bacterial and archaeal amoA, denitrifying nirK and nirS genes in response to N additions (0 and 100 μg NH₄ ⁺–N g^?1 dry soil) and two soil moisture levels (40 and 80 % water holding capacity) using microcosms.

Results and discussion

Soil moisture rather than N availability significantly increased the nitrification rate in the Duolun soil but not in the Canterbury soil. Moreover, N addition promoted denitrification enzyme activities in the Canterbury soil but not in the Duolun soil. The abundance of bacterial and archaeal amoA genes significantly increased as soil moisture increased in the Duolun soil, whereas in the Canterbury soil, only the abundance of bacterial amoA gene increased. The increase in N₂O flux induced by N addition was significantly greater in the Duolun soil than in the Canterbury soil, suggesting that nitrification may have a dominant role in N₂O emission for the Duolun soil, while denitrification for the Canterbury soil.

Conclusions

Microbial processes controlling N₂O emission differed in grassland soils, thus providing important baseline data in terms of global change.

     

An overlooked nitrogen loss linked to anaerobic ammonium oxidation in estuarine sediments in China

Purpose

Despite its importance, anammox (anaerobic ammonium oxidation) in estuarine sediment systems remains poorly understood, particularly at the continental scale. This study aimed to understand the abundance, diversity, and activity of anammox bacteria and to determine the main factors influencing the anammox process in estuarine sediments in China.

Materials and methods

Estuarine sediments were collected from 18 estuaries spanning over 4000 km. Experiments using an ¹⁵ N–tracer, quantitative PCR, and clone library construction were used to determine the activity, abundance, and diversity of anammox bacteria. The impact of environmental factors on anammox processes was also determined.

Results and discussion

The abundance of the anammox-specific hydrazine synthase (hzsB) gene ranged from 1.8 × 10⁵ ± 3.4 × 10⁴ to 3.6 × 10⁸ ± 7.5 × 10⁷ copies g^?1 dw. Candidatus Scalindua, Brocadia, Kuenenia, Jettenia, and two novel unidentified clusters were detected, with Scalindua dominating the anammox population. Additionally, the abundances of Scalindua, Kuenenia, and Brocadia were found to be significantly correlated with latitude. The anammox rates ranged from 0.29 ± 0.15 to 13.68 ± 3.98 nmol N g^?1 dw h^?1 and contributed to 2.39–82.61% of total N₂ production. Pearson correlation analysis revealed that the anammox rate was positively correlated with total nitrogen, total carbon, and temperature, and was negatively correlated with dissolved oxygen (DO). The key factors influencing the hzsB gene abundance were ammonium concentration, salinity, and DO. Ammonium concentration, pH, temperature, and latitude were main variables shaping the anammox-associated bacterial community.

Conclusions

Our results suggested that anammox bacteria are ubiquitous in coastal estuaries in China and underline the importance of anammox resulting in N loss at a continental scale.

     

Behavior of chlorpyrifos and its major metabolite TCP (3,5,6-trichloro-2-pyridinol) in agricultural soils amended with drinking water treatment residuals

Purpose

Chlorpyrifos can be effectively adsorbed by drinking water treatment residuals (WTR), ubiquitous and non-hazardous by-products of potable water production. The major metabolite 3,5,6-trichloro-2-pyridinol (TCP) was found to be much more mobile and toxic than its parent chlorpyrifos. To assess the feasibility of WTR amendment for attenuation of chlorpyrifos and TCP pollution, the sorption/desorption and degradation behavior of chlorpyrifos and TCP in WTR-amended agricultural soils was examined in the present study.

Materials and methods

Two representative agricultural soils were sampled from southern and northern China, respectively. The soils were amended with WTR at the rates of 0, 2, 5, and 10 % (w/w). Batch sorption/desorption test were applied to investigate the sorption/desorption characteristics of chlorpyrifos and TCP in WTR-amended soils. The influence of WTR amendment on chlorpyrifos degradation and TCP formation was evaluated using the incubation test, and its effect on the soil bacterial abundance was further studied through DNA extraction and PCR amplification.

Results and discussion

Results showed that WTR amendment (0–10 %, w/w) significantly enhanced the retention capacity of chlorpyrifos and TCP in both soils examined (P < 0.05). Fractionation analyses further demonstrated that the bioavailability of chlorpyrifos was considerably reduced by WTR amendment, resulting in a decreased chlorpyrifos degradation rate. The WTR amendment also significantly reduced the mobility of TCP formed in chlorpyrifos-contaminated soils (P < 0.001). The chlorpyrifos toxicity to soil bacteria community was largely mitigated following WTR amendment, resulting in increased total bacterial abundance.

Conclusions

Results obtained in the present study indicate a great deal of potential for the beneficial reuse of WTR as soil amendments for chlorpyrifos and TCP pollution control.

     

The effects of different irrigation regimes on nitrous oxide emissions and influencing factors in greenhouse tomato fields

Purpose

Intensive agricultural practices have enhanced problems associated with the competing use of limited water resources. Nitrous oxide (N₂O) is a major contributor to global warming. It is important for researchers to ascertain the relationship between irrigation and soil N₂O emissions in order to identify mitigation strategies to reduce nitrous oxide emissions. Different irrigation amounts affect soil water dynamics and nitrogen turnover. The effect of three lower limits of irrigation on soil N₂O emissions, influencing factors, and abundance of genes involved in nitrification and denitrification were investigated in tomato irrigated in a greenhouse.

Materials and methods

Observations were performed between April and August 2015 in a long-term irrigated field subjected to different lower limits of irrigation: 20 kPa (D₂₀), 30 kPa (D₃₀), and 40 kPa (D₄₀) from greenhouse soil during the tomato crop season. Soil N₂O fluxes were monitored using the static chamber-gas chromatograph method. Copy numbers of genes were determined using the real-time quantitative polymerase chain reaction (real-time PCR) technique. Characteristics of soil N₂O emissions were analyzed, and differences between irrigation regimes were determined. The effects of influencing factors on soil N₂O emissions were analyzed, including soil temperature, soil moisture, soil pH, and soil mineral nitrogen, as well as changes in the abundance of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) based on amoA genes and denitrifier genes (nosZ, nirK, and cnorB).

Results and discussion

Our results showed that peaks in N₂O emissions occurred 1–5 days after each irrigation. During the whole tomato growth period, soil N₂O fluxes were lowest under D₃₀ treatment compared with those under D₂₀ and D₄₀ treatments. Soil NO₃ ^?-N concentrations were significantly higher than NH₄ ⁺-N concentrations. Soil N₂O fluxes were significantly related to soil moisture, NH₄ ⁺-N concentrations (P < 0.01), soil pH, and AOA copy numbers (P < 0.05). There was no consistent correlation between soil N₂O emissions, soil temperature, and soil NO₃ ^?-N concentrations. Different irrigation regimes significantly affected AOA copy numbers but did not affect the expression of other genes. AOA copy numbers were higher than those of AOB. Soil N₂O fluxes significantly affected the AOA copy numbers and potential nitrification rates (P < 0.05).

Conclusions

Soil moisture, pH, and NH₄ ⁺-N concentration were important factors affecting soil N₂O emissions. Compared with other genes associated with nitrification and denitrification, AOA plays an important role in N₂O emissions from greenhouse soils. Selecting a lower limit of irrigation of 30 kPa could effectively reduce N₂O emissions from vegetable soils.