Improving dryland maize productivity and water efficiency with heterotrophic ammonia-oxidizing bacteria via nitrification and cytokinin activity

A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield.A heterotrophic ammonia-oxidizing bacteria(HAOB)strain S2_8_1 was used.Six treatments were applied:(1)no irrigation+HAOB strain(DI),(2)no irrigation+blank culture medium(DM),(3)no irrigation control(DCK),(4)irrigation+HAOB(WI),(5)irrigation+blank culture medium(WM),and(6)irrigation control(WCK).Results revealed that HAOB treatment increased maize growth,yield,and water use efficiency over controls,regardless of whether the year was wet or dry.This improvement was attributed to the accelerated nitrification in the rhizosphere soil due to HAOB inoculation,which subsequently led to increased levels of leaf cytokinins.Overall,these findings suggest that HAOB inoculation holds promise as a strategy to boost water use efficiency and maize productivity in dryland agriculture.

Environmental similarity is more important than distance in the community structuring processes of ammonia-oxidizing archaea in agricultural soils

Ammonia-oxidizing archaea(AOA) are important in converting ammonia into nitrate in soils. While many aspects of their community structure have been studied, the relative importance of stochastic versus deterministic processes has poorly been understood. We compared AOA communities across the North China Plain, targeting the amoA gene. A phylogenetic null modelling approach was used to calculate the beta nearest taxon index to quantify the influence of stochastic and deterministic processes. We found that spatial distance between samples predicted the perceived processes involved in community structuring, with stochastic processes dominating at local scales. At greater distances, stochasticity became weaker. However, soil pH, which was also the strongest determinant of AOA community, was a much stronger predictor of community structuring, leaving the distance effect redundant as an explanation of community structuring processes. The communities of AOA differing by less than 1 pH unit differed mainly stochastically in terms of operational taxonomic unit composition. At larger pH differences, deterministic processes based on heterogeneous selection between clades became increasingly dominant. It appears that AOA community composition is largely determined by the environment. However, very similar pH environments are the exception. In environments with very close pH values, stochastic effects dominantly cause differences in community composition, whether spatially near or far.

Functional relationship between ammonia-oxidizing bacteria and ammonia-oxidizing archaea populations in the secondary treatment system of a full-scale municipal wastewater treatment plant

The abundance of ammonia-oxidizing bacteria and archaea and their amo A genes from the aerobic activated sludge tanks,recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant(WWTP)was determined.Polymerase chain reaction and denaturing gradient gel electrophoresis were used to generate diversity profiles,which showed that each population had a consistent profile although the abundance of individual members varied.In the aerobic tanks,the ammonia-oxidizing bacterial(AOB)population was more than 350 times more abundant than the ammonia-oxidizing archaeal(AOA)population,however in the digesters,the AOA population was more than 10 times more abundant.Measuring the activity of the amo A gene expression of the two populations using RT-PCR also showed that the AOA amo A gene was more active in the digesters than in the activated sludge tanks.Using batch reactors and dd PCR,amo A activity could be measured and it was found that when the AOB amo A activity was inhibited in the anoxic reactors,the expression of the AOA amo A gene increased fourfold.This suggests that these two populations may have a cooperative relationship for the oxidation of ammonia.

Abundance and diversity of ammonia-oxidizing archaea in response to various habitats in Pearl River Delta of China, a subtropical maritime zone

Ammonia-oxidizing archaea （AOA） are widely considered key to ammonia oxidation in various environments. However, little work has been conducted to simultaneously investigate the abundance and diversity of AOA as well as correlations between archaeal amoA genotypes and environmental parameters of different ecosystems at one district. To understand the abundance, diversity, and distribution of AOA in Pearl River Delta of China in response to various habitats, the archaeal amoA genes in soil, marine, river, lake, hot spring and wastewater treatment plant （WWTP） samples were investigated using real-time fluorescent quantitative PCR and clone libraries. Our analyses indicated that the diversity of AOA in various habitats was different and could be clustered into five major clades, i.e., estuary sediment, marine water/sediment, soil, hot spring and Cluster 1. Phylogenetic analyses revealed that the structure of AOA communities in similar ecological habitats exhibited strong relation. The canonical correspondence method indicated that the AOA community structure was strongly correlated to temperature, pH, total organic carbon, total nitrogen and dissolved oxygen variables. Assessing AOA amoA gene copy numbers, ranging from 6.84× 10^6 to 9.45 × 10^7 copies/g in dry soil/sediment, and 6.06× 10^6 to 2.41 ×10^7 copies/L in water samples, were higher than ammonia-oxidizing bacteria （AOB） by 1-2 orders of magnitude. However, AOA amoA copy numbers were much lower than AOB in WWTP activated sludge samples. Overall, these studies suggested that AOA may be a major contributor to ammonia oxidation in natural habitats but play a minor role in highly aerated activated sludge. The result also showed the ratio of AOA to AOB amoA gene abundance was positively correlated with temperature and less correlated with other environmental parameters. New data from our study provide increasing evidence for the relative abundance and diversity of ammonia-oxidizing archaea in the global nitrogen cycle.

Abundance and distribution of ammonia-oxidizing archaea in Tibetan and Yunnan plateau agricultural soils of China

As low oxygen and high ultraviolet （UV） exposure might significantly affect the microbial existence in plateau, it could lead to a specialized microbial community. To determine the abundance and distribution of ammonia-oxidizing archaea （AOA） in agricultural soil of plateau, seven soil samples were collected respectively from farmlands in Tibet and Yunnan cultivating the wheat, highland-barley, and colza, which are located at altitudes of 3200-3800 m above sea level. Quantitative PCR （q-PCR） and clone library targeting on amoA gene were used to quantify the abundances of AOA and ammonia-oxidizing bacteria （AOB）, and characterize the community structures of AOA in the samples. The number of AOA cells （9.34 × 10^7-2.32× 10^8 g^-1 soil） was 3.86-21.84 times greater than that of AOB cells （6.91 × 10^6-1.24 × 10^8 g^-1 soil） in most of the samples, except a soil sample cultivating highland- barley with an AOA/AOB ratio of 0.90. Based Kendall＇s correlation coefficient, no remarkable correlation between AOA abundance and the environmental factor was observed. Additionally, the diversities of AOA community were affected by total nitrogen and organic matter concentration in soils, suggesting that AOA was probably sensitive to several environmental factors, and could adjust its community structure to adapt to the environmental variation while maintaining its abundance.

Nitrification intensity and ammonia-oxidizing bacteria and archaea in different wetland plant rhizosphere soils

In order to explore the nitrogen removal process in constructed wetlands(CW s),the moisture,ammonia nitrogen(NH4+-N),nitrate nitrogen(NO3"-N)and nitrification intensity in three wetland plant rhizosphere soils(Acorns calamus,Typha orientalis,Iris pseudacorus)were investigated at a relatively normal temperature range of15to25The relative abundance of ammonia-oxidizing bacteria(AOB)and ammonia-oxidizing archaea(AOA)were also achieved using fluorescence in situ hybridization(FISH).It is found that T.orientalis achieves the highest nitrification intensity of2.03m g(h?kg)while the second is I.pseudacorrs(1.74m g/(h?kg)),and followed by A.calamus(1.65m g/(h?kg))throughout the experiment.FISH reveals that the abundance of bacteria(1010g_1wet soil)is higher than that of archaea(109g_1wet soil),and AOBare the dominant bacteria in the ammonia oxidation process.The abundance of AOB in te rhizosphere soils from high to low T.orientalis(1.88x1010g"1),I pseudacorus(1.23x1010g1),A.calamus(5.07x109g"1)while the abundance of AOA from high to low ae I.pseudacorus(4.00x109g1),A.calamus(3.52x109g"1),T.orientalis(3.48x109g"1).The study provides valuable evidence of plant selection for nitrogen removal in CWs.

Community diversity and distribution of ammonia-oxidizing archaea in marsh wetlands in the black soil zone in North-east China

Since its first detection, ammonia-oxidizing archaea (AOA) have been proven to be ubiquitous in aquatic and terrestrial ecosystems. In this study, two freshwater marsh wetlands- the Honghe wetland and Qixinghe wetland - in the black soil zone in North-east China were chosen to investigate the AOA community diversity and distribution in wetland soils with different vegetation and depth. In the Honghe wetland, two sampling locations were chosen as the dominant plant transited from Deyeuxia to Carex. In the Qixinghe wetland, one sample location that was dominated by Deyeuxia was chosen. Samples of each location were collected from three different depths, and Illumina MiSeq platform was used to generate the AOA amoA gene archive. The results showed that the AOA amoA genes in the soils of the two wetlands were affiliated with three lineages: Nitrososphaera, Nitrosotalea, and Nitrosopumilus clusters. The different dominant status of these AOA lineages indicated their differences in adapting to acidic habitat, oxygenic/hypoxic alternation, organic matter, and other environmental factors, suggesting high diversity among AOA in marsh soils. The main driver of the AOA community was pH, along with organic carbon and ammonium nitrogen, which also played an important role combined with many other environmental factors. Thus, soil physiochemical characteristics, rather than vegetation, were the main cause of AOA community diversity in the wetlands in the black soil zone in China.

Response of ammonia-oxidizing archaea to heavy metal contamination in freshwater sediment

It has been well-documented that the distribution of ammonia-oxidizing bacteria(AOB) and archaea(AOA) in soils can be affected by heavy metal contamination, whereas information about the impact of heavy metal on these ammonia-oxidizing microorganisms in freshwater sediment is still lacking. The present study explored the change of sediment ammonia-oxidizing microorganisms in a freshwater reservoir after being accidentally contaminated by industrial discharge containing high levels of metals. Bacterial amoA gene was found to be below the quantitative PCR detection and was not successfully amplified by conventional PCR. The number of archaeal amoA gene in reservoir sediments were 9.62 × 10~2–1.35 × 10~7 copies per gram dry sediment. AOA abundance continuously decreased, and AOA richness, diversity and community structure also considerably varied with time. Therefore, heavy metal pollution could have a profound impact on freshwater sediment AOA community. This work could expand our knowledge of the effect of heavy metal contamination on nitrification in natural ecosystems.

Characterisation of the Bacteria and Archaea Community Associated with Wild Oysters, At Three Possible Restoration Sites in the North Sea

With 85% of the global oyster reefs destroyed, there is an urgent need for large scale restoration to benefit from the ecosystem services provided by biogenic oyster reefs and their associated biodiversity, including microorganisms that drive marine biogeochemical cycles. This experiment established a baseline for the monitoring of the bacterial and archaeal community associated with wild oysters, using samples from their immediate environment of the Voordelta, with cohabiting Crassostrea gigas and Ostrea edulis, Duikplaats with only C. gigas attached to rocks, and the Dansk Skaldyrcentre, with no onsite oysters. The microbial profiling was carried out through DNA analysis of samples collected from the surfaces of oyster shells and their substrate, the sediment and seawater. Following 16S rRNA amplicon sequencing and bioinformatics, alpha indices implied high species abundance and diversity in sediment but low abundance in seawater. As expected, Proteobacteria, Bacteroidetes, Firmicutes and Thaumarchaeota dominated the top 20 OTUs. In the Voordelta, OTUs related to Colwellia, Shewanella and Psychrobium differentiated the oysters collected from a reef with those attached to rocks. Duikplaats were distinct for sulfur-oxidizers Sulfurimonas and sulfate-reducers from the Sva 0081 sediment group. Archaea were found mainly in sediments and the oyster associated microbiome, with greater abundance at the reef site, consisting mostly of Thaumarchaeota from the family Nitrosopumilaceae. The oyster free site displayed archaea in sediments only, and algal bloom indicator microorganisms from the Rhodobacteraceae, Flavobacteriaceae family and genus [Polaribacter] huanghezhanensis, in addition to the ascidian symbiotic partner, Synechococcus. This study suggests site specific microbiome shifts, influenced by the presence of oysters and the type of substrate.

Effect of soil archaea on N_(2)O emission in alpine permafrost

Soil microbial communities are pivotal in permafrost biogeochemical cycles,yet the variations of abundant and rare microbial taxa and their impacts on greenhouse gas emissions in different seasons,remain elusive,especially in the case of soil archaea.Here,we conducted a study on soil abundant and rare archaeal taxa during the growing and non-growing seasons in the active layer of alpine permafrost in the Qinghai-Tibetan Plateau.The results suggested that,for the archaeal communities in the sub-layer,abundant taxa exhibited higher diversity,while rare taxa maintained a more stable composition from the growing to non-growing season.Water soluble organic carbon and soil porosity were the most significant environmental variables affecting the compositions of abundant and rare taxa,respectively.Stochastic and deterministic processes dominated the assemblies of rare and abundant taxa,respectively.The archaeal ecological network influenced N_(2)O flux through different modules.Rare taxa performed an essential role in stabilizing the network and exerting important effects on N_(2)O flux.Our study provides a pioneering and comprehensive investigation aimed at unravelling the mechanisms by which archaea or other microorganisms influence greenhouse gas emissions in the alpine permafrost.

Effects of Transgenic DREB Soybean Dongnong 50 on the Diversity of Soil Ammonia-oxidizing Bacteria

ObjectiveThe aim was to understand the effects of transgenic DREB soybean on the ammonia-oxidizing bacteria. MethodThe diversity of the cto gene in pot-planted transgenic soybean and near-isogenic non-transgenic soybean under normal water condition and drought stress was analyzed by PCR-DGGE and sequence analysis. ResultRhizosphere community diversity of ammonia-oxidizing bacteria showed no difference between the treatments of transgenic soybean and its non-transgenic isolines, moreover transgenic soybean under normal water condition and drought stress improved the diversity of the ammonia-oxidizing bacteria in the harvest time. The phylogenetic analysis revealed that all the sequences of excised DGGE bands were closely related to members of the genus Nitrosovibrio and Nitrosospira of the β-subclass Proteobacteria. ConclusionTransgenic DREB soybean has no adverse impact on soil ammonia-oxidizing bacteria.

Linking bacterial and archaeal community dynamics to related hydrological,geochemical and environmental characteristics between surface water and groundwater in a karstic estuary

Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play critical roles in biogeochemical transformations in STEs,limited information is available about how their community dynamics interact with hydrological,geochemical and environmental characteristics in STEs.Here,we studied bacterial and archaeal diversities and distributions with 16S rRNA-based Illumina MiSeq sequencing technology between surface water and groundwater in a karstic STE.Principal-coordinate analysis found that the bacterial and archaeal communities in the areas where algal blooms occurred were significantly separated from those in other stations without algal bloom occurrence.Canonical correspondence analysis showed that nutrients and salinity can explain the patterns of bacterial and archaeal community dynamics.The results suggest that hydrological,geochemical and environmental characteristics between surface water and groundwater likely control the bacterial and archaeal diversities and distributions in STEs.Furthermore,we found that some key species can utilize terrestrial pollutants such as nitrate and ammonia in STEs,indicating that these species(e.g.,Nitrosopumilus maritimus,Limnohabitans parvus and Simplicispira limi)may be excellent candidates for in situ degradation/remediation of coastal groundwater contaminations concerned with the nitrate and ammonia.Overall,this study reveals the coupling relationship between the microbial communities and hydrochemical environments in STEs,and provides a perspective of in situ degradation/remediation for coastal groundwater quality management.

Community analysis of ammonia-oxidizing bacteria in activated sludge of eight wastewater treatment systems

We investigated the communities of ammonia-oxidizing bacteria （AOB） in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction （PCR） followed by terminal restriction fragment length polymorphism （T-RFLP）, cloning, and sequencing of the α-subunit of the ammonia monooxygenase gene （amoA）. The T-RFLP fingerprint analyses showed that different wastewater treatment systems harbored distinct AOB communities. However, there was no remarkable difference among the AOB T- RFLP profiles from different parts of the same system. The T-RFLP fingerprints showed that a full-scale wastewater treatment plant （WWTP） contained a larger number of dominant AOB species than a pilot-scale reactor. The source of influent affected the AOB community, and the WWTPs treating domestic wastewater contained a higher AOB diversity than those receiving mixed domestic and industrial wastewater. However, the AOB community structure was little affected by the treatment process in this study. Phylogenetic analysis of the cloned amoA genes clearly indicated that all the dominant AOB in the systems was closely related to Nitrosomonas spp. not to Nitrosospira spp. Members of the Nitrosomonas oligotropha and Nitrosomonas communis clusters were found in all samples, while members of Nitrosomonas europaea cluster occurred in some systems.

Impact of acetochlor on ammonia-oxidizing bacteria in microcosm soils

Acetochlor is an increasingly used herbicide on corn in North China. Currently, the effect of acetochlor on soil ammonia-oxidizing bacteria (AOB) communities is not well documented. Here, we studied the diversity and community composition of AOB in soil amended with three concentrations of acetochlor (50, 150, 250 mg/kg) and the control (0 mg acetochlor/kg soil) in a microcosm experiment by PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and the phylogenetic analysis of excised ...

Distribution of ammonia-oxidizing Betaproteobacteria community in surface sediment off the Changjiang River Estuary in summer

The spatial distribution of ammonia-oxidizing Betaproteobacteria （βAOB） was investigated by FISH （fluorescence in situ hybridization） and DGGE （denaturing gradient get electrophoresis） techniques in the sediment off the Changjiang River Estuary. Sediment samples were collected from eight stations in June before the formation of hypoxia zone in 2006. The abundance of βAOB ranged from 1.87× 10^5 to 3.53×10^5 cells/g of sediment. βAOB abundance did not present a negative correlation with salinity, whereas salinity was implicated as the primary factor affecting nitrification rates. The DGGE profiles of PCR-amplified amoA gene fragments revealed that the βAOB community structure of sample S2 separated from other samples at the level of 40% similarity. The variations in composition ofβAOB were significantly correlated with the salinity, temperature, absorption ability of sediments and TOC. The statistical analysis indicates that theβAOB abundance was a main factor to influence nitrification rates with an influence ratio of 87.7% at the level of 40% biodiversity similarity. Considering the good correlation between βAOB abundance and nitrification estimates, the abundance and diversity of βAOB community could be expected as an indirect index of nitrification activity at the study sea area in summer.

Seasonal and spatial distribution of ammonia-oxidizing microorganism communities in surface sediments from the East China Sea

Ammonia oxidation plays a significant role in the nitrogen cycle in marine sediments. Seasonal and spatial distribution of ammonia-oxidizing archaea （AOA） and betaproteobacteria （13-AOB） in surface sediments from the East China Sea （ECS） were investigated using ammonia monooxygenase ct subunit （amoA） gene. In order to characterize the community of AOA and 13-AOB, real-time quantitative polymerase chain reaction （qPCR） was carried out in this study, along with environmental parameters. The abundance of 13-AOB amoA gene （2.17x 10^6-4.54x10^7 copy numbers per gram wet weight sediment） was always greater than that ofAOA amoA gene （2.18x 105-9.89x 10^6 copy numbers per gram wet weight sediment） in all sampling stations. The qPCR results were correlated with environmental parameters. AOA amoA gene copy numbers in April were positively related to temperature and nitrite concentration （p〈0.05）. 13-AOB amoA gene copy numbers in August correlated negatively with salinity （p〈0.01）, and correlated positively with ammonium concentration （p〈0.05）. With the increase of salinity, the amoA gene copy ratio of AOB to AOA had a tendency to decrease, which suggested 13-AOB dominated in the area of high level ammonium and AOA preferred high salinity area.

Response of ammonia-oxidizing betaproteobacteria to short- term fertilization in a salt marsh in China

This study examines the impacts of short-term(6 months) fertilization on the community structure and abundance of ammonia-oxidizing betaproteobacteria(β-AOB) and the potential nitrification rate in sediment colonized by S uaeda heteroptera in a saltmarsh located in Shuangtai estuary, China. The sediment samples were collected from plots treated with different amounts of an N fertilizer(urea supplied at 0.1, 0.2, 0.4, and 0.8 g/kg(nitrogen content in dry sediment)), and with different forms of N fertilizers(urea,(NH4) 2 SO 4, and NH_4NO_3, each supplied at 0.2 g/kg). The fertilizers were applied 1–4 times during the plant-growing season in May, July, August and September of 2013. Untreated plots were included as a control. As revealed in denaturing gradient gel electrophoresis of the 16 S r RNA gene, the β-AOB community responded to both the amount and form of N. Real-time quantitative PCR indicated that both abundance and potential nitrification rate of β-AOB increased after N addition, regardless of concentration and form(except NH_4NO_3). These results provide evidence that short-term N application influences the sediment β-AOB community, β-AOB abundance and potential nitrification rate in a saltmarsh ecosystem.

Compositional Shifts in Ammonia-Oxidizing Microorganism Communities of Eight Geographically Different Paddy Soils —Biogeographical Distribution of Ammonia-Oxidizing Microorganisms

Soil nitrification is mediated by ammonia-oxidizing archaea (AOA) and bacteria (AOB), which occupy different specialized ecological niches. However, little is known about the diversification of AOA and AOB communities in a large geographical scale. Here, eight paddy soils collected from different geographic regions in China were selected to investigate the spatial distribution of AOA and AOB, and their potential nitrification activity (PNA). The result showed that the abundance of AOA was predominant over AOB, indicating that the rice fields favor the growth of AOA. PNA highly varied from 0.43 to 3.57 μg NOX-N·g·dry·soil·h-1, and was positively related with soil NH3 content, the abundance of AOA community, and negatively related with the diversity of AOB community (P amoA genes revealed remarkable differences in the compositions of AOA and AOB community. Phylogenetic analyses of amoA genes showed that Nitrosospiracluster-3-like and Nitrosomonas cluster 7-like AOB extensively dominated the AOB communities, and 54d9-like AOA within the soil group 1.1b predominated in AOA communities in paddy soils. Redundancy analysis suggested that the spatial variations of AOA community structure were influenced by soil TN content (P < 0.01), while no significant correlation between AOB community structure and soil properties was found. Findings highlight that ammonia oxidizers exhibit spatial variations in complex paddy fields due to the joint influence of soil variables associated with N availability.

Characterization of Bacterial Community,Ammonia-Oxidizing Bacteria,and Nitrospira During the Operation of a Commercial-Scale Recirculating Aquaculture System for Culturing Pufferfish Takifugu rubripes

We investigated the changes in communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira during the operation of a pufferfish Takifugu rubripes recirculating aquaculture system by using high-throughput DNA sequencing.Differences in bacterial communities were observed at days 1-32,47-62 and 78-93 of biofilm development by using 16S rRNA gene pyrosequencing.The relative abundance of Proteobacteria(Gammaproteobacteria)increased,while that of Bacteroidetes(Flavobacteria)decreased.The proportions of Nitrosomonas and Nitrospina ranged from 0.02%to 0.30%and from 0.02%to 0.83%,respectively.Ammonia monooxygenase gene pyrosequencing revealed that the top three operational taxonomic units were related to Nitrosomonas aestuarii(17.5%-61.1%),uncultured beta proteobacterium clone B67S-54(1.9%-45.2%),and uncultured bacterium clone AZPa8(3.6%-24.7%).Nitrite oxidoreductase gene pyrosequencing revealed that the relative abundance of the dominant strain Nitrospira sp.Ecomares 2.1 increased,but that of the abundant species Nitrospira marina decreased.Our results demonstrated that the communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira were changing during the operation of the pufferfish recirculating aquaculture system.

Distribution characteristics of ammonia-oxidizing bacteria in the Typha latifolia constructed wetlands using fluorescent in situ hybridization(FISH)

A molecular biology method, fluorescent in situ hybridization（FISH）, in which the pre-treatment was improved in allusion to the media of the constructed wetlands（CW）, e.g. the soil and the grit, was used to investigate the vertical distribution characteristics of ammonia-oxidizing bacteria（AOB） quantity and the relation with oxidation-reduction potential（ORP） in the Typha latifolia constructed wetlands under three different Ioadings in summer from May to September. Results showed that the quantity of the AOB decreased in the Typha latifolia CW with the increase of vertical depth. However, the AOB quantity was 2-4 times the quantity of the control in the root area. Additionally, ORP in the rhizosphere was found to be higher than other areas, which showed that Typha latifolia CW was in an aerobic state in summer when using simulated non-point sewage at the rural area of Taihu Lake in China and small town combined sewage.