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.

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.

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.

Use of mesophilic and thermophilic bacteria for the improvement of copper extraction from a low-grade ore

Bioleaching was examined for copper extraction from a low grade ore using mesophilic and moderate thermophilic bacteria. Five equal size columns were used for the leaching of the ore. Sulfuric acid solution with a flow rate of 3.12 L·m-2·h-1 and pH 1.5 passed through each column continuously for 90 d. In the first and the second column, bioleaching was performed without agglomeration of the ore and on the agglomerated ore, respectively. 28wt% of the copper was extracted in the first column after 40 d, while this figure was 38wt% in the second column. After 90 d, however, the overall extractions were almost the same for both of them. Bioleaching with mesophilic bacteria was performed in the third column without agglomeration of the ore and in the fourth column on the agglomerated ore. After 40 d, copper extractions in the third and the fourth columns were 62wt% and 70wt%, respectively. Copper extractions were 75wt% for both the columns after 90 d. For the last column, bioleaching was performed with moderate thermophilic bacteria and agglomerated ore. Copper extractions were 80wt% and 85wt% after 40 and 90 d, respectively. It was concluded that crushing and agglomeration of the ore using bacteria could enhance the copper extraction considerably.

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 ...

Thermodynamic properties of bioleaching liquid mixtures with and without mesophilic bacteria at different temperatures

In this work, low-grade copper sulfide mine has been treated by the bioleaching process using native cultures of Acidithiobacillus ferrooxidans. The bioleaching experiments were carded out in shake flasks at pH 2.0, 180 r.min^-1 and 30℃ for mesophilic bacteria The conductivity of copper bioleaching liquid was determined by the electric conductivity method at temperatures ranging from 298 K to 313 K. The ionic activity coefficients were estimated using Debye-Hucker and Osager-Falkenlagen equations. Meanwhile, the effects of temperature and concentrtion on the mean ionic activity coefficients were discussed. The relative partial molar free energies, enthalpies and entropies of copper teaching solution at above experimental temperatures were calculated. The behaviors of change of relative partial molar quantities were discussed on the basis of electrolytic solution theory. Simultaneously, the thermodynamic characters of bioleaching solution with and without mesophilic bacteria were compared. The existence of mesophilic bacteria changed the Fe^3＋/Fe^2＋ ratio, which resulted in the difference of ionic interaction. The experimental data show that the determination of the thermodynamic properties during the bioleaching processes should be important.

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.

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.

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.

Variation of Potential Nitrification and Ammonia-Oxidizing Bacterial Community with Plant-Growing Period in Apple Orchard Soil

In this study, we investigated the potential nitrification and community structure of soil-based ammonia-oxidizing bacteria （AOB） in apple orchard soil during different growth periods and explored the effects of environmental factors on nitrification activity and AOB community composition in the soil of a Hanfu apple orchard, using a culture-dependent technique and denaturing gradient gel electrophoresis （DGGE）. We observed that nitrification activity and AOB abundance were the highest in November, lower in May, and the lowest in July. The results of statistical analysis indicated that total nitrogen （N） content, NH4＋-N content, NO3-N content, and pH showed significant correlations with AOB abundance and nitrification activity in soil. The Shannon-Winner diversity, as well as species richness and evenness indices （determined by PCR-DGGE banding patterns） in soil samples were the highest in September, but the lowest in July, when compared to additional sampled dates. The DGGE fingerprints of soil-based 16S rRNA genes in November were apparently distinct from those observed in May, July, and September, possessing the lowest species richness indices and the highest dominance indices among all four growth periods. Fourteen DGGE bands were excised for sequencing. The resulting analysis indicated that all AOB communities belonged to the 13-Proteobacteria phylum, with the dominant AOB showing high similarity to the Nitrosospira genus. Therefore, soil-based environmental factors, such as pH variation and content of NHa＋-N and NO3--N, can substantially influence the abundance of AOB communities in soil, and play a critical role in soil-based nitrification kinetics.

Autotrophic potential in mesophilic heterotrophic bacterial isolates from Sino-Pacific marine sediments

Microbial carbon fixation is a paramount process in the ocean especially below the photic zone both in water and sedimentary ecosystems. Autotrophic microbes that fix carbon dioxide are renowned. However, the question whether heterotrophs can also fix carbon is intriguing. Ten heterotrophically grown, identified bacterial isolates from the Sino-Pacific marine sediments were tested for autotrophic uptake potential with and without addition of electron donors. Nine of the ten isolates showed carbon uptake capacity without addition of any substrate at very low rates in the order of 10^（-8） to 10^（-4） fmol/（cell·h）. The addition of manganese and ammonium at 1 mmol/L final concentration enhanced the uptake potential. Addition of 1 mmol/L final concentrations of reduced iron（10^（-6） to10^（-5） fmol/（cell·h） and sulfide（10^（-5） fmol/（cell·h） decreased the uptake potential significantly at p〈0.1. Bacterial tolerance to formaldehyde suggested propensities of anaplerotic chemical reactions that form metabolic intermediates of C-1 metabolism pathways. The isolates displayed high metabolic flexibility. With the changes in electron donors, the isolates metabolically toggled between relatively anoxic reductive iron/sulfur cycles and the oxidative cycles of manganese/ammonium and vice-versa. This property makes these microbes successful survivors in the highly dynamic Sino-Pacific sediments.

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.

Abundance and Community Composition of Ammonia-Oxidizers in Paddy Soil at Different Nitrogen Fertilizer Rates

Ammonia oxidation, the first and rate-limiting step of nitrification, is carried out by both ammonia-oxidizing bacteria （AOB） and ammonia-oxidizing archaea （AOA）. However, the relative importance of AOB and AOA to nitrification in terrestrial ecosystems is not well understood. The aim of this study was to investigate the effect of the nitrogen input amount on abundance and community composition of AOB and AOA in red paddy soil. Soil samples of 10-20 cm （root layer soil） and 0-5 cm （surface soil） depths were taken from a red paddy. Rice in the paddy was fertilized with different rates of N as urea of N1 （75 kg N ha＂ yr-1）, N2 （150 kg N ha~ yrl）, N3 （225 kg N ha1 yrl） and CK （without fertilizers） in 2009, 2010 and 2011. Abundance and community composition of ammonia oxidizers was analyzed by real-time PCR and denaturing gradient gel electrophoresis （DGGE） based on amoA （the unit A of ammonia monooxygenase） gene. Archaeal amoA copies in N3 and N2 were significantly （P〈0.05） higher than those in CK and N1 in root layer soil or in surface soil under tillering and heading stages of rice, while the enhancement in bacterial amoA gene copies with increasing of N fertilizer rates only took on in root layer soil. N availability and soil NO3--N content increased but soil NH4＋-N content didn＇t change with increasing of N fertilizer rates. Otherwise, the copy numbers of archaeal amoA gene were higher （P〈0.05） than those of bacterial amoA gene in root lary soil or in surface soil. Redundancy discriminate analysis based on DGGE bands showed that there were no obvious differs in composition of AOA or AOB communities in the field among different N fertilizer rates. Results of this study suggested that the abundance of ammonia-oxidizers had active response to N fertilizer rates and the response of AOA was more obvious than that of AOB. Similarity in the community composition of AOA or AOB among different N fertilizer rates indicate that the community composition of ammonia-oxidizers was relatively stable in the paddy soil at least in short term for three years.

Isolation and characterization of facultative mixotrophic ammonia-oxidizing bacteria from constructed wetlands

Autotrophic ammonia-oxidizing bacteria （AOB） have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands, and then cultivated in a mixotrophic medium containing ammonium and acetic acid. A molecular characterization was accomplished using ITS-PCR amplification, and phylogenetic analysis based on 16S rRNA gene Sequences. Results showed the presence of 35 bacteria, among 400 initially heterotrophic isolates, that were able to remove ammonia. These 35 isolates were classified into 10 genetically different groups based on ITS pattern. Then, a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies （ARE≥ 80%） and their phylogenetic diversity. In conditions of mixotrophy, these strains were shown to be able to grow （increase of optical density OD660 during incubation with assimilation of nitrogen into cellular biomass） and to oxidize ammonia （important ammonia oxidation efficiencies, AOE between 79% and 87%）. Among these facultative mixotrophic AOB, four isolates were genetically related to Firmicutes （Bacillus and Exiguobacterium）, three isolates were affiliated to Actinobacteria （Arthrobacter） and three other isolates were associated with Proteobacteria （Pseudomonas, Ochrobactrum and Bordetella）.

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.

Active ammonia-oxidizing bacteria and archaea in wastewater treatment systems

Ammonia-oxidizing bacteria(AOB)and archaea(AOA)are two microbial groups mediating nitrification,yet little is presently known about their abundances and community structures at the transcriptional level in wastewater treatment systems(WWTSs).This is a significant issue,as the numerical abundance of AOA or AOB at the gene level may not necessarily represent their functional role in ammonia oxidation.Using amo A genes as molecular markers,this study investigated the transcriptional abundance and community structure of active AOA and AOB in 14 WWTSs.Quantitative PCR results indicated that the transcriptional abundances of AOB amo A(averaged:1.6×10^(8)copies g^(-1)dry sludge)were higher than those of AOA(averaged:3.4×10^(7)copies g^(-1)dry sludge)in all WWTSs despite several higher abundances of AOA amo A at the gene level.Moreover,phylogenetic analysis demonstrated that Nitrosomonas europaea and unknown clusters accounted for 37.66%and 49.96%of the total AOB amo A transcripts,respectively,suggesting their dominant role in driving ammonia oxidation.Meanwhile,AOA amo A transcripts were only successfully retrieved from 3 samples,and the Nitrosospaera sister cluster dominated,accounting for 83.46%.Finally,the substrate utilization kinetics of different AOA and AOB species might play a fundamental role in shaping their niche differentiation,community composition,and functional activity.This study provides a basis for evaluating the relative contributions of ammonia-oxidizing microorganisms(AOMs)to nitrogen conversions in WWTSs.

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.

铁矿区内重金属对土壤氨氧化微生物群落组成的影响

以密云水库上游某铁矿区为研究对象，采用荧光定量PCR和变性梯度凝胶电泳（DGGE）分析了矿区内不同采样点的土壤中氨氧化微生物的数量和群落结构的变化，结果表明，土样中氨氧化细菌（AOB）和氨氧化古菌（AOA）的数量变化范围分别为3.01×107～1.08×109copies/g干土和8.65×107～2.69×109copies/g 干土.重金属含量与氨氧化微生物数量的相关性分析以及氨氧化微生物群落结构的冗余分析结果表明，该矿区内重金属污染改变了土壤中的氨氧化微生物的数量和结构.Cu污染对AOA的数量起到了显著抑制作用（r=-0.653＊， P〈0.05），但是对AOB则没有明显作用；Zn污染对尾矿库区域土壤的AOA/AOB比值影响显著（r=-0.606＊， P〈0.05）；Cd污染改变了AOB的种群分布，降低了AOB 的多样性水平.土壤中 Cr 长期干扰并没有改变氨氧化微生物的数量和结构，但是明显得抑制了氨氧化速率，表明重金属污染在一定程度上也影响了土壤生态系统的氮循环.

微生物菌液的界面特性

借助界面扩张流变测量方法,研究了2种微生物菌液体系与油相形成的界面特性。结果表明,微生物细胞及其代谢产物具有良好的表面活性,能够有效地降低油水界面张力。菌液与油相形成界面后,菌液内活性物质在界面上吸附和富集,界面张力迅速降低并达到较低的平衡值。微生物细胞的表面活性存在一个最佳温度范围,可能与微生物的最佳生长代谢温度相关。微生物细胞具有降低粘弹模量的作用,并改变了扩张粘度模量和扩张弹性模量对粘弹模量的贡献比例。

不同菌种对雌黄的生物浸出机理

分别采用嗜酸氧化亚铁硫杆菌(At.f)、嗜酸氧化硫硫杆菌(At.t)以及中度嗜热西伯利亚硫杆菌(S.s)对雌黄进行浸出。结果表明:嗜酸氧化硫硫杆菌浸出雌黄的效果较好,浸出过程是细菌直接与矿物作用;中温菌浸出雌黄则主要通过将Fe2+氧化成Fe3+,再由Fe3+氧化溶解矿物,浸出后期在矿物表面生成覆盖层;嗜酸氧化亚铁硫杆菌很难在雌黄矿浆中生长;添加适量表面活性剂Tween-80能有效改善矿物表面润湿性,促进Fe3+对砷的浸出,但对细菌生长会造成影响,抑制细菌直接浸出。