Diversity of anammox bacteria and abundance of functional genes for nitrogen cycling in the rhizosphere of submerged macrophytes in a freshwater lake in summer

Purpose

Submerged plants make an important contribution to nitrogen cycling in lakes including in the rhizosphere microenvironment through microbial activities. The main objective of this study was to investigate the abundance of functional genes for nitrogen cycling and the ecological relationship between these genes in the rhizosphere sediment of a freshwater lake in summer.

Materials and methods

Sediment from the rhizosphere of four submerged macrophytes (Ceratophyllum demersum, Hydrilla verticillata, Potamogeton maackianus, and Vallisneria spiralis) was sampled in Lake Liangzi, China, in summer. The anammox bacteria community structure and abundance of five functional genes for nitrogen cycling, ammonia monooxygenase (amoA) of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA), anammox 16S rRNA, and nitrite reductase genes (nirK and nirS) in the sampled sediment, were determined.

Results and discussion

A total of 100 anammox gene sequences were grouped into eight operational taxonomic units (OTUs) and genus Ca. Kuenenia was the dominant species in Lake Liangzi in summer. Quantitative polymerase chain reaction (qPCR) revealed that gene copies of AOA amoA (2.42?×?10⁶ copies g^?1) were more than one order of magnitude higher than those of AOB amoA (1.98?×?10⁵ copies g^?1). The nirS gene (4.13?×?10⁸ copies g^?1) was more abundant than the nirK gene (7.28?×?10⁷ copies g^?1). There was no significant difference in the abundance of the AOB amoA gene among the rhizosphere of the four macrophytes. Redundancy analysis (RDA) showed a positive correlation between the abundance of the anammox 16S rRNA gene, AOA amoA and AOB amoA, which suggested two of these microbes may have provided a substrate for anammox bacteria in summer.

Conclusions

The diversity of anammox in the rhizosphere of submerged macrophytes of the freshwater lake in summer was very low, but the plant species could affect the abundance of most nitrogen circulating bacteria, especially for anammox bacteria. Anammox 16S rRNA gene was positively correlated with four other functional genes, indicating that all four genes had significant effects on anammox bacteria.

     

Community structure of microeukaryotes in a rice rhizosphere revealed by DNA-based PCR-DGGE

Rice roots provide a specific habitat for microorganisms in the rhizosphere of a submerged field through supply of oxygen and organic matter. Many studies have focused on the microbial community in the rice rhizosphere, but less is still known about the microeukaryotic community structure of rice rhizosphere. This study explored the microeukaryotic community structure of a rice rhizosphere through denaturing gradient gel electrophoresis (DGGE) targeting 18S rRNA gene. The rice roots and the rhizosphere soil samples, which were collected from a field under rice-wheat rotation system, were separately analyzed. To characterize the rice rhizosphere-specific community, the bulk soil of rice field and the wheat rhizosphere samples were also examined. DGGE fingerprints showed that the microeukaryotic community of rice roots were distinct from the community of the bulk soil and showed a temporal shift with the growth stage. The rhizosphere soil community was distinct from the root and bulk soil communities, but this could be explained by that the root and bulk soil communities were shared in the rhizosphere. The rice rhizosphere community was also distinct from those in the wheat rhizosphere. Microeukaryotes that characterized the rice rhizosphere (roots and the rhizosphere soil) community could be affiliated to Polymyxa, flagellates, and oomycetes, which suggested that microeukaryotes with various ecological roles, e.g., parasites, bacterial grazers, and decomposers, inhabit the rice rhizosphere. The results showed that the rice root and its growth stages are key factors shaping the microeukaryotic community structure in the rhizosphere.     

Oxygen and redox potential gradients in the rhizosphere of alfalfa grown on a loamy soil

Oxygen (O₂) supply and the related redox potential (E_H) are important parameters for interactions between roots and microorganisms in the rhizosphere. Rhizosphere extension in terms of the spatial distribution of O₂ concentration and E_H is poorly documented under aerobic soil conditions. We investigated how far O₂ consumption of roots and microorganisms in the rhizosphere is replenished by O₂ diffusion as a function of water/air‐filled porosity. Oxygen concentration and E_H in the rhizosphere were monitored at a mm‐scale by means of electroreductive Clark‐type sensors and miniaturized E_H electrodes under various matric potential ranges. Respiratory activity of roots and microorganisms was calculated from O₂ profiles and diffusion coefficients. pH profiles were determined in thin soil layers sliced near the root surface. Gradients of O₂ concentration and the extent of anoxic zones depended on the respiratory activity near the root surface. Matric potential, reflecting air‐filled porosity, was found to be the most important factor affecting O₂ transport in the rhizosphere. Under water‐saturated conditions and near field capacity up to –200 hPa, O₂ transport was limited, causing a decline in oxygen partial pressures (pO₂) to values between 0 and 3 kPa at the root surface. Aerobic respiration increased by a factor of 100 when comparing the saturated with the driest status. At an air‐filled porosity of 9% to 12%, diffusion of O₂ increased considerably. This was confirmed by E_H around 300 mV under aerated conditions, while E_H decreased to 100 mV on the root surface under near water‐saturated conditions. Gradients of pO₂ and pH from the root surface indicated an extent of the rhizosphere effect of 10–20 mm. In contrast, E_H gradients were observed from 0 to 2 mm from the root surface. We conclude that the rhizosphere extent differs for various parameters (pH, Eh, pO₂) and is strongly dependent on soil moisture.     

Analysis of bacterial communities on alkaline phosphatase genes in soil supplied with organic matter

Abstract

We studied the effects of the application of organic matter (OM) and chemical fertilizer (CF) on soil alkaline phosphatase (ALP) activity and ALP-harboring bacterial communities in the rhizosphere and bulk soil in an experimental lettuce field in Hokkaido, Japan. The ALP activity was higher in soils with OM than in soils with CF, and activity was higher in the rhizosphere for OM than in the bulk soil. Biomass P and available P in the soil were positively related to the ALP activity of the soil. As a result, the P concentration of lettuce was higher in OM soil than in CF soil. We analyzed the ALP-harboring bacterial communities using polymerase chain reaction based denaturing gradient gel electrophoresis (DGGE) on the ALP genes. Numerous ALP genes were detected in the DGGE profile, regardless of sampling time, fertilizer treatment or sampled soil area, which indicated a large diversity in ALP-harboring bacteria in the soil. Several ALP gene fragments were closely related to the ALP genes of Mesorhizobium loti and Pseudomonas fluorescens. The community structures of the ALP-harboring bacteria were assessed using principal component analysis of the DGGE profiles. Fertilizer treatment and sampled soil area significantly affected the community structures of ALP-harboring bacteria. As the DGGE bands contributing to the principal component were different from sampling time, it is suggested that the major bacteria harboring the ALP gene shifted. Furthermore, there was, in part, a significant correlation between ALP activity and the community structure of the ALP-harboring bacteria. These results raise the possibility that different ALP-harboring bacteria release different amounts and/or activity of ALP, and that the structure of ALP-harboring bacterial communities may play a major role in determining overall soil ALP activity.     

水稻根际和周围土壤中微生物功能基因丰度与碳、氮状况及其通量之间的关系

Rapid nitrogen（N） transformations and losses occur in the rice rhizosphere through root uptake and microbial activities. However,the relationships between rice roots and rhizosphere microbes for N utilization are still unclear. We analyzed different N forms（NH＋4,NO-3, and dissolved organic N）, microbial biomass N and C, dissolved organic C, CH4 and N2O emissions, and abundance of microbial functional genes in both rhizosphere and bulk soils after 37-d rice growth in a greenhouse pot experiment. Results showed that the dissolved organic C was significantly higher in the rhizosphere soil than in the non-rhizosphere bulk soil, but microbial biomass C showed no significant difference. The concentrations of NH＋4, dissolved organic N, and microbial biomass N in the rhizosphere soil were significantly lower than those of the bulk soil, whereas NO-3in the rhizosphere soil was comparable to that in the bulk soil. The CH4 and N2O fluxes from the rhizosphere soil were much higher than those from the bulk soil. Real-time polymerase chain reaction analysis showed that the abundance of seven selected genes, bacterial and archaeal 16 S rRNA genes, amoA genes of ammonia-oxidizing archaea and ammonia-oxidizing bacteria, nosZ gene, mcrA gene, and pmoA gene, was lower in the rhizosphere soil than in the bulk soil, which is contrary to the results of previous studies. The lower concentration of N in the rhizosphere soil indicated that the competition for N in the rhizosphere soil was very strong, thus having a negative effect on the numbers of microbes. We concluded that when N was limiting, the growth of rhizosphere microorganisms depended on their competitive abilities with rice roots for N.     

Root exudate effects on the bacterial communities, CO2 evolution, nitrogen transformations and ATP content of rhizosphere and bulk soils

The ATP content, soil respiration, bacterial community composition, and gross N mineralization and immobilization rates were monitored under laboratory condition at 25 °C for 28 d in a model system where low molecular weight root exudates (glucose and oxalic acid) were released by a filter placed on the surface of a forest soil also treated with ¹⁵N, so as to simulate rhizosphere conditions. Periodically, the soil was sampled from two layers, 0-2 and 6-14 mm below the filter's surface, which were indicated as rhizosphere and bulk soils, respectively. The isotope dilution technique was used to determine the effect of these low molecular weight organic compounds (LMWOCs) on gross N mineralization and immobilization rates. From 0 to 3 d both glucose and oxalic acid amended soils showed a rapid evolution of CO₂, more pronunced in the latter treatment together with a decrease in the amount of mineral N of the rhizosphere soil, probably due to N immobilization. Nevertheless, these changes were accompanied by a very small increase in the net ATP content probably because the low C application rate stimulated microbial activity but microbial growth only slightly. A positive ‘priming effect’ probably developed in the oxalic acid amended soil but not in the glucose amended soil. Gross N mineralization and immobilization rates were only observed in the rhizosphere soil, probably due to the greater C and N concentrations and microbial activity, and were a little higher in both amended soils than in the control soil, only between 1 and 7 d. Both glucose and oxalic acid influenced the bacterial communities of the rhizosphere soil, as new bands in the DGGE profiles appeared at 3 and 7 d. Glucose induced lower changes in the bacterial community than oxalic acid, presumably because the former stimulated a larger proportion of soil microorganisms whereas the latter was decomposed by specialized microorganisms. Peaks of net daily soil respiration and net ATP content and the appearence of new dominant bacterial populations were shifted in time, probably because there was less ATP synthesis and DGGE patterns changed after complete substrate mineralization.     

Analysis of bacterial communities in soil by PCR-DGGE targeting protease genes

We studied the effects of the application of organic (OM) and inorganic fertilizer (CF) on soil protease activity and proteolytic bacterial communities in rhizosphere and bulk soil on an experimental lettuce field in Hokkaido, Japan. The protease activity always was higher in soils of the OM than with the CF treatment, and also higher in the rhizosphere than in the bulk soil. We analyzed proteolytic bacterial communities by denaturing gradient gel electrophoresis (DGGE) of the alkaline metalloprotease (apr) and neutral metalloprotease (npr) genes. Most apr forms detected were closely related to apr of Pseudomonas fluorescens, and all npr variants closely resembled the gene of Bacillus megaterium. These results were consistent with findings from tests using cultured bacterial communities, indicating a high specificity of our PCR-DGGE for amplifying apr and npr genes. The community compositions of proteolytic bacteria were assessed by principal component analysis of the DGGE profiles. There were significant differences in the effects of CF and OM on the community compositions of apr- and npr-expressing bacteria, and the communities of the two types of bacteria played different roles in rhizosphere and bulk soil. We found significant correlations between the protease activity and the communities of the two types of bacteria. The results indicate that different proteolytic bacteria release different amounts or activities of protease, and that the composition of proteolytic bacterial communities may play a major role in determining overall soil protease activity.     

Detection of aluminium tolerance plasmids and microbial diversity in the rhizosphere of plants grown in acidic volcanic soil

The rhizosphere is considered as a hot-spot for gene exchange among bacteria in terrestrial ecosystems. Chilean volcanic soils are characterized by low pH and high concentrations of aluminium (Al) in the soil solution, thus Al tolerance could be important for the survival of microorganisms in these soils; loss of genes encoding for Al tolerance may affect competitiveness particularly in the rhizosphere where competition is strong. The occurrence of Al-tolerance plasmids was investigated in the rhizospheres of pasture and crop plants growing in acidic volcanic soils from southern Chile. Al tolerance plasmids were captured by biparental mating. Two types of Al tolerance plasmids could be distinguished, based on their endonuclease restriction pattern. One plasmid of each group (denoted as pRPA21 and pOPA21) was selected for further studies. The plasmids showed a high stability in presence and absence of Al. Additionally, microbial community composition in the rhizosphere soils was assessed by denaturing gradient gel electrophoresis (DGGE). Sequencing of DGGE bands revealed among others, members of the bacterial phylum Gemmatimonadetes and archaeal phylum Crenarchaeota. The present study shows that the rhizosphere of pasture and crop plants growing in Chilean volcanic soil harbors genetic mobile elements which could play a role in the adaptation of bacterial populations to environmental stressors, such as Al-toxicity.     

Impact of irrigation water quality on soil nitrifying and total bacterial communities

Disturbance induced by two contrasting irrigation regimes (groundwater versus urban wastewater) was evaluated on a sandy agricultural soil through chemical and microbial analyses. Contrary to wastewater, groundwater displayed very high nitrate contents but small amounts of ammonium and organic matter. Despite these strong compositional shifts, soil organic carbon and nitrogen, nitrate and ammonium contents were not significantly different in both types of irrigated plot. Moreover, neither microbial biomass nor its activity, determined as fluorescein diacetate hydrolysis activity, was influenced by irrigation regimes. Bacterial community structure, assessed by denaturing gradient gel electrophoresis (DGGE) of 16S ribosomal DNA fragments, was also weakly impacted as molecular fingerprints shared an overall similarity of 85%. Ammonia-oxidizing bacterial community (AOB) was monitored by DGGE of the functional molecular marker amoA gene (alpha subunit of the ammonia monooxygenase). Surprisingly, no amoA signals were obtained from plots irrigated with groundwater, whereas signal intensities were high in all plots under wastewater. Among the last, compositional shifts of the AOB community were weak. Overall, impact of irrigation water quality on soil chemistry could not be evidenced, whereas effects were low on the total bacterial compartment but marked on the AOB community.     

Diversity and antagonistic potential of Pseudomonas spp. associated to the rhizosphere of maize grown in a subtropical organic farm

Pseudomonas spp. are one of the most important bacteria inhabiting the rhizosphere of diverse crop plants and have been frequently reported as biological control agents (BCAs). In this work, the diversity and antagonistic potential of Pseudomonas spp. in the rhizosphere of maize cultivars Nitroflint and Nitrodent grown at an organic farm in Brazil was studied by means of culture-dependent and -independent methods, respectively. Sampling of rhizosphere soil took place at three different stages of plant development: 20, 40 and 106 days after sowing. A PCR-DGGE strategy was used to generate specific Pseudomonas spp. fingerprints of 16S rRNA genes amplified from total community rhizosphere DNA. Shifts in the relative abundance of dominant populations (i.e. PCR-DGGE ribotypes) along plant development were detected. A few PCR-DGGE ribotypes were shown to display cultivar-dependent relative abundance. No significant differences in diversity measures of DGGE fingerprints were observed for different maize cultivars and sampling times. The characterisation and assessment of the antagonistic potential of a group of 142 fluorescent Pseudomonas isolated from the rhizosphere of both maize cultivars were carried out. Isolates were phenotypically and genotypically characterised and screened for in vitro antagonism towards three phytopathogenic fungi and the phytopathogenic bacterium Ralstonia solanacearum. Anti-fungal activity was displayed by 13 fluorescent isolates while 40 isolates were antagonistic towards R. solanacearum. High genotypic and phenotypic diversity was estimated for antagonistic fluorescent Pseudomonas spp. PCR-DGGE ribotypes displayed by antagonists matched dominant ribotypes of Pseudomonas DGGE fingerprints, suggesting that antagonists may belong to major Pseudomonas populations in the maize rhizosphere. Antagonists differing in their genotypic and phenotypic characteristics shared the same DGGE electrophoretic mobility, indicating that an enormous genotypic and functional diversity might be hidden behind one single DGGE band. Cloning and sequencing was performed for a DGGE double-band which had no corresponding PCR-DGGE ribotypes among the antagonists. Sequences derived from this band were affiliated to Pseudomonas stutzeri and P. alcaligenes 16S rRNA gene sequences. As used in this study, the combination of culture-dependent and -independent methods has proven to be a powerful tool to relate functional and structural diversity of Pseudomonas spp. in the rhizosphere.     

Fluctuations in the abundance of ammonia oxidizers and the contents of inorganic nitrogen in solarized soil

Solarization makes a great impact on the abundance of ammonia oxidizers and nitrifying activity in soil. To elucidate fluctuations in the abundance of ammonia oxidizers and nitrification in solarized soil, copy numbers of amoA gene of ammonia-oxidizing bacteria (AOB) and archaea (AOA), viable number of ammonia oxidizers and inorganic nitrogen contents were investigated in greenhouse experiments. The copy number of amoA gene and the viable number of ammonia oxidizers were determined by the quantitative polymerase chain reaction and most probable number methods, respectively. Abundance of AOB based on the estimation of amoA gene copy numbers and viable counts of ammonia oxidizers was decreased by the solarization treatment and increased during the tomato (Solanum lycopersicum L.) cultivation period following the solarization. Effect of solarization on the copy number of amoA gene of AOA was less evident than that on AOB. The proportion of nitrate in inorganic nitrogen contents was declined by the solarization and increased during the tomato cultivation period following the solarization. Positive correlations were found between the proportion of nitrate in inorganic nitrogen content and the copy number of bacterial or archaeal amoA gene or the viable number of ammonia oxidizers; the copy number of bacterial amoA gene showed a strong correlation with the viable number of ammonia oxidizers. The present study revealed influences of solarization on the fluctuation in the abundance of ammonia oxidizers and dynamics of inorganic nitrogen contents in soil and the results indicate that the determination of amoA gene of AOB is possibly a quick and useful diagnostic technique for evaluating suppression and restoration of nitrification following solarization.     

Hydrocarbon degradation potential and activity of endophytic bacteria associated with prairie plants

Phytoremediation systems for organic compounds such as petroleum hydrocarbons rely on a synergistic relationship between plants and their root-associated microbial communities. To determine the probable role of endophytic bacterial communities in these systems, this study examined both rhizosphere and endophytic communities of five different plant species at a long-term phytoremediation field site. Hydrocarbon degradation potential and activity were assessed using MPN assays, PCR analysis of catabolic genes associated with hydrocarbon degradation, and mineralization assays with C-14 labeled hydrocarbons. Microbial community structure in each niche was assessed by DGGE analysis of 16S rRNA gene fragments and subsequent band sequencing. Both endophytic degrader populations and endophytic degrader activity showed substantial inter-species variation, largely independent of that shown by the respective rhizosphere populations. Endophytic hydrocarbon degradation was linked to dominant bacterial endophytes. Pseudomonas spp. dominated endophytic communities exhibited increased alkane hydrocarbon degradation potential and activity, while Brevundimonas and Pseudomonas rhodesiae dominated endophytic communities were associated with increased PAH degradation potential and activity. In one plant species, Lolium perenne, increased endophytic alkane hydrocarbon degradation was associated with increased rhizosphere alkane degradation and decreased rhizosphere PAH degradation. Our results show that diverse plant species growing in weathered-hydrocarbon contaminated soil maintain distinct, heterogeneously distributed endophytic microbial populations, which may impact upon the ability of plants to promote the degradation of specific types of hydrocarbons.     

不同生物硝化抑制剂对红壤性水稻土N2O排放的影响及其机制

为揭示不同生物硝化抑制剂(BNIs)对红壤性水稻土N₂O排放的影响差异及作用机制,通过21 d的土柱淹水培养试验,比较了三种BNIs 1,9-癸二醇(1,9-D)、亚麻酸(LN)和3-(4-羟基苯基)丙酸甲酯(MHPP)与化学合成硝化抑制剂双氰胺(DCD)对土壤N₂O排放及相关硝化、反硝化功能基因的影响。结果表明:不同BNIs(1,9-D、LN、MHPP)可以显著平均降低土壤N₂O日排放峰值40.1%;1,9-D和MHPP可分别抑制N₂O排放总量44.5%和43.9%,而DCD和LN对N₂O排放总量没有显著影响。1,9-D和MHPP对AOA(氨氧化古菌)、AOB(氨氧化细菌)硝化菌和nirS、nirK型反硝化菌的调控均有所不同,1,9-D可以同时抑制AOA、AOB和nirS微生物的生长;MHPP仅可以抑制AOA的生长;其中,AOA-amoA和nirS基因丰度与土壤N₂O的排放呈显著正相关关系。同时,1,9-D和MHPP均增加了nosZ基因丰度及其与AOA-...     

Etude integree de sols a pseudo-gley sous vegetation naturelle du plateau des Hautes Fagnes (Belgique): Partie I le milieu de pedogenese, la morphologie et le fonctionnement actuel des profils

This work is an attempt to establish the morphogenesis of four hydromorphic profiles (Plateau des Fagnes, Eastern part of the Belgian Ardennes). Various scientifical approaches were used, namely geomorphology, mineralogy and palynology.The aim was to show that very sharp discordances may appear between “inherited” profile morphology and the present state of hydromorphy characterized by vegetation, humus forms, and physico-chemical measurements, such as redox potential and oxygen diffusion flux intensity (see partie II)     

Are the biogeochemical cycles of carbon,nitrogen, sulfur,and phosphorus driven by the “Fe<Superscript>III</Superscript>–Fe<Superscript>II</Superscript> redox wheel” in dynamic redox environments?

Purpose  

Iron’s fluctuation between the II (ferrous) and III (ferric) oxidation states has been coined as the “Fe^III–Fe^II redox wheel.” Numerous studies have coupled the “iron redox wheel” with the biogeochemical cycle of carbon (C), nitrogen (N), sulfur (S), or phosphorus (P) individually in soils or sediments, but evidence suggests that the Fe^III–Fe^II redox wheel drives the biogeochemical cycles interactively in a fluctuating redox microenvironment. The interactions of the Fe^III–Fe^II redox wheel with the biogeochemical cycles of C, N, S, and P in the fluctuating redox environments were reviewed in this paper.     

Microbial response to exudates in the rhizosphere of young beech trees (Fagus sylvatica L.) after dormancy

Plants act as an important link between atmosphere and soil: CO2 is transformed into carbohydrates by photosynthesis. These assimilates are distributed within the plant and translocated via roots into the rhizosphere and soil microorganisms. In this study, 3 year old European beech trees (Fagus sylvatica L.) were exposed after the chilling period to an enriched ¹³C–CO₂ atmosphere (δ¹³C = 60‰ – 80‰) at the time point when leaves development started. Temporal dynamics of assimilated carbon distribution in different plant parts, as well as into dissolved organic carbon and microbial communities in the rhizosphere and bulk soil have been investigated for a 20 days period. Photosynthetically fixed carbon could be traced into plant tissue, dissolved organic carbon and total microbial biomass, where it was utilized by different microbial communities. Due to carbon allocation into the rhizosphere, nutrient stress decreased; exudates were preferentially used by Gram-negative bacteria and (mycorrhizal) fungi, resulting in an enhanced growth. Other microorganisms, like Gram-positive bacteria and mainly micro eucaryotes benefited from the exudates via food web development. Overall our results indicate a fast turnover of exudates and the development of initial food web structures. Additionally a transport of assimilated carbon into bulk soil by (mycrorhizal) fungi was observed.     

抗真菌转基因水稻根际土壤真菌群落结构的动态变化

以非转基因水稻"七丝软粘"为对照,采用传统平板计数法和变性梯度凝胶电泳技术,研究了抗真菌转基因水稻"转品1"和"转品8"生长周期内对根际土壤中可培养真菌数和真菌群落结构的影响。结果显示,相同生育期转基因水稻根际土壤可培养真菌数量与其非转基因对照水稻相比较无显著性差异,表明转基因水稻的种植没有对根际土壤真菌数量产生明显影响;18S rRNA真菌群落DGGE图谱分析显示,相同生育期转基因水稻与其非转基因对照水稻的根际土壤真菌DGGE条带数量和条带位置均无显著性差异,表明转基因水稻的种植没有对根际土壤真菌群落结构产生明显影响。进一步分析相同生育期转基因水稻与其非转基因对照水稻的根际土壤真菌群落香农多样性指数(Shannon diversity index)和均匀度指数(Evenness index)的动态变化,发现两者均没有显著性差异。以上研究结果表明,外源抗真菌基因的导入对水稻根际土壤中真菌群落数量和群落结构均没有明显影响。此外,将不同位置的真菌DGGE条带切胶回收,克隆、测序后,进行系统进化树分析,结果表明根际土壤真菌群落主要归属为子囊菌门(Ascomycota)、担子菌门(Basidiomycota)、壶菌门(Chytridiomycota)、接合菌门(Zygomycota)和未知真菌(unknown fungi)5个类群。     

Molecular characterization of methane-oxidizing bacteria associated with rice straw decomposition in a rice field

Abstract

Methane-oxidizing bacteria (MOB) are crucial to the reduction of CH₄ emitted to the atmosphere. However, it is unclear how MOB in rice straw are affected by straw decomposition processes. In a Japanese rice field, a year-round experiment was set up to study the effects of agricultural practice (rice cultivation/winter fallow), straw parts (leaf sheath/blade) and the site of straw placement (plow layer/soil surface) on MOB communities in rice straw using denaturing gradient gel electrophoresis (DGGE) and DNA sequencing analyses of key MOB functional genes (pmoA and amoA). Thirty-eight different DGGE bands were observed over the entire investigation period. Principal component analysis of DGGE pattern suggested that agricultural practice is the key factor regulating the MOB communities. Sequencing of dominant DGGE bands showed that: (1) during the rice cultivation period, methanotrophs (particularly type I methanotrophs) dominated the MOB community, (2) during the winter fallow season both type I and type II methanotrophs were dominant in sheath segments placed both on the soil surface and in the plow layer, whereas ammonia oxidizers seemed to dominate blade segments placed in the plow layer. Alignment of diagnostic amino acid sequences of MOB suggested the presence of novel ammonia oxidizers in rice straw in rice fields.     

Characterization of nitrifying bacteria communities of soils from different ecological regions of China by molecular and conventional methods

Soil nitrification rate is very different among soil types, as a result of differences in physical and chemical properties. Little is known about the composition of the nitrifying bacteria community. In this investigation, three soils (fluvo-aquic soil, permeable paddy soil and red earth) from different geo-ecological regions in China were characterized for their nitrification activities and their nitrifying bacteria communities determined either by molecular approaches or by conventional culture methods. A 28-day long-term soil incubation showed that the maximum nitrification potential was found in the fluvo-aquic soil with almost 100% of inorganic N present as NO₃^–-N, while the minimum nitrification potential was in red earth with only a 4.9% conversion rate from ammonium into nitrate. There was no relationship between nitrification potential and numbers of nitrifiers in the soil. The conventional most probable number (MPN) method could enumerate ammonia oxidizers, but failed in enumerating nitrite oxidizers. Therefore, we used an MPN-PCR procedure which gave a convincing nitrite oxidizer count result, instead of MPN-diphylamine. Soils were characterized by denaturing gradient gel electrophoresis (DGGE) of DNA extracted from soils and amplified using a primer specific for the 16S rRNA gene and/or for the amoA gene. The DGGE columns of the three soils differed from each other. There were two similar bands present in DGGE columns of the fluvo-aquic and permeable paddy soils, but no similar band was found in DGGE columns of the red earth. The sequence of amoA indicated that all ammonia oxidizers in these soils were grouped into Nitrosospira clusters 1 and 3, and each soil had a common band similar to the other soils and a special band which differed from the other soils.     

The dynamics of oxygen concentration, pH value, and organic acids in the rhizosphere of Juncus spp.

A novel type of planar optodes for simultaneous optical analysis of pH and oxygen dynamics in the rhizosphere is introduced. The combination of the optical, non-invasive measurement of these parameters with sterile sampling of rhizosphere solution across and along growing roots by use of a novel type of rhizobox provides a methodical step forward in the investigation of the physicochemical dynamics of the rhizosphere and its underlying matter fluxes between roots and soil. In this study, this rhizobox was used to investigate the effect of oxygen releasing roots of three Juncus species on the amount and distribution of organic acids in reductive, oxygen-deficient soils of different pH (pH 3.9-pH 5.9). Pronounced diurnal variations of oxygen concentration and pH along the roots, particularly along the elongation zone were observed. Long-term records over more than eight weeks revealed considerable spatial and temporal patterns of oxygen over a range of almost 200 μmol O₂ L⁻¹ and pH dynamics of ±1.4 pH units in the rhizosphere. A strong effect of oxidative acidification due to oxygen release by the plant roots was clearly visible for Juncus effusus, whereas the roots of Juncus articulatus alkalinized the rhizosphere. In contrast, roots of Juncus inflexus induced no effects on rhizospheric pH. Only four different organic acids (oxalate, acetate, formate and lactate) were detectable in all soil solutions. Maximal concentration of all organic acids occurred at pH 3.9, whereas the lowest concentration of each organic acid was found at pH 5.9. Hence, considering the pH-dependence of the redox potential, the acid soil provided increased reductive conditions leading to slower anaerobic degradation of organic acids to CO₂ or methane (CH₄). The concentration of organic acids decreased by up to 58% within a distance of only 4 mm from the bulk soil to the root surface, i.e. reciprocal to the pronounced O₂-gradient. The decreasing presence of organic acids toward the oxygen releasing roots is possibly due to a change in the composition of the microbial community from anaerobic to aerobic conditions. The present study highlights the dynamic interplay between O₂ concentration, pH and organic acids as key parameters of the physicochemical environment of the rhizosphere, particularly for wetland plants growing in oxygen-deficient waterlogged soils.