Impacts of 3 years of elevated atmospheric CO2 on rhizosphere carbon flow and microbial community dynamics

Carbon (C) uptake by terrestrial ecosystems represents an important option for partially mitigating anthropogenic CO₂ emissions. Short‐term atmospheric elevated CO₂ exposure has been shown to create major shifts in C flow routes and diversity of the active soil‐borne microbial community. Long‐term increases in CO₂ have been hypothesized to have subtle effects due to the potential adaptation of soil microorganism to the increased flow of organic C. Here, we studied the effects of prolonged elevated atmospheric CO₂ exposure on microbial C flow and microbial communities in the rhizosphere. Carex arenaria (a nonmycorrhizal plant species) and Festuca rubra (a mycorrhizal plant species) were grown at defined atmospheric conditions differing in CO₂ concentration (350 and 700 ppm) for 3 years. During this period, C flow was assessed repeatedly (after 6 months, 1, 2, and 3 years) by ¹³C pulse‐chase experiments, and label was tracked through the rhizosphere bacterial, general fungal, and arbuscular mycorrhizal fungal (AMF) communities. Fatty acid biomarker analyses and RNA‐stable isotope probing (RNA‐SIP), in combination with real‐time PCR and PCR‐DGGE, were used to examine microbial community dynamics and abundance. Throughout the experiment the influence of elevated CO₂ was highly plant dependent, with the mycorrhizal plant exerting a greater influence on both bacterial and fungal communities. Biomarker data confirmed that rhizodeposited C was first processed by AMF and subsequently transferred to bacterial and fungal communities in the rhizosphere soil. Over the course of 3 years, elevated CO₂ caused a continuous increase in the ¹³C enrichment retained in AMF and an increasing delay in the transfer of C to the bacterial community. These results show that, not only do elevated atmospheric CO₂ conditions induce changes in rhizosphere C flow and dynamics but also continue to develop over multiple seasons, thereby affecting terrestrial ecosystems C utilization processes.     

Plants‐associated microflora and the remediation of oil‐contaminated soil

The use of plants and their rhizospheric microorganisms is a promising emerging technology for remediating contaminated soils. The degradation of total petroleum hydrocarbon (TPH) in the rhizospheric and nonrhizospheric soil of three domestic plants, namely, alfalfa (Medicaga sativa) broad beans (Vicia faba) and ryegrass (Lolium perenne) was investigated. The experimental data from the studies of plantmicrobe‐soil interactions implicated the enhancement of TPH degradation by the rhizospheric microbial community. Although the three domestic plants exhibited normal growth in the presence of ～1.0% TPH, the degradation was more profound in the case of leguminous plants. The TPH degradation in the soil cultivated with broad beans and alfalfa was 36.6 and 35.8%, respectively, compared with 24% degradation in case of ryegrass. Such a high correlation between plant type and TPH degradation rates indicate that selection for enhanced rhizosphere degradation may be accomplished by selecting leguminous plants.     

Protein profile variation in cultivated and native freshwater microorganisms exposed to chemical environmental pollutants

Assimilation of ³⁵S-precursors into microbial proteins was used to investigate toxicity and adaptational responses that occur in nutrient enriched and natural freshwater samples experimentally contaminated with benzene, toluene, trichloroethylene (TCE), or xylene. Experiments were conducted to analyze (1) the potential of using microbial community protein profiles for responsive identification of chemical pollutant exposure, (2) the inhibition of microbial productivity through reduction in rate of protein synthesis caused by specific chemical pollutants, and (3) whether selection of subpopulations in freshwater microbial communities challenged with chemical pollutants leads to adaptive strategies mediated by production of particular polypeptides. The results show that distinct banding patterns of polypeptides in the range of 30 to 100 kilodaltons that were obtained following collective cultivation of freshwater microorganisms differ with each chemical pollutant. Protein yield and radioisotope incorporation were reduced within ten minutes of microbial exposure to chemical pollutants in the following order: xylene < toluene < benzene < TCE. Adaptation of the freshwater microbial community to chemical pollutants prior to radioisotope incorporation produced differences in polypeptide profiles, in the banding patterns of radioactive polypeptides, and in the rate of radioisotope incorporation. The rate of radioisotope incorporation by freshwater microorganisms pre-adapted to chemical pollutants was lowest with xylene (88.1% reduction), followed by TCE (84.0% reduction), toluene (67.3% reduction), and benzene (43.5% reduction). In long-term radioisotope incorporation experiments, protein yield and polypeptide radioactivity was higher in the presence of chemical pollutants than in uncontaminated control samples, suggesting increased metabolic productivity attributable to the chemical pollutants. Correspondence to: O.A. Ogunseitan.     

Identification of active denitrifiers in full‐scale nutrient removal wastewater treatment systems

Denitrification is essential to the removal of nitrogen from wastewater during treatment, yet an understanding of the diversity of the active denitrifying bacteria responsible in full‐scale wastewater treatment plants (WWTPs) is lacking. In this study, stable‐isotope probing (SIP) was applied in combination with microautoradiography (MAR)‐fluorescence in situ hybridization (FISH) to identify previously unrecognized active denitrifying phylotypes in a full‐scale WWTP with biological N and P removal. Acknowledging that different denitrifiers will have specific carbon source preferences, a fully ¹³C‐labelled complex substrate was used for SIP incubations, under nitrite‐reducing conditions, in order to maximize the capture of the potentially metabolically diverse denitrifiers likely present. Members of the Rhodoferax, Dechloromonas, Sulfuritalea, Haliangium and Thermomonas were represented in the 16S rRNA gene clone libraries from DNA enriched in ¹³C, with FISH probes optimized here for their in situ characterization. FISH and MAR confirmed that they were all active denitrifiers in the community. The combined approach of SIP and MAR‐FISH represents an excellent approach for identifying and characterizing an un‐described diversity of active denitrifiers in full‐scale systems.     

Ethanol degradation and the benefits of incremental priming in pilot-scale constructed wetlands

There has been significant global growth in the use of constructed wetlands for wastewater treatment. The fundamental microbial processes involved in the biodegradation of organic wastewater pollutants determine the range of design and operational parameters relevant to individual constructed wetlands. In this study, the biodegradation and mineralization of ethanol by acclimated and non-acclimated microbial populations in pilot-scale constructed wetlands were compared. By increasing the pollutant concentration at incremental intervals (incremental priming), the biodegradative capacity of a sand-filled constructed wetland was significantly enhanced. At an influent COD concentration of 15,800 mg L⁻¹, no volatile fatty acids were detected in the effluent of an incrementally primed system and the maximum effluent COD concentration was 180 mg L⁻¹. In contrast, an identical, unprimed system, amended with a lower concentration of COD (7587 mg L⁻¹), exhibited a maximum effluent COD concentration of 1400 mg L⁻¹, with the anaerobic metabolites, butyrate and propionate accounting for up to 83% of the effluent COD. It was demonstrated that the use of incremental priming, together with a vertical subsurface flow mode of operation enhanced long-term function of constructed wetlands. Future research should focus on determining the concentration gradients and incremental intervals necessary for optimal microbial acclimation to a range of organic pollutants and/or wastewaters, in order to minimize start-up times without significantly impairing the benefits derived from incremental priming.     

The catabolic pathways of in situ rhizosphere PAH degraders and the main factors driving PAH rhizoremediation in oil-contaminated soil

Rhizoremediation is a potential technique for polycyclic aromatic hydrocarbon (PAH) remediation; however, the catabolic pathways of in situ rhizosphere PAH degraders and the main factors driving PAH rhizoremediation remain unclear. To address these issues, stable-isotope-probing coupled with metagenomics and molecular ecological network analyses were first used to investigate the phenanthrene rhizoremediation by three different prairie grasses in this study. All rhizospheres exhibited a significant increase in phenanthrene removal and markedly modified the diversity of phenanthrene degraders by increasing their populations and interactions with other microbes. Of all the active phenanthrene degraders, Marinobacter and Enterobacteriaceae dominated in the bare and switchgrass rhizosphere respectively; Achromobacter was markedly enriched in ryegrass and tall fescue rhizospheres. Metagenomes of ¹³C-DNA illustrated several complete pathways of phenanthrene degradation for each rhizosphere, which clearly explained their unique rhizoremediation mechanisms. Additionally, propanoate and inositol phosphate of carbohydrates were identified as the dominant factors that drove PAH rhizoremediation by strengthening the ecological networks of soil microbial communities. This was verified by the results of rhizospheric and non-rhizospheric treatments supplemented with these two substances, further confirming their key roles in PAH removal and in situ PAH rhizoremediation. Our study offers novel insights into the mechanisms of in situ rhizoremediation at PAH-contaminated sites.     

Biodegradation of trichloroethylene and toluene by indigenous microbial populations in vadose sediments

The unsaturated subsurface (vadose zone) receives significant amounts of hazardous chemicals, yet little is known about its microbial communities and their capacity to biodegrade pollutants. Trichloroethylene (TCE) biodegradation occurs readily in surface soils; however, the process usually requires enzyme induction by aromatic compounds, methane, or other cosubstrates. The aerobic biodegradation of toluene and TCE by indigenous microbial populations was measured in samples collected from the vadose zone at unpolluted and gasoline-contaminated sites. Incubation at field moisture levels showed little activity on either TCE or toluene, so samples were tested in soil suspensions. No degradation occurred in samples suspended in water or phosphate buffer solution; however, both toluene and TCE were degraded in samples suspended in mineral salts medium. TCE degradation depended on toluene degradation, and little loss occurred under sterile conditions. Studies with specific nutrients showed that addition of ammonium sulfate was essential for degradation, and addition of other mineral nutrients further enhanced the rate. Additional studies with vadose sediments amended with nutrients showed similar trends to those observed in sediment suspensions. Initial rates of biodegradation in suspensions were faster in uncontaminated samples than in gasolinecontaminated samples, but the same percentages of chemicals were degraded. Biodegradation was slower and less extensive in shallower samples than deeper samples from the uncontaminated site. Two toluene-degrading organisms isolated from a gasoline-contaminated sample were identified as Corynebacterium variabilis SVB74 and Acinetobacter radioresistens SVB65. Inoculation with 10⁶ cells of C. variabilis ml^–1 of soil solution did not enhance the rate of degradation above that of the indigenous population. These results indicate that mineral nutrients limited the rate of TCE and toluene degradation by indigenous populations and that no additional benefit was derived from inoculation with a toluene-degrading bacterial strain. Correspondence to: K.M. Scow     

Unraveling uncultivable pesticide degraders via stable isotope probing (SIP)

Uncultivable microorganisms account for over 99% of all species on earth, playing essential roles in ecological processes such as carbon/nitrogen cycle and chemical mineralization. Their functions remain unclear in ecosystems and natural habitats, requiring cutting-edge biotechnologies for a deeper understanding. Stable isotope probing (SIP) incorporates isotope-labeled elements, e.g. ^13?C, ^18?O or ^15?N, into the cellular components of active microorganisms, serving as a powerful tool to link phylogenetic identities to their ecological functions in situ. Pesticides raise increasing attention for their persistence in the environment, leading to severe damage and risks to the ecosystem and human health. Cultivation and metagenomics help to identify either cultivable pesticide degraders or potential pesticide metabolisms within microbial communities, from various environmental media including the soil, groundwater, activated sludge, plant rhizosphere, etc. However, the application of SIP in characterizing pesticide degraders is limited, leaving considerable space in understanding the natural pesticide mineralization process. In this review, we try to comprehensively summarize the fundamental principles, successful cases and technical protocols of SIP in unraveling functional-yet-uncultivable pesticide degraders, by raising its shining lights and shadows. Particularly, this study provides deeper insights into various feasible isotope-labeled substrates in SIP studies, including pesticides, pesticide metabolites, and similar compounds. Coupled with other techniques, such as next-generation sequencing, nanoscale secondary ion mass spectrometry (NanoSIMS), single cell genomics, magnetic-nanoparticle-mediated isolation (MMI) and compound-specific isotope analysis (CSIA), SIP will significantly broaden our understanding of pesticide biodegradation process in situ.     

黄河三角洲滨海湿地微生物多样性及其驱动因子

微生物在湿地的生物地球化学循环和生态功能调节中发挥着重要作用,对全球气候变化具有重大影响,对维持全球生态系统的健康至关重要。以黄河三角洲滨海湿地为研究对象,通过采集代表性植被群落的土壤表层和部分植物根系,探究土壤微生物群落组成、根际微生物、环境因子及其内在的关联性和影响机制。研究结果表明不同植被覆盖地区微生物多样性存在差异,芦苇区和柽柳区微生物丰度高于泥滩区、碱蓬区和棉田,海漫滩微生物丰度高于河漫滩地和泥滩。土壤微生物菌群结构和多样性显著高于根际：土壤细菌的香农指数约为4-5.5,根际微生物的香农指数约为0-4。土壤细菌主要为厚壁菌门、变形菌门、拟杆菌门和放线菌门,占样品总数的90%以上;而根际细菌主要是蓝藻门、变形菌门和放线菌门,二者在属水平上的菌群结构差异更加明显。环境因子的含量与生境类型有关,SO₄^2-和NO₃^-的相关性最高,植被覆盖区土壤中Mn⁴⁺、Fe³⁺和水解氮的含量低于滩涂裸地。冗余分析（RDA）表明,pH值在小空间尺度上对湿地土壤中细菌群落的影响较小,环境因子在门和属水平的解释率分别为89.7%和86.8%,其中K（23.4%）、NO₂^-（11.8%）、Mn⁴⁺（9.8%）和Na（8.0%）是解释门水平微生物区系结构变化和组成的主要因子。研究为理解湿地微生物多样性与湿地生态系统功能之间的影响机制提供了一个生态学视角,有助于了解黄河三角洲滨海湿地土壤和植物根际的细菌分布特征,对黄河三角洲退化滨海湿地的生物修复具有重要的指导意义。     

不同药用植物根际土壤原核微生物多样性研究

为研究不同药用植物根际土壤中的原核微生物多样性,分别采集白术(Atractylodes macrocephala)、白芍(Paeonia sterniana)、牡丹(Paeonia suffruticosa)、玄参(Scrophularia ningpoensis)四种药用植物的根际土壤以及非种植区的土壤,针对16S rRNA基因的V3～V4区进行测序,分析土壤细菌群落的组成。结果表明,药用植物根际土壤中的细菌群落多样性指数显著高于非种植区土壤。五组样本的优势类群差异不大,总体相对丰度较高的有变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、芽单胞菌门(Gemmatimonadetes)、绿弯菌门(Chloroflexi)等,药用植物根际中的放线菌相对丰度高于非种植区。属水平上四种药用植物根际细菌和非种植区的群落结构有较大差异,四种中药材的根际土壤中各自富集了特异性的有益细菌属。药用植物根际土壤中的NMD1、Dongia、Gaiella、Streptomyces等相对丰度高于非种植区,而非种植区土壤中Lysoba...     

Changes of Bt Toxin in the Rhizosphere of Transgenic Bt Cotton and its Influence on Soil Functional Bacteria

Summary The concentrations of Bacillus thuringensis (Bt) toxin released from root exudation of Bt cotton were measured by an enzyme-linked immunosorbent assay (ELISA), and its impacts on the numbers of culturable functional bacteria in the rhizosphere were determined by cultivation. No Bt toxin was found in the rhizosphere of non-Bt cotton (SHIYUAN321), but varying levels of Bt toxin were present in the rhizosphere of two Bt cotton varieties (NuCOTN99^B and SGK321) during the entire growth period. The levels of Bt toxin in the rhizosphere of NuCOTN99^B were significantly higher (p<0.05) than those of SGK321 within all sampling dates except on June 17th in the whole growth season. Significant differences (p<0.05) were found in the numbers of the three functional bacteria between SHIYUAN321 and NuCOTN99^B within each sampling day from May 27th to October 27th. No significant differences were found in the numbers of functional bacteria among three cultivars after growth season. Fortification of pure Bt toxin into rhizospheric soil did not result in significant changes in the numbers of culturable functional bacteria, except the nitrogen-fixing bacteria when the concentration of Bt toxin was higher than 500 ng/g. The results indicated that Bt toxin was not the direct factor causing decrease of the numbers of bacteria in the rhizosphere, and other factors may be involved.     

A Method for Selection and Characterization of Rhizosphere-Competent Bacteria of Chickpea

A greenhouse assay was developed to evaluate the root-colonizing capability of the native chickpea rhizospheric bacterial population. In this assay system, screening time was reduced on two counts. First, spontaneous chromosomal rifampicin-resistant (Rif^r) strains were directly inoculated to seeds without any check for the stability of the mutation, and second, no attempts were made to taxonomically identify all the strains being screened for chickpea rhizosphere competence. Only two chickpea rhizosphere-competent Rif^r strains from the group of six good chickpea rhizosphere colonizers forming 10⁷ to 10⁸ colony-forming units (cfu)/g root were taxonomically identified as Pseudomonas fluorescens NB13R and Pseudomonas spp. NB49R, after screening 49 bacteria. Both the strains showed no difference from their corresponding wild-type strains P. fluorescens NB13 and Pseudomonas spp. NB49 in terms of chickpea rhizosphere competence. Isogenic or equally rhizospheric competitive second non-isogenic bacterial isolate, when present in tenfold higher amount, pre-empted the colonization of the soil by the bacterium, which was present in smaller ratio. These findings indicate that the isogenic or equally rhizospheric competitive second non-isogenic Rif^r strains should be compared for their survival and competition with that of the isogenic parent and with each other for specific ecological niche, before using a mixture of isolates, for stable and consistent biological seed treatment to control soilborn pathogens or pests or to promote plant growth. Received: 31 May 1996 / Accepted: 5 July 1996     

Dynamics of Arsenic Species in Laboratory‐Scale Horizontal Subsurface‐Flow Constructed Wetlands Treating an Artificial Wastewater

Knowledge regarding the dynamics of arsenic species and their interactions under gradient redox conditions in treatment wetlands is still insufficient. The aim of this investigation was to gain more information on the biotransformation of As and the dynamics of As species in horizontal subsurface‐flow constructed wetlands. Experiments were carried out in laboratory‐scale wetland systems, two planted with Juncus effusus and one unplanted, using an As‐containing artificial wastewater under defined organic C‐ and SO₄^2–‐loading conditions. Immobilization of As was found in all systems under conditions of limited C, mainly due to adsorption and/or co‐precipitation. The removal efficiencies were substantially higher in the planted systems (60–70 %) as compared to the unplanted system (37 % on average). Immobilization under the conditions mentioned above appeared to decrease over time in all systems. At the beginning, the dosage of organic carbon immediately caused intensive microbial dissimilatory sulfate reduction in all systems (in the range of 85–95 %) and highly efficient removal of total arsenic (81–96 % on average). Later on, in this operation period, the intensity of sulfate reduction and simultaneous removal of As decreased, particularly in the planted wetlands (ranging from 30–46 %). One reason could be the re‐oxidation of reduced compounds due to oxygenation of the rhizosphere by the emergent wetland plants (helophytes). A significant amount of reduced As [As(III)] was found in the planted systems (> 75 % of total As) during the period of efficient microbial sulfate reduction, compared to the unplanted system (> 25 % of total As). The immobilization of arsenic was found to behave more stably in the planted beds than in the unplanted bed. Both systems (planted and unplanted) were suitable to treat wastewater containing As, particularly under sulfate reducing conditions. The unplanted system seemed to be more efficient regarding the immobilization of As, but the planted systems showed a better stability of immobilized As.     

Kinetics of carbon sharing in a bacterial consortium revealed by combining stable isotope probing with fluorescence‐activated cell sorting

Aims: To determine the kinetics of substrate fluxes in a microbial community in order to elucidate the roles of the community members. Methods and Results: The kinetics of substrate sharing in a bacterial consortium were measured by a new analytical approach combining immunostaining, stable isotope probing and fluorescence‐activated cell sorting (FACS). The bacterial consortium, consisting of four strains and growing on 4‐chlorosalicylate (4‐CS), was pulse‐dosed with the degradation intermediate [U‐¹³C]‐4‐chlorocatechol (4‐CC). Cells were stained with strain‐specific antibodies sorted by FACS and the ¹³C‐incorporation into fatty acids of the two most abundant members of the community was determined by isotope ratio mass spectrometry. From the two most abundant strains, the primary degrader Pseudomonas reinekei MT1 incorporated the labelled substrate faster than strain Achromobacter spanius MT3 but the maximal incorporation in strain MT3 was almost three times higher than in MT1. Conclusions: It has been reported that strain MT1 produces 4‐CC as an intermediate but has a lower LD₅₀ for it than strain MT3; therefore, MT3 still degrades 4‐CC when the concentrations of 4‐CC are already too toxic, even lethal, for MT1. By degrading 4‐CC, produced by MT1, MT3 protects the entire community against this toxin. The higher affinity but lower tolerance of strain MT1 for 4‐chlorocatechol compared to strain MT3 explains the complementary function these two strains have in the consortium adding exceptional stability to the entire community. Significance and Impact of the Study: The novel approach can reveal carbon fluxes in microbial communities generating quantitative data for systems biology of the microbial community.     

Diversity of five anaerobic toluene-degrading microbial communities investigated using stable isotope probing

Time-series DNA-stable isotope probing (SIP) was used to identify the microbes assimilating carbon from [(13)C]toluene under nitrate- or sulfate-amended conditions in a range of inoculum sources, including uncontaminated and contaminated soil and wastewater treatment samples. In all, five different phylotypes were found to be responsible for toluene degradation, and these included previously identified toluene degraders as well as novel toluene-degrading microorganisms. In microcosms constructed from granular sludge and amended with nitrate, the putative toluene degraders were classified in the genus Thauera, whereas in nitrate-amended microcosms constructed from a different source (agricultural soil), microorganisms in the family Comamonadaceae (genus unclassified) were the key putative degraders. In one set of sulfate-amended microcosms (agricultural soil), the putative toluene degraders were identified as belonging to the class Clostridia (genus Desulfosporosinus), while in other sulfate-amended microcosms, the putative degraders were in the class Deltaproteobacteria, within the family Syntrophobacteraceae (digester sludge) or Desulfobulbaceae (contaminated soil) (genus unclassified for both). Partial benzylsuccinate synthase gene (bssA, the functional gene for anaerobic toluene degradation) sequences were obtained for some samples, and quantitative PCR targeting this gene, along with SIP, was further used to confirm anaerobic toluene degradation by the identified species. The study illustrates the diversity of toluene degraders across different environments and highlights the utility of ribosomal and functional gene-based SIP for linking function with identity in microbial communities.     

Degradation of toluene by ortho cleavage enzymes in Burkholderia fungorum FLU100

Burkholderia fungorum FLU100 simultaneously oxidized any mixture of toluene, benzene and mono‐halogen benzenes to (3‐substituted) catechols with a selectivity of nearly 100%. Further metabolism occurred via enzymes of ortho cleavage pathways with complete mineralization. During the transformation of 3‐methylcatechol, 4‐carboxymethyl‐2‐methylbut‐2‐en‐4‐olide (2‐methyl‐2‐enelactone, 2‐ML) accumulated transiently, being further mineralized only after a lag phase of 2 h in case of cells pre‐grown on benzene or mono‐halogen benzenes. No lag phase, however, occurred after growth on toluene. Cultures inhibited by chloramphenicol after growth on benzene or mono‐halogen benzenes were unable to metabolize 2‐ML supplied externally, even after prolonged incubation. A control culture grown with toluene did not show any lag phase and used 2‐ML as a substrate. This means that 2‐ML is an intermediate of toluene degradation and converted by specific enzymes. The conversion of 4‐methylcatechol as a very minor by‐product of toluene degradation in strain FLU100 resulted in the accumulation of 4‐carboxymethyl‐4‐methylbut‐2‐en‐4‐olide (4‐methyl‐2‐enelactone, 4‐ML) as a dead‐end product, excluding its nature as a possible intermediate. Thus, 3‐methylcyclohexa‐3,5‐diene‐1,2‐diol, 3‐methylcatechol, 2‐methyl muconate and 2‐ML were identified as central intermediates of productive ortho cleavage pathways for toluene metabolism in B. fungorum FLU100.     

Evaluating the biodegradation of aromatic hydrocarbons by monitoring of several functional genes

Various microbial activities determine the effectiveness of bioremediation processes. In this work, we evaluated the feasibility of gene array hybridization for monitoring the efficiency of biodegradation processes. Biodegradation of ¹⁴C-labelled naphthalene and toluene by the aromatic hydrocarbon-degrading Pseudomonas putida F1, P. putida mt-2 and P. putida G7 was followed in mixed liquid culture microcosm by a preliminary, nylon membrane-based gene array. In the beginning of the study, toluene was degraded rapidly and increased amount of toluene degradation genes was detected by the preliminary gene array developed for the study. After toluene was degraded, naphthalene mineralization started and the amount of naphthalene degradation genes increased as biodegradation proceeded. The amount of toluene degradation genes decreased towards the end of the study. The hybridization signal intensities determined by preliminary gene array were in good agreement with mineralization of naphthalene and toluene and with the amount of naphthalene dioxygenase and toluene dioxygenase genes quantified by dot blot hybridization. The clear correlation between the results obtained by the preliminary array and the biodegradation process suggests that gene array methods can be considered as a promising tool for monitoring the efficiency of biodegradation processes.     

Characteristics of the microbial community in rhizosphere of <Emphasis Type="Italic">Camptotheca acuminata</Emphasis> cultured with exotic invasive plant <Emphasis Type="Italic">Eupatorium adenophorum</Emphasis>

The traditional culture-dependent plate counting and culture-independent small-subunit-ribosomal RNA gene-targeted molecular techniques, Single-Strand Conformation Polymorphism (SSCP) and terminal Restriction Fragment Length Polymorphism (tRFLP) combined with 16S rDNA clone library were adopted to investigate the impacts of secretion from Camptotheca acuminata (abbreviated to Ca) roots on the quantities and structure of eukaryotic microbes and bacteria in the rhizosphere, and the possibility that Ca controls exotic invasive plant Eupatorium adenophorum (Ea). The counting results indicated that the number of bacteria increased in turn in rhizospheres of Ea, Ca-Ea mixed culture and Ca, while that of eukaryotic microbes decreased. PCR-SSCP profiles showed eukaryotic microbial bands (corresponding to biodiversity) in rhizosphere of Ea were more complex than those of Ca and CE. Meristolohmannia sp., Termitomyces sp. and Rhodophyllus sp. were the dominant populations in the rhizosphere of Ca. Bacterial terminal restriction fragments (TRFs) profiles showed no difference among three kinds of rhizospheres, and the sequences of the 16S rDNA clone library from Ca rhizospheres were distributed in 10 known phyla, in which phylum Proteobacteria were the absolute dominant group and accounted for 24.71% of the cloned sequences (δ-Proteobacteria accounted for up to 17.65%), and phyla Acidobacteria and Bacteroidetes accounted for 16.47% and 10.59% of the cloned sequences, respectively. In addition, high performance liquid chromatography detected a trace amount of camptothecin and hydroxycamptothecin in the rhizospheric soil of Ca and CE, but examined neither camptothecin nor hydroxycamptothecin in rhizospheric soil of Ea. Therefore, invasion and diffusion of Ea evidently depended on distinguishing the eukaryotic community structure, but not on that of the bacterial pattern. Ca was able to alter the eukaryotic community structure of invasive Ea by secreting camptothecin and hydroxycamptothecin into rhizospheres, and may benefit the control of overspread of Ea. This study provided theoretical evidence for rhizospheric microbial aspects on substituting Ca for Ea.     

Redox Dynamics of Arsenic Species in the Root‐Near Environment of Juncus effusus Investigated in a Macro‐Gradient‐Free Rooted Gravel Bed Reactor

In the framework of investigating the dynamics of As species within the planted soil beds of treatment wetlands, the redox dynamics of As species particularly in the root‐near environment of the rhizosphere were investigated. For this purpose, long‐term experiments were carried out using a specially designed macro‐gradient‐free rooted gravel bed reactor, planted with Juncus effusus to treat an artificial wastewater containing As (200 μg As/L). The exceptional quality of the biofilm processes at the helophyte root‐surfaces in treatment wetlands were of special importance in this investigation. The results showed that under C‐deficient conditions, a highly efficient As immobilization (> 85 %), obviously due to adsorption and/or co‐precipitation, was attained. The addition of organic carbon immediately caused an elevated As concentration and enrichment of As(III) (nearly 80 % of total As) in the reactor. Increasing the SO₄^2– concentration in the artificial wastewater inflow facilitated a high As immobilization (> 82 %) under sulfate reducing condition. In principle, a highly efficient microbial dissimilatory sulfate reduction contributed to S^2– formation and a greater As immobilization (most likely as As₂S₃) under C surplus and reducing conditions. Significant differences in As immobilization were observed by varying the inflow of the SO₄^2– concentration (0.2, 5, 10, 25 S/L) under C surplus conditions. More As(III) precipitates (15 % less in the outflow) when the inflow of the SO₄^2– concentration was decreased from 25 mg S/L to 10 mg S/L. Immobilized As showed greater instability by releasing As(V) (up to 85 % of total As) due to changes in the dynamic redox conditions inside the reactor. Re‐oxidation of reduced sulfur into other S species (e.g. S⁰, SO₄^2–) due to plant‐root mediated O₂ release probably caused an oxidative dissolution of already precipitated insoluble As (e.g. As₂S₃) and as a consequent As remobilization. The findings of this study highlighted the significance of SO₄^2– in relation to organic C supply in planted soil beds treating As‐contaminated wastewater under constructed wetland conditions.     

焦化废水O/H/O生物处理工艺二级好氧生物反应器的微生物结构和功能

【背景】焦化废水O/H/O生物处理工艺的二级好氧生物反应器O₂具有剩余污染物矿化和完全硝化功能,对废水的达标排放有重要作用。【目的】阐明O₂生物反应器的微生物结构和功能。【方法】利用16S rRNA基因测序,研究O₂生物反应器的微生物多样性和组成并进行功能预测,揭示其共现性特征和环境影响因子。【结果】O₂的优势菌门以变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、绿菌门(Chlorobi)为主。主要菌属中红游动菌属(Rhodoplanes)、溶杆菌属(Lysobacter)、硫杆菌属(Thiobacillus)等参与化学需氧量(chemical oxygen demand,COD)、酚类(phenols)和硫氰酸盐(thiocyanate,SCN^-)等剩余污染物的去除,亚硝化弧菌属(Nitrosovibrio)和硝化螺菌属(Nitrospira)分别作为氨氧化细菌(ammonia-oxidizing bacteria,AOB)和主要的亚硝酸盐氧化细菌(nitrite-oxidizing bacteria,NOB)。功能预测结果显示苯甲酸酯降解、氨基苯甲酸酯降解、氯烷烃和氯烯烃的降解、氟代苯甲酸酯降解和硝基甲苯降解是外源物质生物降解和代谢的前五大通路,广泛分布在主要菌属中,验证了微生物降解剩余污染物的作用。基因pmoA/B/C-amoA/B/C、hao和nxrA/B编码相关的酶,组成了完整的硝化途径。共现网络结果揭示溶杆菌属、Candidatus Solibacter和红游动菌属在O₂生态中的重要地位。通过冗余分析(redundancy analysis,RDA)表明COD和NH₃是影响O₂微生物群落的主要因素。【结论】红游动菌属和溶杆菌属是O₂中最核心的功能菌属,在污染物矿化和维持群落生态稳定上有重要作用。亚硝化弧菌属和硝化螺菌属是硝化作用的核心菌属。O₂中的代谢通路以剩余污染物矿化和完全硝化为主,微生物群落主要受COD和NH₃的影响。本研究阐明了O₂的微生物结构与功能,为焦化废水O/H/O生物处理工艺的改进提供了微生物学上的依据。