Microbial functional genes reveal selection of microbial community by PAHs in polluted soils

This report shows an increase of PAH-related microbial functional genes with PAH concentration in soils. Adaptation of microbial communities to organic pollutants such as polycyclic aromatic hydrocarbons (PAHs) is a crucial issue. However, there is little knowledge on mechanisms ruling microbial community selection. Here, we studied microbial functional genes in soils contaminated by PAHs. We used GeoChip, an advanced functional gene array for gene analysis. Soil PAH concentrations were measured and microbial functional genes were categorized. PAH-related microbial functional genes, bph, nah, nidA, phd, dfb, and qor, were quantitatively expressed. Total microbial functional genes and PAH-related microbial functional genes were compared with PAH concentration by cluster analysis and curve-fitting analysis. We found that the average abundance of PAH-related microbial functional genes increased from 0.13 to 0.33, whereas that of total microbial functional genes decreased from 0.22 to 0.10 when PAHs concentration increased from 1.01 to 164.28 mg kg^?1. It was also found that the classification of microbial community structure characteristics based on PAH-related microbial functional genes was closely similar to the classification based on PAHs concentration. Findings reveal that PAH stress promotes the dominance of PAH-related microbial communities.     

枯草芽孢杆菌Bs-15对板栗土壤微生物种群数量及功能多样性的影响

土壤微生物是维持土壤质量的重要组成部分,是土壤中生物活性的具体体现。土壤微生物多样性的变异可反映其对环境的响应与适应,能敏感反映生态系统的功能演变和生态环境的变化。本研究采用固体平板法研究了枯草芽孢杆菌（Bacillus subtilis）Bs-15对板栗（Castanea mollissimaBL）土壤微生物种群数量的影响,并通过BIOLOG ECO微孔板法分析Bs-15对板栗土壤功能多样性的影响。结果表明,接种Bs-15后,土壤中细菌数量比对照略有增加,但差异不显著;接种后放线菌的数量与对照相比有所减少,第7天达到极显著（p〈0.01）差异,之后差异变小,第14 d开始,处理与对照之间放线菌数量基本持平;真菌数量则先增加后减少,第7天开始接种后的真菌数量与对照相比达极显著（p〈0.01）差异。BIOLOG ECO微孔板分析显示,Bs-15使得土壤中微生物的AWCD值变大,72小时以后,AWCD值与对照相比差异达到极显著（p〈0.01）水平;接种Bs-15后增大了土壤微生物多样性指数,其中Shannon多样性指数、Simpson多样性指数和McIntosh多样性指数分别增加了4.09%、6.01%和7.20%,对对照相比差异均达到极显著水平（p〈0.01）,Simpson均匀度和McIntosh均匀度分别增加了2.07%和2.53%,与对照相比差异均达到显著水平（p〈0.05）。本研究结果表明,Bs-15不但提高了板栗土壤微生物的整体活性,丰富了土壤微生物种群,有利于保持和促进土壤肥力和健康状况;还提高了板栗土壤微生物功能多样性,使板栗土壤微生态系统功能更加稳定。     

Influence of invasive plant Coreopsis grandiflora on functional diversity of soil microbial communities

Biological invasions are increasingly attracting the ecologists' attention. Invasive plants threaten the natural ecosystems not only by competing with the native plants, but also by altering the structure and function of soil microbial communities belowground. In this study, we studied the effects of the invasive plant Coreopsis grandiflora (C. grandiflora) on the functional diversity of soil microbial communities in Laoshan mountain in the province of Shandong, North of China. We sampled soil from plots that were invaded or not invaded by C. grandiflora. The functional diversity of microbial communities in the sampled soils was assessed by the Biolog procedure test. By the ANOVA analysis of average well color development (AWCD), Shannon index (H'), Shannon evenness (E), principle components analysis of the level physiological profiles (CLPP) and correlation analysis between the studied parameters, we found that the invasive species C. grandiflora enhanced the functional diversity of soil microbial communities where the habitat was invaded by the C. grandiflora. The study indicated thatthe successful invasive plants have profound effects on the function of soil microbial communities.     

Microbial community and functional genes in the rhizosphere of alfalfa in crude oil-contaminated soil

A rhizobox system constructed with crude oil-contaminated soil was vegetated with alfalfa (Medicago sativa L.) to evaluate the rhizosphere effects on the soil microbial population and functional structure, and to explore the potential mechanisms by which plants enhance the removal of crude oil in soil. During the 80-day experiment, 31.6% of oil was removed from the adjacent rhizosphere (AR); this value was 27% and 53%higher than the percentage of oil removed from the far rhizosphere (FR) and from the non-rhizosphere (NR), respectively. The populations of heterotrophic bacteria and hydrocarbon-degrading bacteria were higher in the AR and FR than in the NR. However, the removal rate of crude oil was positively correlated with the proportion of hydrocarbon-degrading bacteria in the rhizosphere. In total, 796, 731, and 379 functional genes were detected by microarray in the AR, FR, and NR, respectively. Higher proportions of functional genes related to carbon degradation and organic remediation, were found in rhizosphere soil compared with NR soil, suggesting that the rhizosphere selectively increased the abundance of these specific functional genes. The increase in water-holding capacity and decrease in pH as well as salinity of the soil all followed the order of AR>FR>NR. Canonical component analysis showed that salinity was the most important environmental factor influencing the microbial functional structure in the rhizosphere and that salinity was negatively correlated with the abundance of carbon and organic degradation genes.     

分子技术在湿地微生物群落解析中的应用

人工湿地的研究和应用近年来受到了广泛重视。微生物是人工湿地系统的重要组成部分,其群落结构对于湿地的净化功能的发挥具有重要影响。与传统的微生物分析技术相比,分子生物学技术在解析人工湿地微生物群落结构时无需纯培养,具有高效、快速、简便的特点,使其广泛应用于环境微生物的研究。文章综述了近年来在聚合酶链反应技术（Polymerase Chain Reaction,PCR）基础上发展起来的几种新的分子生物技术,包括PCR-DGGE、LH-PCR、T-RFLP、PCR-SSCP和ARDRA,以及其在人工湿地微生物研究中的应用现状。通过这些分子技术,可以分析湿地处理特定废水过程中微生物的数量、丰度、多样性及优势种;鉴定湿地中特定功能菌群（如氨氧化细菌、反硝化细菌、除硫菌等）的数量、活动分布、空间变更及与污染物去除的关系;判断各种系统条件（如不同基质、植物、水力负荷等）的设置对微生物多样性和稳定性的影响。最后,对分子技术在湿地领域的未来发展进行了展望。     

Spider, bee, and bird communities in cities are shaped by environmental control and high stochasticity

Spatially organized distribution patterns of species and communities are shaped by both autogenic processes (neutral mechanism theory) and exogenous processes (niche theory). In the latter, environmental variables that are themselves spatially organized induce spatial structure in the response variables. The relative importance of these processes has not yet been investigated in urban habitats. We compared the variance explained by purely spatial, spatially structured environmental, and purely environmental components for the community composition of spiders (Araneae), bees (Apidae), and birds (Aves) at 96 locations in three Swiss cities. Environmental variables (topography, climate, land cover, urban green management) were measured on four different radii around sampling points (< 10 m, 50 m, 250 m, 1000 m), while Moran's eigenvector maps (MEMs) acted as spatial variables. All three taxonomic groups showed weak spatial structure. Spider communities reacted to very fine-scaled environmental changes of lawn and meadow management and climate. Bird community composition was determined by woody plants as well as solar radiation at all radii, the scale of the influence varying among species. Bee communities were weakly explained by isolated variables only. Our results suggest that the anthropogenic structuring of urban areas has disrupted the spatial organization of environmental variables and inhibited the development of biotic spatial processes. The near absence of spatial structure may therefore be a feature typical of urban species assemblages, resulting in urban community composition mainly influenced by local environmental variables. Urban environments represent a close-knit mosaic of habitats that are regularly disturbed. Species communities in urban areas are far from equilibrium. Our analysis also suggests that urban communities need to be considered as being in constant change to adapt to disturbances and changes imposed by human activities.     

桑园土壤微生物群落功能多样性对PAHs污染的响应

为了探明桑园土壤微生物群落功能多样性与多环芳烃(PAHs)污染的关系,采用Biolog检测法研究某路域桑园土壤微生物功能多样性对PAHs污染的响应。结果表明,不同区域PAHs污染程度由大到小依次为区域2(位于区域1和3之间)、3(离公路最远)和1(离公路最近);Biolog分析发现不同区域土壤微生物对碳源利用的平均颜色变化率(AWCD)由大到小依次为区域2、3和1;区域2的Shannon指数和Gini指数均显著高于区域1和3,而区域1的Shannon均匀度显著低于区域2和3,区域2土壤微生物活性最强且群落结构最丰富,其后依次为区域3和1。主成分分析结果显示,3个区域微生物群落的生理功能差异明显,主要表现在对糖类和氨基酸类物质的代谢上。冗余分析表明,区域2土壤微生物与PAHs含量关系最密切,说明较高的PAHs含量更能激发桑园土壤微生物的生理活性,微生物群落代谢功能因PAHs污染而有所提高。     

恩诺沙星对土壤微生物群落代谢功能多样性的影响（Effects of Enrofloxacin on the Functional Diversity of Soil Microbial Communities）

为了评价恩诺沙星(Enrofloxacin)对土壤微生物群落的影响,借助BIOLOG检测法比较了不同浓度恩诺沙星影响下的土壤微生物的群落特征.结果表明,加药组土壤微生物的丰富度指数和多样性指数显著低于空白对照组,且药物浓度越高丰富度和多样性越小.用药后第3d、14d,恩诺沙星含量0.1～100μg·g-1使土壤微生物群落代谢功能多样性显著降低(p<0.05);第35d,恩诺沙星含量10～100μg·g-1使土壤微生物群落代谢功能多样性显著降低,随着药物作用的时间延长,药物含量0.01～1μg·g-1组土壤微生物群落代谢功能多样性与空白对照组之间的差异变小.     

Identifying human-induced influence on microbial community: A comparative study in the effluent-receiving areas in Hangzhou Bay

Microbial compositions showed high differences in two study areas. COD was the key anthropogenic indicator in the coastal wastewater disposal area. Distinctive microbes capable of degrading toxic pollutants were screened. Microbial communities in effluent-receiving areas followed “niche theory”. Microbial community structure is affected by both natural processes and human activities. In coastal area, anthropegenetic activity can usually lead to the discharge of the effluent from wastewater treatment plant (WWTP) to sea, and thus the water quality chronically turns worse and marine ecosystem becomes unhealthy. Microorganisms play key roles in pollutants degradation and ecological restoration; however, there are few studies about how the WWTP effluent disposal influences coastal microbial communities. In this study, sediment samples were collected from two WWTP effluent-receiving areas (abbreviated as JX and SY) in Hangzhou Bay. First, based on the high-throughput sequencing of 16S rRNA gene, microbial community structure was analyzed. Secondly, several statistical analyses were conducted to reveal the microbial community characteristics in response to the effluent disposal. Using PCoA, the significant difference of in microbial community structure was determined between JX and SY; using RDA, water COD and temperature, and sediment available phosphate and ammonia nitrogen were identified as the key environmental factors for the community difference; using LDA effect size analysis, the most distinctive microbes were found and their correlations with environmental factors were investigated; and according to detrended beta-nearest-taxon-index, the sediment microbial communities were found to follow “niche theory”. An interesting and important finding was that in SY that received more and toxic COD, many distinctive microbes were related to the groups that were capable of degrading toxic organic pollutants. This study provides a clear illustration of eco-environmental deterioration under the long-term human pressure from the view of microbial ecology.     

Mapping the niche space of soil microorganisms using taxonomy and traits

The biodiversity of microbial communities has important implications for the stability and functioning of ecosystem processes. Yet, very little is known about the environmental factors that define the microbial niche and how this influences the composition and activity of microbial communities. In this study, we derived niche parameters from physiological response curves that quantified microbial respiration for a diverse collection of soil bacteria and fungi along a soil moisture gradient. On average, soil microorganisms had relatively dry optima (0.3 MPa) and were capable of respiring under low water potentials (-2.0 MPa). Within their limits of activity, microorganisms exhibited a wide range of responses, suggesting that some taxa may be able to coexist by partitioning the moisture niche axis. For example, we identified dry-adapted generalists that tolerated a broad range of water potentials, along with wet-adapted specialists with metabolism restricted to less-negative water potentials. These contrasting ecological strategies had a phylogenetic signal at a coarse taxonomic level (phylum), suggesting that the moisture niche of soil microorganisms is highly conserved. In addition, variation in microbial responses along the moisture gradient was linked to the distribution of several functional traits. In particular, strains that were capable of producing biofilms had drier moisture optima and wider niche breadths. However, biofilm production appeared to come at a cost that was reflected in a prolonged lag time prior to exponential growth, suggesting that there is a trade-off associated with traits that allow microorganisms to contend with moisture stress. Together, we have identified functional groups of microorganisms that will help predict the structure and functioning of microbial communities under contrasting soil moisture regimes.     

醌类图谱分析在环境微生物生态测定中的应用

近年来，随着人们对各种非培养微生物测定法的重视，醌类图谱分析作为微生物群体指标已在各种环境中得到了应用．首先是样品的分析测定技术得到了提高，如用硅胶柱代替薄层色谱．醌类图谱分析的特点是能够同时对应微生物生态评价的3个重要指标，即微生物群体结构、生物量及微生物多样性．在结合统计分析的基础上，在土壤、污泥及堆肥等不同环境样品及不同微生物反应过程中，已通过醌类图谱分析对这3个指标在微生物生态系中的变动进行了成功的评价．另外，对C-14同位素标记醌类图谱分析法及醌类图谱分析法在污染物降解中的应用等最新进展也进行了简介．图4表3参43     

肠道微生物受环境污染的影响及其对宿主疾病的调控作用

宿主肠道内存在的大量微生物与其健康状况直接相关,这些微生物是人和动物健康成长不可或缺的。肠道微生物通过多种途径调节人体生理功能的同时也受到人体内外环境的影响。因此,分析建立肠道微生物、相关疾病的产生原因和作用机制以及环境影响因子之间的联系具有重要意义。本文首先针对肠道微生物对人体的物质与能量代谢、先天和获得性免疫、胃肠道功能等方面的影响进行综述。然后重点分析了近年来有关肠道微生物对环境污染所致健康效应的影响及作用机制的研究进展。以期加深肠道微生物与人类健康之间相互作用机理的理解,并为环境毒理学与肠道微生物之间关系的研究提供新的思路。     

Environmental controls on denitrifying communities and denitrification rates: insights from molecular methods.

The advent of molecular techniques has improved our understanding of the microbial communities responsible for denitrification and is beginning to address their role in controlling denitrification processes. There is a large diversity of bacteria, archaea, and fungi capable of denitrification, and their community composition is structured by long-term environmental drivers. The range of temperature and moisture conditions, substrate availability, competition, and disturbances have long-lasting legacies on denitrifier community structure. These communities may differ in physiology, environmental tolerances to pH and O2, growth rate, and enzyme kinetics. Although factors such as O2, pH, C availability, and NO3- pools affect instantaneous rates, these drivers act through the biotic community. This review summarizes the results of molecular investigations of denitrifier communities in natural environments and provides a framework for developing future research for addressing connections between denitrifier community structure and function.     

油藏微生物群落分布及提高采收率的模拟实验研究

对取自大庆油田各类油藏的126个地层油水样品中部分微生物进行了培养计数和分析,检测出大庆油田地层水中含有好氧和厌氧微生物9类.在此基础上,进行了微生物提高石油采收率的物理模拟试验.结果表明,通过激活油藏地层水中的微生物,可以使油藏压力上升0.2MPa,并提高采收率幅度9%以上.     

鸭粪和环丙沙星对潮土微生物群落功能多样性的影响

为了评价鸭粪中残留的环丙沙星(ciprofloxacin,CIP)对土壤微生物群落功能多样性的影响,布置培养试验,设五个处理,分别是Ⅰ:CK(对照)、Ⅱ:CIP(人工添加的CIP)、Ⅲ:DF(添加不含抗生素的鸭粪)、Ⅳ:DF+CIP(在Ⅲ的基础上再外源添加CIP)、Ⅴ:DF(CIP)(粪源CIP即含有CIP的鸭粪),培养81 d。借助Biolog技术研究了不同来源的CIP对土壤微生物功能多样性的影响。结果表明,培养1 d后,反映土壤微生物活性的平均颜色变化率(AWCD)表现为DFDF+CIPDF(CIP)CKCIP,说明环丙沙星对土壤微生物活性具有一定的抑制作用,且随着培养时间的延长其抑制作用减弱;添加鸭粪显著增强了土壤微生物活性和群落功能多样性;与人工添加进鸭粪中的环丙沙星相比,粪源环丙沙星对微生物活性的抑制作用更强,并显著降低了土壤中利用碳源微生物的物种丰富度指数(H)、均匀度指数(E)、优势度指数(Ds)和碳源利用丰富度指数(S);在培养过程中,土壤微生物利用的主要碳源种类随培养时间延长有所变化,初期利用的主要是糖类、羧酸类、聚合物类、胺类以及氨基酸类,后期主要是糖类和羧酸类。     

三江源区不同建植年代人工草地群落演替与土壤养分变化

研究了了三源区不同建植期人工修复草地在不同演替阶段毒杂草[主要是甘肃马先蒿(Pedicularis kansuensis)]的入侵规律、数量特征,植物群落物种组成、生物苗和草地质最以及土壤养分、微生物活性的变化规律.结果表明,不同建植期人工修复草地植物群落的种类组成、植物功能群组成和群落数量特征存在显著差异.随着演替时间的推移,人工草地群落盖度、高度、物种数、生物最和多样性指数均表现出"V"字型变化规律,杂类草--甘肃马先蒿的数量特征变化尤为明显,在4 a的人工草地群落中开始局部入侵,在5～6 a的人工草地群落中大面积入侵,其入侵速度、入侵面积达到高峰期.土壤的含水量、容重、土壤中有机质、氮素和磷素在演替过程(7 a、9 a草地)中逐渐降低,到一定时期又逐步增加;随着演替的进行,不同建植期人工草地的土壤微牛物生物量碳和酶活性均呈"V"字型,变化.对于退化生态系统的恢复首先是植被恢复,其次是土壤肥力的恢复.土壤有机质等养分的积累、微生物活性的改善不仅能使土壤-植物复合系统的功能得以恢复,同时也能促进物种多样性的形成,有利于人工草地群落稳定性的提高.在试验区尽管植被恢复演替进行得比较缓慢,但从土壤发展的角度看,仍属进展演替.所以,在退化高寒草甸的恢复过程中,若降低和有效控制外界的干扰(如围栏封育),可为退化草地恢复提供繁殖体与土壤环境,实现人工草地逐步向恢复(正向)演替进行.图3表6参34     

Fourth-corner generation of plant functional response groups

Plant functional response groups (PFGs) are now widely established as a tool to investigate plant—environment relationships. Different statistical methods to form PFGs are used in the literature. One way is to derive emergent groups by classifying species based on correlation of biological attributes and subjecting these groups to tests of response to environmental variables. Another way is to search for associations of occurrence data, environmental variables and trait data simultaneously. The fourth-corner method is one way to assess the relationships between single traits and habitat factors. We extended this statistical method to a generally applicable procedure for the generation of plant functional response groups by developing new randomization procedures for presence/absence data of plant communities. Previous PFG groupings used either predefined groups or emergent groups i.e. classifications based on correlations of biological attributes (Lavorel et al Trends Ecol Evol 12:474–478, 1997), of the global species pool and assessed their functional response. However, since not all PFGs might form emergent groups or may be known by experts, we used a permutation procedure to optimise functional grouping. We tested the method using an artificial test data set of virtual plants occurring in different disturbance treatments. Direct trait-treatment relationships as well as more complex associations are incorporated in the test data. Trait combinations responding to environmental variables could be clearly distinguished from non-responding combinations. The results are compared with the method suggested by Pillar (J Veg Sci 10:631–640) for the identification of plant functional groups. After exploring the statistical properties using an artificial data set, the method is applied to experimental data of a greenhouse experiment on the assemblage of plant communities. Four plant functional response groups are formed with regard to differences in soil fertility on the basis of the traits canopy height and spacer length.     

Microbial diversity and ecological networks as indicators of environmental quality

Evaluating the quality of ecosystems in terms of biological patrimony and functioning is of critical importance in the actual context of intensified human activities. Microbial diversity is commonly used as a bioindicator of ecosystems functioning. However, there is a lack of sensitivity of microbial diversity indicators in the case of moderate and chronic environmental degradation, such as atmospheric deposition of pollutants, agricultural practices, diffuse pollution by wastewater and climate change. As a consequence, there is a need for alternative bioindicators of soils and water quality. Here, we discuss the interest of adopting a more integrative approach based on biotic interaction networks beyond the simple diversity indicators. We review how the various biotic interactions can be integrated in the various microbial networks such as trophic, mutualistic and co-occurrence networks. Then we discuss the efficiency of microbial networks and associated metrics to detect changes in microbial communities. We conclude that the connectance, the number of links and the average degree of co-occurrence networks could vary from 10 to 50% in response to minor perturbations when microbial diversity parameters remain stable. Finally, we analyze studies that aimed at linking microbial networks and activity to evaluate the potential of such networks for providing simple and operational indicators of ecosystem quality and functioning.     

Microbiota associated with the migration and transformation of chlorinated aliphatic hydrocarbons in groundwater

Pollution of groundwater with chlorinated aliphatic hydrocarbons (CAHs) is a serious environmental problem which is threatening human health. Microorganisms are the major participants in degrading these contaminants. Here, groundwater contaminated for a decade with CAHs was investigated. Numerical simulation and field measurements were used to track and forecast the migration and transformation of the pollutants. The diversity, abundance, and possible activity of groundwater microbial communities at CAH-polluted sites were characterized by molecular approaches. The number of microorganisms was between 5.65E+05 and 1.49E+08 16S rRNA gene clone numbers per liter according to quantitative real-time PCR analysis. In 16S rRNA gene clone libraries constructed from samples along the groundwater flow, eight phyla were detected, and Proteobacteria were dominant (72.8 %). The microbial communities varied with the composition and concentration of pollutants. Meanwhile, toluene monooxygenases and methane monooxygenases capable of degradation of PCE and TCE were detected, demonstrating the major mechanism for PCE and TCE degradation and possibility for in situ remediation by addition of oxygen in this study.