中国喀斯特地区微生物群落结构和功能的研究进展

频繁的人类生产活动使植被遭到破坏,造成基岩裸露的石漠化现象,严重制约了喀斯特地区自然和社会经济的发展,随着生态修复工程的大力开展,微生物群落的结构和功能在生态修复中逐渐受到重视,因为微生物作为喀斯特生态系统的重要组成部分,不仅在物质循环过程中起着重要作用,也在喀斯特生态系统修复中占有十分重要的地位。所以微生物群落结构和功能的研究可以作为衡量生态系统稳定性的重要指标。就中国喀斯特地区的典型植被恢复的不同阶段、成土过程、不同的土地利用方式、矿山修复过程以及不同水域中的微生物群落结构及功能等方面的研究状况进行系统梳理,结合实例综合论述喀斯特地区生态修复过程中微生物作用的研究进展,以期为喀斯特地区生态修复提供参考。     

矿山废水中微生物生态多样性研究

从湖北铜绿山铜矿和山西中条山铜矿两地采集了矿山废水样品, 用RFLP方法(限制性片段长度多态性方法)分析不同矿山废水中微生物多样性及其群落结构组成变化。结果发现：通过16S rDNA基因序列的系统发育树分析, 在5个矿山废水样品中检测到的细菌主要分为4大类：即Proteobacteria纲、Nitrospira纲、Firmicutes纲和Bacteroidetes纲。通过对古菌16S rDNA 的PCR分析发现, 取自湖北铜绿山铜矿的样品中检测到古菌的存在, 而取自山西中条山铜矿的样品中没有检测到古菌的存在。检测到的古菌主要是属于Ferroplasma属和Thermoplasma属。结合5个矿山废水样品的化学元素分析结果和细菌群落结构数据, 进行PCA分析(主成分分析), 发现不同矿山废水样品的生物地球化学性质及其微生物组成存在很大差异, pH值、温度、不同浓度的元素如S、Cu、Ni和Co可能是形成细菌种群结构差异的关键因素。     

DGGE/TGGE技术及其在微生物分子生态学中的应用

变性梯度凝胶电泳(DGGE)和温度梯度凝胶电泳(TGGE)是近些年微生物分子生态学研究中的热点技术之一。由于DGGE／TGGE技术具有可靠性强、重现性高、方便快捷等优点,被广泛地应用于微生物群落多样性和动态性分析。文章对DGGE／TGGE技术原理与关键环节、局限性和应用前景进行了综述。     

Microbial ecology of submerged marine caves and holes characterized by high levels of hydrogen sulphide

Submarine caves, cavities and niches characterised by H₂S elevated concentrations are particularly interesting for their inhabiting microflora as well as for the overall chemical, geological and biological parameters. These ecosystems are usually populated by well adapted living forms, physically distributed following the in situ concentration and gradient of micronutrients, O₂ and H₂S, and also according to the values of temperature and pH. The biota is primarily characterised by prokaryotes (both autotrophic and heterotrophic) adapted to anoxic and/or microaerophilic condition and capable to form extensive biofilms on the rocky surfaces and even on the bottom sediment. These habitats can be defined as extreme, because the scarcity or absence of solar irradiation, the chemo-physical traits and the fact that specialised prokaryotes are often the only inhabitants. This review is focused on the microbial ecology of marine caves and holes characterised by high levels of hydrogen sulphide. Ecological and geological data are already available but very few insights as far as regard microbiology were achieved in order to describe these fascinating habitats. The autochthonous mesophilic and thermotolerant microorganisms living in these caves may have interesting physiological traits and eventually may lead to potential application in biotechnological processes.     

Role of Nematodes in Soil Health and Their Use as Indicators

The composition of nematode communities (plant-parasitic and free-living) may be used as bioindicators of soil health or condition because composition correlates well with nitrogen cycling and decomposition, two critical ecological processes in soil. Maturity and trophic diversity indices withstand statistical rigor better than do abundances, proportions, or ratios of trophic groups. Maturity indices respond to a variety of land-management practices, based largely on inferred life history characteristics of families. Similarity indices may be more useful than diversity indices because they reflect taxon composition. Improving existing indices or developing alternative indices refined by a greater understanding of the biology of key taxa may enhance the utility of nematodes as bioindicators.     

The Effects of Sample Storage Conditions on the Microbial Community Composition in Hydraulic Fracturing Produced Water

The petroleum industry has an increasing interest in understanding the microbial communities driving biofouling and biocorrosion in reservoirs, wells, and infrastructure. However, sampling of the relevant produced fluids from subsurface environments for microbiological analyses is often challenged by high liquid pressures, workplace regulations, operator liability concerns, and remote sampling locations. These challenges result in infrequent sampling opportunities and the need to store and preserve the collected samples for several days or weeks. Maintaining a representative microbial community structure from produced fluid samples throughout storage and handling is essential for accurate results of downstream microbial analyses. Currently, no sample handling or storage recommendations exist for microbiological analyses of produced fluid samples. We used 16S rRNA gene sequencing to monitor the changes in microbial communities in hypersaline produced water stored at room temperature or at 4?°C for up to 7 days. We also analyzed storage at ?80?°C across a 3-week period. The results suggest ideal handling methods would include placing the collected sample on ice as soon as possible, but at least within 24?h, followed by shipping the samples on ice over 2–3?days, and finally, long-term storage in the ?20?°C or ?80?°C freezer.     

Responses of Microbial Communities to Light-Hydrocarbon Microseepage and Novel Indicators for Microbial Prospecting of Oil/Gas in the Beihanzhuang Oilfield,Northern Jiangsu,China

Previous studies on microbial prospecting of oil/gas only focused on the anomalies of light hydrocarbon-oxidizing microbes as main exploratory indicators and their exploration applications. In this study, we investigated the responses of microbial communities to light-hydrocarbon microseepage in the Beihanzhuang Oilfield, eastern China using denaturing gradient gel electrophoresis (DGGE) analysis and by comparing the difference of two-type areas with high- and low-flux light-hydrocarbon seepages. The results showed that the high-flux light-hydrocarbon seepage favored the growth of Nocardioides, Aciditerrimonas, sulphate-reducing bacteria (SRBs) related to Desulfosporosinus and Desulfovibrio, and Chloroflexi bacteria (b-7), implying that their anomalies might be adopted as novel subsidiary indicators for microbial prospecting of oil/gas in the Beihanzhuang Oilfield. Based on the newly obtained results, we have proposed a general strategy for microbial prospecting of oil/gas, i.e., to determine the anomalies of light hydrocarbon-oxidizing microbes, to select subsidiary indicators for microbial prospecting of oil/gas based on an assessment of the responses of microbial communities to light-hydrocarbon microseepage, to quantitatively measure subsidiary indicators and delimit their anomalies, to comprehensively interpret all microbial anomalies, and to make a suggestion for oil/gas prospecting. This general strategy with novel indicators may provide a more comprehensive evaluation for light-hydrocarbon microseepage and the corresponding anomalies, thereby reducing the exploration risk of oil/gas.     

Geochemistry and Microbial Populations in Sediments of the Northern Baffin Bay,Arctic

The Northern Baffin Bay between Greenland and Canada is a remote Arctic area restricted in primary production by seasonal ice cover, with presumably low sedimentation rates, carbon content and microbial activities in its sediments. Our aim was to study the so far unknown subseafloor geochemistry and microbial populations driving seafloor ecosystems. Shelf sediments had the highest organic carbon content, numbers of Bacteria and Archaea, and microcosms inoculated from Shelf sediments showed highest sulfate reduction and methane production rates. Sediments in the central deep area and on the southern slope contained less organic carbon and overall lower microbial numbers. Similar 16S rRNA gene copy numbers of Archaea and Bacteria were found for the majority of the sites investigated. Sulfate in pore water correlated with dsrA copy numbers of sulfate-reducing prokaryotes and differed between sites. No methane was found as free gas in the sediments, and mcrA copy numbers of methanogenic Archaea were low. Methanogenic and sulfate-reducing cultures were enriched on a variety of substrates including hydrocarbons. In summary, the Greenlandic shelf sediments contain vital microbial communities adapted to their specific environmental conditions.     

Plants and fertilisers as drivers of change in microbial community structure and function in soils

Here we report on a range of studies designed to understand the link between diversity and function in soils and in particular how plants and fertilisers might interact with microbial community dynamics in soils. The data presented indicate that although plants and fertilisers do impact on microbial community structure, the relationship between diversity, community structure and function remains complex and difficult to interpret using currently available chemical and molecular fingerprinting techniques. The paper assesses plants and management practices as drivers of change in soil and argues that whilst understanding diversity per se is unlikely to contribute to our understanding of function, an appreciation of what causes communities to change and also the relative importance of such drivers, could lead to new insights into the sustainable management and conservation of soils and natural resources.     

DGGE分析东江流域农村饮用水源中微生物多样性及其与环境因子相关性

为评价东江流域农村饮用水源中微生物多样性及其与环境因子的相关性, 分别采集了集中式供水井、塘坝型水井、猪场附近水井、普通村落水井、水库水5种水样, 进行基因组总DNA的提取和主要理化指标的测定, 运用DGGE技术分析各水样总DNA的PCR产物。UPGMA聚类分析DGGE指纹图谱结果表明, 相同类型水样的微生物群落结构相似性较高, 聚集到一个分支上; 典型相关性分析(CCA)结果表明, 水体中总磷(TP)和总氮(TN)的浓度与微生物群落结构的关联度最高, 即磷和氮两种生命过程的基本元素对微生物群落影响最大; 序列分析表明农村饮用水源中微生物群落结构丰富, 包含了螺旋体门(Spirochaetes)、蓝藻门(Cyanobacteria)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)5个门的细菌, 且每类水样拥有各自的优势菌。     

Feasibility of Water Efficiency and Reuse Technologies as Demand‐Side Strategies for Urban Water Management

Potable residential water efficiency and reuse technologies have seen increasing adoption in recent years and have been estimated to reduce demands by up to 50%. In this work, we used an engineering economic model to estimate the technically feasible levelized cost of water provided by seven above‐code water efficiency (i.e., beyond that required by building code) and reuse technologies within the Lower Colorado River Authority (LCRA) in central Texas. Unlike other demand‐side studies of residential water use, we model uncertainty and variation in technology adoption cost and performance; include reuse technologies; and differentiate between new construction and retrofits. We developed a conservation supply curve to compare the levelized cost of efficiency and reuse technologies with conventional supply‐side water management strategies. We estimate that efficiency and reuse in the residential sector can meet 85% of 50‐year projected needs (the difference between projected demand and estimated supplies) for the LCRA service area. We also estimate lower levelized costs for immediate retrofits of most technologies, promoting incentives for early technology adoption. However, efficiency and reuse technology performance demonstrates considerable uncertainty and variability. The fraction of demands met by demand‐side strategies range from around 60% to 100%. Occupancy drives much of the variability because it significantly affects demand. These results promote designing incentives for adoption of water efficiency and reuse technologies based upon use. We find that water‐efficient showerheads and bathroom faucet aerators perform well over a variety of assumptions, indicating that these technologies should be a priority for municipalities seeking water demand reductions.     

钱江源国家公园体制试点区日间声景观的质量评价及其空间格局特征

声景观在现行国家公园景观评价体系中,是一个十分重要但又薄弱的环节,关系着国家公园的精细化管理水平。以钱江源国家公园体制试点区为例,采用调查问卷法对国家公园日间的53种声景观进行主观愉悦度评价,考察并选取描述声景观质量的客观心理物理学指标,通过多元线性回归方法建立二者主客观关系,并得到国家公园声景观质量评价模型。结果表明：（1）不同类型声景观表现为不同的声学特征,并在响度、尖锐度、粗糙度指标上具有显著性差异。虫鸣声响度最大（5.829）、尖锐度最大（2.667）、粗糙度最大（0.018）,鸟鸣声的尖锐度与其余六种声景观均显著差异。（2）采用响度、尖锐度、粗糙度、波动度作为描述声景观质量的客观心理物理学指标具有适宜性。其中,响度是影响公众对国家公园声景观主观愉悦度认知的主要因素,二者呈负相关关系。（3）采用K-均值聚类等级划分发现,国家公园声景观质量处于较高水平。其中,虫鸣声质量最高（3.86）、鸟鸣声其次（3.58）、交谈声最低（2.43）。自然声景观质量整体高于人工声景观。（4）国家公园声景观质量在空间上具有南部高、北部其次、中部低的格局特征,且与区域生态保护等级水平存在较强空间重叠性。研究结果为国家公园体制试点结束后,未来声景观科学管理与规划设计提供重要参考,同时为其它类型自然保护地的声景观评价提供方法借鉴。     

Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors.The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis.     

Aeromonas Hydrophila and its Relation with Drinking Water Indicators of Microbiological Quality in Argentine

In Argentine, water municipal supplies disinfection is carried out by chlorine. We have isolated Aeromonas hydrophilafrom a chlorinated water supply in Buenos Aires that fulfilled Argentinean microbiological quality standards. It is an aquatic organism that could produce cytotoxins and enterotoxins associated with acute gastroenteritis and wound infections in human and hemorrhagic septicaemia of fish, reptiles and amphibians. The isolated strain produced protein bands at isoelectric point in the range of 4.6–5.3 lightly labile at 56°C and heat labile at 100°C non cholera antitoxin reactive in electrofocusing assay (IEF) and showed hemolytic activity thermolabile at 56°C and 100°C in 5% rabbit erythrocytes in phosphate buffered saline. A cytotoxic effect thermolabile at 56°C and 100°C and a cytotonic activity were demonstrated in Vero cell cultures. Survival assay of A. hydrophilaand indicator organisms would show no correlation between their contamination sources. Biofilms production could explain the persistence of this gram negative pathogen organism in chlorinated tap water. A water system supply maintenance program and pasteurisation may be performed in spite of water chlorination.     

新疆叶城两种主栽红枣土壤养分、微生物多样性与营养品质的比较分析

比较新疆叶城两种主栽红枣—骏枣与灰枣的土壤养分、微生物多样性和枣营养品质之间的差异,分析土壤养分、微生物与枣营养品质之间的相关性,从土壤养分、微生物角度研究枣营养品质的影响因素。通过化学分析法测定叶城骏枣与灰枣土壤养分、枣营养品质指标,高通量测序分析土壤微生物多样性,将土壤养分和微生物优势门属微生物进行冗余分析,研究枣营养品质指标与土壤养分、微生物功能类群的相关性。结果显示,新疆叶城两种主栽红枣土壤的全氮、速效氮、有机质含量显著不同;细菌群落变化的主要原因是土壤TN(全氮)值,真菌群落变化的主要原因是土壤AN(速效氮)值。两种主栽红枣土壤微生物多样性显示,骏枣土壤微生物在细菌水平上具有更高的物种丰富性和多样性,薄壁杆菌属(Gracilibacillus)、芽胞杆菌目(Bacillales)等溶磷微生物、固氮微生物等功能土壤微生物相对丰度和占比较高,差异显著;枣营养品质,尤其是不可溶膳食纤维、Fe、K、Zn的含量差异明显。而不可溶膳食纤维、K、蔗糖含量与TP(全磷)、OM(有机质)存在显著负相关;Fe、Zn含量与薄壁杆菌属、芽胞杆菌目等九类功能微生物存在显著正相关;Mg、可溶性糖、可溶性...     

Bacterial diversity in the bulk soil and rhizosphere fractions of Lolium perenne and Trifolium repens as revealed by PCR restriction analysis of 16S rDNA

The rhizosphere of Trifolium repens and Lolium perenne was divided into three fractions: the bulk soil, the soil adhering to the roots and the washed roots (rhizoplane and endorhizosphere). After isolation and purification of DNA from these fractions, 16S rDNA was amplified by PCR and cloned to obtain a collection of 16S rRNA genes representative of the bacterial communities of these three fractions. The genes were then characterized by PCR restriction analysis. Each different profile was used to define an operational taxonomic unit (OTU). The numbers of OTUs and the numbers of clones among these OTUs allowed to calculate a diversity index. The number of OTUs decreased as root proximity increased and a few OTUs became dominant, resulting in a lower diversity index. In the root fraction of T. repens, the restriction profile of the dominant OTU matched the theoretical profile of the 16S rRNA gene of Rhizobium leguminosarum. This study showed that plant roots create a selective environment for microbial populations.     

Spatial and Temporal Diversity Patterns of Planktonic Rotifer Assemblages in Water Bodies of the Floodplain Gemenc (Duna‐Dráva National Park,Hungary)

The Gemenc floodplain of the Danube possesses numerous side arms and backwaters. The aim of my study was to explore connections between hydrological events and diversity patterns of the planktonic rotifer assemblages of these water bodies. During the study period (2002–2008) 75 taxa of rotifers were found. Generally, the water bodies of the floodplain had more diverse rotifer assemblages than the main arm. At low and medium water levels diversity was highest in the parapotamal water bodies. This result may be explained by the intermediate disturbance hypothesis. At high water levels the highest biodiversities shifted to the plesiopotamal. Biodiversity, dominance, and evenness correlated with water flow and temperature. Patterns of biodiversity seem to be determined by the water level of the main arm; therefore the water regime appears to provide pronounced temporal variability in the hydro‐ecology of the floodplain. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

乌梁素海冰封期浮游植物分布特征及其与水质指标的关系北大核心CSCD

为揭示寒旱区冰封期富营养化湖泊水体中浮游植物群落结构及其与水质指标的响应关系,该研究以乌梁素海为对象,于2019年1月在湖区设立12个采样点采集水样及浮游植物,通过对浮游植物定性定量检测和水体理化性质测定分析,以明确冰封期乌梁素海浮游植物群落结构空间变化特征及主要水质指标的分布规律;结合RDA分析和Pearson相关性分析揭示了浮游植物与水质指标的响应关系,为水体富营养化程度评估及防控提供理论依据。结果显示:(1)冰封期乌梁素海12个样点的水质指标特征差异明显,各水质指标从北到南具有不同的变化趋势。(2)冰封期乌梁素海共检出浮游植物61种,其中隐藻门的丰度最高(4.76×10^(6)个·L^(-1)),甲藻门的生物量最高(18.09 mg·L^(-1)),但湖区不同位置的优势浮游植物类群有所差异,北湖区蓝隐藻和伪鱼腥藻丰度明显高于南湖区。(3)不同种类的浮游植物在空间分布上存在明显的差异,且在P3样点的多样性最低,P8样点的多样性最高,并发现中、下湖区物种类型多且组成较为均匀。(4)浮游植物的丰度与TP呈显著正相关关系,生物量与TP呈极显著正相关关系,多样性指数与TP呈正相关关系。研究表明,冰封期乌梁素海水体处于中等营养水平,水体中总磷(TP)含量是影响浮游植物物种丰度和生物量的主要影响因子;寒旱区冰封期富营养化湖泊浮游植物分布特征为北湖区隐藻门、蓝藻门和甲藻门占优势;南湖区中绿藻门丰度最高。