生物技术在分子微生物生态学上的应用

分子生物学及其技术的发展和变革开辟了生态学研究的新途径。分子生物学与生态学交叉的研究,越来越引起人们的重视,于是分子生态学便应运而生了。而分子微生物生态学则为其中的重要分支,它是运用分子的方法和技术,在基因水平上估计种的个体丰度,查明种的变异情况以及探究群落中微生物间相互关系的科学。生物技术的应用,使我们不必培养微生物,而直接通过对环境中的遗传物质的研究来达到目的,它为微生物生态学的研究开辟了新的途径。１用于分子微生物生态学的主要生物技术１．１核酸探针检测技术探针是能与特定核苷酸序列发生特异性互…     

基因工程微生物生态学研究进展

基因工程微生物(genctically enginccred microorganism,GEM)生态学的研究已成为微生物分子生态学的一项主要研究内容之一．随着分子标记和分子生物学检测手段的引入,传统的微生物生态学研究被注入了新的活力,在分子水平上探讨基因工程微生物与环境及环境中土著生物之间的关系已成为可能．基因工程微生物生态学是一门内容涉及分子生物学、微生物学、生态学等诸多学科的新型交叉边缘学科．本文提出加紧进行转基因生物生态学和转基因生物的风险评价的研究工作,建立适合中国国情的检测手段和评价标准,有助于我国基因工程微生物生态学的健康发展．     

分子生态学技术及其在环境微生物研究中的应用

分子生态学作为一门新兴的学科已经成为国内外科学家关注和研究的热点。目前的分子生态学技术主要有核酸杂交分析技术、特异性PCR扩增技术、DNA序列分析、基因芯片技术等。这些技术在环境微生物研究中的应用主要包括对微生物多样性的研究、种群结构和动力学的研究、代谢活性的研究以及在全球气候变化中对微生物影响的研究。最后,对环境微生物的分子生态学研究进行了展望。     

微生物分子生态学技术在湖泊微生物多样性研究中的应用

微生物是湖泊生物圈物质循环和能量流动的主要参与者,在湖泊的生态系统中起着重要的作用。但是,湖泊中存在着大量不可培养的细菌,利用传统的培养技术,无法对湖泊微生物的多样性进行深入而全面的研究,而不依赖培养的分子生物学技术的发展为此方面研究开辟了新的路径。微生物分子生态学作为分子生物学与微生物生态学交叉产生的学科,在研究湖泊微生物多样性方面已经得到了广泛的应用。主要综述了变性梯度凝胶电泳(PCR-DGGE)技术,末端限制性酶切片段长度多态性技术(T-RFLP),16SrDNA克隆文库技术等微生物分子生态学技术在研究湖泊微生物多样性方面的应用情况。     

海洋微生物多样性及其分子生态学研究进展

海洋微生物多样性的深入研究将有助于微生物资源更好的开发和利用,海洋微生物多样性有很大的研究价值和研究空间。海洋中大多数微生物处于未可培养状态,在分子生态学基础上对海洋未可培养微生物进行研究是当今微生物多样性研究的主要方向。近年来相关研究进展迅速,研究方法不断更新。主要从分子生态学角度对微生物多样性研究现状进行概述并详细分析探讨了相关的研究方法,而且从分子生态学与海洋微生物多样性研究相结合的层面,对本领域的研究进行展望。旨在为海洋微生物多样性的研究及海洋资源的可持续开发与利用提供参考。     

分子生物学技术在生物修复中的应用研究进展

随着分子生物学技术如分子杂交、PCR、电泳技术等的发展,微生物学研究领域发生了深刻的变革,灵敏的检测和精确的细菌鉴定成为可能.微生物分子生态学作为分子生物学与微生物生态学交叉而形成的学科,在生物修复方面得到广泛应用.从分子生物学实验技术角度综述了各种微生物分子生物学技术在生物修复中的应用研究情况.     

PCR-DGGE技术及其在微生物生态学中的应用

现代分子生物学技术PCR-DGGE是一种分析微生物群落的有效工具,可以用于研究生态系统中微生物多样性和群落动态性。本文简要介绍了PCR-DGGE技术原理及其在微生物生态学领域的应用,并对该技术的局限性进行了评价。     

微生物分子生态学技术及其在环境污染研究中的应用

较为系统地概述了核酸探针检测技术、利用引物的PCR技术、DNA序列分析技术和电泳分离及显示技术在国内外的研究进展,并探讨了这些技术在环境污染研究中的应用及其方向。结果表明,这些被认为是重要的微生物分子生态学技术,在探索微生物与污染环境之间的相互关系中发挥了重要作用。促进了污染环境的微生物遗传适应进化机制的研究,污染物的微生物降解有关基因的定位及微生物工程菌的构建等方面的工作,从而推进了污染环境微生物修复的分子生态学的发展。     

FISH技术在微生物生态学中的研究及进展

分子生物学技术在微生物生态学研究中具有灵敏、精确和快速的优势,但不能提供微生物的形态学、数量性状、空间分布等信息。荧光原位杂交技术结合了分子生物学的精确性和显微镜的可视性信息,可以在自然生境中监测和鉴定不同的微生物个体,尤其是对难培养和未被培养的微生物进行检测。荧光原位杂交技术被广泛用于微生物群落结构诊断和评价,现已成为微生物分子生态学研究中的热点技术。对荧光原位杂交技术的发展和在微生物分子生态学中的应用进行了综述,探讨了该技术应用中存在的问题和发展前景。     

rRNA技术及其在分子微生物生态上的应用

传统的基于微生物培养与纯种分离的技术所具有的局限性,以及分子生物学及其有关技术的长足进展,使微生物生态学的研究进入了分子的阶段.其中rRNA技术的建立、发展及其成功应用,为分子微生物生态和微生物系统分类学的研究掀开了崭新的一页.对rRNA分子技术的研究进展、以之为基础的主要方法及其在环境微生物研究中的应用,以及应用过程中所存在的一些潜在问题及其解决办法等作了详细综述.     

Wastewater treatment: a model system for microbial ecology

Biological wastewater treatment is among the most important biotechnological applications and, as drivers of the key processes, microorganisms are central to its success. Therefore, the study of wastewater microorganisms has obvious applied significance; however, the importance of wastewater treatment reactors as model systems for microbial ecology is often overlooked. Modern molecular techniques, including environmental genomics, have identified unexpected microbial key players for nutrient removal and sludge bulking and/or foaming, and provided many exciting insights into the diversity, functions and niche differentiations of these predominantly uncultivated microorganisms. It is now time for wastewater microbiology to be recognized as a mature and dynamic discipline in its own right, offering much toward a deeper understanding of life in complex microbial communities. Here, we consider selected key findings to illustrate the past and future roles of molecular ecophysiology and genomics in the development of wastewater microbiology as an important subdiscipline of microbial ecology.     

Quantitative real-time PCR approaches for microbial community studies in wastewater treatment systems: Applications and considerations

Quantitative real-time PCR (qPCR) has been widely used in recent environmental microbial ecology studies as a tool for detecting and quantifying microorganisms of interest, which aids in better understandings of the complexity of wastewater microbial communities. Although qPCR can be used to provide more specific and accurate quantification than other molecular techniques, it does have limitations that must be considered when applying it in practice. This article reviews the principle of qPCR quantification and its applications to microbial ecology studies in various wastewater treatment environments. Here we also address several limitations of qPCR-based approaches that can affect the validity of quantification data: template nucleic acid quality, nucleic acid extraction efficiency, specificity of group-specific primers and probes, amplification of nonviable DNA, gene copy number variation, and limited number of sequences in the database. Even with such limitations, qPCR is reportedly among the best methods for quantitatively investigating environmental microbial communities. The application of qPCR is and will continue to be increasingly common in studies of wastewater treatment systems. To obtain reliable analyses, however, the limitations that have often been overlooked must be carefully considered when interpreting the results.     

Microbial communities involved in enhanced biological phosphorus removal from wastewater--a model system in environmental biotechnology

Enhanced biological phosphorus removal (EBPR) is one of the most advanced and complicated wastewater treatment processes applied today, and it is becoming increasingly popular worldwide as a sustainable way to remove and potentially reuse P. It is carried out by complex microbial communities consisting primarily of uncultured microorganisms. The EBPR process is a well-studied system with clearly defined boundaries which makes it very suitable as a model ecosystem in microbial ecology. Of particular importance are the transformations of C, N, and P, the solid-liquid separation properties and the functional and structural stability. A range of modern molecular methods has been used to study these communities in great detail including single cell microbiology, various -omics methods, flux analyses, and modeling making this one of the best studied microbial ecosystems so far. Recently, an EBPR core microbiome has been described and we present in this article some highlights and show how this complex microbial community can be used as model ecosystem in environmental biotechnology.     

Bacterial communities in full-scale wastewater treatment systems

Bacterial metabolism determines the effectiveness of biological treatment of wastewater. Therefore, it is important to define the relations between the species structure and the performance of full-scale installations. Although there is much laboratory data on microbial consortia, our understanding of dependencies between the microbial structure and operational parameters of full-scale wastewater treatment plants (WWTP) is limited. This mini-review presents the types of microbial consortia in WWTP. Information is given on extracellular polymeric substances production as factor that is key for formation of spatial structures of microorganisms. Additionally, we discuss data on microbial groups including nitrifiers, denitrifiers, Anammox bacteria, and phosphate- and glycogen-accumulating bacteria in full-scale aerobic systems that was obtained with the use of molecular techniques, including high-throughput sequencing, to shed light on dependencies between the microbial ecology of biomass and the overall efficiency and functional stability of wastewater treatment systems. Sludge bulking in WWTPs is addressed, as well as the microbial composition of consortia involved in antibiotic and micropollutant removal.     

Molecular biology techniques used in wastewater treatment: An overview

Identification of microorganisms by conventional methods requires the isolation of pure cultures followed by laborious characterization experiments. These procedures are therefore inadequate for study of the biodiversity of a natural or engineered ecosystem. A new set of molecular techniques developed during the 1990s revolutionized microbial ecology research. Among these techniques, cloning and the creation of a gene library, denaturant gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization with DNA probes (FISH) stand out. Cloning provides very precise taxonomical information, but is time consuming and requires specialized personnel and so its introduction in wastewater treatment has been slow. DGGE is a rapid and simple method that provides characteristic band patterns for different samples, allowing quick sample profiling, while retaining the possibility of a more thorough genetic analysis by sequencing of particular bands. FISH makes possible to identify microorganisms at any desired taxonomical level, depending on the specificity of the probe used. It is the only quantitative molecular biology technique, although quantification is either complex or tedious and subjective. Combination with a confocal laser-scanning microscope allows the visualization of three-dimensional microbial structures (granules, biofilms). The methods discussed have deepened our understanding of the microbiology of biological wastewater treatment. PCR-based methods (cloning and DGGE) have proved suitable for identifying the microorganisms that form the sludge. Both DGGE and FISH have been extensively employed. FISH is currently being used for elucidation of the composition, quantification and distribution of different bacterial groups in granules and biofilms, as well as their structure and architecture.     

光照混菌培养应用研究进展

光合细菌与其他微生物在光照条件下混合培养是近年来的研究热点。综述了光照混菌培养的特点和目前光照混菌培养在水体净化、生物制氢和高价值物质生产方面的应用,并对影响混合菌株生长代谢与繁殖的因素做了总结。分析表明菌株之间存在的相互协同共生作用能促进微生物的生长繁殖,使底物被充分利用,提高物质产率。光照混菌培养工艺简单、成本较低,在水体净化、生物制氢、高价值物质生产方面的应用具有相当好的效果。在影响因素中对混合培养影响最大的因素是菌株接种量、接种比和培养基pH。在总结光照混菌培养应用现存不足的基础上,对其发展前景作出展望。     

微生物生态学理论框架

微生物是生态系统的重要组成部分,直接或间接地参与所有的生态过程。微生物生态学是基于微生物群体的科学,利用微生物群体DNA/RNA等标志物,重点研究微生物群落构建、组成演变、多样性及其与环境的关系,在生态学理论的指导和反复模型拟合下由统计分析得出具有普遍意义的结论。其研究范围从基因尺度到全球尺度。分子生物学技术的发展,使人们可以直接从基因水平上考查其多样性,从而使得对微生物空间分布格局及其成因的深入研究成为可能。进而可以从方法学探讨微生物生物多样性、分布格局、影响机制及其对全球变化的响应等。在微生物生态学研究中,群落构建与演化、分布特征(含植物-微生物相互关系)、执行群体功能的机理(生物地球化学循环等)、对环境变化的响应与反馈机理是今后需要关注的重点领域。概述了微生物生态学的概念,并初步提出其理论框架,在对比宏观生态学基础理论和模型的基础上,分析微生物多样性的研究内容、研究方法和群落构建的理论机制,展望了今后研究的重点领域。     

人工湿地氮去除关键功能微生物生态学研究进展

人工湿地是一种能有效处理水体氮素污染的生态技术,其中微生物是驱动人工湿地系统中氮素去除的重要引擎。近20年来,随着分子生物学技术的广泛应用,有关人工湿地氮去除功能微生物生态学方面研究取得了一些重要进展。以硝化-反硝化作用和厌氧氨氧化作用这两种重要的人工湿地微生物脱氮途径为主,针对氨氧化细菌/古菌、厌氧氨氧化菌和反硝化菌等关键脱氮功能微生物的研究,重点归纳总结了目前有关这几类关键功能菌群在人工湿地中的丰度、活性、多样性、分布特征与影响因素,及其对废水中氮去除的作用,并在此基础上对今后的重点研究工作提出了展望。面向未来人工湿地氮去除关键功能微生物的研究应侧重其在污水净化和温室气体减排等方面的生态功能研究,同时加强其代谢过程与机制以及不同功能菌群间的关联研究。     

A review and update of the microbiology of enhanced biological phosphorus removal in wastewater treatment plants

Enhanced biological phosphorus removal (EBPR) from wastewater can be more-or-less practically achieved but the microbiological and biochemical components are not completely understood. EBPR involves cycling microbial biomass and influent wastewater through anaerobic and aerobic zones to achieve a selection of microorganisms with high capacity to accumulate polyphosphate intracellularly in the aerobic period. Biochemical or metabolic modelling of the process has been used to explain the types of carbon and phosphorus transformations in sludge biomass. There are essentially two broad-groupings of microorganisms involved in EBPR. They are polyphosphate accumulating organisms (PAOs) and their supposed carbon-competitors called glycogen accumulating organisms (GAOs). The morphological appearance of microorganisms in EBPR sludges has attracted attention. For example, GAOs as tetrad-arranged cocci and clusters of coccobacillus-shaped PAOs have been much commented upon and the use of simple cellular staining methods has contributed to EBPR knowledge. Acinetobacter and other bacteria were regularly isolated in pure culture from EBPR sludges and were initially thought to be PAOs. However, when contemporary molecular microbial ecology methods in concert with detailed process performance data and simple intracellular polymer staining methods were used, a betaproteobacteria called ‘Candidatus Accumulibacter phosphatis’ was confirmed as a PAO and organisms from a novel gammaproteobacteria lineage were GAOs. To preclude making the mistakes of previous researchers, it is recommended that the sludge ‘biography’ be well understood – i.e. details of phenotype (process performance and biochemistry) and microbial community structure should be linked. This revised version was published online in August 2006 with corrections to the Cover Date.