水稻土中脂肪酸互营氧化的研究进展

水稻田是温室气体甲烷(CH4)的重要释放源之一,有机质在水稻土中通过厌氧分解途径最终产生CH4和CO2.短链脂肪酸互营氧化是水稻土有机质降解的关键环节,但是由于互营微生物独特的生理生态特性,目前人们对于参与该过程的微生物群落及功能了解甚少.稳定同位素探针(SIP)技术被认为是实现环境中参与物质转化微生物种类与功能相耦合的有力工具.本文首先讨论互营过程的热力学基础和互营微生物的种间相互作用模式,然后简要讨论了互营过程的环境影响因子,最后详细综述稳定同位素探针技术在水稻土短链脂肪酸互营氧化过程中的相关研究.目前的研究表明:参与水稻土脂肪酸互营氧化过程的互营细菌种类丰富、多样性高;除已知互营细菌的作用外,大量未培养、功能未知的细菌类型也可能参与短链脂肪酸的互营氧化;对于互营细菌的伙伴而言,新型产甲烷胞菌属(Methanocella)类型的古菌在不同脂肪酸互营降解过程中均起主要作用,揭示了这类产甲烷古菌在水稻土厌氧产甲烷过程中的重要作用.     

厌氧氨氧化菌群体感应系统研究

厌氧氨氧化(Anammox)是以铵为电子供体将亚硝酸盐转化为氮气的生物过程。厌氧氨氧化菌(AAOB)生理代谢和细胞结构均十分特殊,且在氮素循环中起着十分重要的作用。厌氧氨氧化已成为环境学、微生物学、海洋学等领域的研究热点。但是,至今人们未能对厌氧氨氧化菌进行纯培养,这严重限制了对厌氧氨氧化菌的深入研究。群体感应是一种普遍存在于微生物细胞之间的通讯机制,它具有根据菌群密度和周围环境变化调节基因表达,以控制细菌群体行为的功能。厌氧氨氧化菌活性的细胞密度效应和生物团聚行为与细菌中普遍存在的群体感应现象相符。探讨了厌氧氨氧化菌群体感应系统存在的可能性、工作机制及其生态学意义,以期为厌氧氨氧化菌的分离培养、团聚体培育等提供理论指导。     

地下水脱卤过程中的微生物种间代谢互作：提高原位氯代烯烃厌氧脱氯效能的有效途径

有机卤呼吸细菌(organohalide-respiring bacteria, OHRB)在氯代烯烃污染地下水的原位生物修复中扮演着关键性的角色,提高其丰度及活性对氯代烯烃的完全去除具有重要意义。在实际环境中,有机卤呼吸细菌往往与多种微生物共存,微生物种间代谢互作现象十分普遍,有机污染物的完全无害化往往需要通过微生物菌群的协同代谢作用来实现。因此,本文围绕微生物种间代谢互作进行综述,对目前获得的脱氯微生物菌种资源及脱氯机理进行了回顾,重点阐述了专性OHRB、非专性OHRB和非OHRB的种间代谢互作行为及机制,并提出以种间代谢互作为指导进行合成微生物群落的构建来有效提高氯代烯烃厌氧生物降解效率,为实现环境氯代烯烃类有机污染物的快速、彻底无害化提供理论指导。     

互营氧化产甲烷微生物种间电子传递研究进展

甲烷是重要的温室气体,也是典型的可再生性生物质能源。目前约70%的大气甲烷排放来源于产甲烷微生物过程。在产甲烷环境中,产甲烷菌与互营细菌形成互营关系,从而克服有机质厌氧分解反应的热力学能垒,实现短链脂肪酸和醇类物质的互营氧化产甲烷过程。该过程中,种间电子传递是关键步骤。本文首先概述了甲烷的研究意义及微生物互营降解有机质产甲烷的过程,然后分别综述了种间H2转移、种间甲酸转移和种间直接电子传递这3种种间电子传递机制的起源、发展、研究现状和未来所需要解决的研究问题。     

暗黑菌(Atribacteria)的环境分布和功能特征

暗黑菌门包括OP9和JS1两大分支,成员大多为未培养微生物,在自然环境中广泛分布,并且在部分环境如厌氧海洋沉积物、地热环境以及油藏中为优势微生物。基于基因组信息的研究表明,暗黑菌为严格的厌氧微生物,同时具有降解糖类、小分子酸、短链正构烷烃的能力,在地球碳循环过程中可能扮演着重要的角色。然而,由于缺乏相应代表性的纯培养菌株,对于暗黑菌的生理生化功能推测仍有待验证。文章概述了暗黑菌的发现及发展历史,分析了其环境分布和多样性,简述了目前提出的三种代谢方式,提出了未来暗黑菌的研究发展方向。     

典型有机固废厌氧消化微生物研究现状与发展方向

经过人工富集和驯化的兼性和严格厌氧微生物是厌氧消化工艺的核心。不同厌氧消化体系中存在的问题大多可以通过改变微生物群落的代谢活性来得到有效改善。得益于微生物组学检测技术的快速发展,对厌氧消化系统中微生物多样性的认识获得了极大的拓展,同时在微生物类群间、微生物与环境的互作关系研究方面也取得了一系列新的进展。然而,有机固废厌氧消化系统中,各种微生物以及微生物和物质的相互作用构成了更为复杂的代谢网络,所以目前对这些互作关系的解析尚不完善。本文重点关注了厌氧消化过程中的典型菌群互作关系,阐述了典型有机固废厌氧消化系统中存在的问题及微生物在其中发挥的作用,最后,立足于现有组学技术推动的微生物组研究进展,对未来有机固废厌氧消化系统微生物组的研究提出展望。     

隐藏嗜酸菌Acidiphilium cryptum的研究进展

分离自酸性矿坑水的隐藏嗜酸菌Acidiphilium cryptum具有化能兼养型细菌的典型特征,在缺乏有机质的环境中能启用体内硫氧化代谢途径营化能自养,进行硫化矿物的分解;而在有机质存在时该菌营异养生长,并在体内合成聚β羟基丁酸酯(polyhydroxybutyrate,PHB),此外在酸性条件下对Fe(III)和Cr(VI)等金属离子具有还原作用,并具有发展微生物燃料电池的潜力.该菌因其独特的生理特性及在浸矿微生物群落中的显著生态地位而在生物冶金、环境修复、生物材料等领域具有重要的经济和社会意义.对隐藏嗜酸菌的生物学特性、分离培养方法、重要特性的研究进展以及应用等方面的国内外研究现状进行综述,并对其应用前景和研究趋势进行展望.     

马里亚纳海沟共培养细菌多样性动态变化及潜在微生物互作关系

马里亚纳海沟是世界已知最深的海沟,其寡营养、高压、低温、低氧等极端的深海环境孕育出独特的细菌群落结构及多样性特征。选取寡营养培养基对马里亚纳海沟海水及表层沉积物分别进行液体共培养,并在不同培养阶段取样进行高通量测序,分析细菌群落结构组成及其多样性的动态变化,探讨微生物之间可能的互作关系。研究结果表明：液体共培养样品中一共检测到19个门、34个纲、76个目、131个科、227个属的细菌,其中变形菌门（Proteobacteria）和拟杆菌门（Bacteroidetes）为优势菌群,其次为厚壁菌门（Firmicutes）;与其他样品相比,1000米海水样品中细菌群落的多样性最高,并且蓝细菌门（Cyanobacteria）具有更高的相对丰度。共培养样品中细菌丰富度、多样性、群落结构均随培养时间而改变,其中共培养中期样品的细菌多样性较高;表层沉积物样本中,盐单胞菌属（Halomonas）可能由于较强的竞争能力在共培养后期占据优势地位。基因功能预测与代谢通路富集结果显示,随着共培养时间的增加,微生物生长相关的代谢通路丰度明显下降,而与互作相关的代谢通路丰度明显增加。共培养样品检测到的细菌多样性远高于单独分离培养的多样性,仅有少量菌属为单独分离培养与共培养样品均检测到的共有属。综上所述,马里亚纳海沟细菌群落中存在竞争、互利共生的相互作用,共培养法有利于揭示细菌间的互作关系。研究为深渊及深海等极端环境下微生物生态系统组成及维持奠定了理论基础,也为进一步研究极端微生物的生存策略提供了科学指导。     

厌氧氨氧化细菌Candidatus Kuenenia stuttgartiensis铁的吸收利用研究进展

铁是影响微生物生长代谢的关键元素,它与蛋白质结合,起催化、氧化还原或调节作用.厌氧氨氧化(ANAMMOX)细菌的生长代谢严重依赖铁,尤其是含铁蛋白.ANAMMOX细菌的厌氧生活方式和厌氧氨氧化体的存在使其对铁代谢的模式不同于其他微生物.弄清ANAMMOX细菌的铁吸收代谢模式,可为获得其纯培养物奠定基础,有利于促进其在环...     

污水脱氮功能微生物的组学研究进展

生物脱氮是污水处理厂的核心,掌握生物脱氮过程相关微生物代谢特性,对于探索微生物资源和提高污水处理厂脱氮性能具有重要意义。近年来,分子生物学方法不断发展和改进,已被广泛应用于揭示脱氮微生物群落多样性、组成结构和潜在功能等方面,大幅提升了研究者们对污水生物脱氮系统中微生物,尤其是不可培养微生物的代谢机理、抑制调控原理及新型生物脱氮工艺途径的认识。本文对流行的分子生物学方法(16S rRNA基因测序、实时荧光定量PCR技术、宏基因组学、宏转录组学、宏蛋白质组学和代谢组学)进行了介绍,综述了其在硝化细菌、反硝化细菌、完全氨氧化细菌、厌氧氨氧化细菌、厌氧铁氨氧化细菌、硫酸盐型厌氧氨氧化细菌及亚硝酸盐/硝酸盐型厌氧甲烷氧化微生物等方面的研究进展,阐明了这些氮素转化微生物在氮循环过程的代谢途径和酶促反应,并从标准测定方法构建、不同方法的联用及跨学科结合和检测方法的简易化这3个方面展望了分子生物学方法的技术突破及其在污水生物处理系统中的应用前景。本综述从系统角度全面认识脱氮微生物群落及其结构,为未来污水处理生物脱氮微生物的研究提供了新方向。     

Growth and methane oxidation rates of anaerobic methanotrophic archaea in a continuous-flow bioreactor

Anaerobic methanotrophic archaea have recently been identified in anoxic marine sediments, but have not yet been recovered in pure culture. Physiological studies on freshly collected samples containing archaea and their sulfate-reducing syntrophic partners have been conducted, but sample availability and viability can limit the scope of these experiments. To better study microbial anaerobic methane oxidation, we developed a novel continuous-flow anaerobic methane incubation system (AMIS) that simulates the majority of in situ conditions and supports the metabolism and growth of anaerobic methanotrophic archaea. We incubated sediments collected from within and outside a methane cold seep in Monterey Canyon, Calif., for 24 weeks on the AMIS system. Anaerobic methane oxidation was measured in all sediments after incubation on AMIS, and quantitative molecular techniques verified the increases in methane-oxidizing archaeal populations in both seep and nonseep sediments. Our results demonstrate that the AMIS system stimulated the maintenance and growth of anaerobic methanotrophic archaea, and possibly their syntrophic, sulfate-reducing partners. Our data demonstrate the utility of combining physiological and molecular techniques to quantify the growth and metabolic activity of anaerobic microbial consortia. Further experiments with the AMIS system should provide a better understanding of the biological mechanisms of methane oxidation in anoxic marine environments. The AMIS may also enable the enrichment, purification, and isolation of methanotrophic archaea as pure cultures or defined syntrophic consortia.     

Growth and Methane Oxidation Rates of Anaerobic Methanotrophic Archaea in a Continuous-Flow Bioreactor

Anaerobic methanotrophic archaea have recently been identified in anoxic marine sediments, but have not yet been recovered in pure culture. Physiological studies on freshly collected samples containing archaea and their sulfate-reducing syntrophic partners have been conducted, but sample availability and viability can limit the scope of these experiments. To better study microbial anaerobic methane oxidation, we developed a novel continuous-flow anaerobic methane incubation system (AMIS) that simulates the majority of in situ conditions and supports the metabolism and growth of anaerobic methanotrophic archaea. We incubated sediments collected from within and outside a methane cold seep in Monterey Canyon, Calif., for 24 weeks on the AMIS system. Anaerobic methane oxidation was measured in all sediments after incubation on AMIS, and quantitative molecular techniques verified the increases in methane-oxidizing archaeal populations in both seep and nonseep sediments. Our results demonstrate that the AMIS system stimulated the maintenance and growth of anaerobic methanotrophic archaea, and possibly their syntrophic, sulfate-reducing partners. Our data demonstrate the utility of combining physiological and molecular techniques to quantify the growth and metabolic activity of anaerobic microbial consortia. Further experiments with the AMIS system should provide a better understanding of the biological mechanisms of methane oxidation in anoxic marine environments. The AMIS may also enable the enrichment, purification, and isolation of methanotrophic archaea as pure cultures or defined syntrophic consortia.     

Biogeochemistry and microbial ecology of methane oxidation in anoxic environments: a review

Evidence supporting a key role for anaerobic methane oxidation in the global methane cycle is reviewed. Emphasis is on recent microbiological advances. The driving force for research on this process continues to be the fact that microbial communities intercept and consume methane from anoxic environments, methane that would otherwise enter the atmosphere. Anaerobic methane oxidation is biogeochemically important because methane is a potent greenhouse gas in the atmosphere and is abundant in anoxic environments. Geochemical evidence for this process has been observed in numerous marine sediments along the continental margins, in methane seeps and vents, around methane hydrate deposits, and in anoxic waters. The anaerobic oxidation of methane is performed by at least two phylogenetically distinct groups of archaea, the ANME-1 and ANME-2. These archaea are frequently observed as consortia with sulfate-reducing bacteria, and the metabolism of these consortia presumably involves a syntrophic association based on interspecies electron transfer. The archaeal member of a consortium apparently oxidizes methane and shuttles reduced compounds to the sulfate-reducing bacteria. Despite recent advances in understanding anaerobic methane oxidation, uncertainties still remain regarding the nature and necessity of the syntrophic association, the biochemical pathway of methane oxidation, and the interaction of the process with the local chemical and physical environment. This review will consider the microbial ecology and biogeochemistry of anaerobic methane oxidation with a special emphasis on the interactions between the responsible organisms and their environment. This revised version was published online in August 2006 with corrections to the Cover Date.     

六氯-1,3-丁二烯的微生物降解研究进展

六氯-1,3-丁二烯(hexachlorobutadiene,HCBD)是一种有毒有害的脂肪族氯代烃,曾经作为杀虫剂、除草剂、变压器油和传热流体等化学工业产品的重要成分被广泛应用于生产生活。HCBD因满足《关于持久性有机污染物的斯德哥尔摩公约》中风险筛选标准(如毒性、持久性、远距离环境迁移和生物累积性等),缔约方于2015年第七次会议中将其增列为持久性有机污染物,2017年又将其列入该公约的附件Ｃ以控制其环境排放量。目前关于HCBD的环境归趋仍是研究热点,但是对于HCBD的微生物降解转化机制尚缺乏深入研究。鉴于此,本文重点回顾并讨论了地下水、底泥等厌氧环境中已报道的HCBD微生物降解转化途径、速率及机制,并从热力学角度阐述HCBD及其降解产物作为电子受体通过还原性脱氯反应被厌氧脱卤微生物代谢转化的可行性。最后,本文根据现有研究结果,提出微生物厌氧降解HCBD的研究展望,包括多组学技术解析HCBD降解功能菌群结构和潜在互作机制、HCBD厌氧降解微生物的分离与纯化,以及HCBD厌氧降解菌剂的开发与污染场地原位生物修复应用等。     

Anaerobic free-living protozoa: growth efficiencies and the structure of anaeorobic communities

Abstract Most anoxic environments host populations of phagotrophic, eukaryote microorganisms. Many physiological properties of these anaerobic eukaryotes are still incompletely understood and their role in communities of anaerobic microorganisms has so far drawn little attention. Here we present theoretical considerations and experimental evidence to show that the net growth efficiency ([assimilated C]/[assimilated C + dissimilated C]) and gross growth efficiency (yield = [assimilated C]/[consumed C]) of anaerobic protozoa are about 20% and about 25%, respectively of those of their aerobic counterparts. This accords with the observation that the biomass ratios of predators and their prey is about one fourth of that foundin oxic environments. These fiedl data also suggest that bacterial numbers are controlled by protozoa grazing in at least some anoxic environments. Finally, the results explain whe phagotrophic food chains are short and eukaryote diversity is low in anaerobic habitats.     

Stable-isotope probing of microorganisms thriving at thermodynamic limits: syntrophic propionate oxidation in flooded soil

Propionate is an important intermediate of the degradation of organic matter in many anoxic environments. In methanogenic environments, due to thermodynamic constraints, the oxidation of propionate requires syntrophic cooperation of propionate-fermenting proton-reducing bacteria and H(2)-consuming methanogens. We have identified here microorganisms that were active in syntrophic propionate oxidation in anoxic paddy soil by rRNA-based stable-isotope probing (SIP). After 7 weeks of incubation with [(13)C]propionate (<10 mM) and the oxidation of approximately 30 micromol of (13)C-labeled substrate per g dry weight of soil, we found that archaeal nucleic acids were (13)C labeled to a larger extent than those of the bacterial partners. Nevertheless, both terminal restriction fragment length polymorphism and cloning analyses revealed Syntrophobacter spp., Smithella spp., and the novel Pelotomaculum spp. to predominate in "heavy" (13)C-labeled bacterial rRNA, clearly showing that these were active in situ in syntrophic propionate oxidation. Among the Archaea, mostly Methanobacterium and Methanosarcina spp. and also members of the yet-uncultured "rice cluster I" lineage had incorporated substantial amounts of (13)C label, suggesting that these methanogens were directly involved in syntrophic associations and/or thriving on the [(13)C]acetate released by the syntrophs. With this first application of SIP in an anoxic soil environment, we were able to clearly demonstrate that even guilds of microorganisms growing under thermodynamic constraints, as well as phylogenetically diverse syntrophic associations, can be identified by using SIP. This approach holds great promise for determining the structure and function relationships of further syntrophic or other nutritional associations in natural environments and for defining metabolic functions of yet-uncultivated microorganisms.     

Synergistic interactions in the microbial world

After several decades of microbiological research has focused on pure cultures, synergistic effects between different types of microorganisms find increasing interest. Interspecies interactions between prokaryotic cells have been studied into depth mainly with respect to syntrophic cooperations involved in methanogenic degradation of electron-rich substrates such as fatty acids, alcohols, and aromatics. Partners involved in these processes have to run their metabolism at minimal energy increments, with only fractions of an ATP unit synthesized per substrate molecule metabolized, and their cooperation is intensified by close proximity of the partner cells. New examples of such syntrophic activities are anaerobic methane oxidation by presumably methanogenic and sulfate-reducing prokaryotes, and microbially mediated pyrite formation. Syntrophic relationships have also been discovered to be involved in the anaerobic metabolization of amino acids and sugars where energetical restrictions do not necessarily force the partner organisms into strict interdependencies. The most highly developed cooperative systems among prokaryotic cells appear to be the structurally organized phototrophic consortia of the Chlorochromatium and Pelochromatium type in which phototrophic and chemotrophic bacteria not only exchange metabolites but also interact at the level of growth coordination and tactic behaviour. This revised version was published online in August 2006 with corrections to the Cover Date.     

Syntrophic oxidation of propionate in rice field soil at 15 and 30°C under methanogenic conditions

Propionate is one of the major intermediary products in the anaerobic decomposition of organic matter in wetlands and paddy fields. Under methanogenic conditions, propionate is decomposed through syntrophic interaction between proton-reducing and propionate-oxidizing bacteria and H(2)-consuming methanogens. Temperature is an important environmental regulator; yet its effect on syntrophic propionate oxidation has been poorly understood. In the present study, we investigated the syntrophic oxidation of propionate in a rice field soil at 15°C and 30°C. [U-(13)C]propionate (99 atom%) was applied to anoxic soil slurries, and the bacteria and archaea assimilating (13)C were traced by DNA-based stable isotope probing. Syntrophobacter spp., Pelotomaculum spp., and Smithella spp. were found significantly incorporating (13)C into their nucleic acids after [(13)C]propionate incubation at 30°C. The activity of Smithella spp. increased in the later stage, and concurrently that of Syntrophomonas spp. increased. Aceticlastic Methanosaetaceae and hydrogenotrophic Methanomicrobiales and Methanocellales acted as methanogenic partners at 30°C. Syntrophic oxidation of propionate also occurred actively at 15°C. Syntrophobacter spp. were significantly labeled with (13)C, whereas Pelotomaculum spp. were less active at this temperature. In addition, Methanomicrobiales, Methanocellales, and Methanosarcinaceae dominated the methanogenic community, while Methanosaetaceae decreased. Collectively, temperature markedly influenced the activity and community structure of syntrophic guilds degrading propionate in the rice field soil. Interestingly, Geobacter spp. and some other anaerobic organisms like Rhodocyclaceae, Acidobacteria, Actinobacteria, and Thermomicrobia probably also assimilated propionate-derived (13)C. The mechanisms for the involvement of these organisms remain unclear.     

Ecophysiology of syntrophic communities that degrade saturated and unsaturated long-chain fatty acids

Syntrophic relationships are the key for biodegradation in methanogenic environments. We review the ecological and physiological features of syntrophic communities involved in the degradation of saturated and unsaturated long-chain fatty acids (LCFA), as well as their potential application to convert lipids/fats containing waste to biogas. Presently, about 14 species have been described with the ability to grow on fatty acids in syntrophy with methanogens, all belonging to the families Syntrophomonadaceae and Syntrophaceae . The principle pathway of LCFA degradation is through β-oxidation, but the initial steps in the conversion of unsaturated LCFA are unclear. Communities enriched on unsaturated LCFA also degrade saturated LCFA, but the opposite generally is not the case. For efficient methane formation, the physical and inhibitory effects of LCFA on methanogenesis need to be considered. LCFA adsorbs strongly to biomass, which causes encapsulation of active syntrophic communities and hampers diffusion of substrate and products in and out of the biomass. Quantification of archaea by real-time PCR analysis suggests that potential LCFA inhibitory effect towards methanogens might be reversible. Rather, the conversion of adsorbed LCFA in batch assays was shown to result in a significant increase of archaeal cell numbers in anaerobic sludge samples.