藻际菌胶团的特征、调节机制与生态功能

藻类对促进海洋物质循环、维持水生环境的生态平衡具有重要作用。在藻菌关系中,基于微生物的多样性和重要性,藻类与细菌之间的相互关系成为了研究的热点。藻际(phycosphere)环境是藻菌共生的一种典型生态位(niche),在这一生境中微生物形成的菌胶团(zoogloea consortium)是调控藻菌关系的重要组成部分。以往的研究证明菌胶团具有结构多样性和功能多样性,在维持藻际环境稳定、物质循环和信息交流方面具有重要的生态意义。本文中,笔者以藻际菌胶团为重点,综述其物质基础(物种组成、胞外聚合物类别)、结构特征(生物被膜)以及生态功能,并从化学信号的角度探讨了群体感应信号(quorum sensing)对菌胶团的调节机制。论文旨在梳理菌胶团的形成特点及其在营养摄取、环境抗性、功能维持上的最新进展,并在此基础上提出展望,为未来深入理解藻际菌胶团的生态学机制提供化学生态学思路。     

海洋中藻菌相互关系及其生态功能

海洋中藻类与细菌密不可分,具有错综复杂的互作关系(如互利共生、敌对拮抗或竞争抑制等),共同构成了海洋生态系统结构与功能的重要调控者。在藻类细胞周围往往存在着特殊的藻际微环境,其中生存着独特的微生物群落,因此藻际环境成为藻菌相互作用的主战场。藻际环境中细菌群落的构建具有一定的规律。在自然生态系统中,藻菌互作影响赤潮生消动态过程,并在水质修复中具有重要作用潜力。同时,藻类和细菌作为驱动海洋固碳与储碳的主要生物因子,在海洋碳循环中具有尤为重要的作用。本文对海洋中藻菌互作关系的研究现状进行了综述,并在此基础上,对未来研究提出了几点展望。例如,目前对海洋中藻菌关系受病毒的调控作用了解甚少,值得未来深入研究。     

海水中藻菌共培养体系对碳氮磷的吸收转化

海洋环境中,细菌和微藻之间的物质交换是生源要素在自然界中迁移转化的重要方式。为进一步了解生源要素的生物地球化学循环,在实验室模拟条件下,研究了共培养体系中营养盐和有机物在细菌和微藻之间的转换。通过纯培养中肋骨条藻(Skeletonema costatum)、东海原甲藻(Prorocentrum donghaiense)、天然海水中的细菌以及藻菌混合培养,分析了营养盐和有机物随藻菌生物量的变化情况,并计算了溶解有机碳(DOC)和溶解有机氮(DON)的浓度比值[(DOC/DON)a]。结果发现,在共培养体系中,细菌对中肋骨条藻的生长有抑制作用,对东海原甲藻影响不明显;中肋骨条藻有利于细菌生长,东海原甲藻抑制细菌生长,这种不同可能与微藻的粒径有关。海洋细菌在2种藻的指数生长均期均会促进微藻吸收氨氮(NH_4-N),但在生长末期NH_4-N以释放为主。硝氮(NO_3-N)的浓度与藻的生长呈负相关,但在衰亡期NO_3-N略有增加,表明NO_3-N再生所需时间较长。细菌对硝氮的吸收量较少,但对其再生有贡献。细菌和中肋骨条藻对磷酸盐(PO_4-P)的吸收存在竞争,但与东海原甲藻的竞争关系不明显。不同培养体系中DOC浓度变化不同,在藻菌共培养体系中增加较快,纯藻培养体系中增加缓慢,在纯菌培养体系中缓慢减少。通过对DOC与DON浓度比值的分析,发现用判断颗粒有机碳(POC)来源的方法可以分析DOC的来源。     

微生物在藻际环境中的物质循环作用

浮游植物作为海洋初级生产力的主要驱动者,其功能的发挥与共生微生物密不可分.藻类(甲藻、硅藻或蓝藻)的栖息环境中存在多样的共生细菌,各类细菌拥有不同的组成比例,但某些异养细菌在藻际环境中总是占据优势地位,如变形杆菌、黄杆菌及放线菌等.基于微生物在调节微食物网、促进物质循环和维持生态系统平衡中的重要意义,本文主要以赤潮事件的藻际环境为例,尝试梳理上述主导性“常驻微生物”在“藻-菌”共生体物质转化中的作用.特别是针对近些年来倍受关注的黄杆菌和玫瑰杆菌,着重例述了它们在物质代谢中的行为与生态策略,以更好地理解常驻物种在藻际生态位中的生态行为与协同进化.     

菌藻共生提高小球藻生物量和产油率

微藻规模化养殖常伴随着细菌的影响,存在于微藻藻际的细菌对微藻生长的影响及藻菌共生的机理尚缺乏深入研究。为建立有益的菌藻共生体系和提高微藻生物质产量,以埃氏小球藻(Chlorella emersonii)为试材,分离藻际微环境的菌群,并运用16S rDNA测序进行鉴定。通过藻菌(1∶1)共培养筛选优势促生菌。人工构建不同比例的菌藻共培养体系,分析优势促生菌对微藻生长和生物质产量的影响。结果显示,从埃氏小球藻藻株SXND-25藻际分离到6个菌种,属于菠萝泛菌属(Pantoea)、假单胞菌属(Pseudomonas)、鹑鸡肠球菌属(Enterococcus gallinarum)和大肠杆菌属(Escherichia coli)四个菌属。其中假单胞菌(Pseudomonas)和菠萝泛菌(Pantoea)为优势促生菌。与其他不同比例菌藻共培养相比,埃氏小球藻与菠萝泛菌1∶5共培养的促生效果突出,埃氏小球藻在第8天生物量达5.86 g/L,藻细胞含油量为26.88%,总油脂产量为1.575 g/L且单不饱和脂肪酸(MUFA)高达554-564mg/L。另一优异组合为埃氏小球藻与假单胞菌1∶1共培养,埃氏小球藻第8天生物量为4.12 g/L,藻细胞含油量达29.50%,总油脂产量提高到1.215 g/L,但MUFA含量低(168-175 mg/L)。研究表明在埃氏小球藻培养过程中,适量添加促生菌,可同时提高埃氏小球藻生物质和油脂产量,这为探究藻菌互作效应以及有益藻菌共生体系应用于微藻规模化生产提供参考依据。     

微藻与细菌作用关系的研究进展

藻类是水生环境中的初级生产者,它的生长常常伴随着细菌并受菌的影响。有研究者指出藻类和细菌有着密不可分的关系。一些研究表明与藻相关的主体细菌是特定的细菌群体,特别是α-变形菌频繁地发现,说明这类菌可能能够开启和维持共生关系。最近的研究提出了营养物质交换是菌藻共生的基础,这类相关化合物是复杂的和特定的分子,可能参与信号处理和监控作用,而不只是被动扩散。同时,这种作用很明显不是静态的,它的开启和终止可能是对环境和发育的响应。需要指出的是明确菌藻关系的作用机理还有待于进一步的深入研究,本篇综述结合新提出的理论,对细菌与微藻作用关系的研究进展进行总结,概括了微藻与菌的作用关系(进化关系,营养依赖,代谢互补和协作生物合成),这种作用关系涉及到的菌的分类(膜菌和藻际微环境菌,促生菌PGPB和溶藻菌)以及菌藻作用的应用(废水处理和生物燃料生产)的情况,并对菌藻关系的未来发展做了展望。     

藻际环境微生态结构与功能的研究进展

藻际环境是以藻类分泌物为骨架构成的微型生态结构,包含多样的生物和非生物,主要是微生物、胞外多糖、蛋白质和核酸等物质。藻际环境为复杂的藻菌互动提供平台,也为藻际系统的物质代谢、能量流动和信息交流提供基础。藻际环境不仅可以形成特殊的生态位,而且藻菌作用对多种生源要素循环起到了关键的作用;同时,藻际多糖的沉降为碳封存做出了重要贡献;再者,藻类在应激的时候,可以通过藻际环境形成对种群的保护。基于藻际环境结构和功能多样化的特点,解析其中的生态过程,对未来阐释生态现象的发生(如藻华)和生态修复都具有重要意义。该文综述了藻际环境的特性、影响因素和功能,旨在更好地认识藻际生态位,为藻类系统生态学发展提供理论依据。     

海洋细菌生态学的若干前沿课题及其研究新进展

海洋细菌在海洋生态系统中的重要作用随着微食物环的提出被深入认识和充分肯定。本文概述了海洋细菌在微食物环中的重要生态作用及微食物环的研究进展,海洋细菌在碳的生物地球化学循环中的重要性,海洋细菌的活性及其群落结构与功能,分析了藻际环境特性和藻际微生物在赤潮多发海域的生态作用,提出了我国海洋细菌生态学研究的若干新思考与新任务,强调了基于"以菌治藻"的新理念,开展针对于赤潮灾害防除的"微食物环-赤潮-关键微生物菌群"耦合互作这一重要科学问题研究的必要性及紧迫性。     

塔玛亚历山大藻藻际细菌溶藻过程

海洋微藻在生长过程中向周围环境分泌多种胞外产物,形成细菌自由生长的藻际环境,藻际细菌对微藻的生长有一定的调控作用。在指数生长期的塔玛亚历山大藻培养液中加入φ为1%的2216E培养基,在加入2216E后16h内藻细胞全部裂解。用数码显微镜记录了藻细胞形态变化,分别用DAPI法和荧光模拟底物法测定了细菌数量、胞外酶活性变化,结果表明:在溶藻过程中细菌数量、胞外酶活性在第6小时到第10小时增加了50～100倍。塔玛亚历山大藻藻际细菌主要分布在藻细胞表面,其群落结构改变和数量剧增是溶藻的主要原因,细菌分泌的β-葡萄糖苷酶和几丁质酶可能在溶藻过程中起重要作用。     

杜氏盐藻促生菌株的分离与鉴定

拟通过探究藻际微生物对微藻生长及代谢产物积累的影响,筛选出促进微藻生长的促生菌株。以杜氏盐藻(Dunaliella salina)Ds-SXYC-2为试材,分离鉴定盐藻藻际环境中的共生菌株,进一步构建藻菌(1∶1)共培养体系、测试盐藻生长及代谢产物积累等表型。结果显示,从杜氏盐藻藻际环境分离获得5株共生菌株,经16S rDNA分子鉴定,属于3个菌属。菌株B1与B2为涅斯捷连科氏菌(Nesterenkonia),菌株B3与B4为盐单胞菌(Halomonas),菌株B5为海杆菌(Marinobacter)。5株共生菌株对杜氏盐藻的生长均有促进作用,菌株B3能显著促进杜氏盐藻生长及代谢产物的积累。共培养15 d后,杜氏盐藻生物量达到2.3 g/L,比对照组增加了28.9%,叶绿素a的含量达到4.61 mg/L,比对照组增加了36.3%,β-胡萝卜素比对照组提高了56.4%。盐藻多糖、蛋白质、总脂含量分别比对照组增加了34.8%、71.2%和37.6%。菌株B3盐单胞菌可以作为促进杜氏盐藻生长及代谢产物累积的优势菌株,进一步构建共培养体系可应用于杜氏盐藻的商业生产。     

Production of macroaggregates from dissolved exopolymeric substances (EPS) of bacterial and diatom origin

Exopolymeric substances (EPS) isolated from a pure culture of the marine bacterium Marinobacter sp. and the marine diatom Skeletonema costatum (axenic) were partially purified, chemically characterized and used as dissolved organic matter (DOM) for the production of macroaggregates. The role of organic particles such as transparent exopolymeric particles (TEP) and Coomassie stained particles (CSP) in the production of macroaggregates was experimentally assessed. Three experimental rolling tanks containing sterile medium with: (1) EPS, (2) EPS + live diatom cells and (3) EPS + killed bacteria, and three control tanks without any added EPS were used for macroaggregate production. Changes in abundance and average size of macroaggregates were monitored using image analysis, whereas TEP and CSP were enumerated microscopically. In the presence of microbial EPS, macroaggregates of a size of 23-35 mm(2) were produced. Aggregate size and abundance considerably varied with both time and source of EPS. No correlation was observed for macroaggregate size and abundance with either TEP or CSP. One-way ANOVA demonstrated significant differences in the variance of particle abundance and size in tanks having only EPS or EPS in combination with live diatom cells. Our data suggest that production of macroaggregates was influenced by polymer chemistry and surface properties of colliding particles, whereas TEP and CSP concentrations were influenced by molecular weight of EPS and the presence of growing cells. Interestingly, macroaggregates were formed in the near absence of TEP and CSP, highlighting the role of other unknown processes in the transformation of DOM to particulate organic matter (POM) in aquatic environments.     

Shaping the phycosphere: Analysis of the EPS in diatom-bacterial co-cultures

The phycosphere is a unique niche that fosters complex interactions between microalgae and associated bacteria. The formation of this extracellular environment, and the associated bacterial biodiversity, is heavily influenced by the secretion of extracellular polymers, primarily driven by phototrophic organisms. The exopolysaccharides (EPS) represent the largest fraction of the microalgae-derived exudates, which can be specifically used by heterotrophic bacteria as substrates for metabolic processes. Furthermore, it has been proposed that bacteria and their extracellular factors play a role in both the release and composition of the EPS. In this study, two model microorganisms, the diatom Phaeodactylum tricornutum CCAP 1055/15 and the bacterium Pseudoalteromonas haloplanktis TAC125, were co-cultured in a dual system to assess how their interactions modify the phycosphere chemical composition by analyzing the EPS monosaccharide profile released in the culture media by the two partners. We demonstrate that microalgal–bacterial interactions in this simplified model significantly influenced the architecture of their extracellular environment. We observed that the composition of the exo-environment, as described by the EPS monosaccharide profiles, varied under different culture conditions and times of incubation. This study reports an initial characterization of the molecular modifications occurring in the extracellular environment surrounding two relevant representatives of marine systems.     

The Fate of Marine Bacterial Exopolysaccharide in Natural Marine Microbial Communities

Most marine bacteria produce exopolysaccharides (EPS), and bacterial EPS represent an important source of dissolved organic carbon in marine ecosystems. It was proposed that bacterial EPS rich in uronic acid is resistant to mineralization by microbes and thus has a long residence time in global oceans. To confirm this hypothesis, bacterial EPS rich in galacturonic acid was isolated from Alteromonas sp. JL2810. The EPS was used to amend natural seawater to investigate the bioavailability of this EPS by native populations, in the presence and absence of ammonium and phosphate amendment. The data indicated that the bacterial EPS could not be completely consumed during the cultivation period and that the bioavailability of EPS was not only determined by its intrinsic properties, but was also determined by other factors such as the availability of inorganic nutrients. During the experiment, the humic-like component of fluorescent dissolved organic matter (FDOM) was freshly produced. Bacterial community structure analysis indicated that the class Flavobacteria of the phylum Bacteroidetes was the major contributor for the utilization of EPS. This report is the first to indicate that Flavobacteria are a major contributor to bacterial EPS degradation. The fraction of EPS that could not be completely utilized and the FDOM (e.g., humic acid-like substances) produced de novo may be refractory and may contribute to the carbon storage in the oceans.     

Bacterial Activity and Preservation of Sedimentary Organic Matter: The Role of Exopolymeric Substances

Although exopolymeric substances (EPS) are associated with the microorganisms contributing to the production/degradation of sedimentary organic matter, their role in theses processes have so far never been mentioned. Using high-resolution microscopical tools (scanning and transmission electron microscopy, atomic force microscopy), fossil organic matter in the Miocene Monterey Formation (California) and Kimmeridgian laminites (France) has been compared with its present-day analogs, i.e., respectively sulphuroxidizing bacteria and cyanobacterial biofilms. This comparison shows that, particularly in the case of Kimmeridgian cyanobacterial mats deposited in a shallow back-reef environment, organic matter preservation is conditioned by exopolymeric substances secreted by bacteria. A model is proposed for the evolution through time of exopolymeric substances in relation to the mechanical constrains they have been exposed to, during lithification and diagenesis. This model is based on the microscopical observation of sulphuroxidizing bacteria and could explain the morphology of fossil organic matter usually referred to as “amorphous” in standard light microscopy. The highly hydrated nature of exopolymeric substances helps to protect organic matter from degradation and remineralization. These substances can be observed only in microscopy and are undetectable through organic geochemical methods, hence the need to combine these two methods in organic matter studies. Consequently, exopolymeric substances must be considered as an important contributing agent to organic matter preservation. These results confirm the complexity of the bacterial role in geoenvironments and add a new parameter in the productivity-vs-preservation debate.     

Algae-bacteria interactions and their effects on aggregation and organic matter flux in the sea

Aggregation of algae, mainly of diatoms, is an important process in marine pelagic systems, often terminating phytoplankton blooms and leading to the sinking of particulate organic matter in the form of marine snow. This process has been studied extensively, but the specific role of heterotrophic bacteria has largely been neglected, mainly because field studies and most experimental work were performed under non-axenic conditions. We tested the hypothesis that algae-bacteria interactions are instrumental in aggregate dynamics and organic matter flux. A series of aggregation experiments has been carried out in rolling tanks with two marine diatoms typical of temperate regions (Skeletonema costatum and Thalassiosira rotula) in an axenic treatment and one inoculated with marine bacteria. Exponentially growing S. costatum and T. rotula exhibited distinctly different aggregation behavior. This was reflected by their strikingly different release of dissolved organic matter (DOM), transparent exopolymer particles (TEP) and protein-containing particles (CSP), as well as their bacterial biodegradability and recalcitrance. Cells of S. costatum aggregated only little and their bacterial colonization remained low. Dissolved organic matter, TEP and CSP released by this alga were largely consumed by free-living bacteria. In contrast, T. rotula aggregated rapidly and DOM, TEP and CSP released resisted bacterial consumption. Experiments conducted with T. rotula cultures in the stationary growth phase, however, showed rapid bacterial colonization and decomposition of algal cells. Our study highlights the importance of heterotrophic bacteria to control the development and aggregation of phytoplankton in marine systems.     

细菌生物被膜基质的研究进展

细菌生物被膜(biofilm)附着在生物或者非生物表面,由细菌及其分泌的糖、蛋白质和核酸等多种基质组成的细菌群落,是造成病原细菌持续性感染、毒力和耐药性的重要原因之一.细菌的生物被膜基质由复杂的胞外聚合物(extracellular polymeric substances,EPS)构成,影响生物被膜的结构和功能.本文...     

藻类胞外聚合物的产生因素及其在污水处理和生物絮凝方面的应用

藻类胞外聚合物(extracellular polymeric substances, EPS)是一种复杂的高分子聚合物,主要由多糖、蛋白质等物质组成。由于EPS具有独特的结构、大的比表面积及含有大量官能团等物理-化学特性,使其在污水处理及微藻生物质的絮凝回收等方面都有着非常重要的作用。本文系统介绍了EPS的组成及特性,重点论述了影响藻类EPS产生的生物因素及非生物因素,如光照、营养盐、pH及温度等,并对EPS在污水处理及生物絮凝方面的应用进行了总结。对藻类EPS产生机制及机理的深入研究有望为微藻提供更广阔的应用前景。     

电活性微生物胞外聚合物的特征与应用

电活性微生物是一类能够通过直接接触、导电菌毛或氧化还原介质与电极或者其他细胞进行胞外电子传递的微生物。而在这个过程中,胞外聚合物(extracellular polymeric substances, EPS)扮演着重要的角色。EPS是微生物生长过程中通过细胞裂解、水解分泌的高分子聚合物的混合物,主要由蛋白质、多糖和腐殖质等物质组成。来自电活性微生物的EPS的不同组成成分和特性会对EPS的电活性以及电活性微生物胞外电子传递产生一定的影响,同时在环境应用方面发挥重要作用。因此,为了更全面了解电活性微生物EPS的电活性及其对电活性微生物胞外电子传递的作用,本文总体介绍了电活性微生物EPS的电活性的直接表征方法,再从组成成分、化学性质、物理性质和空间分布4个方面综述了其对EPS电活性的影响及其在电子传递中的作用,介绍了当前电活性微生物EPS在染料废水脱色、重金属吸附、有机污染物的生物转化和渗滤液管理等方面的环境应用,并从表征方法、试验规模和互作机理研究等角度展望了未来的研究方向。     

Sedimentation of ballasted cells‐free EPS in meromictic Fayetteville Green Lake

Fayetteville Green Lake (FGL) is a recognized, extensively studied present‐day model of the stratified Proterozoic ocean. Nonetheless, biomass sedimentation in FGL remains hard to explain: while virtually all sediment pigments belong to photosynthetic sulfur bacteria from a chemocline, the isotopic carbon signature of the bulk organic matter suggests its epilimnetic phytoplankton origin. To explain the epilimnetic origin of sedimented carbon, we studied the dominant Synechococci, isolated from FGL. Here, we present experimental evidence that FGL Synechococci produce copious extracellular polysaccharides (EPS) especially when availability of inorganic carbon (C_i) is high relative to availability of other macronutrients, for example phosphorus. The accumulating EPS become impregnated with calcium, magnesium, and sodium cations and are released to the environment as ballasted cell coverings. Sedimentation of these cell‐free EPS can constitute the bulk of pigment‐free organic material in FGL sediment. Because increased availability of C_i specifically stimulates production of EPS and the accumulated EPS adsorb cations and become ballasted, we propose the universal role of cyanobacterial EPS in biomass sedimentation in the high‐C_i Paleoproterozoic ocean as well as in modern aquatic systems like FGL.