基于PCR-DGGE技术的红树林区微生物群落结构

【目的】为了解红树林沉积物中细菌的群落结构特征。【方法】应用PCR-DGGE技术对福建浮宫红树林的16个采样站位样品细菌的群落结构进行了研究。根据DGGE指纹图谱,对它们的遗传多样性进行了分析。【结果】各站位样品细菌多样性指数(H)、丰度(S)和均匀度(EH)均有所不同,这些差异与它们所处站位的不同有关,红树林区细菌多样性高于非红树林区细菌多样性。对不同站位细菌群落相似性分析,它们的相似性系数也存在一定的规律,同一断面的细菌群落结构相近性较高。对DGGE的优势条带序列分析,同源性最高的微生物分别属于变形菌门(Proteobacteria)、酸菌门(Acidobacteria)和绿菌门(Chlorobi),它们均为未培养微生物,分别来自于河口海岸沉积物。【结论】应用PCR-DGGE技术更能客观地反映红树林沉积物中真实的细菌群落结构信息。另外,研究也表明红树林区微生物多样性丰富,在红树林区研究开发未知微生物资源具有巨大的潜力。     

青藏高原淡水湖普莫雍错和盐水湖阿翁错湖底沉积物中细菌群落的垂直分布

【目的】湖泊沉积物中存储着大量独特的微生物,这些微生物在湖泊生态系统生物地球化学循环中扮演着非常重要的角色。然而,很少有研究报道微生物群落在湖泊沉积物中的垂直分布。本文比较研究青藏高原淡水湖普莫雍错和盐水湖阿翁错沉积物在不同深度下细菌的丰度和群落结构。【方法】利用定量PCR(q PCR)和变性梯度凝胶电泳(DGGE)技术分别测定细菌群落的丰度与群落结构。【结果】定量PCR结果显示,湖泊沉积物中细菌丰度均随深度增加而降低,盐水湖阿翁错和淡水湖普莫雍错的细菌丰度分别从1011数量级降到108数量级,从1012数量级降到1010数量级。在相对应的沉积物层,淡水湖沉积物的细菌丰度比盐水湖高1-2个数量级。变性梯度凝胶电泳(DGGE)指纹图谱的分析表明,淡水湖沉积物细菌群落的DGGE条带数(丰富度)显著高于盐水湖(P=0.014);淡水与盐水湖泊沉积物细菌群落结构明显不同,同时在同一湖泊沉积物中上层(0-6 cm)和下层(7-20 cm)细菌群落结构也呈明显分异。系统发育分析表明,盐水湖阿翁错沉积物特有菌门为Gamma-变形菌、拟杆菌门、蓝细菌和栖热菌门,而淡水湖普莫雍错沉积物中特有菌门为Delta-和Beta-变形菌、酸杆菌和绿弯菌门。【结论】青藏高原淡水与盐水湖泊沉积物细菌丰度与群落结构具有明显的差异;同时,细菌群落结构在沉积物的不同深度也表现出差异。这些结果可为进一步阐明青藏高原湖泊生态系统中微生物对气候环境变化的响应提供科学依据。     

重度霾污染时空气细菌群落特征及与关键环境因子的相关性

【背景】近年来频繁发生的霾污染事件严重威胁着人类的正常生活和社会经济发展。目前对霾污染颗粒物的重要组成部分生物气溶胶的关注较少。【目的】明确霾污染时空气微生物的特征,为进一步研究空气微生物对霾污染的形成及反馈机制奠定基础。【方法】采集了北京地区重度霾污染的样品,测定颗粒物的化学组分,通过高通量测序技术分析空气细菌群落结构特征,并明确其与环境因子的相关性。【结果】霾污染显著改变空气细菌群落结构、降低α多样性。霾污染使空气中变形菌门相对丰度显著上升,放线菌门相对丰度下降。颗粒物浓度及化学组分是重度霾污染中影响空气细菌群落结构的关键环境因子。与气态污染物SO_2和NO_2相比,水溶性离子硫酸盐、硝酸盐和铵盐对空气细菌群落结构的影响更大。【结论】解析了重度霾污染时空气细菌群落特征和关键环境因子之间的相关性,为大气环境微生物的研究、空气质量的评价及霾污染的有效防治提供了数据参考。     

马里亚纳海沟不同深度水样微生物分离与多样性分析

【背景】马里亚纳海沟挑战者深渊是地球表面最深点,认识其微生物群落结构和分离培养的微生物对挖掘新的微生物资源具有重要意义。【目的】对马里亚纳海沟不同深度水样进行细菌分离培养,并与这些样品的微生物群落结构进行比较,认识进一步要分离培养的微生物类型。【方法】采用不同培养基对马里亚纳海沟两个站位不同深度水样进行细菌分离培养,并通过Illumina MiSeq高通量测序分析各个水样的细菌和古菌的群落结构。【结果】从来自两个站位不同深度的6个水样样品中分离获得783株细菌,属于4个门6个纲28个属。其中,变形菌门占主导地位,67.8%的菌株属于γ-变形菌纲。分离获得的菌株主要属于亚硫杆菌属、假单胞菌属和交替假单胞菌属,它们在这些样品中广泛分布,且在高通量测序结果中也能检测到。高通量测序结果表明除浅层样品优势微生物为蓝细菌外,其他样品以变形菌门占主导;不同深度样品的微生物群落结构存在较大差异。【结论】马里亚纳海沟不同深度水样中不仅分离培养出了相对丰度较高的一些细菌属,也分离得到一些相对丰度较低的微生物类型。从马里亚纳海沟水样中分离培养获得的细菌菌株资源将用于功能微生物和功能酶挖掘等相关研究,这有利于深渊微生物资源挖掘。     

异育银鲫养殖环境典型病原微生物检测和细菌群落解析

【背景】水产养殖病害是限制淡水渔业发展的一个重要因素,养殖环境的微生物状况与鱼类健康紧密相关,已引起人们广泛关注。【目的】阐明养殖水体和沉积物中病原微生物丰度和细菌群落多样性变化特征。【方法】利用荧光定量PCR方法和末端限制性片段长度多态性(Terminal restriction fragment length polymorphism,T-RFLP)技术,对异育银鲫养殖环境水体和沉积物中的典型病原微生物进行定量检测,以及细菌群落多样性分析。【结果】养殖过程中病原菌丰度呈现不同的变化趋势,嗜水气单胞菌(Aeromonas hydrophila)、温和气单胞菌(A. sobria)和维氏气单胞菌(A. veronii)与鲤疱疹病毒Ⅱ型丰度变化呈显著正相关。水体中病原菌丰度受pH、温度和溶解氧等环境因子的影响显著。T-RFLP分析表明沉积物样品较水体样品细菌群落组成复杂,并且沉积物细菌群落动态变化幅度高于水体。水体和沉积物中细菌T-RFs数量范围分别在11-29和20-32之间,Shannon-wiener指数在1.44-2.87和2.44-3.25之间。【结论】养殖水体中的病原菌丰度高于沉积物,而沉积物细菌群落丰富度和多样性高于水体。病原菌丰度和细菌群落结构变化与环境因子密切相关,明确养殖池塘环境中病原微生物的生态分布和丰度变化,将为指导养殖过程中病原性疾病发生早期预警提供理论依据。     

东太平洋多金属结核区两个站位深海沉积物细菌多样性北大核心CSCD

【目的】为了解东太平洋中国多金属结核勘探合同区西区2个站位(WBC1305和WBC1316A)深海沉积物细菌群多样性。【方法】直接提取环境样品总基因组,通过PCR和TA克隆策略构建了2个站位6个层次16S r RNA基因文库,对2个站位沉积物表层泥样中细菌多样性和群落结构特征进行分析,并通过构建系统发育树,进行系统发育学分析。【结果】2个站位6个文库共获得有效克隆533个,其中472个克隆包括α-变形菌纲、β-变形菌纲、γ-变形菌纲、δ-变形菌纲、浮霉菌门、酸杆菌门、硝化螺旋菌门、放线菌门、绿弯菌门、厚壁菌门、拟杆菌门、迷踪菌门、芽单胞菌门、Hydrogenedentes、Chlorobi和Nitrospinae16个细菌类群,而另外61个克隆为不可分类细菌类群。【结论】结果表明γ-变形菌纲和厚壁菌门分别是WBC1305和WBC1316A站位的优势种群;WBC1316A站位细菌群落结构更加丰富和复杂。     

基于高通量测序技术的不同年代公园绿地土壤细菌多样性

【背景】细菌多样性对绿地土壤生态功能有重要作用,但不同年代公园绿地土壤的细菌多样性尚未见相关报道。【目的】研究北京市不同年代公园绿地土壤细菌多样性和群落结构特征。【方法】利用IlluminaMiSeq测序技术,分别对北京市代表性古典公园和现代公园绿地土壤细菌群落多样性进行分析。【结果】北京市公园绿地土壤细菌群落共划分为45个已知的菌门,其中变形菌门、酸杆菌门、绿弯菌门和放线菌门为优势细菌群。土壤细菌群落α多样性分析结果表明,古典公园和现代公园的土壤细菌多样性存在差异,表现为古典公园的丰富度和多样性都高于现代公园。此外,土壤细菌群落相似性分析和主坐标分析都表明古典公园和现代公园的土壤细菌群落结构存在显著差异。冗余分析表明,对土壤微生物群落结构产生显著影响的环境因子分别为土壤含水量、有机质和全氮,其它土壤环境因子无统计学意义。首次引入公园年代作为影响因子进行冗余分析的研究结果表明,公园年代为影响公园细菌群落多样性的重要因子。【结论】不同年代公园绿地土壤细菌群落结构和物种多样性具有显著差异,随着公园年代的增加,土壤肥力和微生物多样性增加,绿地生态系统更稳定,可通过制定不同的绿地管理措施改变公园绿地土壤环境,进而优化土壤细菌群落结构,促进土壤碳氮养分循环,提高土壤肥力。     

海水混养池塘虾蛤肠道与养殖环境的微生物多样性

【背景】海水混养池塘环境微生物以及动物肠道微生物的群落结构已有研究,但对混养环境中多品种动物肠道与环境微生物群落的关系尚未见报道。【目的】研究海水虾蛤混养环境中微生物多样性以及与养殖动物健康之间的关系。【方法】采用Illumina高通量测序技术测定冬季莆田市北江养殖区2个混养池塘中水体、底泥以及虾蛤肠道的菌群结构。【结果】同一池塘水体与底泥之间、不同池塘水体或底泥之间的微生物结构存在一定的差异;同一养殖区2个混养池塘虾与蛤肠道微生物结构之间具有极高的相似性,与养殖环境存在显著的差异。微生物多样性和丰富度差异很大,表现出底泥水体肠道;虾蛤肠道微生物以厚壁细菌和γ-变形细菌为主;池塘水体以放线菌、α-变形细菌以及拟杆菌为主,底泥以γ-变形细菌和δ-变形细菌为主。养殖动物肠道微生物主要优势种为乳球菌属和假单胞菌属,池塘环境内存在较高丰度的黄杆菌类潜在致病菌,而在虾和蛤的肠道中基本未检出。2个池塘底泥硫还原细菌含量较高,增加了底质产生硫化氢等有害物质的风险。【结论】比较混养池塘中水体、底泥以及虾蛤肠道三者之间微生物群落结构的差异,揭示虾、贝混养模式微生物与养殖环境的关系,为池塘养殖虾、贝疾病防治和混养结构的优化提供参考。     

新疆低阶煤本源生物甲烷转化中微生物群落组成及变化

【目的】探究新疆低阶煤生物甲烷转化过程微生物群落组成及多样性。【方法】采用厌氧培养方法和末端限制性片段长度多态性技术(Terminal restriction fragment length polymorphism,T-RFLP)分析新疆低阶煤本源微生物对甲烷转化及有机酸含量的影响,分析新疆哈密大南湖长焰煤生物甲烷转化过程中微生物群落动态变化。【结果】研究表明长焰煤和褐煤对本源微生物产甲烷影响较小,随着低阶煤生物甲烷转化时间的延长,甲烷产量呈上升趋势,转化60 d后长焰煤甲烷产量高达10.28 m L/g,挥发性有机酸(VFA)浓度则最低;微生物多样性指数变化不明显,不同转化时间微生物主要类群为放线菌门(Actinobacteria),拟杆菌门(Bacteroidetes),厚壁菌门(Firmicutes),变形菌门(Proteobacteria);甲烷菌的群落结构相对于细菌较简单,在整个低阶煤生物转化产甲烷过程中共有古菌类群为甲烷八叠球菌属(Methanosarcina)、甲烷盐菌属(Methanohalobium)、甲烷叶菌属(Methanolobus)、甲烷食甲基菌属(Methanomethylovorans),它们是构成群落结构的基本菌群。【结论】低阶煤生物甲烷转化过程微生物群落具有丰富的多样性,且不同时期多样性有较大差异。甲烷菌群落结构相对于细菌较简单,共有类群明显。     

模拟增温对青海湖鸟岛土壤微生物的影响

【背景】土壤微生物对其生存的微环境变化极为敏感,鸟岛作为湖滨湿地,对气候变化具有敏感性,但目前关于青海湖鸟岛的土壤微生物鲜有研究。【目的】探究气候变暖后青海湖鸟岛土壤微生物群落特征的变化。【方法】利用开顶箱模拟增温,通过高通量测序方法了解温度升高后土壤细菌及真菌的群落结构以及多样性的变化情况。【结果】温度的升高并未改变青海湖鸟岛土壤微生物的优势菌群,细菌优势菌群为变形菌门和酸杆菌门;真菌优势菌门为子囊菌门,优势菌纲为座囊菌纲。但增温改变了土壤微生物的群落结构,显著升高了拟杆菌门、蓝细菌门、Patescibacteria及球囊菌纲的相对丰度,显著降低了锤舌菌纲的相对丰度。土壤微生物群落的多样性指数也发生了变化,温度上升后微生物的ACE指数及Chao1指数均降低,细菌的Simpson指数及真菌的Shannon指数降低。【结论】青海湖鸟岛土壤微生物对温度升高的响应明显,增温改变了土壤细菌拟杆菌门、蓝细菌门、Patescibacteria的相对丰度及真菌的球囊菌纲、锤舌菌纲的相对丰度,降低了土壤微生物的多样性。     

青海湖岸带土壤与沉积物的地化特征与细菌群落对水位上升的响应

【目的】探究青海湖岸带土壤与沉积物的地化特征与细菌群落对水位扩张的响应。【方法】从岸上至岸下沿垂直青海湖岸带方向,采集距离湖面不同高度土壤(土壤：S1、S2)、岸边不同水深表层沉积物(过渡区：E0、E6、E17)及湖心表层沉积物(沉积物：D1、D2)样品,土壤与沉积物水深(土壤水深表示为负数)从小到大的变化表征岸边土壤被淹水转变为沉积物的过程。采用地球化学分析和16SrRNA基因高通量测序技术,探究岸带土壤与沉积物样品中的地化特征与微生物群落构成。【结果】青海湖水位上升导致的生境转变对岸带土壤与沉积物的理化性质、营养水平、有机碳类型等地化特征产生显著影响。具体表现为,随着水位升高,岸带土壤与沉积物的pH、矿物结合态有机碳含量显著升高,而碳氮比值、可溶性有机碳(dissolved organic carbon,DOC)、颗粒态有机碳含量显著下降。随着水位上升,青海湖岸带被淹没土壤的细菌群落多样性下降,且群落结构发生明显变化。这种变化与环境因子变化密切相关,具体表现为,细菌群落物种丰富度指数和香农多样性指数随着水位上升呈下降趋势;活性金属结合态有机碳含量与细菌群落多样性的变化密切相关;理化...     

Evidence for indigenous Streptomyces populations in a marine environment determined with a 16S rRNA probe.

A 16S rRNA genus-specific probe was used to determine whether Streptomyces populations are an indigenous component of marine sediment bacterial communities. Previous debates have suggested that marine Streptomyces isolates are derived not from resident populations but from spores of terrestrial species which have been physically transported to marine ecosystems but remain dormant until isolation. Rigorously controlled hybridization of rRNA extracted from coastal marsh sediments with the genus-specific probe indicated that Streptomyces rRNA accounted for 2 to 5% of the sediment community rRNA and that spores are not the source of the hybridization signal. Streptomyces populations must therefore be at least the 26th most abundant genus-level source of bacterial rRNA. the relative amounts of rRNAs from Streptomyces spp. and members of the Bacteria (69 to 79%) and Archaea (4 to 7%) domains were highly consistent in these marine sediments throughout an annual cycle, indicating that the species composition of sediment bacterial communities may be more stable than recent studies suggest for marine planktonic bacterial communities. Laboratory studies designed to investigate the possible functional roles of Streptomyces populations in coastal sediments demonstrated that population levels of this genus changed relatively rapidly (within a time frame of 6 weeks) in response to manipulation of substrate availability. Amendments of intact sediment cores with two compounds (vanillic acid and succinic acid) consistently resulted in Streptomyces populations contributing an increased percentage of rRNA (6 to 15%) to the total bacterial rRNA pool.     

Spatial distribution patterns of benthic microbial communities along the Pearl Estuary,China

In the present study, benthic microbial communities along the Pearl Estuary, a typical subtropical estuary in China subjected to extensive anthropogenic disturbance, were investigated using 16S rRNA gene-based pyrosequencing. The results showed that microbial communities in freshwater samples were clearly distinct from those in saltwater samples, since the relative sequence abundances of Deltaproteobacteria, Thermoplasmata and Marine Group I (MG-I) were higher in saltwater sediments, whereas Chloroflexi, Spirochaetes, Betaproteobacteria and methanogens were more prevalent in freshwater sediments. In addition, bacterial communities showed vertical stratifications in saltwater sediments, but remained constant with depth in freshwater sediments. The total organic carbon and carbon/nitrogen ratio in sediments correlated significantly with the overall community variations. The predominance of various microorganisms in specific niches led to efforts to identify their functional couplings by exploring their co-occurrence patterns. Using network analysis, strong positive correlations were observed between sulfate-reducing bacteria (SRB) and sulfur-oxidizing bacteria, and between SRB and nitrite-oxidizing bacteria, indicating the potential interactions of intra-sulfur cycle processes, as well as sulfur and nitrogen cycles, in coastal sediments. Archaeal clades revealed strong and wide correlations between the Miscellaneous Crenarchaeotal Group (MCG) and other groups, suggesting a central role of MCG in the coastal benthic environment. Inversely, MG-I displayed negative correlations with other clades, which might indicate that the lifestyles of heterotrophic and autotrophic clades were mutually exclusive. This study presented a detailed outline of the biogeographic patterns of benthic microbial communities along the Pearl Estuary and provided new information regarding the potential interactions of various biogeochemical cycles in coastal sediments.     

Effects of Spilled Oil on Bacterial Communities of Mediterranean Coastal Anoxic Sediments Chronically Subjected to Oil Hydrocarbon Contamination

The effects of spilled oil on sedimentary bacterial communities were examined in situ at 20 m water depth in a Mediterranean coastal area. Sediment collected at an experimental site chronically subjected to hydrocarbon inputs was reworked into PVC cores with or without a massive addition of crude Arabian light oil (∼20 g kg⁻¹ dry weight). Cores were reinserted into the sediment and incubated in situ at the sampling site (20 m water depth) for 135 and 503 days. The massive oil contamination induced significant shifts in the structure of the indigenous bacterial communities as shown by ribosomal intergenic spacer analysis (RISA). The vertical heterogeneity of the bacterial communities within the sediment was more pronounced in the oiled sediments particularly after 503 days of incubation. Response to oil of the deeper depth communities (8–10 cm) was slower than that of superficial depth communities (0–1 and 2–4 cm). Analysis of the oil composition by gas chromatography revealed a typical microbial alteration of n-alkanes during the experiment. Predominant RISA bands in oiled sediments were affiliated to hydrocarbonoclastic bacteria sequences. In particular, a 395-bp RISA band, which was the dominant band in all the oiled sediments for both incubation times, was closely related to hydrocarbonoclastic sulfate-reducing bacteria (SRB). These bacteria may have contributed to the main fingerprint changes and to the observed biodegradation of n-alkanes. This study provides useful information on bacterial dynamics in anoxic contaminated infralittoral sediments and highlights the need to assess more precisely the contribution of SRB to bioremediation in oil anoxic contaminated areas.     

Composition and variation of sediment bacterial and nirS-harboring bacterial communities at representative sites of the Bohai Gulf coastal zone,China

With rapid urbanization, anthropogenic activities are increasingly influencing the natural environment of the Bohai Bay. In this study, the composition and variation of bacterial and nirS-harboring bacterial communities in the coastal zone sediments of the Bohai Gulf were analyzed using PCR-based clone libraries. A total of 95 genera were detected in the bacterial communities, with Proteobacteria (72.1 %), Acidobacteria (10.5 %), Firmicutes (1.7 %), Bacteroidetes (1.4 %), Chloroflexi (0.7 %) and Planctomycetes (0.7 %) being the dominated phyla. The NirS sequences were divided into nine Clusters (A–I). Canonical correlation analysis showed that the bacterial or denitrifying communities were correlated with different environmental factors, such as total organic carbon, total nitrogen, ammonium, sulfate, etc. Furthermore, bacterial communities’ composition and diversity are influenced by oil exploration, sewage discharge and other anthropogenic activities in the coastal area of the Bohai Sea. Thus, this study provided useful information on further research on regional or global environmental control and restore.     

Contrasting distributions of bacteriophages and eukaryotic viruses from contaminated coastal sediments

Viruses are ubiquitous, abundant and play an important role in all ecosystems. Here, we advance understanding of coastal sediment viruses by exploring links in the composition and abundance of sediment viromes to environmental stressors and sediment bacterial communities. We collected sediment from contaminated and reference sites in Sydney Harbour and used metagenomics to analyse viral community composition. The proportion of phages at contaminated sites was significantly greater than phages at reference sites, whereas eukaryotic viruses were relatively more abundant at reference sites. We observed shifts in viral and bacterial composition between contaminated and reference sites of a similar magnitude. Models based on sediment characteristics revealed that total organic carbon in the sediments explained most of the environmental stress-related variation in the viral dataset. Our results suggest that the presence of anthropogenic contaminants in coastal sediments could be influencing viral community composition with potential consequences for associated hosts and the environment.