大兴安岭典型永久冻土土壤细菌群落组成和多样性

【背景】土壤微生物是土壤生物中的重要成分,参与了土壤生态系统中关键的生物化学循环过程。但是关于寒温带多年冻土土壤微生物的研究还比较薄弱。【目的】探究大兴安岭多年冻土土壤中微生物的多样性和种群结构。【方法】利用MiSeq高通量测序技术对黑龙江大兴安岭地区呼中保护区落叶松冻土和樟子松林冻土土壤样品进行测序。【结果】在落叶松冻土和樟子松林冻土土壤中,相对丰度最高的优势菌群的组成基本一致,在门水平有疣微菌门(Verrucomicrobia)、变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、浮霉菌门(Planctomycetes)、绿菌门(Chlorobi)、Parcubacteria、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、芽单胞菌门(Gemmatimonadetes)10个细菌门类,其中,疣微菌门(Verrucomicrobia)在樟子松林土壤中的相对丰度较多,变形菌门(Proteobacteria)在落叶松林土壤中的相对丰度较多。通过α多样性分析可知,落叶松冻土土壤微生物的群落多样性高于樟子松林冻土,而且两者的细菌群落组成与结构差异性较大。【结论】为深入认识大兴安岭多年冻土区的土壤微生物群落结构组成以及影响因素提供数据支撑。     

Activity and composition of the denitrifying bacterial community respond differently to long-term fertilization

The objective of this study was to explore the long-term effects of different organic and inorganic fertilizers on activity and composition of the denitrifying and total bacterial communities in arable soil. Soil from the following six treatments was analyzed in an experimental field site established in 1956: cattle manure, sewage sludge, Ca(NO3)2, (NH4)2SO4, and unfertilized and unfertilized bare fallow. All plots but the fallow were planted with corn. The activity was measured in terms of potential denitrification rate and basal soil respiration. The nosZ and narG genes were used as functional markers of the denitrifying community, and the composition was analyzed using denaturing gradient gel electrophoresis of nosZ and restriction fragment length polymorphism of narG, together with cloning and sequencing. A fingerprint of the total bacterial community was assessed by ribosomal intergenic spacer region analysis (RISA). The potential denitrification rates were higher in plots treated with organic fertilizer than in those with only mineral fertilizer. The basal soil respiration rates were positively correlated to soil carbon content, and the highest rates were found in the plots with the addition of sewage sludge. Fingerprints of the nosZ and narG genes, as well as the RISA, showed significant differences in the corresponding communities in the plots treated with (NH4)2SO4 and sewage sludge, which exhibited the lowest pH. In contrast, similar patterns were observed among the other four treatments, unfertilized plots with and without crops and the plots treated with Ca(NO3)2 or with manure. This study shows that the addition of different fertilizers affects both the activity and the composition of the denitrifying communities in arable soil on a long-term basis. However, the treatments in which the denitrifying and bacterial community composition differed the most did not correspond to treatments with the most different activities, showing that potential activity was uncoupled to community composition.     

长期施肥制度对稻田土壤反硝化细菌群落活性和结构的影响

以中国科学院桃源农业生态试验站长期定位施肥试验为平台,研究了3种长期施肥制度(对照不施肥-CK,化学施肥-NPK,化学施肥+有机肥-NPKOM)下土壤反硝化速率的差异.同时,以硝酸还原酶基因(narG)作为反硝化细菌的功能标志物,分析了施肥对反硝化细菌群落结构和多样性的影响.结果表明,长期施用有机肥的土壤反硝化速率,反硝化菌多样性都高于对照和施用化肥处理.从3个处理的土壤样品中共获得35个narG基因的可操作分类单元(OTU)主要分布在两个簇,与变形菌门(Proteobacteria)和放线菌门(Actinobacteria)的反硝化细菌有一定的亲缘关系,均为首次从土壤中克隆.Shannon多样性指数显示,NPKOM处理的narG基因多样性最高,CK处理次之,NPK处理最低.LUBSHUFF软件对narG基因群落组成的分析显示,施有机肥后含narG基因的细菌群落组成与CK之间有显著性差异(P<0.05),而化肥(NPK)没有产生显著影响.实验结果为进一步研究亚热带地区水稻土反硝化作用及反硝化功能菌提供了重要的依据.     

喀斯特原生土壤与退化生态系统土壤细菌群落结构

运用PCR-RFLP技术,对桂西北喀斯特原生土壤和退化生态系统土壤细菌16S rDNA基因多样性及系统发育关系进行了研究.结果表明：原生土壤比退化生态系统土壤具有更丰富的16S rDNA基因型和更高的多样性指数,两样地共有的基因型仅有2个.从每种基因型中随机选择一个克隆子作为代表进行测序分析,所有序列与GenBank 数据库中序列的同源性为87%～100%,且两样地中均有超过一半的基因型序列与数据库中已知序列同源性低于97%,属于分类在“种”地位上的新发现细菌;通过系统发育研究将两样地的细菌分为10大类群,两样地共同拥有5大类群,但两样地的细菌优势类群明显不同,原生土壤为Proteobacteria,含39种基因型,占总克隆子数的58.0%,退化生态系统土壤为Acidobacteria和Proteobacteria,分别含19种和15种基因型,占总克隆子数的32.5%和30.5%;与原生土壤细菌类群相比,退化生态系统土壤Proteobacteria类群明显减少,Acidobacteria类群明显增加.土壤理化性质及土壤环境因素的差异是引起两类型土壤细菌多样性差异的原因.     

若尔盖高原湿地不同退化阶段的土壤细菌群落多样性

采用传统分离培养和基于16S rDNA的变性梯度凝胶电泳(Denaturing gradient gel electrophoresis, DGGE)的方法, 研究了若尔盖高原湿地不同退化阶段土壤类型(泥炭土、沼泽土、草甸土、风沙土)和土壤深度(0 cm?20 cm、20 cm?40 cm、40 cm?60 cm)中微生物的数量、细菌群落结构及其多样性。结果表明: 微生物数量总体上不仅随着土壤类型的改变而减少(泥炭土>草甸土>沼泽土>风沙土)也随着土壤深度的增加而递减(0 cm?20 cm>20 cm?4     

祁连山中部4种典型植被类型土壤细菌群落结构差异

土壤微生物参与土壤生态过程,在土壤生态系统的结构和功能中发挥着重要作用。2013年7月采集了祁连山中段4种典型植被群落(垫状植被、高寒草甸、沼泽草甸和高寒灌丛)的表层土壤,分析了表层土壤微生物生物量碳氮和采用Illumina高通量测序技术研究了土壤细菌群落结构及多样性,并结合土壤因子对土壤细菌群落结构和多样性进行了相关性分析。结果表明:(1)土壤微生物生物量碳氮的大小排序为:沼泽草甸高寒草甸高寒灌丛垫状植被;(2)土壤细菌群落相对丰度在5%以上的优势类群是放线菌门、酸杆菌门、α-变形菌、厚壁菌门和芽单胞菌门5大门类;(3)沼泽草甸土壤细菌α多样性(物种丰富度和系统发育多样性)显著高于其它3种植被类型(P0.05),而垫状植被土壤细菌α多样性最低;(4)冗余分析和Pearson相关性分析表明,土壤pH、土壤含水量、土壤有机碳和总氮是土壤细菌群落结构和α多样性的主要影响因子。研究结果可为祁连山高寒生态系统稳定和保护提供理论依据。     

三江平原沼泽湿地垦殖及自然恢复对土壤细菌群落多样性的影响

为量化垦殖对我国东北沼泽湿地的影响程度,于2015年6月基于Illumina Miseq PE300第二代高通量测序平台,对黑龙江省洪河国家级自然保护区的原始沼泽、耕地、退耕湿地的土壤细菌16S r RNA基因高变区域进行测序,并分析沼泽湿地土壤细菌群落多样性和结构组成与垦殖的关系。结果表明:共获得358737条修剪序列且被划分为36个已知的菌门,在11个主要的土壤细菌门类中(相对丰度1%),芽单胞菌门(P0.01)、拟杆菌门(P0.01)、厚壁菌门(P0.01)和绿菌门(P0.01)在不同生境类型样地差异显著,土壤细菌群落alpha多样性由高到低排序依次是:耕地、退耕湿地、原始沼泽。结合相关性分析和冗余分析可以证明,长期的耕作,特别是旱田耕作对沼泽湿地土壤细菌群落组成产生显著的影响,土壤p H、含水量、全碳、有机碳、可溶有机碳、碱解氮、微生物量碳、微生物量氮对土壤细菌的多样性和群落组成产生影响。总之,研究结果发现自然恢复可以显著促进土壤细菌群落多样性的恢复,但是沼泽湿地土壤细菌群落结构一旦被垦殖干扰改变将很难恢复到原始状态,强调了有效地利用土壤细菌群落对维护土壤生态系统平衡具有重要的意义。     

Bacterial diversity and community composition from seasurface to subseafloor

We investigated compositional relationships between bacterial communities in the water column and those in deep-sea sediment at three environmentally distinct Pacific sites (two in the Equatorial Pacific and one in the North Pacific Gyre). Through pyrosequencing of the v4–v6 hypervariable regions of the 16S ribosomal RNA gene, we characterized 450 104 pyrotags representing 29 814 operational taxonomic units (OTUs, 97% similarity). Hierarchical clustering and non-metric multidimensional scaling partition the samples into four broad groups, regardless of geographic location: a photic-zone community, a subphotic community, a shallow sedimentary community and a subseafloor sedimentary community (⩾1.5 meters below seafloor). Abundance-weighted community compositions of water-column samples exhibit a similar trend with depth at all sites, with successive epipelagic, mesopelagic, bathypelagic and abyssopelagic communities. Taxonomic richness is generally highest in the water-column O₂ minimum zone and lowest in the subseafloor sediment. OTUs represented by abundant tags in the subseafloor sediment are often present but represented by few tags in the water column, and represented by moderately abundant tags in the shallow sediment. In contrast, OTUs represented by abundant tags in the water are generally absent from the subseafloor sediment. These results are consistent with (i) dispersal of marine sedimentary bacteria via the ocean, and (ii) selection of the subseafloor sedimentary community from within the community present in shallow sediment.     

生姜根系不同生态位细菌群落多样性特征、组成及结构差异

生姜作为常见的调味品和传统中药材,是我国重要的经济作物之一。作为取食部分的生姜块茎与根系直接相连,其产量、品质与根相关细菌群落密切相关。然而,关于生姜根系微环境中细菌群落的特点仍鲜有报道,土壤环境能否衍生出宿主特异性内生菌群落尚不清楚。以生姜根系不同生态位细菌群落为研究对象,采用高通量测序技术,对非根际、根际及根内细菌进行16S rRNA基因测序。结果表明,不同生态位细菌群落多样性存在显著差异,其中非根际及根际细菌群落多样性(Shannon index, Observed species, Faith′s PD)显著高于内生菌群落。同时,各生态位共现网络稳定性和复杂度表现为非根际>根际>根内细菌群落。而在组成上,细菌群落在不同生态位差异显著(R²=0.57,P=0.001)。其中变形菌门(Proteobacteria)是根内的优势门,该门类下假单胞菌属(Pseudomonas)、短波单胞菌属(Brevundimonas)、寡养单胞菌属(Stenotrophomonas)及泛菌属(Pantoea)在根内显著富集。在根际细菌中,拟杆菌门(Bacteroid...     

毛竹种植对土壤细菌和真菌群落结构及多样性的影响

为揭示天然林改为毛竹林过程中土壤微生物变化规律,在浙江省湖州市安吉县和长兴县两地选择不同种植历史的粗放经营毛竹林,分层采集0～20和20～40 cm的混合土壤样品,应用PCR-DGGE技术分析土壤细菌和真菌群落结构及多样性变化.结果表明: 在马尾松林改种毛竹林或毛竹林入侵杂灌阔叶林形成毛竹纯林过程中,土壤细菌和真菌的群落结构均发生明显变化,且细菌结构对毛竹种植的响应更敏感;随着毛竹生长时间的延长,表层土壤细菌群落表现出抵抗干扰、最后向改种毛竹之前状态恢复的趋势.毛竹种植时间、样地和土层均对土壤细菌和真菌多样性产生显著影响,其中样地和土层的影响明显大于种植时间.土壤性质和细菌、真菌结构的冗余分析结果表明,不同地点、不同土层驱动土壤微生物结构随时间变化的主要因子没有一致规律,且第1、2轴对样地变化的解释率大多低于65.0%,说明除本研究分析的5个土壤化学指标外,可能还有其他土壤理化性质共同驱动微生物结构的变化.     

Light availability affects stream biofilm bacterial community composition and function,but not diversity

Changes in riparian vegetation or water turbidity and browning in streams alter the local light regime with potential implications for stream biofilms and ecosystem functioning. We experimented with biofilms in microcosms grown under a gradient of light intensities (range: 5–152 μmole photons s^?1 m^?2) and combined 454‐pyrosequencing and enzymatic activity assays to evaluate the effects of light on biofilm structure and function. We observed a shift in bacterial community composition along the light gradient, whereas there was no apparent change in alpha diversity. Multifunctionality, based on extracellular enzymes, was highest under high light conditions and decoupled from bacterial diversity. Phenol oxidase activity, involved in the degradation of polyphenolic compounds, was twice as high on average under the lowest compared with the highest light condition. This suggests a shift in reliance of microbial heterotrophs on biofilm phototroph‐derived organic matter under high light availability to more complex organic matter under low light. Furthermore, extracellular enzyme activities correlated with nutrient cycling and community respiration, supporting the link between biofilm structure–function and biogeochemical fluxes in streams. Our findings demonstrate that changes in light availability are likely to have significant impacts on biofilm structure and function, potentially affecting stream ecosystem processes.     

Symbiont community diversity is more variable in corals that respond poorly to stress

Coral reefs are declining globally as climate change and local water quality press environmental conditions beyond the physiological tolerances of holobionts—the collective of the host and its microbial symbionts. To assess the relationship between symbiont composition and holobiont stress tolerance, community diversity metrics were quantified for dinoflagellate endosymbionts (Family: Symbiodiniaceae) from eight Acropora millepora genets that thrived under or responded poorly to various stressors. These eight selected genets represent the upper and lower tails of the response distribution of 40 coral genets that were exposed to four stress treatments (and control conditions) in a 10‐day experiment. Specifically, four ‘best performer’ coral genets were analyzed at the end of the experiment because they survived high temperature, high pCO₂, bacterial exposure, or combined stressors, whereas four ‘worst performer’ genets were characterized because they experienced substantial mortality under these stressors. At the end of the experiment, seven of eight coral genets mainly hosted Cladocopium symbionts, whereas the eighth genet was dominated by both Cladocopium and Durusdinium symbionts. Symbiodiniaceae alpha and beta diversity were higher in worst performing genets than in best performing genets. Symbiont communities in worst performers also differed more after stress exposure relative to their controls (based on normalized proportional differences in beta diversity), than did best performers. A generalized joint attribute model estimated the influence of host genet and treatment on Symbiodiniaceae community composition and identified strong associations among particular symbionts and host genet performance, as well as weaker associations with treatment. Although dominant symbiont physiology and function contribute to host performance, these findings emphasize the importance of symbiont community diversity and stochasticity as components of host performance. Our findings also suggest that symbiont community diversity metrics may function as indicators of resilience and have potential applications in diverse disciplines from climate change adaptation to agriculture and medicine.     

Seasonal variability in bacterial and fungal diversity and community composition of the near-surface atmosphere in coastal megacity

Bacteria and fungi are ubiquitous in the near-surface atmosphere where they may impact on the surrounding environment and human health, especially in coastal megacities. However, the diversity, composition, and seasonal variations of these airborne microbes remain limited. This study investigated the airborne microbes of the near-surface atmosphere in coastal megacity Qingdao over one year. It was found that the sample in summer displayed the highest bacterial and fungal diversity, while sample in winter exhibited the lowest bacterial and fungal diversity. Proteobacteria was the dominating bacteria, and Dothideomycetes was the most dominating fungi in the near-surface atmosphere, which took up 53–76 and 49–78% relative abundance, respectively. However, the bacterial diversity and community composition had significant seasonal variations. These data suggest that a complex set of environmental factors, including landscaping ratio, solar radiation temperature, and marine microorganisms, can affect the composition of airborne microbes in the near-surface atmosphere in coastal megacity. The analysis of the pathogenic microorganisms or opportunistic pathogenic microorganisms existed in the near-surface atmosphere revealed that the relative abundance of pathogenic microorganisms in autumn was the highest. The main pathogenic microorganisms or opportunistic pathogenic microorganisms were Acinetobacter baumannii (accounting for up to 9.93% relative abundance), Staphylococcus epidermidis (accounting for up to 11.26% relative abundance), Mycobacterium smegmatis (accounting for up to 3.68% relative abundance), Xanthomonas oryzae pv. oryzae (accounting for up to 5.36% relative abundance), which may be related to certain human or plant diseases in specific environments or at certain seasons. Therefore, the investigation of airborne microbial communities of near-surface atmosphere in coastal megacities is very important to both the understanding of airborne microbes and public health.     

施用化肥对黑土地区线虫群落组成及多样性产生的影响

本文针对玉米生长期内施用化肥对东北黑土地区线虫群落组成及多样性产生的影响进行了研究,共获得14科,16属,其中头叶科（Cephalobidae),小杆科（Rhobditidae) ,真滑刃属（Aphelenchus),垫刃属（Tylenchus)为优势科／优势度指数分析比较了施肥和不施肥试验小区土壤线虫群落多样性分布规律,其中Shannon多样性指数和丰富度指数是评价线虫群薄对化肥投入最敏感的指数。     

炭基肥对植烟黄壤细菌、真菌群落结构和多样性的影响

【目的】探究施用炭基肥对植烟黄壤质量影响的微生物学机制,为应用炭基肥培育植烟黄壤肥力提供科学依据。【方法】2016年,以烤烟品种云烟87和贵州黄壤为供试材料,通过大田试验,设置不施肥(NF)、常规肥(CF,条施有机肥和烤烟专用基肥,穴施烤烟专用追肥)和炭基肥(BF,条施炭基有机肥和炭基复混肥,穴施烤烟专用追肥) 3个处理,2年后采集样品,采用Illumina Miseq高通量测序技术,剖析土壤细菌、真菌群落结构和多样性的响应差异,揭示影响微生物的主要土壤因子。【结果】与NF处理和CF处理比较,BF处理烤烟产量提高,土壤碱解氮和有效磷含量显著提高。BF处理土壤细菌OTUs数量最大(1592),显著大于其余两处理;土壤细菌丰富度和多样性指数最高。BF处理土壤真菌OTUs数量最小(280);土壤真菌丰富度和多样性指数最低。BF处理土壤细菌群落物种最多样,属于25个门,CF处理属于23个门,NF处理仅属于22个门。细菌门水平,BF处理拟杆菌门(Bacteroidetes)相对丰度(1.50%)提高,Latescibacteria相对丰度(0.11%)降低;与NF处理比较,拟杆菌门、Latesc...     

矿区高硫煤覆盖对土壤细菌群落组成和多样性的影响

【背景】土壤微生物生态研究对于矿区污染生态影响调查和矿区生态恢复都具有非常重要的意义,高硫煤覆盖对其下方土壤微生物群落的影响目前尚不清楚。【目的】探究煤矿区长期高硫煤覆盖对其下方土壤细菌群落组成和多样性产生的影响。【方法】从高硫煤矿附近煤场采集3种类型土壤样品(高硫煤覆盖层、高硫煤下方土壤、对照土壤),通过测定土壤理化性质和运用高通量测序技术研究高硫煤覆盖对土壤性质和细菌群落产生的影响。【结果】与对照土壤相比,高硫煤覆盖使其下方土壤pH值降低,硫酸盐、有机质、有效氮、有效磷含量升高。高硫煤覆盖使其下方土壤中细菌的多样性指数降低,细菌群落组成发生了较大改变。在属水平上,与对照土壤相比,高硫煤下方土壤中Bacillus的相对丰度较高;Acidiphilium与Sulfobacillus在3个样本组中的相对丰度呈现为高硫煤覆盖层>高硫煤下方土壤>对照土壤,硫氧化菌在高硫煤中的优势地位及其对下方土壤的影响由此可以得到证实。共现性网络分析表明,Acidiphilium和Sulfobacillus在高硫煤覆盖区域的土壤细菌群落中占有重要地位,对其他细菌类群影响非常大。【结论】高硫煤覆盖及硫氧化菌的优势存在,对高硫煤下方土壤的理化性质及细菌群落产生明显影响。研究结果有助于增进对矿区土壤微生物生态的认识,为高硫煤矿区生态恢复治理提供微生物学理论基础。     

Spatial scales of bacterial diversity in cold-water coral reef ecosystems

Background

Cold-water coral reef ecosystems are recognized as biodiversity hotspots in the deep sea, but insights into their associated bacterial communities are still limited. Deciphering principle patterns of bacterial community variation over multiple spatial scales may however prove critical for a better understanding of factors contributing to cold-water coral reef stability and functioning.

Methodology/Principal Findings

Bacterial community structure, as determined by Automated Ribosomal Intergenic Spacer Analysis (ARISA), was investigated with respect to (i) microbial habitat type and (ii) coral species and color, as well as the three spatial components (iii) geomorphologic reef zoning, (iv) reef boundary, and (v) reef location. Communities revealed fundamental differences between coral-generated (branch surface, mucus) and ambient microbial habitats (seawater, sediments). This habitat specificity appeared pivotal for determining bacterial community shifts over all other study levels investigated. Coral-derived surfaces showed species-specific patterns, differing significantly between Lophelia pertusa and Madrepora oculata, but not between L. pertusa color types. Within the reef center, no community distinction corresponded to geomorphologic reef zoning for both coral-generated and ambient microbial habitats. Beyond the reef center, however, bacterial communities varied considerably from local to regional scales, with marked shifts toward the reef periphery as well as between different in- and offshore reef sites, suggesting significant biogeographic imprinting but weak microbe-host specificity.

Conclusions/Significance

This study presents the first multi-scale survey of bacterial diversity in cold-water coral reefs, spanning a total of five observational levels including three spatial scales. It demonstrates that bacterial communities in cold-water coral reefs are structured by multiple factors acting at different spatial scales, which has fundamental implications for the monitoring of microbial diversity and function in those ecosystems.