普洱地区茶叶内生细菌与根际土壤细菌群落结构分析

[目的]茶叶内生细菌、根际土壤细菌在普洱茶的发酵中起着重要的作用,还可以促进茶树生长,诱导茶树抗病性.研究其群落结构组成及相互关系可为微生物资源开发利用提供理论依据.[方法]本研究以普洱地区茶树叶片和根际土壤为材料,采用高通量测序技术,对茶叶及根际土壤细菌的16S核糖体RNA基因(16S rRNA)进行测序,比较分析茶...     

宁夏不同地区盐碱化土壤细菌群落多样性分布特征及其影响因子

为了解宁夏不同地区盐碱化土壤细菌群落多样性的分布特征及其关键影响因子,选择贺兰县红星村、惠农县黄渠拐子、燕子墩、庙台、平罗县银星村、分水闸、侯家梁、西大滩为研究样点,采集宁夏地区8个典型盐碱化土壤样品,0—2 cm和2—25 cm土层,共48份。利用Illumina Hiseq高通量测序技术解析不同采样点土壤细菌群落结构的特征,同时分析了土壤理化因子与细菌群落结构的关系。试验结果表明:变形菌门、拟杆菌门和放线菌门为8个采样点土壤细菌群落的优势种类。其中,变形菌门占总量的24.69%—56.44%;γ-变形菌纲相对丰度在燕子墩土壤中显著高于其他样点(P<0.05)。0—2 cm土层,变形菌门相对丰度在燕子墩、红星村和分水闸这3个样点较高;拟杆菌门的相对丰度在分水闸样点土壤中最高;2—25 cm土层,变形菌门与表层土具有相同趋势;放线菌门的相对丰度在西大滩样点土壤中最高。在属水平,芽孢杆菌属是所有样点的优势属种。α多样性指数结果显示:燕子墩的土壤细菌群落多样性和丰富度都显著低于其他样点。土壤理化性质测定结果表明:在0—2 cm,全氮、有机碳、碱解氮、速效钾在黄渠拐子土壤中含量最高;含...     

舟山群岛不同功能区划海域细菌群落结构分析

浮游细菌在海洋生态系统中不可或缺,在海洋生物地球化学循环过程中起着关键性作用。【目的】为了解舟山群岛不同功能区划海域细菌群落结构及丰度变化,探索海洋生态因子对细菌群落结构的影响。【方法】于2016年夏季(8月)在舟山群岛不同功能区划海域共设置8个典型站位采集表层海水,基于细菌16S rRNA基因进行高通量测序;利用流式细胞术揭示各海域细菌丰度;利用典范对应分析(Canonical correspondence analysis,CCA)探讨海洋生态因子与细菌多样性之间的关系。【结果】共获取到305487条原始序列,基于97%相似性水平进行OTU(Operational Taxonomic Units)聚类分析,共得到1088个OTUs,包括29个门、62个纲、138个目、239个科、416个属。细菌群落组成在各个站位之间不尽相同,但都主要包括Flavobacteria、Alphaproteobacteria、Gammaproteobacteria三大优势菌纲。CCA结果表明细菌群落结构和多样性情况与站位分布和所在站位的环境因子息息相关,Cyanobacteria受硝酸盐影响显著,Parcubacteria受温度影响最大,而磷酸盐对本实验海域菌群影响甚微。对海洋菌群潜在功能进行预测的结果显示,各海域菌群在氨基酸代谢、碳水化合物代谢、膜运输等方面功能较为突出,为今后舟山海洋微生物研究提供了新的方向。【结论】高通量测序分析可以更精确地揭示海洋菌群的群落结构信息。该研究为细菌群落结构与环境因素的关联提供参考。本研究所取得的大量数据既可以作为对舟山市海洋功能区划施行情况的响应,又将为舟山及其邻近海域浮游细菌群落结构的进一步研究奠定基础。     

基于PCR-DGGE和高通量测序分析白云边酒窖泥细菌群落结构与多样性

【目的】探索白云边酒不同窖龄窖泥细菌群落结构及其多样性,分析窖泥细菌群落特征对兼香型白酒风格形成的影响。【方法】分别提取2年和23年窖泥样品总DNA,采用PCR-DGGE、基因克隆以及高通量测序技术,研究窖泥细菌的分布情况。【结果】白云边窖泥细菌归属于变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)4个菌门。2、23年窖泥共同的优势菌属(≥1.0%)包括棒状杆菌属(Corynebacterium)、香味菌属(Myroides)、鞘氨醇杆菌属(Sphingobacterium)、乳杆菌属(Lactobacillus)、梭菌属(Clostridium)、醋杆菌属(Acetobacter)、产碱杆菌属(Alcaligenes)、肠杆菌属(Enterobacter)和不动杆菌属(Acinetobacter)。2年窖泥特有优势菌属为Dysgonomonas、Fluviicola、变形杆菌属(Proteus)和Wohlfahrtiimonas,而23年窖泥特有优势菌属为葡萄球菌属(Staphylococcus)、Hazenella、魏斯氏菌属(Weissella)、葡糖醋杆菌属(Gluconacetobacter)和摩根氏菌属(Morganella)。【结论】2年窖泥细菌群落多样性高于23年窖泥。比较了白云边两种窖龄窖泥主要细菌组成情况,为研究微生物对白云边酒浓酱兼香型独特风味形成的影响提供依据。     

纳米Fe3O4对生菜生长及土壤细菌群落结构的影响

纳米Fe₃O₄的广泛应用增加了其暴露到农田环境的可能性,因此亟待研究纳米Fe₃O₄对农业生态环境的影响.本研究采用盆栽试验方式,研究不同浓度纳米Fe₃O₄颗粒(1、10、100 mg·kg^-1)对生菜生长及土壤细菌群落的影响,并与相应浓度的普通Fe₃O₄处理进行对比.通过测定植物光合速率常数、植株Fe含量来评价植物生长;采用高通量测序技术研究土壤细菌群落结构及组成.结果表明: 不同浓度纳米Fe₃O₄的影响不同.低浓度纳米Fe₃O₄能提高植物生物量,增强植物叶片光合速率,增加土壤中黄单胞菌目的相对丰度,降低蓝细菌、鞘脂杆菌纲的相对丰度,但对群落多样性指数影响不显著.高浓度纳米Fe₃O₄抑制作物生长,提高植株中Fe积累及土壤电导率,降低细菌群落系统发育多样性,降低黄单胞菌目、鞘脂杆菌纲相对丰度,增加蓝细菌相对丰度.此外,一些土壤功能微生物对纳米Fe₃O₄及普通Fe₃O₄处理的响应也存在差异,说明不同粒径及浓度的Fe₃O₄均会对土壤微生物群落产生影响,并可能影响地上部分植物性状.因此,在评估纳米颗粒的生物学效应时需较多关注土壤微生物.     

山西左云县采煤区人工湿地冬季沉积物细菌群落多样性

【背景】人工湿地处理煤炭矿井废水效果显著,而关于人工湿地沉积物的细菌群落结构和特征研究较少。【目的】阐明人工湿地净化采煤区废水的效应和细菌群落结构及特征。【方法】对山西省左云县采煤区人工湿地不同采样点水质进行监测,利用高通量测序技术对人工湿地沉积物中细菌16S rRNA基因V3-V4区进行测序,分析沉积物中细菌群落组成。【结果】采煤区废水流经人工湿地后水质得到有效改善,其中生化需氧量(Biochemical oxygen demand,BOD5)、化学需氧量(Chemical oxygen demand,CODCr)、总氮(Total nitrogen,TN)、总磷(Total phosphorus,TP)去除率分别达到76.2%、93.4%、73.4%和99.3%;测序共获得2 832个操作分类单元(Operational taxonomic units,OTUs),共753个属,分布于51个门的150个纲;4个采样点中优势菌门为变形菌门(Proteobacteria,50%-64.7%)和拟杆菌门(Bacteroidetes,15.8%-21.2%),变形菌门中优势菌纲为β-变形菌纲和γ-变形菌纲,其中M1、M2、M3中富含氨氮氧化细菌的β-变形菌纲和γ-变形菌纲丰度最高;M4沉积物细菌OTU最多,多样性最高;沉积物特有优势菌属Sulfurimonas、Sulfuricurvum的相对丰度与NH4+-N的含量显著负相关,Azoarcus菌属相对丰度与TN存在显著正相关,Novosphingobium菌属相对丰度与TP含量存在显著正相关。【结论】煤矿废水流经人工湿地可得到有效的净化,细菌多样性在人工湿地生态功能中起到了决定作用。     

锦鸡儿植物对盐碱地土壤理化性质和细菌群落的影响

锦鸡儿属的多种植物在干旱/半干旱区域水土保持等方面具有显著的作用效果,该研究以柠条锦鸡儿、中间锦鸡儿和小叶锦鸡儿为材料,选择宁夏银北西大滩核心试验站的盐渍化荒地为研究对象,分析了3种锦鸡儿建植1年对盐碱地土壤理化性质及土壤细菌群落的影响,以了解该类植物在盐碱地改良中的作用及生态效应。结果显示:(1)种植锦鸡儿1年能够显著降低盐碱地土壤全盐含量和pH,提高土壤全氮和全磷含量。(2)种植锦鸡儿不影响其非根际土壤细菌群落的α多样性,但小叶锦鸡儿非根际土壤细菌群落的香浓指数最低。(3)不同种锦鸡儿对盐碱地细菌群落结构的影响有所不同,小叶锦鸡儿根际/非根际细菌群落的结构分别与对照土壤以及柠条、中间锦鸡儿根际土壤存在明显差异;在属水平,包括假黄色单胞菌属、新鞘氨醇菌属及根瘤菌属在内的12个优势细菌属的相对丰度在小叶锦鸡儿非根际土壤中显著增加。(4)db-RDA分析表明,盐碱地土壤全盐、全氮及全磷含量分别解释了锦鸡儿非根际细菌群落结构变异信息的23.5%、25.4%和22.2%,3种因子累计共解释48.5%的细菌群落变异(P0.05),但这些因子对锦鸡儿根际细菌群落的变化无显著影响。研究表明,短期种植锦鸡儿属植物对盐碱地土壤理化性质具有一定改良效果,且不影响非根际土壤细菌群落的α多样性,但不同种锦鸡儿对土壤细菌群落结构的影响存在差异。     

基于16S rRNA基因测序分析微生物群落多样性

微生物群落多样性的研究对于挖掘微生物资源,探索微生物群落功能,阐明微生物群落与生境间的关系具有重要意义。随着宏基因组概念的提出以及测序技术的快速发展,16S rRNA基因测序在微生物群落多样性的研究中已被广泛应用。文中系统地介绍了16S rRNA基因测序分析流程中的四个重要环节,包括测序平台与扩增区的选择、测序数据预处理以及多样性分析方法,就其面临的问题与挑战进行了探讨并对未来的研究方向进行了展望,以期为微生物群落多样性相关研究提供参考。     

石油污染对土壤微生物群落多样性的影响

土壤中的微生物主要有细菌、放线菌、真菌三大类群,微生物在石油污染的土壤中发挥着维持生态平衡和生物降解的功能。文中以四川省遂宁市射洪县某废弃油井周围不同程度石油污染土壤为供试土壤,首先对各组供试土壤的基本理化性质进行测定分析;然后采用平板菌落计数法测定了供试土壤中三大类微生物数量的变化,结果表明:相比未被污染的对照土壤,石油污染的土壤中细菌、放线菌、真菌数量均减少,并且土壤中可培养微生物的数量与土壤含水量呈正相关;再采用454焦磷酸测序技术对土壤中的细菌群落多样性及变化进行16S rRNA基因分析。在所有供试的4个土壤样品中,共鉴定出不少于23 982个有效读取序列和6 123种微生物,相比于未被污染的对照土壤,石油污染土壤中细菌的种类更加丰富,主要优势门类为酸杆菌门、放线菌门、拟杆菌门、绿弯菌门、浮霉菌门和变形菌门。但不同土壤样品中优势菌群的群落结构有所差异,石油污染的土壤中,酸杆菌门、放线菌门和变形菌门的数量最多,未被石油污染的土壤中,放线菌门、拟杆菌门和变形菌门的数量最多。     

Diversity of endophytic bacterial communities in poplar grown under field conditions

Bacterial endophytes may be important for plant health and other ecologically relevant functions of poplar trees. The composition of endophytic bacteria colonizing the aerial parts of poplar was studied using a multiphasic approach. The terminal restriction fragment length polymorphism analysis of 16S rRNA genes demonstrated the impact of different hybrid poplar clones on the endophytic community structure. Detailed analysis of endophytic bacteria using cultivation methods in combination with cloning of 16S rRNA genes amplified from plant tissue revealed a high phylogenetic diversity of endophytic bacteria with a total of 53 taxa at the genus level that included Proteobacteria, Actinobacteria, Firmicutes and Bacteroidetes. The community structure displayed clear differences in terms of the presence and relative proportions of bacterial taxa between the four poplar clones studied. The results showed that the genetic background of the hybrid poplar clones corresponded well with the endophytic community structure. Out of the 513 isolates and 209 clones identified, Actinobacteria, in particular the family Microbacteriaceae, made up the largest fraction of the isolates, whereas the clone library was dominated by Alpha- and Betaproteobacteria. The most abundant genera among the isolates were Pseudomonas and Curtobacterium, while Sphingomonas prevailed among the clones.     

N2O emission associated with shifts of bacterial communities in riparian wetland during the spring thawing periods

Soil freeze–thaw processes lead to high nitrous oxide (N₂O) emissions and exacerbate the greenhouse effect. The wetlands of the Inner Mongolia Plateau are in the pronounced seasonal freeze–thaw zone, but the effect of spring thaw on N₂O emissions and related microbial mechanisms is still unclear. We investigated the effects of different periods (freeze, freeze–thaw, and thaw) on soil bacterial community diversity and composition and greenhouse gas emissions during the spring freeze–thaw in the XiLin River riparian wetlands in China by amplicon sequencing and static dark box methods. The results showed that the freeze–thaw periods predominantly impact on the diversity and composition of the bacterial communities. The phyla composition of the soil bacteria communities of the three periods is similar in level, with Proteobacteria, Chloroflexi, Actinobacteria, and Acidobacteria dominating the microbial communities. The alpha‐diversity of bacterial communities in different periods varies that the freezing period is higher than that of the freeze–thaw period (p < .05). Soil total carbon, soil water content, and microbial biomass carbon were the primary factors regulating the abundance and compositions of the bacterial communities during spring thawing periods. Based on functional predictions, the relative abundance of nitrification and denitrification genes was higher in the freezing period than in the thawing period, while the abundance was lowest in the freeze–thawing period. The correlation results found that N₂O emissions were significantly correlated with amoA and amoB in nitrification genes, indicating that nitrification may be the main process of N₂O production during spring thaw. This study reveals potential microbial mechanisms of N₂O emission during spring thaw and provides data support and theoretical basis for further insight into the mechanism of N₂O emission during spring thaw.     

不同土壤深度对宁夏石嘴山盐碱地细菌菌群多样性的影响

宁夏自治区是我国受到土壤次生盐渍化危害的重点区域,然而对该地区盐碱化形成机制及影响因素的研究资料较少。在宁夏石嘴山地区采集土壤样本,采用基于16S rRNA的PCR-DGGE技术对不同深度土壤的细菌群落多样性和优势种群进行分析,以期从土壤生态角度探索该地盐渍化成因及改良措施。带谱相似性和UPGMA聚类结果表明,表层(D20 cm)土壤和底层(D80 cm)土壤样本中细菌菌群相似程度较高;而中间层(20 cmD80 cm)土壤样本中细菌菌群相似程度很低。多样性指数分析显示,随着地层深度的增加,菌落的丰富度和多样性均趋于下降;而均匀程度变化幅度不大。对其中5个优势条带进行测序比对的结果表明,该地区土壤可识别优势细菌菌群为变形细菌门Proteobacteria和拟杆菌门Bacteroidetes,但大部分条带未可识别。结果表明该地区盐碱土壤中的细菌菌群多样性程度较高,且优势菌群不尽相同;土壤深度与细菌菌落多样性在一定程度上存在线性关系。为深入研究次生盐渍化土壤的形成机制以及影响因素提供理论参考。     

Shifts of tundra bacterial and archaeal communities along a permafrost thaw gradient in Alaska

Understanding the response of permafrost microbial communities to climate warming is crucial for evaluating ecosystem feedbacks to global change. This study investigated soil bacterial and archaeal communities by Illumina MiSeq sequencing of 16S rRNA gene amplicons across a permafrost thaw gradient at different depths in Alaska with thaw progression for over three decades. Over 4.6 million passing 16S rRNA gene sequences were obtained from a total of 97 samples, corresponding to 61 known classes and 470 genera. Soil depth and the associated soil physical–chemical properties had predominant impacts on the diversity and composition of the microbial communities. Both richness and evenness of the microbial communities decreased with soil depth. Acidobacteria, Verrucomicrobia, Alpha‐ and Gamma‐Proteobacteria dominated the microbial communities in the upper horizon, whereas abundances of Bacteroidetes, Delta‐Proteobacteria and Firmicutes increased towards deeper soils. Effects of thaw progression were absent in microbial communities in the near‐surface organic soil, probably due to greater temperature variation. Thaw progression decreased the abundances of the majority of the associated taxa in the lower organic soil, but increased the abundances of those in the mineral soil, including groups potentially involved in recalcitrant C degradation (Actinomycetales, Chitinophaga, etc.). The changes in microbial communities may be related to altered soil C sources by thaw progression. Collectively, this study revealed different impacts of thaw in the organic and mineral horizons and suggests the importance of studying both the upper and deeper soils while evaluating microbial responses to permafrost thaw.     

Culture-independent study of bacterial communities in tropical river sediment

Ubiquitous microbial communities in river sediments actively govern organic matter decomposition, nutrient recycling, and remediation of toxic compounds. In this study, prokaryotic diversity in two major rivers in central Thailand, the Chao Phraya (CP) and the Tha Chin (TC) distributary was investigated. Significant differences in sediment physicochemical properties, particularly silt content, were noted between the two rivers. Tagged 16S rRNA sequencing on a 454 platform showed that the sediment microbiomes were dominated by Gammaproteobacteria and sulfur/sulfate reducing Deltaproteobacteria, represented by orders Desulfobacteriales and Desulfluromonadales together with organic degraders Betaproteobacteria (orders Burkholderiales and Rhodocyclales) together with the co-existence of Bacteroidetes predominated by Sphingobacteriales. Enrichment of specific bacterial orders was found in the clayey CP and silt-rich TC sediments, including various genera with known metabolic capability on decomposition of organic matter and xenobiotic compounds. The data represent one of the pioneered works revealing heterogeneity of bacteria in river sediments in the tropics.     

转几丁质酶和葡聚糖酶双价基因棉花对土壤细菌种群多样性的影响

以转几丁质酶和葡聚糖酶双价基因棉花为研究对象,非转基因受体棉花为对照,通过比较可培养细菌数量和基于16S rRNA克隆文库细菌种群分析,评价外源双价基因的导入在苗期、蕾期、花铃期和吐絮期对棉花根际细菌群落多样性的影响。结果表明,可培养细菌的数量不受外源双价基因的影响,随着棉花生育期的交替而变化,以代谢旺盛的花铃期最多。构建的转基因和非转基因不同生育期根际土壤细菌16S rRNA文库容量为2400个克隆,涵盖了细菌的283个属。其中,Acidobacterium是最大优势类群,共包括624个克隆,其次为未知细菌种群和Flavisolibacter。比较转基因和非转基因棉花根际土壤细菌的种群结构,结果显示,同一生育期内前者种群的多样性显著低于后者,二者的共有类群随着生长发育的进行而增多。研究结果说明几丁质酶基因和葡聚糖酶基因对棉花根际细菌种群多样性有着不同程度的削减作用,但是随着种植时间的延长,该差异呈现逐渐缩小的趋势。     

利用小亚基核糖体RNA 技术分析温室黄瓜近根土壤古菌和真菌多样性

土壤古菌和真菌在温室生态系统是仅次于细菌的微生物,具有类似于细菌的重要生态功能。通过构建古菌16S rRNA和真菌18S rRNA基因克隆文库,分析温室黄瓜近根土壤古菌和真菌群落结构组成,为开发利用温室这一特殊的生态环境中丰富的微生物资源以及理解微生物与植物间的互作提供参考依据。采用研磨-冻融-溶菌酶-蛋白酶K-SDS热处理以及CTAB处理等理化方法,提取和纯化微生物总DNA,构建古菌16S rRNA和真菌18S rRNA基因克隆文库。利用DOTUR软件将古菌和真菌序列按照相似性97％的标准分成若干个可操作分类单元 (OTUs)。土壤古菌克隆文库主要包括泉古菌门和未分类的古菌两大类,并有少部分广域古菌类群,所有泉古菌均属于热变形菌纲,共45个OTUs;真菌克隆文库包括真菌门的大多数亚门真菌,共24个OTUs,未发现担子菌亚门真菌。古菌多样性比较丰富,且发现少量的广域古菌 (甲烷菌),这一情况可能与温室长期高温高湿,高有机质含量,土壤处于缺氧环境有关;土壤真菌的优势种群为子囊菌,占到土壤真菌的80％以上,这可能与绝大多数植物真菌性病害属于土传病害,通过菌丝体、菌核或子囊壳在土壤病残体中越冬有一定的关系。     

Soil parent material is a key determinant of the bacterial community structure in arable soils

The bacterial community composition in soil and rhizosphere taken from arable field sites, differing in soil parent material and soil texture, was analyzed using terminal restriction fragment length polymorphism (T-RFLP) of 16S rRNA genes. Nine sandy to silty soils from North-East Germany could clearly be distinguished from each other, with a relatively low heterogeneity in the community structure within the field replicates. There was a relationship between the soil parent material, i.e. different glacial and aeolian sediments, and the clustering of the profiles from different sites. A site-specific grouping of T-RFLP profiles was also found for the rhizosphere samples of the same field sites that were planted with potatoes. The branching of the rhizosphere profiles corresponded partly with the soil parent material, whereas the effect of the plant genotype was negligible. Selected terminal restriction fragments differing in their relative abundance within the nine soils were analyzed based on the cloning of the 16S rRNA genes of one soil sample. A high phylogenetic diversity observed to include Acidobacteria, Betaproteobacteria, Bacteroidetes, Verrucomicrobia, and Gemmatimonadetes. The assignment of three out of the seven selected terminal restriction fragments to members of Acidobacteria suggested that this group seems to participate frequently in the shifting of community structures that result from soil property changes.     

结合宏基因组末端随机测序和16S rDNA技术分析温室黄瓜根围土壤细菌多样性

土壤细菌在温室土壤环境中具有十分重要的生态功能,与温室作物以及微生物内部存在互作关系。研究土壤细菌的群落结构组成,有助于了解土地利用变化与生态环境效应之间的关系。结合16S rRNA基因克隆文库和宏基因组末端测序对温室黄瓜根围土壤细菌的多样性进行了分析。在16S文库中,根据97%的序列相似性水平划分OTU,共有35个OTU,其中优势菌群是γ-Proteobacteria,其次为Firmicutes,Bacillus为优势细菌。在纲分类水平上,16S文库和宏基因组末端测序结果均包含γ-Proteobacteria、α-Proteobacteria、δ-Proteobacteria、β-Proteobacteria、Actinomycetales和Firmicutes,各纲比例有差别;在优势种群属水平上,末端测序的结果包含的属多于16S文库(4035);在优势细菌种类上,两者反映的结果一致,均为Bacillus。但是,宏基因组末端测序包含了大多数的弱势种群,更能反映细菌多样性的真实水平。与露地土壤细菌16S文库相比较,土壤细菌多样性降低,这可能与温室多年连作,种植蔬菜种类单一直接相关。