不同生境中桂花和夹竹桃叶际细菌的群落结构

植物叶际微生物多样性是目前植物-微生物关系研究的热点之一,但影响叶际微生物群落结构的主要因素目前还存在很大争议.本研究以生长在3处生境的桂花和夹竹桃为对象,基于高通量测序技术,分析2种植物叶际细菌的群落结构,探讨影响植物叶际细菌群落结构的主要因素.结果表明:来自3处生境的2种植物叶际细菌多样性无显著差异,构成叶际细菌群落的优势门主要包括放线菌门、拟杆菌门、衣原体门、蓝细菌门、厚壁菌门和变形菌门,优势属主要包括甲基杆菌属、鞘氨醇单胞菌属、薄层杆属、Polaromonas和无毛螺旋体属.植物种类、生境及二者的交互作用均能显著影响叶际细菌群落结构,其中生境的影响最大.     

重庆烟区“K326”烟草品种靶斑病发生的叶际微生态因子分析

为明确烟草靶斑病在重庆烟区“K326”烟草品种上对叶际微生物群落结构的影响,在重庆市酉阳县分别采集移栽40 d、 60 d、 80 d和100 d后具有烟草靶斑病与健康的烟叶样品,采用高通量测序方法分析样品中细菌群落结构,并结合烟株生长期气象因子和病害发生情况进行分析.移栽后40 d靶斑病开始发生,且发病率和病情指数呈上升趋势,移栽后60～80 d为发病高峰期.发病叶中叶际细菌优势菌门为变形菌门和厚壁菌门,优势菌属为假单胞菌属、微小杆菌属等.健康叶叶际细菌的优势菌门分别为蓝藻门、放线菌门、拟杆菌门等,优势菌属为不动杆菌属、微小杆菌属、芽孢杆菌属、泛细菌属等.健康叶的整体群落丰度大于发病叶,移栽40 d、 60 d和100 d后采集样品的发病叶与健康叶叶际细菌群落结构比较差异具有统计学意义.靶斑病发生较重情况下,发病叶与健康叶叶际细菌群落结构上差异具有统计学意义,提示靶斑病的发生影响了叶际细菌群落结构.     

连作对地黄根际土壤细菌群落多样性的影响

采用末端限制性片段长度多态性(T-RFLP)技术研究地黄连作下根际土壤细菌群落的动态变化.结果表明:地黄根际土壤细菌群落的香农多样性指数、丰富度指数和相似性指数均为对照(CK)1年2年,地黄连作下细菌优势种群的比例显著下降,种植1年土壤中厚壁菌门的芽孢杆菌纲在整个细菌群落中处于主导地位,而种植2年土壤中变形菌门ε-变形菌纲处于主导地位.地黄连作使其根际土壤细菌种类大量减少,群落结构趋于简单.连作后细菌群落多样性水平的变化导致根际土壤微生物群落功能的失调,可能是引发连作障碍的原因之一.     

轮作和连作对胡萝卜根际和非根际细菌多样性的影响

细菌是土壤最重要的组成部分,土壤细菌的群落结构对土壤的健康至关重要,可以直接影响植物的生长。胡萝卜的长年连作导致其产量低、质量差等一系列问题的发生,连作问题已成为不可忽视的问题。本研究通过HiSeq PE250高通量测序和生物信息学方法获得了玉米胡萝卜轮作种植和胡萝卜连作种植中胡萝卜根际和非根际土壤细菌的组成结构。研究结果表明,相较于胡萝卜连作,玉米胡萝卜轮作种植改变了胡萝卜根际土壤细菌的组成和群落多样性。RM.M (玉米胡萝卜轮作根际土壤样品)样品中的优势菌门为放线菌门和厚壁菌门,RM.R (胡萝卜连作根际土壤样品)的优势菌门是变形菌门;有益菌如芽孢杆菌属和根瘤菌属在玉米胡萝卜轮作种植中的胡萝卜根际富集。RM.M与RM.R的细菌类群差异显著,群落多样性较低;NRM.M(玉米胡萝卜轮作非根际土壤样品)和NRM.R (胡萝卜连作非根际土壤样)的细菌类群差异不明显,群落多样性较高。与连作根际土壤相比,轮作根际土壤中有益菌的相对丰度较大,细菌相对丰度明显增加。本研究结果对通过调节根际土壤细菌组成来保证胡萝卜产量和质量的稳定,解决连作问题具有重要的意义。     

生菜连作及生菜-菠菜轮作对土壤细菌群落结构的影响

以生菜连作和生菜-菠菜轮作不同次数的土壤样品为材料,研究比较生菜不同种植方式下土壤细菌群落的差异。采集0-20 cm深度土壤样品,分别提取样品总DNA,采用高通量测序技术分析生菜种植前后土壤样品中细菌多样性。测序结果经过质控过滤,共获得有效序列46 865条,对相似水平为97%的OTU进行生物信息统计分析,共得到OTU 21 692个。Heatmap图和群落组成分析显示,样品共检测到细菌40个门,89个纲,190个目,371个科,693个属,其中变形菌门、厚壁菌门和放线菌门在连作过程中占比逐渐下降,而在轮作过程中较为稳定。另外,壤霉菌属、节细菌属、黄杆菌属、溶杆菌属及微细菌属等有益菌属逐渐成为轮作土壤样品中的优势菌属。主成分分析结果表明,轮作和连作土壤样品差异明显。生菜在连续种植第5次后,产量和过氧化氢酶活性分别下降了24.1%和20.7%,出现连作障碍,而轮作生菜产量和过氧化氢酶活性较连作分别提高了49.6%和10.5%。菠菜-生菜轮作土壤中细菌类群具有丰富的多样性,随着轮作的增加有上升的趋势,可在一定程度上缓解生菜连作过程中出现的问题。     

滇西北兰坪金顶铅锌矿矿区可培养细菌多样性初步研究

目的对兰坪金顶铅锌矿矿区样品中的可培养细菌进行分离并对其多样性进行研究。方法采集云南兰坪金顶铅锌矿矿区土样和矿石样,采用固体肉汤培养基、卯黄培养基及PYGV培养基分离该矿区环境中的可培养细菌,利用16SrRNA基因序列分析构建系统发育树,并统计不同种属细菌的数量,初步评估细菌多样性。结果兰坪金顶铅锌矿矿区环境细菌的主要种群包括放线菌门、变形菌门和厚壁菌门的不同菌属：微球菌属、节杆菌属、假单胞菌属、短波单胞菌属、芽孢杆菌属、类芽孢杆菌属、考克菌属、葡萄球菌属、芽孢八叠球菌及Skermanella属的菌株,其中抗逆性较强的优势菌群为放线菌门的细菌。结论本研究初步证实兰坪金顶铅锌矿矿区可培养细菌种类丰富。     

云南省富宁磁铁矿矿区可培养细菌多样性初步研究

目的对云南富宁磁铁矿矿区样品中可培养细菌进行分离并对其多样性进行了初步研究。方法采集云南富宁磁铁矿矿区土样和矿石样,采用固体肉汤培养基、卵黄培养基及PYGV培养基分离该矿区环境中的可培养细菌,利用16SrRNA基因序列分析构建系统发育树,并统计不同种属细菌的数量,初步评估细菌多样性。结果富宁磁铁矿矿区环境细菌的主要种群包括厚壁菌门、放线菌门和变形菌门的不同菌属：芽孢杆菌属、葡萄球菌属、节杆菌属和假单胞菌属的菌株,其中抗逆性较强的优势菌群为厚壁菌门的芽孢杆菌。结论本研究初步表明富宁磁铁矿矿区可培养细菌种类具有一定的多样性。     

添加高粱根茬根际土对连作黄瓜生长和根际微生物的影响

为了分析添加高粱根茬根际土对连作黄瓜生长和根际微生物群落特征的影响,本研究通过盆栽试验,采用荧光定量PCR和高通量测序技术分析土壤细菌和真菌群落组成的差异。试验共设4个处理: CK(不施肥),T₁(单施化肥),T₂(优化施肥),T₃(优化施肥+高粱根茬根际土)。结果表明: 与其他处理相比,T₃处理促进了黄瓜生长发育,提高了土壤中16S rRNA和ITS rRNA基因数量。与T₁处理相比,T₂和T₃处理明显提高了细菌群落的丰富度和多样性,不同处理间真菌群落的丰富度和多样性差异不明显。添加高粱根茬根际土在一定程度上改变了基于门、属水平上的细菌和真菌群落结构。其中,细菌中提高了酸杆菌门和拟杆菌门的丰度,降低了变形菌门、厚壁菌门、硝化螺旋菌门和芽孢杆菌属的丰度;真菌中提高了担子菌门、木霉菌属和假散囊菌属的丰度,降低了镰刀菌属和绿僵菌属的丰度。冗余分析表明,土壤硝态氮和有机质含量分别是影响细菌和真菌群落结构差异的关键因子。添加高粱根茬根际土不仅提高了连作黄瓜土壤微生物数量和细菌多样性,而且增加了有益菌木霉菌属的丰度,降低了致病镰刀菌的丰度和数量,保障了黄瓜存活率,为缓解黄瓜连作障碍提供了一条切实可行的解决途径。     

冷箭竹根际土壤中可培养细菌的多样性

为了解冷箭竹(Bashania fangiana)根际土壤中可培养细菌的多样性,2006年5月从四川卧龙自然保护区冷箭竹根际土壤中共分离出50株具不同菌落形态的细菌.16S rDNA序列分析结果表明:50株菌分属于10个属26个种.27株属于变形菌门γ亚群(Gammaproteobaeteria)(42.3%)、9株属于厚壁菌门(Firmicutes)(26.9%)、4株属于放线菌门(Actinobacteria)(15.4%)、6株属于变形菌门β亚群(Betaproteobacteria)(7.7%)、3株属于变形菌门α亚群(Alphaproteobacteria)(3.9%),1株与土地杆菌属(Pedobacter)关系密切.假单胞菌属(Pseudomonas)和芽孢杆菌属(Bacillus)为优势菌属.2株菌为可能的新种或属.研究表明冷箭竹根际土壤中含有较为丰富的微生物多样性.     

银北盐渍化土壤中6种耐盐植物根际细菌群落结构及其多样性

土壤微生物对土壤肥力的形成和植物营养的转化起着积极的作用。对盐渍化土壤中植物根际微生物群落组成和多样性进行研究,有助于发现新的重要微生物功能类群或者功能潜力,对于盐碱土壤的植被恢复和生态重建都具有十分重要的意义。通过高通量测序和分离培养方法,对宁夏银北地区盐渍化土壤中的6种耐盐植物根际细菌的群落结构和多样性进行了分析。结果表明:所有土壤样品共检测到31门(亚门) 67纲253科400属细菌类群,不同植物根际微生物群落组成相似,但丰度各有差异,厚壁菌门、变形菌门、放线菌门和拟杆菌门为主要优势类群;在纲水平芽孢杆菌纲相对丰度最高,其次为α-变形菌、γ-变形菌和梭菌纲;在属水平上,芽孢杆菌属以极高的丰度(15.57%—53.85%)占据绝对优势,其次是鞘氨醇单胞菌属、不动杆菌属和节杆菌属。6种植物根际细菌的群落结构组成表现为芨芨草和柽柳相似,柳枝稷、苜蓿与枸杞相似,苦豆子则与另外5种差别最大。分离培养出的110株根际细菌多数具有较强的盐碱耐受性和一种以上生物学活性,分别隶属于芽孢杆菌属、假单胞菌属、鞘氨醇杆菌属、节杆菌属和中华根瘤菌属,在属水平上多样性单一,其中芽孢杆菌属是优势属。传统的分离培养和高通量测序结果都反映出银北地区盐渍化土壤中定殖着许多有重要生态功能的微生物类群,可用于进一步筛选和利用根际微生物改良盐碱土壤。     

Characterization of the microbial diversity in a permafrost sample from the Canadian high Arctic using culture-dependent and culture-independent methods

A combination of culture-dependent and culture-independent methodologies (Bacteria and Archaea 16S rRNA gene clone library analyses) was used to determine the microbial diversity present within a geographically distinct high Arctic permafrost sample. Culturable Bacteria isolates, identified by 16S rRNA gene sequencing, belonged to the phyla Firmicutes, Actinobacteria and Proteobacteria with spore-forming Firmicutes being the most abundant; the majority of the isolates (19/23) were psychrotolerant, some (11/23) were halotolerant, and three isolates grew at -5 degrees C. A Bacteria 16S rRNA gene library containing 101 clones was composed of 42 phylotypes related to diverse phylogenetic groups including the Actinobacteria, Proteobacteria, Firmicutes, Cytophaga - Flavobacteria - Bacteroides, Planctomyces and Gemmatimonadetes; the bacterial 16S rRNA gene phylotypes were dominated by Actinobacteria- and Proteobacteria-related sequences. An Archaea 16S rRNA gene clone library containing 56 clones was made up of 11 phylotypes and contained sequences related to both of the major Archaea domains (Euryarchaeota and Crenarchaeota); the majority of sequences in the Archaea library were related to halophilic Archaea. Characterization of the microbial diversity existing within permafrost environments is important as it will lead to a better understanding of how microorganisms function and survive in such extreme cryoenvironments.     

林下植被管理措施对杉木大径材林土壤细菌群落结构的影响

林下植被是人工林生态系统的重要组成部分。本研究采用高通量测序技术,分析林下植被保留、林下植被去除和林下套种3种林下植被管理措施对杉木大径材人工林土壤细菌多样性、细菌群落结构组成以及相对丰度的影响,并分析土壤理化性质与细菌群落多样性的关系。结果表明: 林下植被保留处理的土壤细菌群落Chao1指数、Ace指数和Shannon指数高于林下植被去除和林下套种;放线菌门、酸杆菌门和绿弯菌门为本研究区杉木人工林土壤主要优势细菌;与林下植被去除和林下套种处理杉木人工林土壤细菌群落相比,林下植被保留处理土壤变形菌门、浮霉菌门、厚壁菌门和疣微菌门等相对丰度较高,而放线菌门、酸杆菌门和绿弯菌门的相对丰度较低;3种林下植被管理措施之间,土壤厚壁菌门、浮霉菌门、疣微菌门、Parcubacteria门和放线菌门等类群相对丰度差异显著;土壤含水率、全氮、全磷、水解氮和速效磷含量是影响细菌群落结构的重要因子,细菌多样性指数与土壤全氮、全磷、全钾、水解氮和速效钾含量呈显著正相关。     

徐州权台煤矿井下700 m水平微生物群落分布特征

煤矿井下与表生环境具有明显差异,研究其微生物群落分布及多样性对深入探讨井下水-岩-气-生作用机制具有重要意义.本研究在徐州权台煤矿井下700 m水平采集了8个沉积物样本,对样品进行了理化指标测试和Miseq高通量测序.结果表明,煤矿井下具有丰富的微生物多样性,8个样本共检测到35个细菌门类,其中变形菌门(Proteob...     

Diversity and structure of bacterial communities in Fildes Peninsula, King George Island

To examine the bacterial community structure in the Fildes Peninsula, King George Island, Antarctica, we examined the bacterial diversity and community composition of samples collected from lacustrine sediment, marine sediment, penguin ornithogenic sediments, and soils using culture-dependent and culture-independent methods. The 70 strains fell into five groups: Actinobacteria, Bacteroidetes, Firmicutes, Gammaproteobacteria, and Betaproteobacteria. Bacterial diversity at the phylum level detected in Denaturing Gradient Gel Electrophoresis (DGGE) profiles comprised Proteobacteria (including the subphyla Alpha-, Beta-, Gamma-, Deltaproteobacteria), Bacteroidetes, Firmicutes, Chlorobi, and Deinococcus-Thermus. Gammaproteobacteria was identified to be the dominant bacterial subphylum by cultivation and DGGE method. By cluster analysis, the overall structure and composition of bacterial communities in the soil and lacustrine sediment were similar to one another but significantly different from bacterial communities in penguin ornithogenic sediment and marine sediment, which were similar to one another. The majority of 16S rDNA sequences from cultured bacteria were closely related to sequences found in cold environments. In contrast, a minority of 16S rDNA sequences from the DGGE approach were closely related to sequences found in cold environments.     

Effects of tobacco–peanut relay intercropping on soil bacteria community structure

A reasonable cultivation pattern is beneficial to maintain soil microbial activity and optimize the structure of the soil microbial community. To determine the effect of tobacco−peanut (Nicotiana tabacum−Arachis hypogaea) relay intercropping on the microbial community structure in soil, we compared the effects of relay intercropping and continuous cropping on the soil bacteria community structure. We collected soil samples from three different cropping patterns and analyzed microbial community structure and diversity using high-throughput sequencing technology. The number of operational taxonomic units (OTU) for bacterial species in the soil was maximal under continuous peanut cropping. At the phylum level, the main bacteria identified in soil were Proteobacteria, Actinobacteria, and Acidobacteria, which accounted for approximately 70% of the total. The proportions of Actinobacteria and Firmicutes increased, whereas the proportion of Proteobacteria decreased in soil with tobacco–peanut relay intercropping. Moreover, the proportions of Firmicutes and Proteobacteria among the soil bacteria further shifted over time with tobacco–peanut relay intercropping. At the genus level, the proportions of Bacillus and Lactococcus increased in soil with tobacco–peanut relay intercropping. The community structure of soil bacteria differed considerably with tobacco–peanut relay intercropping from that detected under peanut continuous cropping, and the proportions of beneficial bacteria (the phyla Actinobacteria and Firmicutes, and the genera Bacillus and Lactococcus) increased while the proportion of potentially pathogenic bacteria (the genera Variibacter and Burkholderia) decreased. These results provide a basis for adopting tobacco–peanut relay intercropping to improve soil ecology and microorganisms, while making better use of limited cultivable land.     

Dominance of endospore-forming bacteria on a Rotating Activated Bacillus Contactor biofilm for advanced wastewater treatment

The bacterial diversity inherent to the biofilm community structure of a modified rotating biological contactor wastewater treatment process, referred to as the Rotating Activated Bacillus Contactor (RABC) process, was characterized in this study, via both culture-dependent and culture-independent methods. On the basis of culture-dependent methods, Bacillus sp. were found to exist in large numbers on the biofilm (6.5% of the heterotrophic bacteria) and the microbial composition of the biofilms was quite simple. Only three phyla were identified-namely, the Proteobacteria, the Actinobacteria (High G+C Gram-positive bacteria), and the Firmicutes (Low G+C Gram-positive bacteria). The culture-independent partial 16S rDNA sequence analysis revealed a considerably more diverse microbial composition within the biofilms. A total of eight phyla were recovered in this case, three of which were major groups: the Firmicutes (43.9%), the Proteobacteria (28.6%), and the Bacteroidetes (17.6%). The remaining five phyla were minor groups: the Planctomycetes (4.4%), the Chlorobi (2.2%), the Actinobacteria (1.1%), the Nitrospirae (1.1%), and the Verrucomicrobia (1.1%). The two most abundant genera detected were the endospore-forming bacteria (31.8%), Clostridium and Bacillus, both of which are members of the Firmicutes phylum. This finding indicates that these endospore-forming bacteria successfully colonized and dominated the RABC process biofilms. Many of the colonies or clones recovered from the biofilms evidenced significantly high homology in the 16S rDNA sequences of bacteria stored in databases associated with advanced wastewater treatment capabilities, including nitrification and denitrification, phosphorus accumulation, the removal of volatile odors, and the removal of chlorohydrocarbons or heavy metals. The microbial community structures observed in the biofilms were found to correlate nicely with the enhanced performance of advanced wastewater treatment protocols.     

Microbial community on healthy and diseased leaves of an invasive plant <Emphasis Type="Italic">Eupatorium adenophorum</Emphasis> in Southwest China

Invasive plants have caused great economic losses and environmental problems worldwide. Eupatorium adenophorum is one of the most invasive weeds in China. To better understand its invasive mechanisms, in the present paper, the microbial communities of healthy and diseased leaves of E. adenophorum were obtained using both culture-independent and -dependent methods and their diversities were compared. The bacteria obtained from culture-independent method belong to Proteobacteria (95.8%), Actinobacteria (2.1%), and Firmicutes (2.1%) and fungi belong to Ascomycota (65.2%) and Basidiomycota (34.8%). Very few overlapped microbial species were found by culture-dependent and -independent methods. Healthy leaves display higher bacterial diversity than diseased leaves. Phylogenetic structures are very different between healthy and diseased phyllosphere microbial communities. Bacteria close to Acinetobacter and Pseudomonas were dominant on healthy leaves, whereas those close to Shigella were dominant on diseased leaves. 52.9% of fungal clones from healthy leaves were Ustilaginomycetes, close to Rhodotorula phylloplana and uncultured basidomycete; by contrast, 60% of clones from diseased leaves were Lecanoromycetes, close to Umbilicaria muehlenbergii. No bacteria but four fungal strains phylogenetically close to Myrothecium sp. and Alternaria alternate were pathogenic to seedlings and detached leaves of the invasive plant. Therefore, this plant may be resistant to pathogens from bacteria but not fungi in its introduced range.     

太子参不同休耕年限土壤理化特征和微生物群落变化

太子参为福建道地中药材,连作效应严重限制其产业发展。休耕可以在一定程度上缓解连作效应,然而休耕对于缓解连作效应的机制尚不清楚。本文通过高通量测序分析不同休耕年限对太子参土壤微生物丰度及群落多样性的影响,并探索土壤理化性质、酚酸类物质与土壤微生物群落之间的关联性。结果表明:与对照太子参土壤相比,休耕土壤真菌多样性降低,细菌多样性增多。在细菌优势菌门中,酸杆菌门相对丰度显著增加,变形菌门与放线菌门相对丰度显著降低;真菌优势菌门相对丰度无显著变化。土壤酸性减弱,有机质含量随休耕年限增加呈现递减模式,酚酸类物质中苯甲酸和水杨酸含量显著降低,而对香豆酸等逐渐积累。综上,太子参土壤休耕改善了土壤微生物的菌群结构、土壤性质,从而缓解了连作效应。     

肠道菌群对凡纳滨对虾健康的指示作用

高密度、集约化的凡纳滨对虾养殖常伴随病害的发生,但还不能确定能否利用对虾的肠道细菌差异来指示其健康状况.因此,我们分别采集了健康和发病养殖塘水样和对虾样品,利用Illumina MiSeq测序技术测定细菌16S rRNA基因,研究细菌的群落结构和多样性.结果表明： 细菌群落结构变异主要受水体中亚硝酸盐、叶绿素a和磷酸盐的影响.发病对虾肠道细菌群落多样性显著低于健康对虾.在健康和发病对虾肠道细菌中,运用响应比分析,我们筛选到了28个丰度差异显著的分类操作单元(OTUs),其中来源于放线菌纲、黄杆菌纲和芽孢杆菌纲的OTUs丰度在发病对虾肠道中显著降低,而隶属于梭状芽孢杆菌纲的OTUs丰度在发病对虾肠道显著增加.此外,我们挑选出61个指示物种,主要属于拟杆菌门、变形菌门、厚壁菌门、浮霉菌门和放线菌门.值得注意的是,它们能够区分样品的来源(水样或虾肠道)和健康状态.本研究为对虾病害预测和益生菌开发提供了理论依据.