青藏高原高寒草地地下生物多样性: 进展、问题与展望

栖息于土壤中的微生物和微型动物种类繁多、数量巨大, 在对地上生物多样性的调控和在生态系统功能与服务的维系中, 具有举足轻重的作用。虽然对土壤微生物以及土壤动物已经开展了广泛的调查, 但是整体上对于地下生物多样性的分布格局、驱动机制及其对全球变化的响应与适应过程, 仍缺乏深刻的认识。青藏高原是全球变化的敏感区域, 其中高寒草地是高原最主要的植被类型, 占高原面积的60%左右, 在高寒生态系统生物多样性维持中具有重要意义。近年来, 已有大量研究关注于高寒草地地下生物多样性, 但是缺乏系统的总结与论述。基于此, 本文从细菌、真菌、古菌、线虫、节肢动物五大土壤生物类群出发, 阐述了青藏高原高寒草地的地下物种丰富度、分布格局及其影响因素, 重点探讨了它们对气候变化和人类活动的响应, 并就未来高寒草地地下生物多样性亟需关注的关键问题进行了展望, 包括: (1)地下各个生物类群的分布格局、各类群之间的联系及驱动机制; (2)地上与地下生物多样性耦联的机制; (3)地下生物多样性对生态系统功能和健康的影响; (4)地下生物多样性的调控实验研究。     

模拟增温对青海湖河源湿地土壤微生物群落特征的影响

湿地生态系统在生物多样性保护等方面有着重要的功能及地位, 不同的湿地生态系统在功能上存在差异, 土壤微生物在湿地生态系统中发挥着重要作用, 但目前对河源湿地的土壤微生物群落开展的研究较少。全球变暖大背景下, 为探究温度升高对河源湿地土壤微生物的影响, 利用高通量测序方法来深入了解模拟增温后土壤细菌及真菌的群落结构及多样性的变化。青海湖河源湿地细菌的优势菌群为变形菌门、酸杆菌门、放线菌门及厚壁菌门, 真菌的优势菌群为子囊菌门、担子菌门。细菌群落对比真菌群落而言对土壤增温的响应更为明显, 细菌菌群的相对丰度呈增加趋势, 真菌群落仅Hypocreales目相对丰度显著增加; 土壤细菌及真菌群落的丰富度均降低, 而群落多样性增加。增温影响了土壤细菌及真菌的群落结构及多样性, 且细菌群落对土壤增温更为敏感。     

真菌群落构建机制研究进展

真菌是真核生物中生物多样性最丰富的类群之一,广泛分布于自然界,在生物多样性的分布与维持、群落构建以及物质与能量循环过程中发挥着重要的生态功能。近年来随着DNA高通量测序技术和生物信息学的发展,极大推动了真菌多样性分布格局与群落构建机制的研究进展。本文概述了基于生态位理论(niche theory)和中性理论(neutral theory)的选择(selection)、扩散(dispersal)、成种(speciation)和漂变(drift)等过程与生态模型的定量分析确定性过程和随机性过程对群落构建贡献的机制,及其在真菌群落构建中应用的最新研究进展。     

土壤微生物多样性海拔格局研究进展

生物多样性的海拔分布格局与维持机制是生物多样性与生态系统功能研究的热点领域。相比动植物多样性海拔分布格局,土壤微生物多样性海拔分布格局的研究还处在起步阶段。近年来,随着以罗氏454、Illumina Mi Seq等为代表的高通量测序平台的发展,土壤微生物海拔梯度分布格局的研究进展较快。对土壤微生物多样性海拔分布格局最新研究综述发现,土壤微生物海拔分布模式并不明确,表现为无趋势、下降、单峰或者下凹型等多种海拔分布模式。这与大型动植物并不相同,暗示其驱动机制可能存在一定的差异。微生物由于其个体微小、扩散能力强以及较高的多样性和个体丰度而在局域尺度上可能更易受到气候环境因素的影响。土壤pH、碳、氮等因子是影响微生物多样性和群落组成在海拔梯度上变异的重要因素。此外,温度和降水也具有重要作用。另外,除微生物自身属性以及取样限制外,测序深度可能是影响土壤微生物物种丰富度海拔分布格局的重要因素。目前,对土壤微生物群落的研究在功能基因、群落构建机制以及生态学理论的验证方面还存在着不足。未来的研究应进一步加大测序深度,增加取样密度,着重关注全球气候变化及生物多样性丧失背景下土壤微生物群落的构建和维持机制及其生态系统功能等方面。     

污水处理过程中微生物群落多样性及其对环境因子响应的研究进展

污水生物处理系统的性能和稳定性与微生物群落结构和动态密切相关。通过深入了解活性污泥中微生物群落结构及其影响因素,有助于提高污水厂污染物的去除效果。在不同污水活性污泥处理系统中细菌群落主要以变形菌、绿弯菌、放线菌、厚壁菌和拟杆菌为功能菌群;活性污泥中寄居的大多数真菌来自于子囊菌门,还有少量担子菌门;古菌以产甲烷菌为主;而病毒中分布最广的噬菌体和致病性病毒是最主要的关注点。本文通过对相关文献分析及总结,综述了进水组成、不同处理工艺、参数(理化参数和运行参数)、地理位置和气候条件等环境因子对活性污泥中细菌、真菌、古菌以及病毒群落组成的影响,尽可能全面地介绍污水厂微生物群落多样性及其对环境因子的响应。同时,对未来研究方向进行探讨,以期能够为活性污泥中功能微生物的应用及调控提供理论和应用基础。     

五台山亚高山土壤真菌海拔分布格局与构建机制

生物多样性的海拔分布格局与维持机制是生物多样性与生态系统功能研究的热点领域。尽管微生物驱动着地球上许多重要的生物地球化学循环,但与大型生物体相比,对微生物多样性海拔梯度分布格局知之甚少。运用Illumina Miseq高通量测序技术,全面分析了五台山亚高山生态系统（海拔2000-3058 m范围内）土壤真菌群落组成和多样性变化特征。结果表明,子囊菌门（Ascomycota）、担子菌门（Basidiomycota）、接合菌门（Zygomycota）、壶菌门（Chytridimycota）为主要的优势菌门。非度量多维尺度分析（NMDS）和相似性分析（ANOSIM）表明真菌群落组成和结构在海拔梯度上存在显著的差异（P<0.05）。典范对应分析（CCA）显示pH、植物丰富度、总碳含量与土壤真菌群落结构存在显著相关性（P<0.05）。局域海拔尺度上,土壤真菌多样性与植物多样性（α和β多样性）呈显著正相关关系（P<0.05）。方差分解分析（VPA）和偏Mantel分析表明土壤真菌群落构建过程中,环境因子和空间变量都起作用,并且环境因子占绝对的优势。土壤真菌群落之间的Bray-Curtis距离与海拔距离呈显著正相关关系（P<0.05）,说明环境选择是亚高山土壤真菌海拔分布格局的决定因素。总之,五台山亚高山沿海拔梯度土壤真菌群落结构和多样性产生明显的变化,群落构建主要由确定性过程和随机过程驱动,但确定性过程占主导地位。土壤pH、植物丰富度、总碳含量是影响土壤真菌群落结构的重要因素。     

微生物性状揭示物种分布格局、群落构建机制和生态系统功能

微生物性状是指与其存活、生长和繁殖紧密相关的一系列核心属性,这些属性能够反映微生物对环境变化的响应,进而影响微生物的物种分布格局、群落构建机制以及相应的生态系统功能。越来越多的研究表明,相比于微生物分类学信息,微生物性状可以在种群、群落和生态系统尺度等视角扩展我们对微生物生态过程的理解,并提供生态模式的机理性解释。本文回顾微生物性状研究的发展历程,总结近年来基于微生物性状研究的前沿科学问题,比如微生物性状的分类和测定方法、基于性状的功能多样性定义及应用、性状与物种分布格局和群落构建机制的关系、性状对生物多样性和生态系统功能的影响以及对环境变化的响应等。尽管微生物性状研究已经延伸到生态学领域的各个方面,有力推动着各个前沿科学问题的研究发展,但是仍然面临很多机遇与挑战。因此,本文也从研究方法和研究方向等方面对未来基于微生物性状的研究提出了展望。     

海南东寨港红树林不同植被土壤微生物群落结构比较

【目的】比较不同植被下红树林土壤细菌和古菌的多样性及群落结构,认识红树林土壤微生物资源多样性。【方法】直接提取红树林土壤总DNA,采用细菌通用引物27F/1492R和古菌通用引物Arch21F/Arch958R进行PCR扩增,构建细菌和古菌16S rRNA基因文库,对海南东寨港自然保护区秋茄林、无瓣海桑林和无红树林裸滩土壤的细菌和古菌多样性和群落结构进行分析和比较。【结果】3种土壤样品的细菌类群包括变形细菌门(Proteobacteria)等16个类群,其中变形细菌门(Proteobacteria)与绿屈挠菌门(Chloroflexi)是优势类群;古菌包括6个嗜泉古菌界(Crenarchaeota)类群和7个广域古菌界(Euryarchaeota)类群,分别以Marine Benthic Group C、Marine Benthic Group D为优势类群。多样性指数(H’)和物种丰富度指数(Schao1)表明,本地种秋茄林下土壤细菌和古菌的多样性指数最高,外来种无瓣海桑显著低于秋茄林,甚至明显低于相邻无红树林裸滩沉积物;不同植被下土壤细菌和古菌群落结构存在显著差异,秋茄林土壤微生物群落结构和无红树林裸滩沉积物更相似。【结论】红树林土壤微生物类群丰富,不同植被下土壤细菌和古菌多样性和群落结构存在显著差异。     

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。     

土壤微生物网络复杂性预测黄土高原造林恢复生态系统多功能性

人类活动导致黄土高原土地退化和生物多样性丧失,进而降低了生态系统功能。人工造林是该区域退化土地恢复的重要措施。现有的生态修复研究通常侧重于微生物群落物种多样性的恢复对单一生态系统功能的影响,而忽略了微生物间存在的相互作用与生态系统多功能性(Ecosystem multifunctionality, EMF)的关系。为探究造林恢复过程中土壤微生物多样性和网络复杂性与EMF的关系,本研究采用时空代换法(space-time substitution method),沿50年造林恢复时间序列,分析了黄土高原地区造林恢复对土壤微生物群落多样性、土壤微生物网络复杂性以及与土壤养分循环相关的10个生态系统功能指标的影响,明确了土壤微生物群落特征与EMF的关系。结果表明,随造林恢复时间序列的增加,土壤微生物群落的综合多样性、网络复杂性和EMF均呈现出显著增加后下降的趋势(P<0.05),其中土壤微生物综合多样性和网络复杂性在第8年达到最高值,EMF在第20年达到最大值。在未控制土壤环境因素时,细菌和古菌多样性与EMF无显著相关性,真菌多样性与EMF呈显著正相关(P<0.001);土壤微生...     

Do bacterial and fungal communities assemble differently during primary succession?

High‐throughput sequencing technologies are now allowing us to study patterns of community assembly for diverse microbial assemblages across environmental gradients and during succession. Here we discuss potential explanations for similarities and differences in bacterial and fungal community assembly patterns along a soil chronosequence in the foreland of a receding glacier. Although the data are not entirely conclusive, they do indicate that successional trajectories for bacteria and fungi may be quite different. Recent empirical and theoretical studies indicate that smaller microbes (like most bacteria) are less likely to be dispersal limited than are larger microbes – which could result in a more deterministic community assembly pattern for bacteria during primary succession. Many bacteria are also better adapted (than are fungi) to life in barren, early‐successional sediments in that some can fix nitrogen and carbon from the atmosphere – traits not possessed by any fungi. Other differences between bacteria and fungi are discussed, but it is apparent from this and other recent studies of microbial succession that we are a long way from understanding the mechanistic underpinnings of microbial community assembly during ecosystem succession. We especially need a better understanding of global and regional patterns of microbial dispersal and what environmental factors control the development of microbial communities in complex natural systems.     

Dispersal alters soil microbial community response to drought

Microbial communities will experience novel climates in the future. Dispersal is now recognized as a driver of microbial diversity and function, but our understanding of how dispersal influences responses to novel climates is limited. We experimentally tested how the exclusion of aerially dispersed fungi and bacteria altered the compositional and functional response of soil microbial communities to drought. We manipulated dispersal and drought by collecting aerially deposited microbes after precipitation events and subjecting soil mesocosms to either filter-sterilized rain (no dispersal) or unfiltered rain (dispersal) and to either drought (25% ambient) or ambient rainfall for 6 months. We characterized community composition by sequencing 16S and ITS rRNA regions and function using community-level physiological profiles. Treatments without dispersal had lower soil microbial biomass and metabolic diversity but higher bacterial and fungal species richness. Dispersal also altered soil community response to drought; drought had a stronger effect on bacterial (but not fungal) community composition, and induced greater functional loss, when dispersal was present. Surprisingly, neither immigrants nor drought-tolerant taxa had higher abundance in dispersal treatments. We show experimentally that natural aerial dispersal rate alters soil microbial responses to disturbance. Changes in dispersal rates should be considered when predicting microbial responses to climate change.     

Diversity and biotechnological potential of the sponge-associated microbial consortia

Sponges are well known to harbor diverse microbes and represent a significant source of bioactive natural compounds derived from the marine environment. Recent studies of the microbial communities of marine sponges have uncovered previously undescribed species and an array of new chemical compounds. In contrast to natural compounds, studies on enzymes with biotechnological potential from microbes associated with sponges are rare although enzymes with novel activities that have potential medical and biotechnological applications have been identified from sponges and microbes associated with sponges. Both bacteria and fungi have been isolated from a wide range of marine sponge, but the diversity and symbiotic relationship of bacteria has been studied to a greater extent than that of fungi isolated from sponges. Molecular methods (e.g., rDNA, DGGE, and FISH) have revealed a great diversity of the unculturable bacteria and archaea. Metagenomic approaches have identified interesting metabolic pathways responsible for the production of natural compounds and may provide a new avenue to explore the microbial diversity and biotechnological potential of marine sponges. In addition, other eukaryotic organisms such as diatoms and unicellular algae from marine sponges are also being described using these molecular techniques. Many natural compounds derived from sponges are suspected to be of bacterial origin, but only a few studies have provided convincing evidence for symbiotic producers in sponges. Microbes in sponges exist in different associations with sponges including the true symbiosis. Fungi derived from marine sponges represent the single most prolific source of diverse bioactive marine fungal compounds found to date. There is a developing interest in determining the true diversity of fungi present in marine sponges and the nature of the association. Molecular methods will allow scientists to more accurately identify fungal species and determine actual diversity of sponge-associated fungi. This is especially important as greater cooperation between bacteriologists, mycologists, natural product chemists, and bioengineers is needed to provide a well-coordinated effort in studying the diversity, ecology, physiology, and association between bacteria, fungi, and other organisms present in marine sponges.     

中高温污泥厌氧消化系统中微生物群落比较

【目的】结合中温与高温消化两者优势的两相厌氧消化工艺可能是推进污泥厌氧消化发展的重要方向,因此,探究和比较中温和高温污泥厌氧消化系统中微生物群落组成的异同具有重要意义。【方法】利用高通量测序技术检测中温和高温厌氧消化系统中细菌与古菌的16S r RNA基因序列信息和真菌的内转录间隔(ITS)序列信息,利用基因芯片(Geo Chip 5.0)检测病毒和病原菌致病基因的信息,以对比中温和高温条件下微生物群落在物种组成和功能基因层面上的异同。【结果】中温和高温条件下细菌和古菌在群落物种组成上存在显著差异,病毒和病原菌毒性基因也显著不同,而两种系统中真菌群落的物种组成相似且丰度相对较低。中温条件下产甲烷古菌和未分类微生物相对丰度较高,而高温条件下产酸及嗜热菌相对丰度较高,且高温消化后病毒和病原菌毒性基因相对丰度下降。微生物群落结构与COD、TS和VS有着显著相关性。【结论】微生物群落组成和功能基因在中高温的污泥厌氧消化系统中显著不同,从而解释了两个系统功能的差异。微生物群落的形成与进水参数相关,说明微生物对进水条件敏感。     

Linking Microbial Community Structure and Function to Seasonal Differences in Soil Moisture and Temperature in a Chihuahuan Desert Grassland

Global and regional climate models predict higher air temperature and less frequent, but larger precipitation events in arid regions within the next century. While many studies have addressed the impact of variable climate in arid ecosystems on plant growth and physiological responses, fewer studies have addressed soil microbial community responses to seasonal shifts in precipitation and temperature in arid ecosystems. This study examined the impact of a wet (2004), average (2005), and dry (2006) year on subsequent responses of soil microbial community structure, function, and linkages, as well as soil edaphic and nutrient characteristics in a mid-elevation desert grassland in the Chihuahuan Desert. Microbial community structure was classified as bacterial (Gram-negative, Gram-positive, and actinomycetes) and fungal (saprophytic fungi and arbuscular mycorrhiza) categories using (fatty acid methyl ester) techniques. Carbon substrate use and enzymic activity was used to characterize microbial community function annually and seasonally (summer and winter). The relationship between saprophytic fungal community structure and function remained consistent across season independent of the magnitude or frequency of precipitation within any given year. Carbon utilization by fungi in the cooler winter exceeded use in the warmer summer each year suggesting that soil temperature, rather than soil moisture, strongly influenced fungal carbon use and structure and function dynamics. The structure/function relationship for AM fungi and soil bacteria notably changed across season. Moreover, the abundance of Gram-positive bacteria was lower in the winter compared to Gram-negative bacteria. Bacterial carbon use, however, was highest in the summer and lower during the winter. Enzyme activities did not respond to either annual or seasonal differences in the magnitude or timing of precipitation. Specific structural components of the soil microbiota community became uncoupled from total microbial function during different seasons. This change in the microbial structure/function relationship suggests that different components of the soil microbial community may provide similar ecosystem function, but differ in response to seasonal temperature and precipitation. As soil microbes encounter increased soil temperatures and altered precipitation amounts and timing that are predicted for this region, the ability of the soil microbial community to maintain functional resilience across the year may be reduced in this Chihuahuan Desert ecosystem.     

Evaluation of the ISO Standard 11063 DNA Extraction Procedure for Assessing Soil Microbial Abundance and Community Structure

Soil DNA extraction has become a critical step in describing microbial biodiversity. Historically, ascertaining overarching microbial ecological theories has been hindered as independent studies have used numerous custom and commercial DNA extraction procedures. For that reason, a standardized soil DNA extraction method (ISO-11063) was previously published. However, although this ISO method is suited for molecular tools such as quantitative PCR and community fingerprinting techniques, it has only been optimized for examining soil bacteria. Therefore, the aim of this study was to assess an appropriate soil DNA extraction procedure for examining bacterial, archaeal and fungal diversity in soils of contrasting land-use and physico-chemical properties. Three different procedures were tested: the ISO-11063 standard; a custom procedure (GnS-GII); and a modified ISO procedure (ISOm) which includes a different mechanical lysis step (a FastPrep ®-24 lysis step instead of the recommended bead-beating). The efficacy of each method was first assessed by estimating microbial biomass through total DNA quantification. Then, the abundances and community structure of bacteria, archaea and fungi were determined using real-time PCR and terminal restriction fragment length polymorphism approaches. Results showed that DNA yield was improved with the GnS-GII and ISOm procedures, and fungal community patterns were found to be strongly dependent on the extraction method. The main methodological factor responsible for differences between extraction procedure efficiencies was found to be the soil homogenization step. For integrative studies which aim to examine bacteria, archaea and fungi simultaneously, the ISOm procedure results in higher DNA recovery and better represents microbial communities.     

Molecular tools for deciphering the microbial community structure and diversity in rumen ecosystem

Rumen microbial community comprising of bacteria, archaea, fungi, and protozoa is characterized not only by the high population density but also by the remarkable diversity and the most complex microecological interactions existing in the biological world. This unprecedented biodiversity is quite far from full elucidation as only about 15-20?% of the rumen microbes are identified and characterized till date using conventional culturing and microscopy. However, the last two decades have witnessed a paradigm shift from cumbersome and time-consuming classical methods to nucleic acid-based molecular approaches for deciphering the rumen microbial community. These techniques are rapid, reproducible and allow both the qualitative and quantitative assessment of microbial diversity. This review describes the different molecular methods and their applications in elucidating the rumen microbial community.     

Burning Fire-Prone Mediterranean Shrublands: Immediate Changes in Soil Microbial Community Structure and Ecosystem Functions

Wildfires subject soil microbes to extreme temperatures and modify their physical and chemical habitat. This might immediately alter their community structure and ecosystem functions. We burned a fire-prone shrubland under controlled conditions to investigate (1) the fire-induced changes in the community structure of soil archaea, bacteria and fungi by analysing 16S or 18S rRNA gene amplicons separated through denaturing gradient gel electrophoresis; (2) the physical and chemical variables determining the immediate shifts in the microbial community structure; and (3) the microbial drivers of the change in ecosystem functions related to biogeochemical cycling. Prokaryotes and eukaryotes were structured by the local environment in pre-fire soils. Fire caused a significant shift in the microbial community structure, biomass C, respiration and soil hydrolases. One-day changes in bacterial and fungal community structure correlated to the rise in total organic C and NO(3)(-)-N caused by the combustion of plant residues. In the following week, bacterial communities shifted further forced by desiccation and increasing concentrations of macronutrients. Shifts in archaeal community structure were unrelated to any of the 18 environmental variables measured. Fire-induced changes in the community structure of bacteria, rather than archaea or fungi, were correlated to the enhanced microbial biomass, CO(2) production and hydrolysis of C and P organics. This is the first report on the combined effects of fire on the three biological domains in soils. We concluded that immediately after fire the biogeochemical cycling in Mediterranean shrublands becomes less conservative through the increased microbial biomass, activity and changes in the bacterial community structure.