近岸海洋线虫与细菌、古菌的共现性及其对碳、氮循环的影响

【目的】初步探究海洋线虫与微生物的相互作用对碳、氮循环的影响。【方法】利用16S r RNA和18S r RNA基因高通量测序方法,对33个近岸沉积物样品中细菌、古菌和真核生物的多样性进行调查;对海洋线虫与细菌、海洋线虫与古菌的共现性进行网络分析,并采用Spearman统计学方法,识别出与海洋线虫共现性呈显著相关性的微生物种类。【结果】在夏季,红树林和潮间带泥滩样品中线虫OTU平均相对丰度基本呈随深度增加而递减趋势;冬季的红树林样品中发现相类似变化规律,只有在冬季潮间带泥滩样品中线虫OTU平均相对丰度在深层较高于表层。相对丰度最高的海洋线虫隶属于单宫目(47%)、色矛目(19%)、刺嘴目(16%)和垫刃目(9%),它们与热源体古菌、深古菌、γ-和δ-变形菌等微生物有显著正/负相关关系。【结论】在香港米埔湿地沉积物中,与相对丰度最高的5种线虫显著相关的几大类微生物均在碳、氮、硫等元素循环方面起十分重要的作用,暗示海洋线虫与微生物潜在的相互作用对元素地球化学循环具有重要影响。研究结果有助于深入了解线虫在生态系统中未被揭示的生态功能,有助于更清晰地认识海洋线虫在底栖生态系统中所扮演的角色。     

海洋古菌多样性研究进展

海洋古菌是海洋微生物中的一个大的类群,然而绝大多数的古菌不能分离培养.近年来分子生物学的方法广泛地应用于微生物多样性的研究中,研究发现,海洋古菌广泛地生活在各类海域环境中,而不仅仅是生活在极端的环境中.海洋古菌为海洋生态系统中主要的原核细胞成分,在海洋生态系统中的物质与能量循环中扮演着重要角色.主要阐述了生活在海洋不同环境中海洋古菌的多样性,有海洋浮游古菌的多样性、海底环境及海洋沉积物中古菌的多样性、附着或寄共生古菌多样性等的研究状况,以及研究海洋古菌多样性的分子生物学的主要方法.     

现代分子生物技术在湖泊沉积物古DNA研究中的应用

分子生物学技术的发展,为从分子生物学角度探讨湖泊生态系统演化提供了一个新窗口。依据湖泊沉积物中古DNA研究,从古微生物、动植物群落结构组成及数量变化等方面开展研究,可进一步反演湖泊及周边的气候环境变化,探讨生物与气候、人类活动之间的关系。本文首先介绍了湖泊沉积物中古DNA来源、保存特点以及提取方法,其次比较分析了现代分子生物技术在湖泊沉积物古DNA中的应用特点,并阐述了湖泊沉积物古DNA在重建古气候、古生态以及人类活动对古环境影响方面的应用,最后对目前湖泊沉积物古DNA研究方法存在的一些问题进行了总结。     

深海微生物多样性

海洋面积约占地球总面积的70％,平均深度3,800 m,海底平均压力38 MPa,海水以下更是包含有物理化学性质迥异的多种地质结构,例如海洋沉积物、洋壳、热液口以及冷泉等.这些性质迥异的地质结构环境造就了丰富的生物多样性,构成了地球上最大的微生物生态系统.深海海水中最主要的微生物类群是α-,γ-变形菌(Alpha-&Gammaproteobacteria),以及海洋古菌群I(Marine Group I).深海沉积物中微生物含量与有机物含量和距离大陆板块的距离相关,以异养微生物为主.深海冷泉区富集了厌氧甲烷氧化古菌ANME和硫酸盐还原菌(Deltaproteobacteria);深海热液区由于具有化学物质的多样性和快速的动态变化而导致形成微生物的高度多样性.洋壳主要由基性、超基性岩构成,含有丰富的矿物,其中不乏参与铁、锰、硫等关键代谢反应的化能自养微生物.同时,由于环境中99％以上的微生物没有已培养的亲缘种,因此对深海微生物的多样性、生理功能特性以及生物地球化学作用的理解和研究仍然存在巨大的挑战.本文将尝试从不同的深海环境分区来综述深海海水、沉积物、洋壳,以及冷泉区和热液口等特殊生态环境中微生物的分布和多样性.     

北部湾红树林沉积物中微生物群落结构的时空变化分析

微生物群落结构对海洋红树林湿地生态系统中的物质循环非常关键。本研究从中国亚热带北部湾典型海洋红树林湿地生态系统中采集沉积物样品,利用宏基因组学技术手段,完成了沉积物样品16S rDNA基因的扩增子的高通量测序,分析了不同时空条件下红树林沉积物的微生物群落结构分布和变化规律。研究结果表明,不管在干季还是湿季,在门分类水平上变形菌门(Proteobacteria)是最丰富的类别。此外,变形菌门(Proteobacteria)与绿弯菌门(Chloroflexi)、拟杆菌门(Bacteroidetes)、酸杆菌门(Acidobacteria)、硝化螺旋菌门(Nitrospirae)和放线菌门(Actinobacteria)一共占据总丰度的80%左右。干季时期的微生物α多样性显著高于湿季,主坐标分析(principal co-ordinates analysis, PCoA)显示干、湿两季的微生物群落结构具有明显差异。干季时期的细菌的相对丰度显著高于湿季,而古菌则相反。不同时空期(干季和湿季)的变形菌门(Proteobacteria)的相对丰度没有显著差异。冗余分析(redundancy analysis, RDA)和相关性分析表明温度和盐度对微生物群落结构具有显著影响。本研究对揭示亚热带海洋红树林湿地生态系统沉积物中的微生物多样性及其群落结构演替规律具有重要的实践意义。     

南海Formosa冷泉区沉积物微生物多样性与分布规律研究

【目的】当前对全球冷泉生态系统微生物生态学研究显示，冷泉生态系统中主要微生物类群为参与甲烷代谢的微生物，它们的分布差异与所处冷泉区生物地球化学环境密切相关。但在冷泉区内也存在环境因子截然不同的生境，尚缺乏比较冷泉区内小尺度生境间微生物多样性和分布规律的研究。本研究旨在分析南海Formosa冷泉区内不同生境间微生物多样性差异，完善和理解不同环境因子对冷泉内微生物群落结构的影响。【方法】对采集自南海Formosa冷泉区不同生境(黑色菌席区、白色菌席区和碳酸盐岩区)沉积物样本中古菌和细菌16S rRNA基因进行测序，结合环境因子，比较微生物多样性差异，分析环境因子对微生物分布的影响。【结果】发现在Formosa冷泉内的不同生境中，甲烷厌氧氧化古菌(anaerobic methanotrophic archaea,ANME)是主要古菌类群，占古菌总体相对丰度超过70%；在菌席区ANME-1b和ANME-2a/b是主要ANME亚群，碳酸盐岩区则是ANME-1b。硫酸盐还原菌(sulfate-reducing bacteria,SRB)和硫氧化菌(sulfur-oxidizing bacteria...     

新疆天山北坡不同盐湖微生物菌群结构及其影响因子

新疆分布的众多湖泊由于干旱气候成盐作用强烈,近半数已演化到盐湖发展阶段,不同盐湖中也因此蕴含着丰富的耐盐及嗜盐微生物资源。为更好的掌握新疆盐湖微生物资源分布规律及对环境因子变化的响应规律,利用高通量测序技术对新疆天山北坡5个不同演化阶段盐湖湖底沉积物中细菌、古菌多样性和菌群结构及其主要驱动因子进行研究,探讨盐湖演化过程中原核微生物群落结构变化规律。分别采集5个盐湖湖底沉积物样本,进行理化因子测试与细菌和古菌16S rRNA扩增子测序分析,比较不同盐湖理化性质和原核微生物菌群差异,并对原核微生物丰度与环境因子进行关联分析。实验结果表明：5个盐湖湖底沉积物总盐和Na⁺含量顺序为：巴里坤湖 > 伊吾湖 > 艾比湖 > 盐湖 > 柴窝堡湖,除艾比湖外其他四个盐湖沉积物均呈碱性。Alpha多样性结果显示5个盐湖细菌richness、chao1、ACE和shannon丰富度指数均大于古菌相应丰富度指数,不同盐湖细菌丰富度指数差异较大,古菌丰富度指数差异相对较小。从5个盐湖湖底沉积物中共检测获得细菌58门、68纲、138目、253科和560属,古菌4门、8纲、12目、21科和60属,细菌以变形菌门为主,古菌以广古菌门为主。不同盐湖细菌和古菌优势属种类均不相同,巴里坤湖主要是一些嗜盐和耐盐细菌属,而伊吾湖主要是嗜盐和耐盐古菌属,PCoA分析结果也表明不同盐湖微生物在OTUs水平有其独特菌群结构类型。RDA和Bioenv分析结果表明,盐湖湖底沉积物中微生物菌群群落结构主要受Na⁺和总盐（TS）浓度的影响,对细菌菌群结构影响较大,而古菌菌群结构可能受多种理化因子共同调节。此外,盐湖特殊卤水成分会对微生物群落结构产生重大影响。     

海州湾近岸海域的微生物群落结构和抗生素抗性基因时空变化

【背景】近岸海域抗生素抗性基因(antibiotic resistance genes, ARGs)的污染和累积将直接影响海产品质量和安全,海州湾作为江苏省的四大渔场之一,是江苏渔业发展的主要载体,有多条大小河流注入,沿岸为重要农业区,对公众健康产生重大影响。【目的】对海州湾夏秋季的水样及沉积物展开微生物及ARGs检测。【方法】基于宏基因组测序技术开展海州湾夏秋两季近岸6个站点中水体和沉积物中ARGs种类和相对丰度以及微生物群落的组成研究。【结果】变形菌门(Proteobacteria)和放线菌门(Actinobacteria)是夏秋季度两种介质中最优势的门类,水样中优势的科级细菌为红细菌科(Rhodobacteraceae),沉积物样品中为脱硫杆菌科(Desulfobacteraceae);夏季水样中的ARGs相对丰度要明显高于秋季,但沉积物中不同季节的ARGs相对丰度未表现出明显的变化趋势;在水样中主要门级微生物群落的抗性机制主要是抗生素靶位替换和抗生素靶位保护,沉积物样品则以抗生素灭活机制为主,而主要科级微生物群落的抗性机制更加多样;冗余分析(redundancyanalysis...     

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。     

河流沉积物氮循环主要微生物的生态特征

微生物驱动的氮循环过程是全球生物地球化学循环的重要组成部分,由于人类活动的影响,氮循环负荷加剧,氮素的生态平衡和微生物的功能特征也相应地受到干扰。河流生态系统是陆地与海洋联系的纽带,因人类活动过量活性氮的输入导致水体富营养化,明显影响着河流的生态功能以及河口沿岸海洋生态系统的平衡。富含微生物的沉积物对氮素的转化和去除起着至关重要的作用。本文主要介绍河流沉积物氮循环主要功能微生物,包括氨氧化细菌、氨氧化古菌、亚硝酸盐氧化菌、反硝化细菌和厌氧氨氧化细菌的群落特征和生态功能,总结氮相关营养盐、溶氧和季节变化等环境因子,以及河道控制管理措施和污水处理厂扰动等条件下氮循环过程主要功能类群的生态特征和响应关系。指出还需深入全面地研究河流沉积物生态系统氮循环过程的驱动机制和微生物的贡献效率,加强城市河流沉积物微生物功能作用的研究及河道生物修复技术的开发。     

The Variation of Microbial Communities in a Depth Profile of Peat in the Gahai Lake Wetland Natural Conservation Area

The Gahai Lake wetland natural conservation area in northwestern China includes peatland that has been accumulating over hundreds of years and is seldom disturbed by industry. Bacteria and archaea in peat soil, which is a reservoir for carbon and water, may influence its ecological function. The objective of this study was to obtain a clearer understanding of peat microbial ecology and its relationship to the environmental conditions of this area. Hence, the microbial community of the peatland ecosystem was investigated by sequencing bacterial and archaeal DNA extracted from samples collected at different peat depths. Results showed that in all samples the dominant bacterial phyla were Proteobacteria (relative abundance 0.39 ± 0.12) and Chloroflexi (0.16 ± 0.09), while the dominant archaeal phyla were Miscellaneous Crenarchaeotic Group (MCG) (0.62 ± 0.21) and Euryarchaeota (0.27 ± 0.16). The diversity and microbial community structure at deeper depths (90 and 120 cm below the peat surface) significantly differ from that at shallower depths (10, 30 and 50 cm deep). In contrast to the shallow layers, the deeper layers became more abundant in the bacterial phyla Chloroflexi, Bacteroidetes, Atribacteria, Aminicenantes, Chlorobi, TA06, Caldiserica and Spirochaetae; and in the archaeal phyla MCG and Miscellaneous Euryarchaeotic Group (MEG). This study revealed a significant shift in microbial community in peat between 50 cm and 90 cm deep, as probably influenced by the oxygen supply at different depths. Furthermore, new insights into the microbial taxa were obtained, thus providing a baseline for future studies of this peat ecosystem.     

Microbial Communities of Deep Marine Subsurface Sediments: Molecular and Cultivation Surveys

Recent molecular analyses show that microbial communities of deep marine sediments harbor members of distinct, uncultured bacterial and archaeal lineages, in addition to Gram-positive bacteria and Proteobacteria that are detected by cultivation surveys. Several of these subsurface lineages show cosmopolitan occurrence patterns; they can be found in cold marine sediments and also in hydrothermal habitats, suggesting a continuous deep subsurface and hydrothermal biosphere with shared microbiota. The physiologies and activities of these uncultured subsurface lineages remain to be explored by innovative combinations of genomic and biogeochemical approaches.     

Microbiological Assessment of Circulation Mud Fluids During the First Operation of Riser Drilling by the Deep-Earth Research Vessel Chikyu

Quality assurance and control (QA/QC) is significant for the scientific drilling in order to accurately characterize physical, geochemical, and biological properties in the cored deep subseafloor materials. To explore the deep subseafloor life and its biosphere, identification and control of microbial contamination in drilling cores is critical for highly sensitive molecular analyses as well as cultivations, especially for the evaluation of low biomass and/or extremely harsh deep environments. Here we report some microbiological characteristics of circulation mud fluids before and after the first riser drilling operation by the newly constructed deep-earth research vessel Chikyu. During the Chikyu shakedown expedition CK06-06 in 2006, we used the riser system for drilling 547 to 647 meter below the seafloor into the sediments offshore the Shimokita Peninsula of Japan. Cultivation experiments showed that no microbial growth was observed in the precirculation mud fluid, while 4 × 10⁵ colonies per 1 ml were observed in the postcirculation mud fluid; all cultured bacterial isolates were found to be Halomonas. Using culture-independent molecular analysis, 16S rRNA gene sequences of Xanthomonas, which is used for industrial production of the mud fluid viscosifier “xanthan gum”, were predominantly detected in the precirculation mud fluid, while Halomonas sequences consistently dominated the clone library constructed from the postcirculation mud fluid. Archaeal 16S rRNA genes were amplified only from the postcirculation mud fluid; these archaeal clone sequences were affiliated to the Marine Crenarchaeota Group I (MGI), Marine Euryarchaeota Group II (MGII), Miscellaneous Crenarchaeotic Group (MCG), South African Gold Mine Euryarchaeotic Group (SAGMEG), Soil Group, and Methanococcus aeolicus. These results suggest that Halomonas contaminated and grew in the tank of circulation mud fluids, and other indigenous deep subseafloor microbial components, especially deep subsurface archaea, were also mixed into the post-circulation mud fluid.     

Thermophilic archaeon orchestrates temporal expression of GDGT ring synthases in response to temperature and acidity stress

Glycerol dibiphytanyl glycerol tetraethers (GDGTs) are unique archaeal membrane-spanning lipids with 0–8 cyclopentane rings on the biphytanyl chains. The cyclization pattern of GDGTs is affected by many environmental factors, such as temperature and pH, but the underlying molecular mechanism remains elusive. Here, we find that the expression regulation of GDGT ring synthase genes grsA and grsB in thermophilic archaeon Sulfolobus acidocaldarius is temperature- and pH-dependent. Moreover, the presence of functional GrsA protein, or more likely its products cyclic GDGTs rather than the accumulation of GrsA protein itself, is required to induce grsB expression, resulting in temporal regulation of grsA and grsB expression. Our findings establish a molecular model of GDGT cyclization regulated by environment factors in a thermophilic ecosystem, which could be also relevant to that in mesophilic marine archaea. Our study will help better understand the biological basis for GDGT-based paleoclimate proxies. Archaea inhabit a wide range of terrestrial and marine environments. In response to environment fluctuations, archaea modulate their unique membrane GDGTs lipid composition with different strategies, in particular GDGTs cyclization significantly alters membrane permeability. However, the regulation details of archaeal GDGTs cyclization in response to different environmental factor changes remain unknown. We demonstrated, for the first time, thermophilic archaea orchestrate the temporal expression of GDGT ring synthases, leading to delicate control of GDGTs cyclization to respond environmental temperature and acidity stress. Our study provides insight into the regulation of archaea membrane plasticity, and the survival strategy of archaea in fluctuating environments.     

Unique Microbial Community in Drilling Fluids from Chinese Continental Scientific Drilling

Circulating drilling fluid is often regarded as a contamination source in investigations of subsurface microbiology. However, it also provides an opportunity to sample geological fluids at depth and to study contained microbial communities. During our study of deep subsurface microbiology of the Chinese Continental Scientific Deep drilling project, we collected 6 drilling fluid samples from a borehole from 2290 to 3350 m below the land surface. Microbial communities in these samples were characterized with cultivation-dependent and -independent techniques. Characterization of 16S rRNA genes indicated that the bacterial clone sequences related to Firmicutes became progressively dominant with increasing depth. Most sequences were related to anaerobic, thermophilic, halophilic or alkaliphilic bacteria. These habitats were consistent with the measured geochemical characteristics of the drilling fluids that have incorporated geological fluids and partly reflected the in-situ conditions. Several clone types were closely related to Thermoanaerobacter ethanolicus, Caldicellulosiruptor lactoaceticus, and Anaerobranca gottschalkii, an anaerobic metal-reducer, an extreme thermophile, and an anaerobic chemoorganotroph, respectively, with an optimal growth temperature of 50–68°C. Seven anaerobic, thermophilic Fe(III)-reducing bacterial isolates were obtained and they were capable of reducing iron oxide and clay minerals to produce siderite, vivianite, and illite. The archaeal diversity was low. Most archaeal sequences were not related to any known cultivated species, but rather to environmental clone sequences recovered from subsurface environments. We infer that the detected microbes were derived from geological fluids at depth and their growth habitats reflected the deep subsurface conditions. These findings have important implications for microbial survival and their ecological functions in the deep subsurface.     

A Window to the Subsurface: Microbial Diversity in Hot Springs of a Sulfidic Cave (Kaklik,Turkey)

Caves are windows to the extreme habitats of deep subsurface, and provide answers of unknowns about the underground life. Furthermore, sulfidic caves are important analogues for the early Earth environments, since some environmental conditions are common, such as high sulfur concentration, high temperature and oxygen-poor conditions. Kaklik Cave (Denizli, Turkey) with its travertine formation, carbonate- and sulfur-rich thermal springs, exhibits a unique ecosystem as a sulfidic cave. This study represents the first molecular survey of the microbial community in the Kaklik Cave, Turkey using high-throughput 16S rRNA gene amplicon sequencing analysis. An average of 859–2,416 operational taxonomic units per sample were observed including 25 bacterial phyla and 3 archaeal phyla. The bacterial diversity profiles were generally dominated by Epsilonproteobacteria and Gammaproteobacteria. At the carbonate-rich hot spring, that formed travertine structure, 9.7% of sequence reads affiliated with Thiofaba spp. In contrast, 38.74% of the total sequence reads at the sulfidic hot spring samples associated with the genus Sulfurimonas and Sulfurovum. In the archaeal community composition, Thermoplasmata was the most abundant group in all sampling areas. The 454-pyrotag results provide leads about ammonia-, nitrite- and sulfur-oxidation as well as sulfur-reduction, carbon dioxide fixation, and nitrogen fixation.     

Insights into the Phylogeny and Metabolic Potential of a Primary Tropical Peat Swamp Forest Microbial Community by Metagenomic Analysis

A primary tropical peat swamp forest is a unique ecosystem characterized by long-term accumulation of plant biomass under high humidity and acidic water-logged conditions, and is regarded as an important terrestrial carbon sink in the biosphere. In this study, the microbial community in the surface peat layer in Pru Toh Daeng, a primary tropical peat swamp forest, was studied for its phylogenetic diversity and metabolic potential using direct shotgun pyrosequencing of environmental DNA, together with analysis of 16S rRNA gene library and key metabolic genes. The community was dominated by aerobic microbes together with a significant number of facultative and anaerobic microbial taxa. Acidobacteria and diverse Proteobacteria (mainly Alphaproteobacteria) constituted the major phylogenetic groups, with minor representation of archaea and eukaryotic microbes. Based on comparative pyrosequencing dataset analysis, the microbial community showed high metabolic versatility of plant polysaccharide decomposition. A variety of glycosyl hydrolases targeting lignocellulosic and starch-based polysaccharides from diverse bacterial phyla were annotated, originating mostly from Proteobacteria, and Acidobacteria together with Firmicutes, Bacteroidetes, Chlamydiae/Verrucomicrobia, and Actinobacteria, suggesting the key role of these microbes in plant biomass degradation. Pyrosequencing dataset annotation and direct mcrA gene analysis indicated the presence of methanogenic archaea clustering in the order Methanomicrobiales, suggesting the potential on partial carbon flux from biomass degradation through methanogenesis. The insights on the peat swamp microbial assemblage thus provide a valuable approach for further study on biogeochemical processes in this unique ecosystem.     

Subsurface and surface halophile communities of the chaotropic Salar de Uyuni

Salar de Uyuni (SdU) is the biggest athalosaline environment on Earth, holding a high percentage of the known world Li reserves. Due to its hypersalinity, temperature and humidity fluctuations, high exposure to UV radiation, and its elevated concentration of chaotropic agents like MgCl₂, LiCl and NaBr, SdU is considered a polyextreme environment. Here, we report the prokaryotic abundance and diversity of 46 samples obtained in different seasons and geographical areas. The identified bacterial community was found to be more heterogeneous than the archaeal community, with both communities varying geographically. A seasonal difference has been detected for archaea. Salinibacter, Halonotius and Halorubrum were the most abundant genera in Salar de Uyuni. Different unclassified archaea were also detected. In addition, the diversity of two subsurface samples obtained at 20 and 80 m depth was evaluated and compared with the surface data, generating an evolutionary record of a multilayer hypersaline ecosystem.     

Colonization of subsurface microbial observatories deployed in young ocean crust

Oceanic crust comprises the largest hydrogeologic reservoir on Earth, containing fluids in thermodynamic disequilibrium with the basaltic crust. Little is known about microbial ecosystems that inhabit this vast realm and exploit chemically favorable conditions for metabolic activities. Crustal samples recovered from ocean drilling operations are often compromised for microbiological assays, hampering efforts to resolve the extent and functioning of a subsurface biosphere. We report results from the first in situ experimental observatory systems that have been used to study subseafloor life. Experiments deployed for 4 years in young (3.5 Ma) basaltic crust on the eastern flank of the Juan de Fuca Ridge record a dynamic, post-drilling response of crustal microbial ecosystems to changing physical and chemical conditions. Twisted stalks exhibiting a biogenic iron oxyhydroxide signature coated the surface of mineral substrates in the observatories; these are biosignatures indicating colonization by iron oxidizing bacteria during an initial phase of cool, oxic, iron-rich conditions following observatory installation. Following thermal and chemical recovery to warmer, reducing conditions, the in situ microbial structure in the observatory shifted, becoming representative of natural conditions in regional crustal fluids. Firmicutes, metabolic potential of which is unknown but may involve N or S cycling, dominated the post-rebound bacterial community. The archaeal community exhibited an extremely low diversity. Our experiment documented in situ conditions within a natural hydrological system that can pervade over millennia, exemplifying the power of observatory experiments for exploring the subsurface basaltic biosphere, the largest but most poorly understood biotope on Earth.