泗顶矿区6种土地利用类型土壤微生物群落结构特征

以广西柳州泗顶铅锌矿区的玉米地、菜地、灌木、尾矿库、森林和柑橘园6种不同利用类型土壤为研究对象,采用高通量测序技术,分析了不同土地利用类型土壤微生物群落结构特征及土壤环境因子对微生物群落结构的影响.结果表明,变形菌门（Proteobacteria）是6种土壤的优势菌门,所占比例为31.34%~53.47%,γ-变形菌纲（Gammaproteobacteria）为优势菌纲.冗余分析表明,在重金属含量最高的灌木区,厚壁菌门（Firmicutes）和拟杆菌门（Bacteroidetes）的丰度与土壤中铅、锌、镉、铜的含量呈现显著正相关;在土壤钠和钾含量较高的玉米地和菜地,浮霉菌门（Planctomycetes）的丰度有所增加,而厚壁菌门和拟杆菌门的丰度则有所下降.在土壤有机质含量较高而总氮、总磷含量较低的柑橘园,酸杆菌门（Acidobacteria）和疣微菌门（Verrucomicrobia）的丰度较高.芽单胞菌门（Gemmatimonadetes）和浮霉菌门的丰度与土壤pH值呈显著正相关.研究结果表明土地利用类型及土壤环境因子对土壤微生物群落结构多样性有显著的影响.     

响应土壤阴离子类型的盐碱土古细菌群落多样性研究

为初步解析我国黑龙江苏打盐碱地（HA）、新疆荒漠盐碱地（XD）、山西平原盐碱地（SY）、江苏滨海盐碱地（JD）和天津滨海盐碱地（TB）5个地区的土壤古细菌群落结构多样性,以及其响应土壤不同阴离子类型的分布特征,采用基于Illumin-Hiseq平台的高通量测序技术获得489~604个古细菌可操纵分类单元（OTU）.OTU多样性分析结果表明XD的古细菌群落物种丰富度最高,SY和HA分别具有最高和最低的古细菌群落多样性和均匀度;OTU的物种注释结果表明HA以奇古菌门（Thaumarchaeota）为优势菌门,其他土壤均以广古菌（Euryarchaeota）为优势;综合5个样品古细菌群落结构、优势古细菌属和土壤阴离子进行的典范对应分析（CCA）结果显示,XD古细菌群落结构和Haloterrigena属（XD中最优势,相对丰度20.2%）明显的响应土壤SO₄^2-浓度;HA古细菌群落结构和Nitrososphaera属（HA中最优势,62.3%）明显的响应土壤HCO₃^-/CO₃^2-浓度和pH;JD、SY和TB古细菌群落结构、Halorubrum属（JD和TB中最优势,24.4%和15.6%）,和Natronomonas属（SY和JD中第二优势,8.1%和9.2%）明显的响应土壤Cl^-浓度.研究结果说明我国不同地域分布的盐碱土受其土壤阴离子类型及其浓度的影响,呈现不同的古细菌群落结构和分布特征.研究结果为我国不同类型盐碱土古细菌资源的挖掘提供依据,也为揭示我国不同类型盐碱土古细菌群落的生态功能提供参考.     

模拟气候升温对湿地土壤微生物群落及磷素形态的影响

通过构建微宇宙湿地柱模拟气候升温的方法,采用高通量测序和核磁共振技术,分别研究了湿地土壤微生物群落和磷素形态对暖化作用的响应特征.结果表明,暖化作用导致了Firmicutes、Clostridia、Clostridiales、Clostridiaceae和Clostridium的相对丰度分别显著下降65%~98%、69%~87%、67%~87%、73%~97%和74%~93%,这表明暖化作用对不同分类水平上的物种Firmicutes到Clostridium具有显著的抑制效应.通过主坐标分析和聚类分析,不同湿地柱采样点的暖化组与对照组样本表现出显著的分离特征,揭示了暖化作用能够诱导微生物群落组成发生显著性变化.磷酸单酯和正磷酸盐是各湿地柱土壤主导的磷素形态,同时暖化作用导致了XX湿地柱采样点的磷酸单酯和正磷酸盐相对丰度分别显著升高275%和下降20%,JH湿地柱采样点的磷酸单酯和多聚磷酸盐相对丰度分别显著升高85%和下降49%,这表明不同磷素形态对暖化作用的响应具有土壤异质性特征.通过典型对应分析,揭示了暖化条件下微生物群落组成的显著变化对磷素形态具有显著的影响效应.     

采用颜色空间表征ANAMMOX启动与冲击过程

厌氧氨氧化(ANAMMOX)是一种高效、节能的脱氮技术,但是其启动过程和稳定状态的表征目前仍缺乏一个简单易行的方法.针对这一问题,在水质分析的基础上,采用颜色空间重点监测ANAMMOX启动过程中污泥的颜色变化,并借助高通量测序技术对微生物菌群的种类和数量等进行分析.结果表明:(1)根据水质特征,整个启动过程可分为活性迟滞期、活性增强期、负荷提高期、稳定运行期.与此同时,HSV、CIELAB颜色空间的指标,整体上呈现先降低后升高的趋势,然后保持稳定.污泥颜色的变化与水质的变化以及分子生物学特征中优势菌群的变迁一致,因此,三者之间具有相关性,进而表明,采用颜色空间可对ANAMMOX启动过程进行准确表征;(2)当遇到高负荷引起的冲击时,H、S、a~*、b~*、C_(ab)~*等指标与总无机氮(TIN)容积去除速率均呈现降低的趋势,而H_(ab)出现了突升,但均能准确反映出系统的冲击特性.提出了基于颜色空间表征ANAMMOX启动各阶段的方法并建立了一套冲击过程指标体系,为颜色空间在ANAMMOX系统中的应用提供了理论基础.     

同步脱氮除磷系统中两种颜色好氧颗粒污泥的微生物群落特征

为研究同步脱氮除磷系统中出现的WG（白色好氧颗粒污泥）外形特点及微生物群落特征,探究其成因,利用SEM（扫描电子显微镜）表征了系统中的WG与YG（黄色好氧颗粒污泥）的微观形态,并采用Illumina HiSeq 2500高通量测序平台对两种好氧颗粒污泥中细菌与真菌的群落组成进行研究.结果表明:WG结构疏松外形不规则,颗粒表面分布大量杆菌;而YG饱满紧实轮廓清晰,颗粒表面分布大量球菌.WG与YG的细菌群落组成相似,但真菌组成差异较大.与YG相比,WG具有更高的细菌和真菌多样性.变形菌门（Proteobacteria）和拟杆菌门（Bacteroidetes）为WG和YG中的细菌优势门,其在WG中的相对丰度分别为68.85%和26.61%,在YG中的相对丰度分别为82.52%和12.30%.Candidatus competibacter、Candidatus accumulibacter和Chiayiivirga为WG中的优势属,相对丰度分别为22.13%、8.95%和7.37%;Candidatus competibacter、Chiayiivirga和Xanthomonas为YG中的优势属,相对丰度分别为47.94%、6.95%和7.06%.子囊菌门（Ascomycota）和Rozellomycota分别为WG与YG中真菌优势门,其在两个样品中的相对丰度分别为50.10%和81.77%.在属水平,WG中存在大量青霉属（Penicillium）和假丝酵母属（Candida）等丝状真菌,为WG的形成提供了框架.研究显示,当YG破碎成为小菌胶团后,附着在真菌框架上,造成了WG的快速形成,同时WG中Candidatus competibacter的相对丰度较低,使其外形疏松、透光性较好,呈现出白色.      

富营养化淡水中黑色金属试样腐蚀加速的微生物学机制研究

目的探究黑色金属在污染淡水中后期腐蚀加速规律的生物学机制。方法采用高通量测序分析A3钢在富营养化水体和清洁水体中腐蚀产物内微生物的种群结构,并结合元素分析和XPS分析。结果获得了A3钢的腐蚀产物内微生物的门水平物种分布柱状图、OTU估计数统计表、OTU韦恩图、硫酸盐还原菌各属的相对丰度、微生物属水平热图等数据。结论在富营养化水体中,硫酸盐还原菌呈现更显著的水平,造成金属的硫化腐蚀更为严重,验证了黑色金属在富营养化污染淡水水体中后期腐蚀速率反转加速的现象。     

A fluorescence-based bioassay for aquatic macrophytes and its suitability for effect analysis of non-photosystem II inhibitors

Background, Goals and Scope During the last years the miniaturization of toxicity test systems for rapid and parallel measurements of large quantities of samples has often been discussed. For unicellular algae as well as for aquatic macrophytes, fluorescence-based miniaturized test systems have been introduced to analyze photosystem II (PSII) inhibitors. Nevertheless, high-throughput screening should also guarantee the effect detection of a broad range of toxicants in order to ensure routinely applicable, high-throughput measuring device experiments which can cover a broad range of toxicants and modes of action others than PSII inhibition. Thus, the aim of this study was to establish a fast and reproducible measuring system for non-PSII inhibitors for aquatic macrophyte species to overcome major limitations for use. Methods A newly developed imaging pulse-amplitude-modulated chlorophyll fluorometer (I-PAM) was applied as an effect detector in short-term bioassays with the aquatic macrophyte species Lemna minor. This multiwell-plate based measuring device enabled the incubation and measurement of up to 24 samples in parallel. The chemicals paraquat-dichloride, alizarine and triclosan were chosen as representatives for the toxicant groups of non-PSII herbicides, polycyclic aromatic hydrocarbons (PAHs) and pharmaceuticals and personal care products (PPCPs), which are often detected in the aquatic environment. The I-PAM was used (i) to establish and validate the sensitivity of the test system to the three non-PSII inhibitors, (ii) to compare the test systems with standardized and established biotests for aquatic macrophytes, and (iii) to define necessary time scales in aquatic macrophyte testing. For validation of the fluorescence-based assay, the standard growth test with L. minor (ISO/DIS 20079) was performed in parallel for each chemical. Results The results revealed that fluorescence-based measurements with the I-PAM allow rapid and parallel analysis of large amounts of aquatic macrophyte samples. The I-PAM enabled the recording of concentration-effect-curves with L. minor samples on a 24-well plate with single measurements. Fluorescence-based concentration-effect-curves could be detected for all three chemicals after only 1 h of incubation. After 4–5 h incubation time, the maximum inhibition of fluorescence showed an 80–100% effect for the chemicals tested. The EC50 after 24 h incubation were estimated to be 0.06 mg/L, 0.84 mg/L and 1.69 mg/L for paraquatdichloride, alizarine and triclosan, respectively. Discussion The results obtained with the I-PAM after 24 h for the herbicide paraquat-dichloride and the polycyclic aromatic hydrocarbon alizarine were in good accordance with median effective concentrations (EC50s) obtained by the standardized growth test for L. minor after 7 d incubation (0.09 mg/L and 0.79 mg/L for paraquat-dichloride and alizarine, respectively). Those results were in accordance with literature findings for the two chemicals. In contrast, fluorescence-based EC50 of the antimicrobial agent triclosan proved to be two orders of magnitude greater when compared to the standard growth test with 7 d incubation time (0.026 mg/L) as well as with literature findings. Conclusion Typically, aquatic macrophyte testing is very time consuming and relies on laborious experimental set-ups. The I-PAM measuring device enabled fast effect screening for the three chemicals tested. While established test systems for aquatic macrophytes need incubation times of ≥ 7 d, the I-PAM can detect inhibitory effects much earlier (24 h), even if inhibition of chemicals is not specifically associated with PSII. Thus, the fluorescence-based bioassay with the I-PAM offers a promising approach for the miniaturization and high-throughput testing of chemicals with aquatic macrophytes. For the chemical triclosan, however, the short-term effect prediction with the I-PAM has been shown to be less sensitive than with long-term bioassays, which might be due to physicochemical substance properties such as lipophilicity. Recommendations and Perspectives The results of this study show that the I-PAM represents a promising tool for decreasing the incubation times of aquatic macrophyte toxicity testing to about 24 h as a supplement to existing test batteries. The applicability of this I-PAM bioassay on emergent and submerged aquatic macrophyte species should be investigated in further studies. Regarding considerations that physicochemical properties of the tested substances might play an important role in microplate bioassays, the I-PAM bioassay should either be accompanied by evaluating physicochemical properties modeled from structural information prior to an experimental investigation, or by intensified chemical analyses to identify and determine nominal concentrations of the toxicants tested. The chemicals paraquat-dichloride, alizarine and triclosan were chosen as representatives for the toxicant groups of non-PSII herbicides, PAHs and PPCPs which are often detected in the aquatic environment. Nevertheless, in order to ensure a routinely applicable measuring device, experiments with a broader range of toxicants and samples of surface and/or waste waters are necessary. ESS-Submission Editor: Dr. Markus Hecker (MHecker@Entrix.com)     

贵州喀斯特地区土壤细菌群落结构特征及变化

为探明贵州喀斯特不同植被演替群落下的土壤细菌群落结构及变化特征,本文利用高通量测序技术对5个主要植被演替群落(稀灌草丛、藤刺灌丛、灌木林、乔灌过渡林、乔木林)的根际(竹叶椒)、非根际土壤细菌群落结构及环境因子进行了分析研究。结果表明:贵州喀斯特高原土壤细菌类群主要为变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、疣微菌门(Verrucomicrobia)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和浮霉菌门(Planctomycetes),其相对丰富度分别为43.35%、12.97%、7.53%、7.12%、6.19%、5.35%、5.05%,未分类类群占7.36%。样品中检测到了较低丰度的广古菌门和泉古菌门。随植被群落演替,根际土壤中变形菌门、厚壁菌门和浮霉菌门丰度逐渐增加;非根际土壤中酸杆菌门和疣微菌门丰度随植被演替逐渐减小。贵州喀斯特高原土壤细菌的影响因子大小为土壤有机碳、土壤总氮、含水量、电导率等,其中土壤有机碳和土壤总氮有显著性影响。     

闽江河口互花米草海向入侵对湿地土壤nirS型反硝化微生物群落结构及多样性的影响

2020年秋季,在闽江河口鳝鱼滩东部的互花米草分布区,由陆向海方向选择互花米草海向入侵前的光滩(MF)、入侵1～2年的互花米草湿地(SAN)和入侵6～7年的互花米草湿地(SA)为研究对象.基于高通量测序技术,探讨了互花米草海向入侵对土壤nir S型反硝化微生物群落结构及多样性的影响.结果表明,变形菌门(Proteobacteria)均是互花米草不同入侵阶段土壤中nir S型反硝化微生物的优势门(90.41%～97.36%),其次是厚壁菌门(Firmicutes)和放线菌门(Actinobacteria),三者在SAN和MF土壤中的丰度均与SA存在显著差异(p<0.05).不同入侵阶段土壤中共有的nir S型反硝化细菌属为假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)、偶氮弧菌属(Azoarcus)、陶厄氏菌属(Thauera)、Sulfurifustis属、副球菌属(Paracoccus)、鲁杰氏菌属(Ruegeria)、红肠命菌属(Rubrivivax)、趋磁螺菌属(Magnetospirillum)和福格斯氏菌属(Vogesella),其中,假单胞菌属是...     

三峡库区消落带落羽杉人工林土壤细菌群落结构多样性及动态变化

消落带是连接陆地与水域的交错地带,具有重要的生态功能,其适生木本植物的种植对消落带土壤的生物地球化学循环起着重要的作用.为探究适生植物对三峡库区消落带环境的适应机制,采用高通量测序技术对消落带退水后适生植物落羽杉(Taxodium distichum)不同生长时期(T1:5月、 T2:7月和T3:9月)的土壤细菌群落组成及多样性进行研究,同时采用PICRUSt2对细菌功能进行预测.结果表明,土壤pH值、硝态氮、铵态氮、土壤蔗糖酶、磷酸酶和脲酶等理化指标随时间变化显著(P<0.05);土壤细菌多样性、丰富度和结构也随时间变化而变化,除Chao1以外,根际土壤细菌的α多样性均表现为：T1>T2>T3,非根际土壤细菌α多样性则表现为：T3>T1>T2. RDA分析表明,影响细菌群落的理化指标主要有土壤pH值、脲酶、铵态氮和硝态氮.所有土壤样本中共检测到细菌60门,其中以变形菌门和酸杆菌门为优势细菌门.根据PICRUSt2预测,代谢是落羽杉土壤细菌群落中普遍存在的基本功能,与C、 N和P有关的各代谢途径存在一定的时间差异.以上研究结果有助于加深对三峡消落带植被修复...