厌氧氨氧化启动过程细菌群落多样性及PICRUSt2功能预测分析

细菌群落是实现厌氧氨氧化系统高效脱氮的核心,而厌氧氨氧化启动过程细菌群落多样性及其功能特征仍未被充分阐明.本研究采用升流式厌氧污泥床（UASB）反应器进行厌氧氨氧化系统启动,利用16S rRNA基因高通量测序技术并结合PICRUSt2功能预测分析,研究启动过程不同时间（d0、d30、d60和d90）细菌群落多样性及功能动态变化特征.结果表明,启动过程共检测到48个门、111个纲、269个目、457个科、840个属和1497个种;Candidatus_Brocadia和Candidatus_Kuenenia为检测到的厌氧氨氧化菌,且它们的相对丰度在启动过程不同时间存在显著差异（P<0.05）.启动过程,细菌群落α多样性指数整体呈现显著的降低趋势（P<0.05）,细菌群落结构呈现出明显的空间分异特征,且差异显著（R=0.846,P<0.01）.PICRUSt2功能预测分析表明,启动过程,细菌群落具有丰富的功能多样性,一级功能层表现为有机系统和代谢方面较为活跃,二级功能层子功能基因丰度在厌氧氨氧化启动过程发生明显变化;细菌群落涉及49个参与氮素代谢的相关功能基因,且不同时间阶段参与硝化、反硝化、厌氧氨氧化、硝酸盐同化/异化还原和亚硝酸盐同化/异化还原过程的相关功能基因丰度发生明显变化.     

Effects of salinity on soil bacterial and archaeal community in estuarine wetlands and its implications for carbon sequestration: verification in the Yellow River Delta

Soils from two typical tidal salt marshes with varied salinity in the Yellow River Delta wetland were analysed to determine possible effects of salinity on soil carbon sequestration through changes in soil microbiology. The mean soil respiration (SR) of the salt water–fresh water mixing zone (MZ) was 2.89 times higher than that of the coastal zone (CZ) (4.73 and 1.63?μmol?m^?2?s^?1, respectively, p?Pseudomonas sp. and Limnobacter sp. that might have led to its higher dehydrogenase activity and respiratory rates. Additionally, the CZ possessed more Halobacteria and Thaumarchaeota with the ability to fix CO₂ than the MZ. Significantly lower soil salinity in MZ (4.25?g?kg^?1) was suitable for β-Proteobacteria, but detrimental for Halobacteria compared with CZ (7.09?g?kg^?1, p?相似文献   

赣江南昌段丰水期细菌群落特征

细菌群落是河流生态系统的重要组成部分,本次研究基于高通量测序技术分析了赣江南昌段丰水期（4～8月）细菌群落特征.结果表明,赣江南昌段细菌优势类群为放线菌门（Actinobacteria, 41.18%）和变形菌门（Proteobacteria, 31.79%）,其次为厚壁菌门（Firmicutes, 10.04%）,拟杆菌门（Bacteroidetes, 7.26%）,蓝藻菌门（Cyanobacteria, 4.01%）.在属分类水平上,相对丰度最高的是hgcI_clade（16.39%）.赣江南昌段细菌丰度和多样性在城区上游、城区中心和城区下游采样点间没有显著差别,在不同月份有显著差别.除变形菌门（Proteobacteria）和疣微菌门（Verrucomicrobia）外,其它门水平分类细菌相对丰度在不同月份都有显著差异;不同采样点中,只有Proteobacteria差异显著（主要是Betaproteobacteria差异显著）,其它门水平分类细菌相对丰度在不同采样点的差异均不显著.温度和流量是影响河流细菌群落的主要因子,其中温度与细菌可操作分类单元（OTU）相关性更高,流量则与门分类水平细菌相关性更高,暴雨径流中Firmicutes取代Actinobacteria和Proteobacteria成为丰度最高的菌群.温度、流量和电导率（EC）是影响OTU的最佳环境因子组合,流量和温度是影响门水平细菌群落的最佳环境因子组合.河水化学指标对细菌群落的影响小于温度、流量等水文气象条件.     

基于Miseq的好氧反硝化菌源水脱氮的种群演变

为了研究好氧反硝化菌源水脱氮过程中水体微生物群落的演变,利用Miseq高通量测序法对投菌和对照两系统水体样本的微生物信息进行统计,并对两组样品进行了优化序列统计,OTU分布统计和分类学分析的基础分析;以及细菌群落结构,PCA,Rank-Abundance,Hcluster,Specaccum和OTU分布的高级分析.结果显示,投加贫营养好氧反硝化菌的源水系统的氮素得到有效去除,脱氮效果明显;层次聚类和主成分分析显示两系统内的群落结构发生变化,投菌系统与对照系统主要表现为变形菌和拟杆菌门;细菌主要门类和水质参数的相关性分析得出,水质指标对两系统群落变化作用明显;与此同时,投菌系统中有关氮循环的细菌有上升的变化过程. Miseq高通量测序研究源水脱氮过程的微生物种群演变可行,为研究原位生物脱氮过程的水体微生物群落演变提供技术支撑.     

堆肥添加剂降低碳氮损失的微生物学机制研究

为了探明添加剂如何影响堆肥微生物优势群落的演替进而影响碳氮损失,以玉米秸秆和鸡粪为原料,添加不同量的生物质炭和凹凸棒作为添加剂,设置5个处理（CK （鸡粪+玉米秸秆）、BC1（鸡粪+玉米秸秆+5%（干重）生物质炭）、BC2（鸡粪+玉米秸秆+10%（干重）生物质炭）、PG1（鸡粪+玉米秸秆+5%（干重）凹凸棒）和PG2（鸡粪+玉米秸秆+10%（干重）凹凸棒））进行堆肥.结果表明,相较于CK,BC1、BC2、PG1、PG2处理的碳和氮损失分别减少了8.60%、12.05%、2.03%、6.14%和14.54%、20.14%、8.40%、11.23%.优势微生物菌群与碳氮损失的冗余分析表明,添加10%生物质炭和添加10%凹凸棒都显著促进了堆肥过程中固氮类细菌相对丰度,而抑制反硝化细菌的相对丰度,且添加10%生物质炭效果更佳;KEGG分析表明,添加10%生物质炭显著影响氨基酸代谢和碳水化合物代谢功能基因,而添加10%凹凸棒显著影响氨基酸代谢功能基因.由此可见,10%添加剂的碳氮损失都低于5%添加剂的处理,添加生物质炭的碳氮损失都低于凹凸棒处理,添加剂通过影响优势微生物群落及其氨基酸代谢和碳水化合物代谢功能基因抑制堆肥过程中的碳氮代谢,从而减少碳氮损失.     

贵州喀斯特山区不同种植年限花椒根际土壤细菌群落结构特征研究

以贵州花江峡谷花椒(Zanthoxylum bungeamun)林为研究对象,采用16S rRNA高通量测序技术,分析种植5、10、20、30 a花椒根际土壤细菌群落结构和多样性特征,探讨不同种植年限花椒土壤理化因子对根际细菌群落分布的影响,为喀斯特石漠化地区花椒农业可持续发展提供有效的理论依据。结果表明,随着花椒种植年限的增加,土壤含水率、pH和有效磷逐渐升高,有机质、铵态氮和硝态氮含量先降低后升高(P<0.05)。根际土壤细菌Shannon、Simpson指数总体呈现上升趋势,OTUs、Chao1指数在10a较低。PCo A分析显示,不同种植年限花椒根际细菌群落组成差异显著,并且随着年限的增加,群落结构趋于相似;差异指示种分析表明,5—30a差异指示种分别为Firmicutes(厚壁菌门)、Cyanobacteria(蓝藻菌门)、Planctomycetes(浮霉菌门)、Entotheonellaeota(肠杆菌门)。RDA分析表明,有机质和含水率与细菌群落分布显著相关(P<0.05)。系统发育多样性分析表明,丰富类群多样性与环境因子存在较多的正相关,稀有类群比丰富类群有更强的发育信号。Bug Base预测分析发现:随着年限增加根际土壤好氧细菌增加,厌氧细菌下降,5、10 a氧化胁迫耐受菌低于20、30 a,致病菌在5、10 a高于20、30 a。综上,种植年限影响了花椒根际土壤细菌群落多样性,不同年限根际土壤肥力不同使得土壤细菌选择性生长,随着一定时间发育花椒抗病能力提高。     

Impact of heavy metals on bacterial communities from mangrove soils of the Mahanadi Delta (India)

This study aimed to assess soil nutrient status and heavy metal content and their impact on the predominant soil bacterial communities of mangroves of the Mahanadi Delta. Mangrove soil of the Mahanadi Delta is slightly acidic and the levels of soil nutrients such as carbon, nitrogen, phosphorous and potash vary with season and site. The seasonal average concentrations (μg/g) of various heavy metals were in the range: 14 810–63 370 (Fe), 2.8–32.6 (Cu), 13.4–55.7 (Ni), 1.8–7.9 (Cd), 16.6–54.7 (Pb), 24.4–132.5 (Zn) and 13.3–48.2 (Co). Among the different heavy metals analysed, Co, Cu and Cd were above their permissible limits, as prescribed by Indian Standards (Co=17 μg/g, Cu=30 μ g/g, Cd=3–6 μ g/g), indicating pollution in the mangrove soil. A viable plate count revealed the presence of different groups of bacteria in the mangrove soil, i.e. heterotrophs, free-living N₂ fixers, nitrifyers, denitrifyers, phosphate solubilisers, cellulose degraders and sulfur oxidisers. Principal component analysis performed using multivariate statistical methods showed a positive relationship between soil nutrients and microbial load. Whereas metal content such as Cu, Co and Ni showed a negative impact on some of the studied soil bacteria.     

嗜盐菌群对菲的降解及萘双加氧酶基因的表达规律

从胜利油田富集了一个能够降解多环芳烃的嗜盐菌群,通过克隆文库技术解析了菌群萘双加氧酶(ndo)基因的多样性.结果表明,该嗜盐菌群有6种ndo基因型,其中3种主要基因型(占总克隆子92.7%)与经典nah-like基因的相似度为89%.采用real-time RT-PCR等技术分析了不同盐度下菲降解过程中ndo基因的表达量、降解速率、生长曲线以及菲的生物可利用度,探索了菌群对菲的降解及ndo基因的表达规律.结果表明,当盐度由10%升高到20%,菌群完全降解100mg/L菲的时间从6d延长到9d,菌群生长的迟滞期由1d延长为3d.溶解菲的浓度在菌群生长过程中呈增加的趋势.Ndo基因的表达量在降解过程中呈先降低后随溶解菲浓度升高而升高的趋势,表明高浓度菲在初始阶段可能抑制ndo基因的表达.单因素方差分析(n=3, P>0.05)表明溶解菲浓度在10%和20%盐度下没有显著差异,表明一定范围的盐度不会影响菌群降解过程中菲的溶解度.菌群ndo基因的相对表达量在10%盐度下较在20%盐度下高,说明盐度对嗜盐菌群功能基因的表达具有抑制作用.     

钻井废弃液中石油污染物及细菌生态多样性

采用GC/MS分析方法,发现平湖油田钻井废弃液中含有烃类、酸类和酯类等23种有机污染物质,含量达到49 800 mg/L。运用16S rRNA基因分析法测得的微生物生态多样性表明,废弃液中微生物种类丰富,包括γ和β变形杆菌,γ-Proteobacteria是优势菌群,占克隆总数78.6%。Oceanospirillales、Pseudomonadales和Enterobacteriales是γ-Proteobacteria纲的主要菌群。     

Development of pretreatment protocol for DNA extraction from biofilm attached to biologic activated carbon (BAC) granules

The biologic activated carbon (BAC) process is widely used in drinking water treatments. A comprehensive molecular analysis of the microbial community structure provides very helpful data to improve the reactor performance. However, the bottleneck of deoxyribonucleic acid (DNA) extraction from BAC attached biofilm has to be solved since the conventional procedure was unsuccessful due to firm biomass attachment and adsorption capacity of the BAC granules. In this study, five pretreatments were compared, and adding skim milk followed by ultrasonic vibration was proven to be the optimal choice. This protocol was further tested using the vertical BAC samples from the full-scale biofilter of Pinghu Water Plant. The results showed the DNA yielded a range of 40 μg·g^-1 BAC (dry weight) to over 100 μg·g^-1 BAC (dry weight), which were consistent with the biomass distribution. All results suggested that the final protocol could produce qualified genomic DNA as a template from the BAC filter for downstream molecular biology researches.