Diversity of methanotrophs in Zoige wetland soils under both anaerobic and aerobic conditions

Zoige wetland is one of the most important methane emission centers in China. The oxidation of methane in the wetland a ects global warming, soil ecology and atmospheric chemistry. Despite their global significance, microorganisms that consume methane in Zoige wetland remain poorly characterized. In this study, we investigated methanotrophs diversity in soil samples from both anaerobic site and aerobic site in Zoige wetland using pmoA gene as a molecular marker. The cloning library was constructed according to the pmoA sequences detected. Four clusters of methanotrophs were detected. The phylogenetic tree showed that all four clusters detected were a liated to type I methanotrophs. Two novel clusters (cluster 1, cluster 2) were found to relate to none of the recognized genera of methanotrophs. These clusters have no cultured representatives and reveal an ecological adaptation of particular uncultured methanotrophs in Zoige wetland. Two clusters were belonging to Methylobacter and Methylococcus separately. Denaturing gradient gel electrophoresis gel bands pattern retrieved from these two samples revealed that the community compositions of anaerobic soil and aerobic soil were di erent from each other while anaerobic soil showed a higher metanotrophs diversity. Real-time PCR assays of the two samples demonstrated that aerobic soil sample in Zoige wetland was 1.5 times as much copy numbers as anaerobic soil. These data illustrated that methanotrophs are a group of microorganisms influence the methane consumption in Zoige wetland.     

天然红树林土壤微生物大片段宏基因组文库的构建

采用改良及优化后的原位裂解法提取红树林土壤环境DNA,通过脉冲场电泳图谱显示所得DNA片段主要集中于23～50 kbp范围, 表明大片段DNA在总DNA中的比率明显提高; 对改良后的原位裂解提取法与不同DNA纯化方法的组合进行比较分析, 确定电洗脱为最佳纯化方法; 运用改良后的原位裂解提取法与电洗脱纯化法联用实验方案,构建了4个季节天然红树林土壤Cosmid宏基因组文库. 随机挑取克隆子,提取质粒后经BamHI酶切验证, 平均插入片段均大于35 kbp,且片段类型各不相同, 推测该文库至少包含了335 Mbp潮间带微生物基因组信息. 该方法的建立使得开发红树林湿地生态系统的特色微生物资源成为可能, 并为探索其它生境的未培养微生物资源提供借鉴.     

红树林湿地中好氧-厌氧反硝化菌脱氮特性及其种群结构分析

随着沿岸水体氮素富营养化的加剧,生物脱氮作用越来越受重视,位于海陆交界的湿地红树林生态系统作为一个自然脱氮体系备受关注.本研究以典型亚热带湿地红树林(香港Mai Po)作为对象,结合传统的富集筛选和分子生物学方法--建立nosZ基因克隆文库和RFLP分析技术对红树林沉积物中反硝化细菌脱氮能力、种群结构和丰度进行研究.从Mai Po红树林沉积物中共筛选到12株好氧反硝化菌和8株厌氧反硝化菌,其中好氧反硝化菌包括Pseudomonas(4株)、Comamonas(2株)和Acinetobacter(2株)等7个菌属,厌氧反硝化菌属于Pseudomonas、Agrobacterium和Uncultured Betaproteobacteria bacterium(2株)等7个菌属.筛选到的好氧和厌氧反硝化菌均具有较高的脱氮能力,大部分在2d内NO3--N去除率达到98%以上.建立Mai Po红树林湿地反硝化细菌的nosZ基因克隆文库的结果表明,反硝化细菌的50个克隆子中有26个克隆子分属于11个未知类群,其余克隆子属于Pennisetum类群(26%),β-proteobacterium类群(10%),Entandrophragma类群(4%),Pseudomonas类群(4%)和denitrifying bacterium类群(4%).可见Mai Po红树林中反硝化细菌具有很高的生物多样性.     

我国东北典型河流冰封期细菌多样性的研究——以松花江为例

在我国东北松花江冰封期时采集了该河流域5个监测断面的河水样本,并分别构建16S rDNA克隆文库,通过16S rDNA序列的系统发育分析,对水样中细菌群落结构和种群多样性进行了研究.试验结果表明,5个采样点的水样pH都呈弱酸性,并且都有不同程度的多环芳烃(PAHs)污染,其中,B(九站)采样点污染最为严重.5个样品中检测到的共同细菌种类有5个:β-变形菌纲(Betaproteobacteria)、γ-变形菌纲(Gammaproteobacteria)、放线菌门(Actinobacteria)、蓝细菌门(Cyanobacteria)和拟杆菌门(Bacteroidetes).5个河水样品中的细菌与许多已知降解菌的亲缘关系较近,并以能降解多环芳烃类有机污染物的微生物种类居多,且在5个样品中均发现了指示河流富营养化的菌种,表明该河流存在普遍的富营养化趋势,特别是D、E段(佳木斯上和佳木斯下)比较严重.     

克隆文库方法分析厌氧氨氧化反应器中细菌群落结构

为了认识低基质浓度污水厌氧氨氧化(ANAMMOX)脱氮过程的生物学机制,为ANAMMOX脱氮工艺的优化提供理论依据,通过构建细菌16S rDNA(约1 500 bp)克隆文库和浮霉菌特有16S rDNA(约830 bp)克隆文库对ANAMMOX脱氮反应器中活性污泥的细菌群落结构进行分析。从细菌16S rDNA克隆文库中随机挑选到160个克隆子,共31个分类单元(OTU),与GenBank数据库比对结果表明,厌氧颗粒污泥中的细菌群落具有丰富的多样性,包括:变形菌门(Proteobacteria)、拟杆菌门(Bacteroidete)、硝化螺旋菌门(Nitrospira)、酸杆菌门(Acidobacteria)、绿弯菌门(Chloroflexi)、Candidate division OP10、浮霉菌门(Planctomycetes)和未知菌,其中,变形菌门和拟杆菌门为优势菌群,分别占41.9%和34.2%。在浮霉菌特有16S rDNA克隆文库的40个克隆子中,34个克隆子属于CandidatusKuenenia属的厌氧氨氧化细菌,它们是ANAMMOX脱氮过程的主要功能菌。     

连云港海底底泥及青海湖底泥细菌多样性研究

通过对不同盐环境连云港海底底泥和青海湖底泥样品构建细菌16S rRNA克隆文库,对2个环境样品中细菌群落的多样性、丰富度和优势度等进行了比较,并对其群落组成结构进行了分析.结果表明,连云港海底底泥文库的多样性指数Shannon diversity（H）达到3.53,青海湖底泥文库的Shannon diversity（H...     

网络环境下图书馆防灾减灾问题的探讨

网络环境下图书馆防灾减灾问题是必须引起人们足够重视的问题。通过对引起图书馆网络灾害的各种因素进行分析,从制度建设、网络业务人员的培训、制定应急预案、加大网络安全建设的投入、加强网络的软硬件管理和创建异地容灾系统等方面进行了网络环境下图书馆防灾减灾问题的探讨。     

北方典型浅水湖泊沉积物有机碳沉积特征研究—以白洋淀为例

湖泊沉积物中有机碳是全球碳库的重要组成部分,为明确浅水湖泊沉积物中有机碳的沉积特征,采集了白洋淀的沉积柱样品,利用连续提取的方法,分析了沉积物自然沉积过程中水溶性有机碳（WSOC）、热解性有机碳（THOC）、酸解性有机碳（HHOC）、碱解性有机碳（AHOC）和不溶性有机碳（WISOC）的含量变化规律、光谱组分特征及影响因素.结果显示,各有机碳组分垂直方向上含量变化规律一致,均随深度增加呈下降趋势,表层20 cm左右的沉积物中有机碳组分含量普遍较高,含量特征呈WISOC>AHOC>HHOC>THOC>WSOC.用紫外光谱的特征参数SUVA₂₅₄、SUVA₂₈₀来分析溶解性有机碳的腐殖程度和分子量大小变化规律,发现腐殖程度与分子量大小变化趋势一致,WSOC和HHOC随深度增加呈增大趋势,THOC随深度增加呈减小趋势,AHOC在0~21 cm阶段随深度逐渐下降,在21~69 cm范围随深度逐渐上升.用A₃₀₀/A₄₀₀表征 有机碳分子构成,发现THOC、HHOC主要由富里酸构成,WSOC、AHOC主要成分为胡敏酸.用A₂₅₃/A₂₀₃表征芳环取代基的种类,发现WSOC、THOC、HHOC苯环上的脂肪链发生聚合反应转化为羰基、羧基等;AHOC的芳香分子上的取代基被微生物代谢分解或转化为脂肪链.相关性分析结果显示,沉积物有机碳及其各组分之间均呈显著正相关,沉积物中的有机碳组分在沉积过程中受到类似的矿化分解作用,且有机碳及其组分含量与C/N值、pH值呈显著负相关.研究成果揭示了白洋淀区域沉积环境中有机碳组分的分布及部分转化规律,可为进一步明确浅水 湖泊的碳源-汇的转换关系结构提供数据支持.     

Abundance and distribution of ammonia-oxidizing archaea in Tibetan and Yunnan plateau agricultural soils of China

As low oxygen and high ultraviolet （UV） exposure might significantly affect the microbial existence in plateau, it could lead to a specialized microbial community. To determine the abundance and distribution of ammonia-oxidizing archaea （AOA） in agricultural soil of plateau, seven soil samples were collected respectively from farmlands in Tibet and Yunnan cultivating the wheat, highland-barley, and colza, which are located at altitudes of 3200-3800 m above sea level. Quantitative PCR （q-PCR） and clone library targeting on amoA gene were used to quantify the abundances of AOA and ammonia-oxidizing bacteria （AOB）, and characterize the community structures of AOA in the samples. The number of AOA cells （9.34 × 10^7-2.32× 10^8 g^-1 soil） was 3.86-21.84 times greater than that of AOB cells （6.91 × 10^6-1.24 × 10^8 g^-1 soil） in most of the samples, except a soil sample cultivating highland- barley with an AOA/AOB ratio of 0.90. Based Kendall＇s correlation coefficient, no remarkable correlation between AOA abundance and the environmental factor was observed. Additionally, the diversities of AOA community were affected by total nitrogen and organic matter concentration in soils, suggesting that AOA was probably sensitive to several environmental factors, and could adjust its community structure to adapt to the environmental variation while maintaining its abundance.     

Effects of cotton straw amendment on soil fertility and microbial communities

Maintaining soil fertility, while controlling pollution from excessive chemical fertilizer application is important for keeping soil productivity of sustainable agriculture. Variety of straws have been used and proven to be good soil amendments for increasing soil organic matter (OM) and a range of additional soil nutrients. However, little is known about the utilization of cotton straw for soil amendment. To better understand the mechanism behind cotton straw soil amendments, investigations were performed upon cucumber seedlings, where changes to soil nutrients and microbial communities were investigated. The results revealed that the cotton straw application promoted the cucumber seedling growth by significantly increasing the soil OM, available nitrogen, available phosphorus, and available potassium. The concentration of cotton straw was positively correlated to both the number of the culturable microorganisms and also the total microbial biomass within soil. Furthermore, assessment of cotton straw application using Biolog metabolic profiling and phospholipid fatty acid analysis revealed that such application increased the microbial community metabolic activity, and markedly changed the structure of microbial community. 16S rRNA gene clone library construction and phylogenetic analysis of soil bacteria revealed γ- Proteobacteria sequences dominated the cotton straw amendment soil, comprising 27.8% of the total number of analyzed sequences, while they were less represented in control soil (13.4%). On the contrary, the Sphingobacteria (7.8%) and Verrucomicrobia (2.4%) in the cotton straw amendment soil decreased after application when compared to the control soil 15.2% and 15.2%.