抚顺西露天油页岩可培养微生物的分离与鉴定

为了解油页岩中可培养微生物的多样性,利用稀释平板培养法对我国抚顺盆地西露天组油页岩中可培养微生物进行分离纯化,并对所获菌株进行16S rDNA(细菌)和rDNA ITS(真菌)序列测定和系统发育树分析.以营养琼脂培养基分离到8株细菌,以马铃薯葡萄糖琼脂培养基分离到4株真菌.16S rDNA序列测定结果表明细菌菌株属于芽孢杆菌属(Bacillus),赖氨酸芽孢杆菌属(Lysinibacillus),短杆菌属(Brevibacillus),类芽胞杆菌属(Paenibacillus)和红球菌属(Rhodococcus); rDNA ITS序列测定结果显示真菌菌株分别属于芽枝霉属(Cladosporium),曲霉属(Aspergillus),犁头霉属(Absidia)和正青霉属(Eupenicillium). 2株细菌(ZK4和ZK5)能够在以油页岩为唯一碳源和氮源的固体培养基上连续生长5代以上,且长势良好, 可作为油页岩生物转化的潜力菌株进行深入研究.     

底泥中红霉素耐受菌群的多样性分析

为了正确评估抗生素残留的环境风险,了解长期受低浓度红霉素污染条件下,水体底泥中红霉素耐受菌群结构的变化特点,采用传统的微生物培养法和分子生物学方法对长期受低浓度红霉素污染的水体底泥中红霉素耐受菌群结构的多样性进行了分析.结果表明:采用传统的培养方法,从底泥中可分离筛选出3株对红霉素有耐受能力的菌株,根据其形态学特征及16S rDNA序列分析,初步鉴定为赖氨酸芽孢杆菌(Lysinibacillussp.)、土壤芽孢杆菌(Solibacillus silvestris)以及蜡样芽孢杆菌(Bacillus cereus),其中敏感菌蜡样芽孢杆菌经低浓度红霉素诱导对红霉素表现出一定的耐药性.进一步构建底泥耐药菌群16S rDNA克隆文库发现,底泥中红霉素耐受菌群可分为三大类群,其中未获培养的细菌(Uncultured bacterium)在整个文库中比例最大,占65.06%,其次为芽孢杆菌纲(Bacilli)和梭菌纲(Clostridia),分别占文库比例33.73%和1.20%.文库中可培养的耐药菌优势类群为芽孢杆菌纲,与传统培养方法得到的结果相一致.上述研究表明,在长期受低浓度红霉素污染的环境中,红霉素耐药菌的存在具有一定的广泛性和潜在性,将可能威胁人类健康和生态系统,应当重视其生态风险评价与管理.     

16S rDNA 克隆文库法分析Biostyr 曝气生物滤池处理城市污水的细菌多样性研究

采用分子生物学手段16SrDNA克隆文库方法,对第三代生物膜法代表工艺Biostyr曝气生物滤池（BAF）中滤料表面细菌进行了多样性研究.从16SrDNA克隆文库中随机挑选了50个克隆子进行序列测定（约1.5kb）,对测序结果进行了BLAST对比.结果表明,BiostyrBAF系统中的细菌群落具有高度多样性,有41个克...     

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.     

小麦与不同作物多样化轮作对土壤真菌群落的影响

为明确华北平原冬小麦与不同作物多样化轮作下的土壤真菌群落差异,为生态可持续种植制度的构建和优化提供理论依据,采用实时荧光定量PCR和高通量测序技术研究了连续冬小麦-夏玉米M、冬小麦-夏花生(夏玉米)PM和冬小麦-夏大豆(夏玉米)SM轮作处理的土壤真菌群落丰度、组成和多样性.结果表明,与连续冬小麦-夏玉米处理相比,轮作花生PM2和大豆SM2处理显著降低了土壤真菌ITS序列拷贝数,轮作花生或大豆处理增加了土壤真菌群落丰富度和多样性.非度量多维尺度分析(NMDS)结果显示,不同轮作茬口之间土壤真菌群落存在明显分离,轮作作物对土壤真菌群落结构的影响达显著水平.华北平原砂壤质潮土中不同作物轮作处理的土壤真菌群落均以Ascomycota为优势菌门,以Sordariomycetes和Eurotiomycetes为优势菌纲.不同轮作茬口土壤真菌群落组成存在差异显著的类群,Neocosmospora、Plectosphaerella和Gibellulopsis等潜在病原真菌在冬小麦-夏花生(夏玉米)轮作处理中显著富集,而Penicillium和Zopfiella等潜在有益真菌在冬小麦-夏大豆(夏玉米)轮...     

Soil fungistasis and its relations to soil microbial composition and diversity: a case study of a series of soils with different fungistasis

Fungistasis is one of the important approaches to control soil-borne plant pathogens.Some hypotheses about the mechanisms for soil fungistasis had been established,which mainly focused on the soil bacterial community composition,structure,diversity as well as function.In this study,the bacterial community composition and diversity of a series of soils treated by autoclaving,which coming from the same original soil sample and showing gradient fungistasis to the target soil-borne pathogen fungi Fusarium gr...     

抚顺盆地西露天组油页岩微生物碳代谢多样性分析

为探讨油页岩微生物的碳代谢功能多样性,以抚顺盆地西露天组油页岩为研究对象,利用Biolog-GN2和GP2微孔板检测法,在连续7d的培养期内,测定油页岩本源微生物群落水平生理图谱(CLPP),并根据化学基团的不同,分析微生物对不同类型碳源的代谢能力和代谢差异.结果显示,微生物对不同类型碳源的代谢程度和代谢速率存在明显的差异.革兰氏阴性和阳性细菌群落均对氨基类碳源的利用较好,其次是羧酸类以及糖和糖衍生物,而对磷酸糖利用较差.连续多点测算Shannon-Wiener,Simpson和McIntosh多样性指数,分析表明微生物群落碳代谢功能多样性总体较高,但是随着培养时间的增加,多样性略有降低.     

应用PCR-DGGE分析三江湿地土壤真菌群落结构的多样性

采用PCR-DGGE技术,研究毛苔草湿地土壤、小叶章湿地土壤(氮肥施用量分别为0、20、40和60 kg/hm2)和岛状林湿地土壤在生长期(2010年7月和2011年6月)和凋落期(2010年10月)的真菌群落结构变化规律. 结果表明:湿地植物生长期和凋落期真菌群落结构差异显著;退化程度较高的岛状林湿地土壤真菌群落的多样性指数最高,2010年7月、10月和2011年6月多样性指数分别为3.22、3.25和3.21;在小叶章湿地土壤中添加氮肥20～40 kg/hm2,2010年7月、10月和2011年6月湿地土壤真菌群落的多样性指数分别升至0.27、0.15和0.05. 对湿地土壤真菌18S rDNA基因测序分析可知,三江湿地土壤真菌类群主要包括外囊菌纲(Taphrinomycetes)、座囊菌纲(Dothideomycetes)和锤舌菌纲(Leotiomycetes),其中锤舌菌纲是该湿地主要的土壤真菌类群.      

Soil fungistasis and its relations to soil microbial composition and diversity: A case study of a series of soils with di erent fungistasis

Fungistasis is one of the important approaches to control soil-borne plant pathogens.Some hypotheses about the mechanisms for soil fungistasis had been established,which mainly focused on the soil bacterial community composition,structure,diversity as well as function.In this study,the bacterial community composition and diversity of a series of soils treated by autoclaving,which coming from the same original soil sample and showing gradient fungistasis to the target soil-borne pathogen fungi Fusarium gr...     

亚硝化/电化学生物反硝化全自养脱氮工艺细菌形态及多样性研究

采用电镜观察和分子生物学手段16S rDNA克隆文库方法对亚硝化/电化学生物反硝化全自养脱氮工艺中的细菌进行了形态和多样性研究,从16S rDNA克隆文库中随机挑选60(61)个克隆子进行序列测定(约500bp),对测序结果进行BLAST比对和系统发育分析.结果表明,亚硝化段内的细菌主要为球状和椭球状的氨氧化细菌,以亚硝酸氮作为进水基质时,电化学反硝化生物段内细菌主要为短杆状和椭球状的脱氮菌.亚硝化/电化学生物反硝化脱氮系统中蕴藏着特有的微生物新资源.亚硝化段细菌类群的优势顺序为β-Proteobacteria类群(60.00%)、Bacteroidetes类群(28.33%)和Chloroflexi类群(11.67%).当电化学生物反硝化段进水氮基质为亚硝氮(429～543mg.L-1)和氨氮(412～525mg.L-1)时,细菌优势类群顺序为β-Proteobacteria(78.33%)类群和ε-Proteobacteria类群(21.67%);当电化学生物反硝化段进水氮基质为亚硝氮(519～578mg.L-1)时,细菌优势类群顺序为β-Proteobacteria类群(81.97%)、ε-Proteobacteria(16.39%)类群和γ-Proteobacteria类群(1.64%);优势类群变化不大,但每种类群中细菌的种类和数量变化较大,这主要是由进水基质变化导致.     

石油污染地下水中细菌多样性研究

采集了某废弃炼油厂的石油污染地下水样品,提取水中微生物总DNA,构建细菌16S rDNA克隆文库,并通过16S rDNA序列的系统发育分析,对样品中的细菌种群多样性以及群落结构进行了研究.结果表明,文库中阳性克隆的16S rDNA序列分属10个细菌类群,分别为γ-Proteobacteria(49.1%)、α-Proteobacteria(12.9%)、β-Proteobacteria(11.1%)、Bacteroidetes(9.2%)、Verrucomicrobia(6.7%)、Acidobacteria(2.5%)、δ-Proteobacteria(1.2%)、Actinobacteria(1.2%)、Planctomycetes(0.6%)、Unidentifiedbacteria(5.5%).在这一生态系统中,γ-Proteobacteria类细菌占据主导地位,接近50%,尤其是假单胞菌属(Pseudomonas)微生物在文库中的比例达35.6%.该石油污染地下水样品中细菌与许多其它已知的降解菌亲缘关系较近,如鞘胺醇单胞菌(Sphingomonas)、红球菌(Rhodococcus)和短波单胞菌(B...     

空心菜浮床对鱼塘水质和微生物多样性的影响

为从环境微生物的角度探讨生物浮床的水质调节机理,采用宏基因组学测序技术和生物信息学手段,分析了环境微生物和根系微生物群落的多样性.研究结果表明,鱼菜共生模式下优势菌门是变形菌门（Proteobacteria）、蓝细菌门（Cyanobacteria）、拟杆菌门（Bacteroidetes）、放线菌门（Actinobacteria）、绿弯菌门（Chloroflexi）和厚壁菌门（Firmicutes）等,主要菌属是聚球藻属（Synechococcus）、hgcI_clade、CL500-29_marine_group、芽孢杆菌属（Bacillus）和分支杆菌属（Mycobacterium）等;空心菜根系微生物群落中最丰富的优势菌门是变形菌门和蓝细菌门,主要菌属是红细菌属（Rhodobacter）、噬氢菌属（Hydrogenophaga）和聚球藻属等.研究结果还表明低覆盖率（3.5%）的空心菜生物浮床能够增加水体的溶解氧含量,降低水体中含氮化合物的浓度和改善养殖环境中的微生态平衡,提高有益菌和氮循环细菌的含量.     

铜离子冲击下活性污泥微生物多样性的分子生态学分析

采用基于 16SrDNA序列分析方法 ,在长度多态 (LPM)和 16SrDNA的GC含量二维水平分析了受重金属污染的活性污泥系统内细菌的优势种类和多样性 .设计的PCR引物可以将细菌分为三大类 :即 1)proteobacterialα&δ(变形杆菌α&δ) ,2 )pro teobacterialβ&γ(变形杆菌 β&γ) ,以及 3)flexibacter(屈桡杆菌 )和革兰氏阳性菌 .分析了未受重金属驯化和受重金属驯化的两类活性污泥系统在重金属作用下优势种和多样性的变化 ,结果显示未驯化的活性污泥系统多样性减少但优势种的变化微小 ,而驯化系统优势种有较大的变化 ,但多样性基本不变 .这一结果证实驯化有助于提高微生物对重金属的抗性     

Conservation for the landscape ecological diversity in Wulingyuan scenic area of China

Wulingyuan is located at the mountainous area of the middle reach of the Yangtze River,it is one of the three nature heritages in China which ranks in the “List of Worls‘s Heritage”by UNESCO.It is characterized by quartz sandstone peaks landform with several landform components(pattern,corridor)and rich in landscape ecological diversity and biodiversity.The main patterns(ecosystem)include mid-height mountain peaks,rift-avlley and streams among peaks,peaks and gullies on slopes,square mountain-platforms and peaks among blind valleys and so on.The corridor system consists of natural corridors and artificial corridors among which the stream corridors account for a major part.The fracturing of habitat is unfavorable for the biodiversity conservation,but meanwhile the habitat diversity leads to an increase in biodiversity.Therefore,it is still rich in landscape ecological diversity in Wulingyuan.The biodiversity at the level of landscape component(ecosystem) and the function of the Wulingyuan complex ecosystem,and the measures for the biodiversity conservation in Wulingyuan ecotourism area are discussed in this paper.     

Microbial community diversity in the profile of an agricultural soil in northern China

The soil microorganisms at different depths play an important role in soil formation,ecosystem biogeochemistry,recycling of nutrients,and degradation of waste products.The aims of this study were to observe the microbial diversity in the profile of an agricultural soil in northern China,and to research the correlation between soil microbes and geochemistry.First,the soil geochemistry of the profile was investigated through 25 chemical elements.Secondly,the various physiological groups of microorganisms w...     

太湖不同湖区真核微型浮游生物基因多样性的研究

用DGGE(denatured gradient gel electrophoresis)和构建18S rDNA克隆文库2种方法对太湖不同湖区的真核微型浮游生物(0.8～20μm)多样性及组成结构进行了研究.DGGE结果表明,不同湖区真核微型浮游生物的DGGE指纹图谱存在明显差异,其中营养水平较低的东太湖和贡湖DGGE条带数最多,分别为23和24,香农多样性指数分别为3.135和3.178,而营养水平较高的梅梁湾和五里湖最少,均为18,香农多样性指数为2.890,表明营养水平较低湖区的多样性高于营养水平较高的湖区.克隆测序结果表明太湖中真核微型浮游生物种类车富,占优势的主要是一些鞭毛藻、异养鞭毛虫、纤毛虫和真菌,而营养水平不同的梅梁湾、湖心、东太湖中真核微型浮游生物组成明显不同.在营养水平较高的梅梁湾,28.6%的OUT(operational taxonomicunit)属于异养鞭毛虫,另外隐藻、金藻分别占22.9%和14.3%;在湖心,金藻的比例最大,占25.7%,另外比较多的是异养鞭毛虫和隐藻,分别为20.0%和14.3%;而营养水平较低的东太湖各类纤毛虫所占比份最大,为26.8%,异养鞭毛虫较少,仅占4.9%,另外真菌含量较高,占12.2%.     

再生水湿地香蒲根内生细菌群落多样性及其水质特征分析

以北京白河人工湿地再生水补水河段香蒲根为研究对象,采用16S rDNA克隆文库技术对香蒲根内生细菌群落多样性进行分析,得出该克隆文库包括4大类群细菌,最优势类群为变形杆菌门Proteobacteria,其中包括Gammaproteobacteria、Betaproteobacteria、Epsilonproteobacteria、Alphaproteobacteria;其次为厚壁菌门Firmicutes、疣微菌门Verrucomicrobia、拟杆菌门Bacteroidetes;另外还包括部分非培养未确定种属的细菌.结合研究区再生水排放口与湿地下游水质数据,得出TN、TP、NO₃^-去除率达到了42.15%、47.34%、28.56%,表明湿地对再生水中的TN、TP、NO₃^-具有明显去除机制,说明香蒲根内生细菌克隆文库中菌株与湿地氮、磷等生物地球循环关系密切.     

东海浮游十足类(Decapods)多样性研究

根据1997~2000年对东海23°30′~33°N、118°30′~128°E海域4个季节海洋调查资料,运用定量、定性方法,探讨了东海浮游十足类的种类组成、时空分布、多样性及与环境的关系。结果显示:东海4季共出现浮游十足类9种,其中秋季4种,冬季5种,夏春季各7种;中型莹虾、细鳌虾、汉森莹虾、刷状莹虾和正型莹虾是常见种,其余4个为稀有种。春季转至冬季,夏季转至秋季、秋季转至冬季,其种类随着季节的变换而出现更替。种数分布在冬春季主要受温度的影响,夏秋季的主要影响因子是盐度。冬春季种类变化主要与台湾暖流与黄海冷水团季节性的进退以及东海气温季节性变化有关,夏季种类分布特征与沿岸流和暖流势力强弱有关。常见种对不同季节的环境适应性较强,其出现频率和个体丰度远大于稀有种。中型莹虾的大量出现是影响东海浮游十足类多样性指数值较低的主要原因。     

Molecular diversity of arbuscular mycorrhizal fungi at a large-scale antimony mining area in southern China

Arbuscular mycorrhizal fungi(AMF) have great potential for assisting heavy metal hyperaccumulators in the remediation of contaminated soils. However, little information is available about the community composition of AMF under natural conditions in soils contaminated by antimony(Sb). The objective of this study was to investigate the characteristics of AMF molecular diversity, and to explore the effects of Sb content and soil properties on the AMF community structure in an Sb mining area. Four Sb mine spoils and one adjacent reference area were selected from around the Xikuangshan mine in southern China. The association of AMF molecular diversity and community composition with the rhizosphere soils of the dominant plant species was studied by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis(PCR-DGGE). Results from all five studied sites showed that the diversity of AMF decreased with increasing Sb concentration. Principal component analysis(PCA) indicated that the AMF community structure was markedly different among these groups. Further redundancy analysis(RDA) showed that Sb contamination was the dominating factor influencing the AMF community structure in the Sb mine area. However, the multivariate analysis showed that, apart from the soil Sb content, extractable nitrogen content and organic matter content also attributed to AMF sequence distribution type. Some AMF sequences were only found in the highly contaminated area and these might be ideal candidates for improving phytoremediation efficiency in Sb mining regions. Gene sequencing analysis revealed that most species were affiliated with Glomus, suggesting that Glomus was the dominant AMF genus in the studied Sb mining area.     

Molecular diversity of arbuscular mycorrhizal fungi at a large-scale antimony mining area in southern China

Arbuscular mycorrhizal fungi (AMF) have great potential for assisting heavy metal hyperaccumulators in the remediation of contaminated soils. However, little information is available about the community composition of AMF under natural conditions in soils contaminated by antimony (Sb). The objective of this study was to investigate the characteristics of AMF molecular diversity, and to explore the effects of Sb content and soil properties on the AMF community structure in an Sb mining area. Four Sb mine spoils and one adjacent reference area were selected from around the Xikuangshan mine in southern China. The association of AMF molecular diversity and community composition with the rhizosphere soils of the dominant plant species was studied by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE). Results from all five studied sites showed that the diversity of AMF decreased with increasing Sb concentration. Principal component analysis (PCA) indicated that the AMF community structure was markedly different among these groups. Further redundancy analysis (RDA) showed that Sb contamination was the dominating factor influencing the AMF community structure in the Sb mine area. However, the multivariate analysis showed that, apart from the soil Sb content, extractable nitrogen content and organic matter content also attributed to AMF sequence distribution type. Some AMF sequences were only found in the highly contaminated area and these might be ideal candidates for improving phytoremediation efficiency in Sb mining regions. Gene sequencing analysis revealed that most species were affiliated with Glomus, suggesting that Glomus was the dominant AMF genus in the studied Sb mining area.